Joint International Conference on Long-term Experiments ...
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<str<strong>on</strong>g>Joint</str<strong>on</strong>g> <str<strong>on</strong>g>Internati<strong>on</strong>al</str<strong>on</strong>g> <str<strong>on</strong>g>C<strong>on</strong>ference</str<strong>on</strong>g> <strong>on</strong><br />
L<strong>on</strong>g-<strong>term</strong> <strong>Experiments</strong>,<br />
Agricultural Research and<br />
Natural Resources<br />
Debrecen-Nyírlugos<br />
31 st May – 1 st June, 2007
JOINT INTERNATIONAL CONFERENCE ON LONG-TERM EXPERIMENTS,<br />
AGRICULTURAL RESEARCH AND NATURAL RESOURCES<br />
Debrecen-Nyírlugos 31 st May – 1 st June, 2007<br />
Organiser:<br />
Research Institute for Soil Science and Agricultural Chemistry of<br />
the Hungarian Academy of Sciences and<br />
the University of Debrecen Faculty of Agriculture<br />
Co-organisers:<br />
University of Oradea Faculty of Envir<strong>on</strong>mental Protecti<strong>on</strong><br />
Town Council Nyírlugos<br />
Editors:<br />
István Láng<br />
János Lazányi<br />
Nicolae Csép<br />
Lectors:<br />
János Lazányi<br />
Nicolae Csép<br />
Published with the financial support of<br />
Westsik Vilmos Foundati<strong>on</strong> for Rural Development in Nyírség Regi<strong>on</strong><br />
4400 Nyíregyháza Westsik Vilmos út 4-6. Hungary<br />
Published by:<br />
University of Debrecen Centre of Agricultural Sciences<br />
4032 Debrecen Böszörményi út 138. Hungary<br />
HU-ISBN: 978-963-473-054-5<br />
Printed in Pentaprint Printing House
CONTENTS<br />
INFLUENCE OF CHEMICAL FERTILIZERS, MANURE AND LIME<br />
APPLIED IN LONG TERM FIELD EXPERIMENTS ON PH-VALUE OF<br />
PRELUVOSOILS FROM NORTH-WEST PART OF ROMANIA<br />
Cristian Hera, Gheorghe Ciobanu, Cornelia Ciobanu,<br />
Cornel Domuţa, A. Vuşcan, Georghe Sarca<br />
EFFECT OF VARIOUS CROP PRODUCTION FACTORS ON THE YIELD<br />
AND YIELD STABILITY OF MAIZE IN A LONG-TERM EXPERIMENT<br />
Zoltán Berzsenyi, Dang Quoc Lap<br />
THE IMPACT OF PLANT BREEDING ON THE IMPROVEMENT OF SEED<br />
OIL CONTENT AND QUALITY IN EVENING PRIMROSE CROPS<br />
Andrew F. Fieldsend<br />
SOME LESSONS LEARNED FROM THE NYÍRLUGOS LONG-TERM FIELD<br />
EXPERIMENT Imre Kádár<br />
ANALYZE OF BIOMASS PRODUCTIVITY BY TIMESERIES<br />
REMOTESENSING DATA IN REGION OF NYÍRLUGOS<br />
János Tamás, Tibor Bíró, Nikolett Szőllősi<br />
RAINFALL, FERTILIZATION AND LIMING RESPONSE ON TRITICALE (X<br />
Triticosecale W.) YIELD IN THE 44 YEAR OLD NYÍRLUGOS FIELD TRIAL<br />
BETWEEN 1999 AND 2006 Márt<strong>on</strong> László<br />
EVALUATION OF THE DATA SET OF THE HUNGARIAN LONG-TERM K-<br />
FERTILIZATION FIELD TRIALS, SET UP BETWEEN 1960 AND 2000<br />
Péter Csathó<br />
LONG-TERM FIELD EXPERIMENTS AS THE BASE OF SITE SPECIFIC<br />
FARMING KESZTHELY LONG-TERM FIELD EXPERIMENTS<br />
Tamás Kismányoky, Zoltán Tóth<br />
LONG-TERM EFFECT OF ORGANIC AND MINERAL FERTILIZATION ON<br />
DIFFERENT SOIL-FERTILITY PARAMETERS<br />
Sándor Hoffmann, Katalin Berecz<br />
GREENHOUSE GAS EMISSION IN LONG-TERM TILLAGE AND<br />
ROTATION EXPERIMENTS<br />
Anita Gál, Péter Hegymegi, Erika Michéli, T<strong>on</strong>y J. Vyn<br />
INFLUENCES OF AMELIORATIVE FERTILIZATION ON YIELD AND<br />
NUTRITIONAL STATUS OF MAIZE<br />
Kovačević Vlado, Josipovic Marko, Stojić Biserka<br />
3<br />
12<br />
20<br />
29<br />
37<br />
44<br />
51<br />
56<br />
62<br />
68<br />
75<br />
83
YIELD AND QUALITY OF WINTER WHEAT GENOTYPES (Triticum<br />
aestivum L.) Desimir Knezevic, Veselinka Zecevic, Danica Micanovic,<br />
Nevena Djukic, Aleksandar Paunovic, Milomirka Madic, Ivica Djalovic<br />
CROP ROTATION AND GREEN MANURE INFLUENCE ON YIELD AND<br />
WATER USE EFFICIENCY IN WHEAT AND MAIZE FROM CRIS PLAIN<br />
Cornel Domuţa, Gheorghe Ciobanu, Maria Şandor, Alina Samuel, Cornelia<br />
Ciobanu, Nicu Cornel Sabău, Viorel Şcheau, Ioana Borza, Cristian Domuţa,<br />
APPLICATION OF A DIGESTATE AS A NUTRIENT SOURCE AND ITS<br />
EFFECT ON SOME CROPS AND SOIL PROPERTIES Marianna Makádi,<br />
Attila Tomócsik, József Lengyel, Zsolt Bogdányi, Árpád Márt<strong>on</strong><br />
SOIL TILLAGE SYSTEMS AND NITROGEN FERTILIZATION FOR<br />
WINTER BARLEY AFTER SOYBEAN<br />
Bojan Stipesevic, Danijel Jug, Miro Stosic, Ivan Zugec, Irena Jug<br />
RESEARCHES REGARDING THE INFLUENCE OF THE CROP ROTATION<br />
ON SOME INDICATORS OF THE WHEAT YIELD QUALITY IN THE<br />
CONDITIONS OF THE CRISURILOR PLAIN Gheorghe. Bandici , Cornel.<br />
Domuţa, Ileana Ardelean, Ioana Borza, Cristian Domuţa<br />
CRISANA -THE FIRST ROMANIAN WHEAT VARIETY WITH<br />
TOLERANCE TO ALUMINIUM IONS TOXICITY FROM ACID SOILS<br />
Gheorghe Bunta, Elena Bucurean<br />
EXPERIMENTS WITH PERENNIAL RYE (SECALE CEREANUM) IN<br />
HUNGARY, AT UNIVERSITY OF DEBRECEN Erika Halász, Tamás Sipos<br />
MYCOTOXINS CONTAMINATION IN FRESH AND SILAGE OF ALFALFA<br />
Branislav Gálik, D. Bíro, M. Juráček, M. Šimko, J. Michálková<br />
USAGE OF NATURAL ADDITIVES FOR BROILER CHICKENS<br />
Erika Horniaková, Ladislav Bušta<br />
PERSPECTIVES ON ROMANIAN-HUNGARIAN CROSS-BORDER<br />
COOPERATION. THE BIHOR - HAJDÚ-BIHAR EUROREGION MODEL<br />
Gabriela Popoviciu, M.C. Neacşu<br />
EFFECT OF BORON FERTILIZATION ON ANNUAL FLUCTUATION OF B<br />
IN SWEET CHERRY LEAVES AND FRUIT QUALITY Péter Tamás Nagy,<br />
Sándor Thurzó, Ida Kincses, Zoltán Szabó, József Nyéki<br />
DRY MATTER PRODUCTIVITY AND NITROGEN UPTAKE OF<br />
PERENNIAL RYEGRASS INFLUENCED BY NITROGEN AND WATER<br />
SUPPLY Imre Vágó, János Kátai, Ida Kincses, Andrea Balla Kovács<br />
4<br />
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95<br />
102<br />
108<br />
113<br />
119<br />
127<br />
132<br />
136<br />
140<br />
149<br />
156
THE EFFECT OF NITROGEN SUPPLY, LIMING AND NICKEL<br />
CONTAMINATION ON THE PRODUCTIVITY OF RYEGRASS (Lolium<br />
perenne L.) IN A GREENHOUSE EXPERIMENT Marianna Sipos, Imre Vágó<br />
TOXIGENIC FUNGI AND MYCOTOXINS IN GRAINS AND BAKERY<br />
PRODUCTS FROM ROMANIA Carmen Puia, Viorel Florian, Alexandra<br />
Suciu, Augusta Lujerdean, Dana Pusta<br />
CORRELATIONS OF SOCIAL AND ECONOMIC FUNCTIONS IN<br />
COMMUNITIES AROUND HORTOBÁGY Erzsébet Csengeri<br />
RIDING TOURISM IN THE NORTHERN GREAT PLAIN REGION<br />
(HUNGARY): STATUS AND POTENTIAL Réka Incze, Gábor Hevessy<br />
TISSUE CULTURE METHODS FOR SCREENING RESISTANCE TO<br />
PECTOBACTERIUM CHRYSANTHEMI IN POTATO<br />
Ildikó Hudák, Mária Hevesi, Judit Dobránszki Katalin Magyar-Tábori<br />
EFFECT OF AUXIN ON CALLUS FORMING CAPACITY OF DIFFERENT<br />
PEA GENOTYPES (PISUM SATIVUM L.) Katalin Magyar-Tábori,<br />
Judit Iszály-Tóth, Judit Dobránszki, Nóra Mendler-Drienyovszki<br />
COMBINING ABILITY STUDIES IN A SEVEN- PARENTAL DIALLEL<br />
CROSS OF DIFFERENT PEAS VARIETES (Pisum sativum L.) Nóra Mendler-<br />
Drienyovszki, Mándi Lajosné, Katalin Magyar-Tábori, Györgyi Cs<strong>on</strong>tos<br />
SUSTAINABLE LAND-USE BASED ON WESTSIK CROP ROTATION<br />
EXPERIMENT János Lazányi<br />
THE EFFECT OF LONG-TERM PHOSPHORUS FERTILIZATION ON THE<br />
TRYPTOPHAN AND ZINC CONTENT OF FEED PEAS<br />
Ágnes Elek, Zoltán Győri, Mária Borbély<br />
THE IMPORTANCE OF CALCIUM AND MAGNESIUM SUPPLY ON THE<br />
ACID BROWN FOREST SOILS IN THE NYIRSÉG REGION<br />
Jakab Loch, János Lazányi, Péter Tamás Nagy and Rita Kremper<br />
COMPARATIVE ANALYSIS OF CHEMICAL AND BIOLOGICAL SOIL<br />
EXAMINATION TO DETERMINE THE PLANT AVAILABLE N CONTENT<br />
OF SOIL IN THE NYÍRLUGOS LONG TERM FIELD EXPERIMENTS<br />
Péter Tamás Nagy, János Lazányi, Jakab Loch<br />
EFFECT OF FERTILIZATION SYSTEMS AND LIMING ON MOBILE<br />
PHOSPHORUS CONTENT IN DIFFERENT SOIL TYPES<br />
Augusta Olivia Lujerdean<br />
COMPOSTED SLAUGHTERHOUSE BY-PRODUCTS EFFECT ON CROP<br />
YIELD Péter Ragályi, Imre Kádár<br />
5<br />
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175<br />
181<br />
191<br />
196<br />
201<br />
207<br />
214<br />
220<br />
226<br />
234<br />
239
THE EFFECT OF BENTONITE ON SPECIFIC SOIL PARAMETERS AND<br />
MICROBIAL CHARACTERISTICS OF THE CARBON CYCLE János Kátai,<br />
Magdolna Tállai, János Lazányi, Edina Veres Lukácsné, Zsolt Sándor<br />
INVESTIGATION OF MINERAL CONTENT IN WINTER WHEAT GRAIN<br />
SAMPLES FROM LONG-TERM FIELD EXPERIMENTS<br />
Zoltán Győri, Árpád Tóth, Diana Ungai<br />
COMPARISON OF SOIL-TEST EXTRACTANTS FOR ZINK AND COPPER<br />
Rita Kremper, Sándor Berényi, Jakab Loch, Andrea Balla Kovács<br />
YIELD OF RYE AS AFFECTED BY THE CROPYEAR IN THE WESTSIK`S<br />
CROP ROTATION LONG-TERM FIELD EXPERIMENT<br />
István Henzsel, Gyuláné Györgyi<br />
RESULT OF ALFALFA CROPPING BY THE BASIC OF TIME- AND<br />
VILLAGE TYPE DATA FOR COUNTY OF SZABOLCS SZATMÁR BEREG<br />
László Nagy<br />
THE IMPORTANCE OF LONG-TERM FIELD EXPERIMENTS IN THE<br />
QUALITY ASSURANCE OF CROP PRODUCTION Éva Széles, Zsuzsanna<br />
Szathmáry, Péter Sipos, Dóra Hovánszki, Csilla Uri, Zoltán Győri<br />
MORPHOLOGICAL AND AFLP VARIATION IN SOME GENOTYPES OF<br />
POA ANGUSTIFOLIA L. AND POA HUMILIS EHRH. EX HOFFM Zsuzsa<br />
Lisztes, Ákos Szabó, A. Zubor, Béla Tóthmérész, Mária Papp, József Prokisch<br />
THE EFFECT OF SOME FOLIAR COMPOSITIONS ON THE PRODUCTION,<br />
NUMBER OF INFLORESCENCES AND FRUIT IN THE TECHNOLOGY OF<br />
CULTIVATING TOMATO IN THE FIELD Popa Alina Grigoriţa<br />
METHODS AND MEASURES FOR AN AGROCHEMICAL OPTIMIZATION<br />
OF THE GREENHOUSE SOIL AT THE TOMATOES CULTURE<br />
Popa Alina Grigoriţa<br />
THE INFLUENCE OF THE SEED’ BIOLOGICAL LINKS ON THE WHEAT<br />
PRODUCTION QUALITY Mariana Popovici<br />
THE INFLUENCE OF SOIL OIL POLLUTION ON AGRICULTURAL CROPS<br />
Maria Şandor, Nicu Cornel Sabău, Cornel Domuţa, Cristian Domuţa,<br />
Radu Brejea<br />
EFFECT OF BIOFERTILIZATION ON PARSLEY YIELD AND N CONTENT<br />
ON TWO SOILS<br />
Ida Kincses, Tibor Filep, Andrea B. Kovács, Péter T. Nagy, Imre Vágó<br />
6<br />
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255<br />
262<br />
269<br />
274<br />
279<br />
284<br />
392<br />
396<br />
300<br />
304<br />
312
THE INFLUENCE OF CHEMICAL AND BIOFERTILIZERS ON THE YIELD<br />
AND NITROGEN CONTENT OF LETTUCE (Lactuca sativa L.)<br />
Andrea Balla Kovács, Ida Kincses, Imre Vágó, Rita Kremper<br />
THE INFLUENCE OF USING BIOLOGICAL ADDITIVES ON THE<br />
NUTRITIONAL QUALITIES OF CORN SILAGE AND ON THE FATTENING<br />
PERFORMANCES OF BULL CALVES<br />
Daniel Mierlita; I. Chereji; Cristina Maerescu<br />
PHYTOPHTORA INFESTANS - ECOLOGICAL CONTROL WITHIN<br />
ROMANIAN (TRANSYLVANIAN PLANE) POTATO CROP<br />
Ioan Oroian, Liviu Hol<strong>on</strong>ec, Viorel Florian, Laura Paulette, Ant<strong>on</strong>ia Odagiu<br />
THE DYNAMICS OF THE POPULATIONS OF CEREAL PESTS AND<br />
PATHOGENS IN THE ROMANIAN WESTERN PLAIN<br />
Elena Bucurean, Nicolae Csép<br />
THE IMPORTANCE OF CONTROLLING FORMALDEHYDE FUMIGATION<br />
Lucian Bara, Camelia Bara<br />
THE CA AND MG SUPPLY OF THE SOIL EXTRACTED WITH DIFFERENT<br />
SOLVENT SOLUTIONS<br />
István Kocsis, Lajos Szabó, Simándi Péter, László Pásztor<br />
BIOLOGICAL INVESTIGATION ON AN EROSION CATENA<br />
B. Szeder, B. Sim<strong>on</strong>, M. Dombos, T. Szegi<br />
THE PRESENT STATE OF THE ROMANIAN AGRO-ALIMENTARY<br />
CONSUMPTION Anca M<strong>on</strong>ica Brata<br />
FROM FARM TO FORK – EVALUATION OF CROPPING FIELDS WITH<br />
REGARD TO FOOD AND FEED SAFETY AFTER LONG-STANDING<br />
CULTIVATION Péter Sipos, Zsuzsanna Szathmáry, Árpád Tóth, Diána<br />
Ungai, Dóra Hovánszki, Zoltán Győri<br />
MICROCLIMATIC STUDY OF THE MAIN WINE-GROWING REGIONS IN<br />
ROMANIA DURING 2006 Dorin Popa<br />
WHEAT GRAINS PROCESSING INFLUENCE AT QUALLITY<br />
PARAMETERS Timar Adrian<br />
OCEAN DYNAMICS: AIR-SEA INTERFACE GASES EXCHANGE<br />
Speranta Coldea<br />
RESEARCHES ON THE QUICK METHOD OF SUGAR PRODUCTIVITY<br />
EVALUATION Gheorghe Sarca<br />
RESEARCH REGARDING THE CONCENTRATION OF THIN JUICE AND<br />
THICK JUICE Gheorghe Sarca<br />
7<br />
319<br />
327<br />
334<br />
342<br />
347<br />
352<br />
357<br />
364<br />
369<br />
376<br />
382<br />
389<br />
397<br />
402
RESEARCH ON SEEDING AGENTS TO EFFECT THE SUGAR<br />
CRYSTALLIZATION Gheorghe Sarca<br />
WATER DIFFUSION IN BREAD DURING BAKING<br />
Ruska L., Chereji Rodica, Purcărea Cornelia, Timar A.<br />
BREAD PROPERTIES AND CRUMB STRUCTURE<br />
Ruska L., Chereji Rodica, Purcărea Cornelia, Timar A.<br />
THE MATHEMATICAL MODELLING OF THE PRESSURE REGULATOR<br />
AND OF THE ELECTROMAGNETIC INJECTION AT ENGINES WITH THE<br />
REVOLUTION AND THE TEMPERATURE OF THE ENVIRONMENT<br />
SURROUNDINGS Vasile Blaga, Nicolae Chioreanu, Adriana Cătaş<br />
THE VARIATION OF EFFECTIVE POWER AND SPECIFIC EFFECTIVE<br />
FUEL CONSUME OF 106-20 ENGINE WITH THE REVOLUTION AND THE<br />
TEMPERATURE OF THE ENVIRONMENT SURROUNDINGS<br />
Vasile Blaga, Nicolae Chioreanu, Adriana Cătaş<br />
THE EFFECT OF THE GENOTYPE AND EXPLANT UPON THE SOMATIC<br />
EMBRYO INDUCTION IN THREE QUERCUS SPECIES<br />
AI. Timofte, M. Palada,Nicolau, M. Ardelean, D. Pamfil,<br />
COMPARATIVE TESTS FOR SEPARATION METHODS OF<br />
ORGANOCHLORINE PESTICIDES IN MILK Adriana Chiş, Vasile Bara<br />
HIGHLIGHTING THE STATE OF FRESHNESS OF THE FRESH RAW<br />
MATERIAL FISH ON THE BASIS OF THE PHYSICAL-CHEMICAL<br />
PROPERTIES Purgea Ram<strong>on</strong>a<br />
ASSESSMENT OF THE HYGIENIC QUALITY OF THE FISH FRESH RAW<br />
MATERIAL BASED ON THE BACTERIOLOGICAL EXAMINATIONS<br />
Purgea Ram<strong>on</strong>a<br />
FORENSIC ASPECTS REGARDING DISASTER MANAGEMENT<br />
Carmen Radu, Camelia Buhaş, Nicolae Csép, Radu Gabriela, Andrei Csép<br />
ENERGY RECOVERY FROM ANIMAL PRODUCTS László Nagypál 470<br />
THE IMPLICATIONS OF MATERNAL TOXOPLASMOSIS INFECTION IN<br />
REPRODUCTION S<strong>on</strong>ia-Maria Drăghici, A. Csép, Ildiko Lenard, Adriana<br />
Jarca, Nicoleta Negruţ<br />
BOTULISM: CLINICAL – EPIDEMIOLOGICAL STUDY Ildiko Lenard,<br />
S<strong>on</strong>ia Draghici, Viorica Coldea, Mirela Indries,Nicoleta Negrut, Andrei Csép,<br />
Timea Lenard<br />
8<br />
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413<br />
420<br />
427<br />
435<br />
441<br />
453<br />
459<br />
462<br />
465<br />
474<br />
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STUDIES REGARDING THE INFLUENCE OF SALICYLIC ACID ON THE<br />
GERMINATION OF MAIZE CARYOPSIS (Zea mays) AND ON THE<br />
PEROXIDASE ACTIVITY IN MAIZE PLANTLETS<br />
Cornelia Purcarea, Dorina Cachită-Cosma<br />
THE INFLUENCE OF SALICYLIC ACID ON CONTENT OF<br />
ASSIMILATORY PIGMENTS IN THE PRIMARY LEAVES OF WHEAT<br />
(Triticum aestivum) PLANTLETS Cornelia Purcarea, Dorina Cachiţă-Cosma<br />
TREE GROWTH EQUATIONS IN DIFFERENT SITES FOR MIXED STANDS<br />
OF SESSILE OAK WITH BEECH FROM THE MIDDLE BASIN OF THE<br />
CRISUL REPEDE RIVER Sorin Dorog<br />
THE SIMULATION AND EVALUATION OF A BREAK IN MIXED STANDS<br />
OF BEECH WITH SESSILE OAK Sorin Dorog<br />
THE FREQUENCY OF DAYS WITH DIFFERENT TEMPERATURES IN THE<br />
CRISUL REPEDE HYDROGRAPHICAL BASIN Ana Cornelia Moza<br />
THE RISK OF INUNDATIONS INDUCED BY THE FLOODS ON THE<br />
RIVERS FROM ALMAS-AGRIJ DEPRESSION AND CLUJ - DEJ HILLS<br />
Alina-Daciana Dumitra<br />
RESEARCH REGARDING MAIN CHEMICAL REACTIONS IN<br />
PREDEFECATION OF DIFFUSION JUICE Gheorghe Sarca<br />
DETERMINATION OF NITROGEN-CONTAINING POLYNUCLEAR<br />
AROMATIC HYDROCARBONS IN SMOKED FOODS BY LIQUID<br />
CHROMATOGRAPHY Lucian Bara<br />
THE IMPORTANCE OF VOLATILE N-NITROSAMINES IN FOOD-<br />
PRODUCTS Lucian Bara, Camelia Bara, Vasile Bara, Dana Marele<br />
LUNG CANCER CAUSED BY SMOKING AND CADMIUM AND<br />
CHROMIUM IN HUMAN LUNG TISSUE Camelia Bara<br />
FORMATION OF AFLATOXIN M- TRIFLUOROACETIC ACID<br />
DERIVATIVE IN OPTIMUM CONDITIONS Lucian Bara<br />
DETERMINATION OF “CARCINOGENESIS” BY ALTERNATIVE<br />
BIOASSAYS Camelia Bara, Lucian Bara<br />
CLINICAL AND EPIDEMIOLOGICAL ASPECTS OF HEPATIC HYDATID<br />
CYST Mirela Indrieş, Viorica Coldea, Ciprian Brisc, Ildiko Lenard, S<strong>on</strong>ia<br />
Drăghici, Andrei Csép, Daciana Sabău, Nicoleta Negruţ<br />
THIN LAYER CHROMATOGRAPHY OF STERIGMATOCYSTIN IN<br />
CHEESE Bara Lucian<br />
9<br />
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492<br />
497<br />
502<br />
509<br />
516<br />
523<br />
528<br />
531<br />
539<br />
542<br />
546<br />
553<br />
561
TISSUE LEVELS AND BIOLOGICAL EFFECTS OF N-<br />
NITROSODIMETHYLAMINE IN MICE DURING CHRONIC LOW OR HIGH<br />
DOSE EXPOSURE WITH OR WITHOUT ETHANOL Bara Camelia<br />
DETERMINATION OF METALLOTHIONEIN, CADMIUM, COPPER AND<br />
ZINC LEVELS IN HUMAN AND RAT TISSUES Camelia Bara<br />
THE ROLE OF WATER QUALITY IN INFANTILE ALIMENTATION<br />
Andrei Csép, S<strong>on</strong>ia Draghici, Ildikó Lenard, Mirela Indries, Nicoleta Negrut<br />
SOME ASPECTS OF SHELTER REGENERATION OF QUERCUS SPP.<br />
FOREST STANDS FROM PRODUCTION UNIT II VALEA LUNGA<br />
(FORESTRY DISTRICT, LUGOJ (TIMIS, COUNTY) Ghiţă C. Crainic<br />
WASTEWATER TREATMENT TEHNOLOGIES OF BEER INDUSTRY OF<br />
ROMANIA<br />
Pantea Emilia Valentina, Mirel I., Romocea Tamara, Mitrasca Mihaela<br />
REGIONAL DEVELOPMENT USING CONSTRUCTED WETLANDS FOR<br />
WASTEWATER TREATMENT IN VALEA IERULUI (ÉRMELLÉK) REGION<br />
Bot<strong>on</strong>d L. Pete<br />
STUDY OF VERTICAL MOVEMENT OF SOME MICROELEMENTS IN<br />
THE SOIL László Szegedi Lajos Szabó Mária Takács Hájos<br />
Descrierea CIP a Bibliotecii Naţi<strong>on</strong>ale a României<br />
INTERNATIONAL CONFERENCE ON LONG-TERM EXPERIMENTS<br />
AGRICULTURAL RESEARCH, NATURAL RESOURCES AND<br />
SUSTAINABLE DEVELOPMENT (5; 2007; Debrecen-Oradea)<br />
<str<strong>on</strong>g>Internati<strong>on</strong>al</str<strong>on</strong>g> <str<strong>on</strong>g>C<strong>on</strong>ference</str<strong>on</strong>g> <strong>on</strong> L<strong>on</strong>g-<strong>term</strong> <strong>Experiments</strong>, Agricultural Research,<br />
Natural Resources and Sustainable Development:<br />
Debrecen-Nyírlugos-Oradea: 31 May-01 June 2007<br />
University of Debrecen, Faculty of Agriculture.<br />
University of Oradea, Faculty for Envir<strong>on</strong>mental Protecti<strong>on</strong><br />
Bibliogr.<br />
Index<br />
ISBN 978-973-759-298-9<br />
63(063)<br />
UNIVERSITY OF ORADEA PUBLISHING HOUSE<br />
ACCREDITED BY CNCSIS BUCHAREST, COD 149<br />
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PREFACE<br />
With the general aims to c<strong>on</strong>tribute to the exchange of opini<strong>on</strong>s and to<br />
the development of cross border cooperati<strong>on</strong> the <str<strong>on</strong>g>Internati<strong>on</strong>al</str<strong>on</strong>g><br />
<str<strong>on</strong>g>C<strong>on</strong>ference</str<strong>on</strong>g> <strong>on</strong> L<strong>on</strong>g-<strong>term</strong> <strong>Experiments</strong>, Agricultural Research and<br />
Natural Resources is dedicated to the 45 th anniversary of the<br />
establishment of the Field Experiment at Nyírlugos.<br />
In this regard, <strong>on</strong>e of the most important tasks is to evaluate results<br />
of l<strong>on</strong>g-<strong>term</strong> research sites, test sustainability of water and nutrient<br />
management systems and identify priorities of natural resource<br />
management. For sustainable use of natural resources, we need research<br />
in the principles of multifuncti<strong>on</strong>al agriculture and educati<strong>on</strong> that will<br />
address the problems of communities. The goal is to bring together<br />
people and resources, to promote an agriculture that is efficient,<br />
profitable, socially acceptable and envir<strong>on</strong>mentally sustainable.<br />
The primary objective is to provide a model, where the total<br />
agricultural system and community are taken into account, and<br />
agriculture is not separated from the natural ecosystem of the regi<strong>on</strong>. The<br />
most critical challenge is to c<strong>on</strong>sider the needs of technological<br />
development and natural ecosystem, and provide an educati<strong>on</strong>al<br />
envir<strong>on</strong>ment for people living in Nyírség regi<strong>on</strong>. The internati<strong>on</strong>al<br />
c<strong>on</strong>ference will also help cross border cooperati<strong>on</strong> <strong>on</strong> the sustainable use<br />
of natural resources. All Hungarian and foreign experts are welcome,<br />
including members and leaders of doctoral schools in the fields of<br />
agriculture, natural and social sciences.<br />
Debrecen-Nyírlugos 31. May 2007<br />
István Láng Tamás Németh<br />
János Nagy János Kátai<br />
Vasile Bara György Hovánszki<br />
11
INFLUENCE OF CHEMICAL FERTILIZERS, MANURE AND LIME APPLIED IN<br />
LONG TERM FIELD EXPERIMENTS ON PH-VALUE OF PRELUVOSOILS<br />
FROM NORTH-WEST PART OF ROMANIA<br />
Cristian Hera*, Gheorghe Ciobanu**, Cornelia Ciobanu**,<br />
Cornel Domuţa**, Adrian Vuşcan***, Georghe Sarca**<br />
* Academy of Agricultural and Forestry Sciences<br />
"GHEORGHE IONESCU-SISESTI" Bucuresti<br />
** University of Oradea, Faculty for Envir<strong>on</strong>mental Protecti<strong>on</strong> Oradea, Romania<br />
***Agricultural Research and Development Stati<strong>on</strong> Oradea<br />
ABSTRACT<br />
In Romania was elaborated since 1968 a stati<strong>on</strong>ary l<strong>on</strong>g <strong>term</strong> experiments with fertilizers<br />
and lime in all the Agricultural Research Stati<strong>on</strong>s bel<strong>on</strong>gs to Research Institute from<br />
Fundulea.<br />
The experiments was set aup using a unitary schem for knowing the evoluti<strong>on</strong> of soil<br />
fertilizers and the influence of fertilizers and lime rates and combinati<strong>on</strong>s <strong>on</strong> level and<br />
quality yield of different crops.<br />
The preluvosoil from North-West part of Romania is a medium soil, provide with the<br />
principale nutritive elements, with a weak acid reacti<strong>on</strong> in the ploughing horiz<strong>on</strong>.<br />
In this paper are presented the results regarding the influence of fertilizers and lime<br />
rates and combinati<strong>on</strong>s <strong>on</strong> pH values of preluvosoil from North-West part of Romania.<br />
L<strong>on</strong>g <strong>term</strong> fertilizati<strong>on</strong> with nitrogen de<strong>term</strong>ined the decrease of pH values and the<br />
increase of mobile aluminium c<strong>on</strong>tent up to the phytotoxicity level.<br />
Lime applicati<strong>on</strong> de<strong>term</strong>ined the increase of pH values and the decrease of mobile<br />
aluminium c<strong>on</strong>tent obtaining positive effects <strong>on</strong> yield.<br />
INTRODUCTION<br />
In Romania acid ploughing soils are spread <strong>on</strong> 2,0 milli<strong>on</strong>s ha which represent 20% from<br />
total agricol land.<br />
The factors which has a negative influence <strong>on</strong> growing plants are: high level<br />
c<strong>on</strong>centrati<strong>on</strong> by H + and Al 3+ , high level soil c<strong>on</strong>tent in Fe + and Mn 2+ and low level soil<br />
c<strong>on</strong>tent in principal nutrients elements, low activity of microorganisms, stagnati<strong>on</strong> of water,<br />
because of unsatisfactory infiltrati<strong>on</strong>.<br />
Much research <strong>on</strong> white luvic soil and preluvosoil c<strong>on</strong>diti<strong>on</strong>s (Bedo and Lang, 1977,<br />
Ciobanu and Nagy 1978, Nemeth 1996, Stefanescu 2003) has shown the negative effect of<br />
l<strong>on</strong>g-<strong>term</strong> applicati<strong>on</strong> of nitrogen as amm<strong>on</strong>ium nitrate <strong>on</strong> soil reacti<strong>on</strong>, which became more<br />
acidic and led to growth of mobile aluminum and manganese soil c<strong>on</strong>tent, which can<br />
de<strong>term</strong>ine phitotoxicity in the first part of vegetative period, with negative influence <strong>on</strong><br />
yield level and quality.<br />
12
For a better knowledge of applicati<strong>on</strong> effect <strong>on</strong> time of chemical fertilizers, manure and lime<br />
<strong>on</strong> soil chemistry was set up in the network of Agricultural Research Stati<strong>on</strong>s from<br />
Romania, l<strong>on</strong>g-<strong>term</strong> field experiments in different pedoclimatic c<strong>on</strong>diti<strong>on</strong>s.<br />
This paper presents the results regarding the influence of NP chemical fertilizers, manure<br />
and lime <strong>on</strong> evoluti<strong>on</strong> of preluvosoil acidity.<br />
MATERIAL AND METHOD<br />
Experimental site: The research data was obtained at the Agricultural and Development<br />
Research Stati<strong>on</strong> Oradea, using a unique design in the all research network of Research<br />
Institute from Fundulea. The investigati<strong>on</strong> has been carried out beginning with the autumn<br />
of 1974 in Oradea, in a flat plain area <strong>on</strong> the third terrace of the Crisul Repede river, whose<br />
geographical coordinates are: 21 0 56’ Eastern l<strong>on</strong>gitude, 47 0 03’ Northern latitude and 136 m<br />
altitude. The solidificati<strong>on</strong> rock c<strong>on</strong>sists of clay loam. The ground water is located at a depth<br />
of 6-8 m. the soil is a brown <strong>on</strong>e with horiz<strong>on</strong> dispositi<strong>on</strong> and the main physical and<br />
chemical characteristics are shown in table 1. The presence of clay migrati<strong>on</strong>, B horiz<strong>on</strong> is<br />
to be remarked noticed <strong>on</strong> the thickness of the soil profile, with high and very high values of<br />
the bulk density and compacti<strong>on</strong> level and low or very low total porosity and hydraulic<br />
c<strong>on</strong>ductivity.<br />
The soil reacti<strong>on</strong> is acid in the ploughing A horiz<strong>on</strong>, then slightly acid. The lack of<br />
CaCO3 in the soil profile is underlined. The mobile Al c<strong>on</strong>tent in the A horiz<strong>on</strong> may cause<br />
poor growth of some crops (clover). The soil is well provided with mobile potassium and<br />
phosphorus. The soil humus medium c<strong>on</strong>tent may not cause distorti<strong>on</strong>s to the neutr<strong>on</strong>ic<br />
de<strong>term</strong>inati<strong>on</strong> of the soil moisture.<br />
Soil depth<br />
cm<br />
Table 1: The main properties of the brown luvic soil from Oradea – Romania<br />
Sand Silt Clay OC Humus<br />
%<br />
Ca CO3<br />
%<br />
Al mobile<br />
mg/100g<br />
soil<br />
PH<br />
1:2<br />
H2O<br />
N<br />
Total<br />
%<br />
P mobile<br />
ppm<br />
Kmobile<br />
ppm<br />
0 - 5 43,5 28,3 28,2 1,25 2.32 0.00 3.68 6,3 0.12 21.8 83.0<br />
5 - 15 41,8 28,4 29,8 1,12 2.28 0.00 2.32 6,4 0.11 22.7 102.1<br />
15 - 30 40,0 28,5 31,5 1,02 1.91 0.00 0.52 6,3 0.09 5.7 112.1<br />
30 - 60 32,0 28,0 40,0 0,99 1.93 0.00 0.77 6,6 0.09 6.1 117.9<br />
60 - 90 24,1 36,7 39,2 0,29 0.00 0.32 6,6<br />
90 - 150 35,1 27,3 37,6 0,17 0.00 0.59 6,5<br />
The experimental factors in field experiments with NP fertilizers, manure and lime was:<br />
1. Field experiment with NP was set up in 1974 was used from plants in follow crop<br />
rotati<strong>on</strong>-pea-winter wheat-sunflower-maize<br />
P rates ware the same for each plant: 0, 40, 80, 120, 160 kg P2O5 ha -1<br />
N rates were differentiated:<br />
For pea-N0, N25, N50, N75, N100,<br />
13
For winter wheat and sunflower- N0, N40, N80, N120, N160,<br />
For maize- N0, N50, N100, N150, N200,<br />
The chemical NP fertilizers were: amm<strong>on</strong>ium nitrates and superphosphate<br />
2.Field experiment with manure and NP fertilizers was set up in 1974.<br />
Was used a short plant rotati<strong>on</strong> winter wheat-maize<br />
The manure was applied <strong>on</strong>ce at four years in autumn for maize using the rates: 0, 20, 40, 60<br />
to/ha.<br />
The NP rates were: N0P0, N50P0, N50P50 and N100P100,<br />
3.Field experiment with lime was set up in 1974 using a crop rotati<strong>on</strong>: pea, winter<br />
wheat, maize, alfalfa.<br />
The lime rates were: 0, 3, 6, 9 to/ha applied <strong>on</strong>ce at 6 years.<br />
NPK rates were differentiated:<br />
Pea: N0P80, N30P80, N120P80, N60P80, N90P80, N90P80K80,<br />
Winter wheat: N0P80, N30P80, N120P80, N160P80, N160P80K80,<br />
Maize: N0P80, N80P80, N160P80, N240P80, N240P80K80,<br />
Alfalfa: N0P100, N40P100, N80P100, N120P100, N120P100K80,<br />
Sampling and analytical method: Soil samples from top soil (0-20cm) were collected<br />
from each experiment plot, in august 2000,after wheat harvesting. All samples were taken to<br />
the laboratory and used for routine soil chemical analysis. pH was de<strong>term</strong>ined in water<br />
suspensi<strong>on</strong>.<br />
RESULTS AND DISCUSSION<br />
Influence of NP fertilizers <strong>on</strong> preluvosoil reacti<strong>on</strong>(figure 1)<br />
As an effect of systematically applying of NP fertilizers were registered significant<br />
modificati<strong>on</strong>s of soil acidity.<br />
The higher influence <strong>on</strong> soil reacti<strong>on</strong> had the nitrogen fertilizers applied <strong>on</strong> different P<br />
backgrounds.<br />
In the case of background P0, applying of N fertilizers lead to decreasing of pH values<br />
from 6,20 to 5,15 when N rate is taking values from 0 to 160 kg N/ha.<br />
In the case of all P backgrounds the decreasing trend of pH values due to N fertilizers<br />
applied is obvious. Based <strong>on</strong> the research data was established the interrelati<strong>on</strong>s existing<br />
between pH values and N rates.<br />
The phosphorus fertilizers applied affected more little pH values but it is noticed a low<br />
trend to increase soil acidity <strong>on</strong>ce with increasing of P rates, because of depleti<strong>on</strong> of bases<br />
as an effect of higher level of yield obtained.<br />
14
pH values<br />
pH values<br />
8<br />
6<br />
pH values<br />
6.50<br />
6.00<br />
5.50<br />
5.00<br />
4.50<br />
4.00<br />
N rates kg/ha<br />
Background P0<br />
y = 7E-06x 2 - 0.0076x + 6.2034<br />
R 2 = 0.9957<br />
N0 N40 N80 N120 N160<br />
4<br />
0 40 80 120 160<br />
8<br />
6<br />
N rates kg/ha<br />
Background P80<br />
y = -3E-05x 2 - 0.0015x + 5.9257<br />
R 2 = 0.9951<br />
4<br />
0 40 80 120 160<br />
N rates kg/ha<br />
pH values<br />
160<br />
8<br />
6<br />
80<br />
0<br />
P rates kg/ha<br />
Background P40<br />
y = -0.006x + 6.06<br />
R 2 = 0.9965<br />
4<br />
0 40 80<br />
N rates kg/ha<br />
120 160<br />
8<br />
6<br />
Background P160<br />
y = -3E-05x 2 - 0.0009x + 6.0611<br />
R 2 = 0.9982<br />
4<br />
0 40 80 120 160<br />
N rates kg/ha<br />
Figure 1: Influence of nitrogen and phosphorus <strong>on</strong> preluvosoil reacti<strong>on</strong> from N-W part<br />
of Romania<br />
The lower values of pH was registered in the case of plots fertilized with P80, when was<br />
obtained the higher level of producti<strong>on</strong>.<br />
15
Influence of manure applied <strong>on</strong> different NP backgrounds <strong>on</strong> preluvosoil reacti<strong>on</strong><br />
(figure 2)<br />
It is well known that the manure applied in acid soil c<strong>on</strong>diti<strong>on</strong>s are increasing the<br />
degree of base saturati<strong>on</strong> in the same time increasing the buffering capacity of the soil. On<br />
this way is possible to avoid unfavorable effect of chemical fertilizers with acid potential.<br />
pH values<br />
7.00<br />
6.00<br />
5.00<br />
Background N0P0<br />
y = 0.0002x 2 - 0.0011x + 6.273<br />
R 2 = 0.9601<br />
4.00<br />
0 20 40 60<br />
pH values<br />
7.00<br />
6.00<br />
5.00<br />
pH values<br />
Manure rates-to/ha<br />
Background N50P50<br />
y = 6E-06x 2 + 0.0061x + 5.8465<br />
R 2 = 0.9971<br />
4.00<br />
0 20 40 60<br />
7<br />
6<br />
5<br />
4<br />
Manure rates -to/ha<br />
60<br />
Manure rates- to/ha<br />
40<br />
20<br />
0<br />
pH values<br />
7.0<br />
6.0<br />
5.0<br />
4.0<br />
pH values<br />
Background N50P0<br />
y = 2E-05x 2 + 0.0052x + 5.609<br />
R 2 = 0.9998<br />
0 20 40 60<br />
7.00<br />
6.00<br />
5.00<br />
Manure rates- to/ha<br />
Background N100P100<br />
y = -4E-05x 2 + 0.0097x + 5.6885<br />
R 2 = 0.9858<br />
4.00<br />
0 20 40 60<br />
Manure rates- to/ha<br />
Figure 2: Influence of manure of preluvosoil reacti<strong>on</strong> from North-West part of Romania<br />
16<br />
N0P0<br />
N50P0<br />
N50P50<br />
N100P100<br />
NP rates kg/ha
In the preluvosoil c<strong>on</strong>diti<strong>on</strong>s the manure applied <strong>on</strong> different NP backgrounds had a<br />
significant positive effect <strong>on</strong> soil acidity. Applying manure in the rates of 20, 40 and 60<br />
to/ha in the lack of N, P fertilizers the pH values are increasing from 6, 29 to 6,76 units. In<br />
the case of the other NP backgrounds the manure de<strong>term</strong>ined an increase of pH values<br />
ranging between 0,4-0,6 units. The negative effect of nitrogen fertilizers applicati<strong>on</strong> is lower<br />
in the case of manure applicati<strong>on</strong>.<br />
Neutralizati<strong>on</strong> of soil acidity and completi<strong>on</strong> of calcium reserve (and magnesium)<br />
trough lime applicati<strong>on</strong> is a essential measure for increasing yield capacity of acid soils.<br />
In the case of preluvosoil applicati<strong>on</strong> of lime in the rate of 3, 6 and 9 to/ha <strong>on</strong>ce at six years<br />
lead to increasing of pH values depending <strong>on</strong> NP background utilized.<br />
When the lime is applied al<strong>on</strong>e pH values are taking values between 6,22 and 7, 08 when<br />
the lime rates are increasing from 0 to 9 to/ha.<br />
Influence of lime applicati<strong>on</strong> <strong>on</strong> preluvosoil reacti<strong>on</strong>(figure .3)<br />
pH values<br />
8<br />
6<br />
4<br />
CaCO3 rates<br />
t/ha<br />
Background N0P80<br />
y = 0.0056x 2 + 0.05x + 6.2<br />
R 2 = 1<br />
0 3 6 9<br />
CaCO3 rates - t/ha<br />
9<br />
6<br />
3<br />
0<br />
N0P80<br />
17<br />
4<br />
N160P80K80<br />
N160P80<br />
N90P80<br />
N80P80<br />
pH values<br />
8<br />
6<br />
4<br />
6<br />
8<br />
pH values<br />
NP rates kg/ha<br />
Background N80P80<br />
y = 0.12x + 5.76<br />
R 2 = 0.9529<br />
0 3 6 9<br />
CaCO3 rates - t/ha
pH values<br />
8<br />
6<br />
4<br />
Background N160P80<br />
y = -0.0028x 2 + 0.1683x + 5.155<br />
R 2 = 0.9581<br />
0 3 6 9<br />
CaCO3 rates - t/ha<br />
pH values<br />
8<br />
6<br />
4<br />
Background N160P80K80<br />
y = -0.0028x 2 + 0.195x + 4.885<br />
R 2 = 0.9966<br />
0 3 6 9<br />
CaCO3 rates - t/ha<br />
Figure 3: Influence of lime <strong>on</strong> preluvosoil reacti<strong>on</strong> from North-West part of Romania<br />
On the other NPK backgrounds the increasing of pH values are taking values between 0,9-<br />
1.42 pH units.<br />
Lime applicati<strong>on</strong> in preluvosoil c<strong>on</strong>diti<strong>on</strong>s is a necessary measure in the case of using NP<br />
fertilizers in high rates.<br />
CONCLUSIONS<br />
The results presented above let us come to the following c<strong>on</strong>clusi<strong>on</strong>s:<br />
1. L<strong>on</strong>g <strong>term</strong> experiments are important tools for examining soil fertility<br />
2. The soil reacti<strong>on</strong> evoluti<strong>on</strong> is depends by fertilizers type and by the rates level applied<br />
3. In the case of preluvosoil from North-West Romania by systematic applicati<strong>on</strong> of<br />
nitrogen fertilizers like as amm<strong>on</strong>ium nitrate leads to a decreasing of pH values from<br />
6.3 to 4.9 as a functi<strong>on</strong> of rates level applied<br />
4. Phosphorus fertilizers applied influence not to str<strong>on</strong>g preluvosoil reacti<strong>on</strong> but it can be<br />
observed a slow decreasing of pH values because of depleti<strong>on</strong> of bases due to yields<br />
spores obtained<br />
5. The manure applied al<strong>on</strong>e or associated with NP fertilizers, favorable influenced soil<br />
reacti<strong>on</strong>, pH values increasing with 0.3-0.4 units if the manure rates applied are 40<br />
to/ha respectively 60 to/ha<br />
6. For to avoid the decreasing pH values due to chemical fertilizers applied in preluvosoil<br />
c<strong>on</strong>diti<strong>on</strong>s is necessary lime applicati<strong>on</strong> for acidity neutralizati<strong>on</strong><br />
7. Lime applicati<strong>on</strong> <strong>on</strong>ce at six years in the rate of 9 to/ha maintain pH values between 6.4-<br />
7.0, which ensure optimal growing and developing c<strong>on</strong>diti<strong>on</strong> for plants<br />
18
REFERENCES<br />
Bedo, Z.- Lang, L. (1977) - Growing and breeding of quality wheat. Agro-21 Fuzetek,<br />
97/14, 8-28<br />
Ciobanu Gh.. (1999) - Influenta ingrasamintelor organice si minerale asupra productiei si<br />
calitatii graului si porumbului. "Agricultura durabila- performanta". Edit. Agric-<br />
Bucuresti.<br />
Hera Cr., Borlan Z., (1980) – Ghid pentru alcatuirea planurilor de fertilizare. Ed. Ceres<br />
Bucuresti.<br />
Hera Cr., Eliade Gh., Ghinea L., (1984) – Asigurarea azotului necesar culturilor agricole.<br />
Ed. Ceres Bucuresti.<br />
Nemeth, T. (1996) - Envir<strong>on</strong>ment friendly fertilizer recommendati<strong>on</strong> for sustainable<br />
agriculture. In: Envir<strong>on</strong>mental Polluti<strong>on</strong>, Ed. B. Nath et. All ECRP, Queen Mary and<br />
Westfield, college, L<strong>on</strong>d<strong>on</strong>, 99-105.<br />
19
EFFECT OF VARIOUS CROP PRODUCTION FACTORS ON THE YIELD<br />
AND YIELD STABILITY OF MAIZE IN A LONG-TERM EXPERIMENT<br />
Zoltán Berzsenyi – Dang Quoc Lap<br />
Agricultural Research Institute of the Hungarian Academy of Sciences, Mart<strong>on</strong>vásár<br />
ABSTRACT<br />
In a l<strong>on</strong>g-<strong>term</strong> experiment set up in 1960 <strong>on</strong> a medium heavy loamy soil, the effect of<br />
five crop producti<strong>on</strong> factors in increasing maize yields was studied in seven treatment<br />
combinati<strong>on</strong>s. The factors studied were soil cultivati<strong>on</strong>, fertilisati<strong>on</strong>, plant density,<br />
variety and weed c<strong>on</strong>trol. All the factors had a minimum and an optimum level. Yield<br />
data recorded over 42 years were evaluated using analysis of variance and stability<br />
analysis. The highest yield (8.59 t ha –1 ) was obtained when all the producti<strong>on</strong> factors<br />
were optimum and lowest (2.09 t ha –1 ) when these factors were at a minimum. When<br />
<strong>on</strong>ly <strong>on</strong>e factor was at a minimum and all the other factors were optimum the following<br />
yields were obtained (t ha –1 ): soil tillage: 8.32, fertilisati<strong>on</strong>: 5.21, genotype: 4.98, plant<br />
density: 6.31, weed c<strong>on</strong>trol: 7.01.The crop producti<strong>on</strong> factors c<strong>on</strong>tributed to the increase<br />
in maize yield in the following ratios (%): fertilisati<strong>on</strong> 30.6, variety 32.6, plant density<br />
20.2, weed c<strong>on</strong>trol 14.2, soil cultivati<strong>on</strong> 2.4. The highest value of CV%, expressing the<br />
deviati<strong>on</strong> of the yield averages, was obtained when all the producti<strong>on</strong> factors were at the<br />
minimum level (45.7%) and when weed c<strong>on</strong>trol or fertilisati<strong>on</strong> were at a minimum<br />
(36.6% and 34.8%, respectively), while the lowest value was recorded when all the<br />
factors were optimum (19.5%). The significant treatment × year interacti<strong>on</strong> could be<br />
attributed principally to treatments in which weed c<strong>on</strong>trol, fertilisati<strong>on</strong>, genotype or all<br />
the factors were at a minimum. The regressi<strong>on</strong> coefficient of linear regressi<strong>on</strong> analysis<br />
provided a satisfactory characterisati<strong>on</strong> of the stability of the treatments in different<br />
envir<strong>on</strong>ments, while the distance between the straight lines expressed the yield<br />
differences between the treatment pairs. The AMMI (Additive Main Effect and<br />
Multiplicative Interacti<strong>on</strong>) model proved to be a valuable approach for understanding<br />
agr<strong>on</strong>omic treatment x envir<strong>on</strong>ment interacti<strong>on</strong>s and assessing yield stability.<br />
Key words: l<strong>on</strong>g-<strong>term</strong> experiment, maize, producti<strong>on</strong> factors, stability analysis, AMMI<br />
model<br />
INTRODUCTION<br />
L<strong>on</strong>g-<strong>term</strong> experiments are often c<strong>on</strong>ducted to compare the l<strong>on</strong>g-<strong>term</strong> effects (e.g.,<br />
sustainability) of various treatments <strong>on</strong> <strong>on</strong>e or more resp<strong>on</strong>se variables. Measurements<br />
made <strong>on</strong> the plots are generally taken each year in the case of crop yield and other plant<br />
measurements. These data are ultimately subjected to some kind of statistical analysis,<br />
often with the goal of understanding something about the potential for different<br />
cumulative effects of treatments over time. As a result, the time x treatment interacti<strong>on</strong><br />
becomes the focus of an analysis.<br />
It is generally accepted that measurements made in field experiments may be<br />
influenced by unc<strong>on</strong>trollable envir<strong>on</strong>mental factors. Aside from random variati<strong>on</strong> there<br />
is also a predictable element, the l<strong>on</strong>g-<strong>term</strong> average of yield in the experiment. The<br />
20
mean (fixed, i.e. systematic effect) and the variance (random element) are the two main<br />
parameters describing the resp<strong>on</strong>se pattern of a cropping system. Variati<strong>on</strong>s in the<br />
measurements or in the treatment differences across years are simultaneously due to<br />
both the fixed and the random effects, and the two sets are often inseparable from each<br />
other.<br />
The improvement in maize producti<strong>on</strong> realized in the fields is the result of the<br />
combined effects of genetic, ecophysiological and technological changes superimposed<br />
<strong>on</strong> short- and l<strong>on</strong>g-<strong>term</strong> climatological variati<strong>on</strong>s. The simple partiti<strong>on</strong>ing of yield<br />
improvement into a genetic and an agr<strong>on</strong>omic comp<strong>on</strong>ent assumes the lack of<br />
interacti<strong>on</strong> between the two comp<strong>on</strong>ents. Although the increase has been the result of<br />
both genetic and agr<strong>on</strong>omic-management improvements, most of this improvement is<br />
the result of the genotype x management interacti<strong>on</strong> (Tollenaar and Lee 2002).<br />
The effect of major crop producti<strong>on</strong> factors has been investigated in l<strong>on</strong>g-<strong>term</strong><br />
experiments in Mart<strong>on</strong>vásár since 1960 (Győrffy 1969, Berzsenyi and Győrffy 1995).<br />
The aim of the present work was to carry out a biometric analysis <strong>on</strong> the 42-year data<br />
series from the experiments in order to de<strong>term</strong>ine how the minimum and optimum<br />
levels of five producti<strong>on</strong> factors (tillage, fertilisati<strong>on</strong>, plant density, variety, weed<br />
c<strong>on</strong>trol) influenced the yield of maize, and to use the single-variable and multi-variable<br />
methods of stability analysis to de<strong>term</strong>ine the stability of these producti<strong>on</strong> factors in<br />
diverse envir<strong>on</strong>ments.<br />
MATERIAL AND METHODS<br />
The effect of five crop producti<strong>on</strong> factors <strong>on</strong> maize yields has been investigated since<br />
1960 in a l<strong>on</strong>g-<strong>term</strong> experiment set up <strong>on</strong> medium heavy loam soil. Each factor is<br />
represented by two treatments, <strong>on</strong>e minimum and <strong>on</strong>e optimum. The treatments applied<br />
for each factor were as follows:<br />
Factor Minimum level Optimum level<br />
Tillage * Shallow ploughing Deep ploughing<br />
to a depth of 12–14 cm to a depth of 24–28 cm<br />
Fertilisati<strong>on</strong> ** No fertilisati<strong>on</strong> 60 t ha –1 farmyard manure<br />
every 4 years;<br />
N140P60K60 annually<br />
Plant density ‡ 35,000 plants ha –1 70,000 plants ha –1<br />
Variety Open-pollinated: Hybrid<br />
Aranyözön and<br />
Mindszentpusztai Sárga<br />
Weed c<strong>on</strong>trol Poor: late thinning and Good: complete freedom<br />
two delayed hoeings from weeds; timely<br />
thinning<br />
* From 1985 <strong>on</strong>wards the optimum level was deep loosening every four years<br />
** From 1980 <strong>on</strong>wards the fertiliser rate was doubled<br />
‡ Until 1980 the minimum plant density was 20,000 plants ha –1 and the optimum 40,000<br />
Seven treatment combinati<strong>on</strong>s were used in the experiment. In Treatment 1 all the<br />
factors were at the minimum level and in Treatment 2 all were optimum. In Treatments<br />
3–5 <strong>on</strong>e of the crop producti<strong>on</strong> factors was at a minimum, while all the others were<br />
21
optimum. The factors at a minimum were tillage in Treatment 3, fertilisati<strong>on</strong> in<br />
Treatment 4, plant density in Treatment 5, variety in Treatment 6 and weed c<strong>on</strong>trol in<br />
Treatment 7.<br />
The experimental results were evaluated using a variety of methods. The<br />
cumulative yield analysis method (Sváb 1981) designed for the evaluati<strong>on</strong> of l<strong>on</strong>g-<strong>term</strong><br />
experiments was used to compare the cumulative yield-enhancing effect of individual<br />
producti<strong>on</strong> factors compared with the minimum combinati<strong>on</strong>. Analysis of variance was<br />
performed by using the “General Analysis of Variance” procedure of the GenStat 9<br />
program. Analysis was c<strong>on</strong>ducted separately for each year and pooled together<br />
afterwards. Stability analysis <strong>on</strong> the experimental treatments was carried out using<br />
single-variable (variance and regressi<strong>on</strong> parameters) and multi-variable (AMMI)<br />
methods. Am<strong>on</strong>g the variance parameters, the ecovalence (W²), the stability variance<br />
(σ²) and the yield stability (YS) parameters were calculated using the STABLE model<br />
proposed by Kang (1995), where the covariant was the annual rainfall quantity from<br />
April to September. The regressi<strong>on</strong> method of stability analysis was applied as<br />
described by Finlay and Wilkins<strong>on</strong> (1963). The year × treatment interacti<strong>on</strong> was<br />
investigated by means of AMMI analysis, a combinati<strong>on</strong> of analysis of variance and<br />
principal comp<strong>on</strong>ent analysis (Crossa 1990).<br />
RESULTS AND DISCUSSION<br />
Annual analysis of variance indicated that the effect of the experimental treatments was<br />
significant at the P=0.1% level every year. Combined ANOVA over the 42 years<br />
dem<strong>on</strong>strated that both the year effect and the treatment effect were significant, but the<br />
MS values showed that the treatment effect was more than 16 times as str<strong>on</strong>g as the<br />
year effect (Table 1).<br />
Table 1. Analysis of variance of the l<strong>on</strong>g-<strong>term</strong> experiment (1960-2001)<br />
Source df SS MS F-value<br />
Total 1175 8540<br />
Envir<strong>on</strong>ments (Years) 41 2091 51.0 92.5 ***<br />
Replicati<strong>on</strong> 3 16.6 5.54 10.1<br />
Year (replicati<strong>on</strong>) 123 67.8 0.55<br />
Treatments 6 5054 842.3 643.9 ***<br />
Residuals 1002 1311 1.31<br />
The cumulative yield-enhancing effect of the producti<strong>on</strong> factors compared with the<br />
minimum combinati<strong>on</strong> is illustrated for the years 1960–2001 in Figure 1. It is clear from<br />
the figure that under Hungarian c<strong>on</strong>diti<strong>on</strong>s deep tillage <strong>on</strong>ly had a slight effect, ranging<br />
from 2.4–3.0% over the period in questi<strong>on</strong>. A pr<strong>on</strong>ounced increase in the fertiliser effect<br />
was observed in the l<strong>on</strong>g-<strong>term</strong> experiment, from 18.6% in the early years to 30.7%. The<br />
effect of an increase in plant density <strong>on</strong> maize yields changed little over the 42 years,<br />
having values of 19.9–23.3%. Apart from fertilisati<strong>on</strong>, the factor with the greatest<br />
influence <strong>on</strong> maize yields was the genotype, the effect of which increased from 26.6%<br />
to 32.6% over the years. The effect of weed c<strong>on</strong>trol <strong>on</strong> the maize yield ranged from 23–<br />
22
29% in the first 15 years of the experiment, but has declined to 14–18% over the last ten<br />
years.<br />
Cumulative yield increase t ha -1<br />
Years<br />
Basic treatment (1)<br />
Figure 1. Cumulative yield increase in various treatments compared to the minimum<br />
combinati<strong>on</strong> of factors (1960-2001)<br />
The effect of the crop producti<strong>on</strong> factors in increasing maize yields is depicted in Figure<br />
2, averaged over the 42 years. The greatest difference in the mean yield resp<strong>on</strong>se was<br />
found between treatments where all the factors were minimum (2.09 t ha –1 ) or optimum<br />
(8.59 t ha –1 ). When <strong>on</strong>ly <strong>on</strong>e factor was at a minimum and all the others optimum, the<br />
depth of soil tillage was found to have the least effect <strong>on</strong> maize yields (mean yield<br />
resp<strong>on</strong>se: 8.32 t ha –1 ), while both fertilisati<strong>on</strong> and genotype had a substantial influence<br />
<strong>on</strong> the yield (mean yield resp<strong>on</strong>se: 5.21 and 4.98 t ha –1 , respectively). The minimum<br />
level of plant density or weed c<strong>on</strong>trol led to yield reducti<strong>on</strong>s of 2.2 and 1.6 t ha –1 (mean<br />
yield resp<strong>on</strong>ses of 6.36 and 7.01 t ha –1 ). Based <strong>on</strong> this 42-year data series, the crop<br />
producti<strong>on</strong> factors were found to make the following c<strong>on</strong>tributi<strong>on</strong>s to yield increases in<br />
maize: variety 32.6%, fertilisati<strong>on</strong> 30.6%, plant density 20.2%, weed c<strong>on</strong>trol 14.2% and<br />
soil tillage 2.4%.<br />
The variance and regressi<strong>on</strong> parameters of stability analysis for each experimental<br />
treatment are presented in Table 2. The CV% value expressing the annual deviati<strong>on</strong> in<br />
yield averages was the greatest when all the producti<strong>on</strong> factors were at a minimum<br />
(45.7%) and smallest when they were all at the optimum level (19.5%). When <strong>on</strong>e<br />
23
factor was at the minimum level and the remainder were optimum the highest values of<br />
CV% were observed for weed c<strong>on</strong>trol and fertilisati<strong>on</strong> (36.6% and 34.8%, respectively).<br />
T F D V W<br />
0 0 0 0 0<br />
1 1 1 0 1<br />
1 0 1 1 1<br />
1 1 0 1 1<br />
1 1 1 1 0<br />
0 1 1 1 1<br />
1 1 1 1 1<br />
Treatments<br />
0 1 2 3 4 5 6 7 8 9<br />
Maize grain yield t ha -1<br />
Figure 2. Effect of crop producti<strong>on</strong> factors <strong>on</strong> maize grain yield increments in a l<strong>on</strong>g<strong>term</strong><br />
experiment (Mart<strong>on</strong>vásár 1960-2001). Treatments : T: Depth of soil tillage, F:<br />
Fertilisati<strong>on</strong>, D: Plant density, V: Variety, W: Weed c<strong>on</strong>trol. Each factor at two levels:<br />
0: Minimum level, 1: Optimum level. Treatments designated with the same letter did<br />
not differ significantly according to Duncan’s test.<br />
The magnitude of the ecovalence (W²) and stability variance (σ²) parameters expresses<br />
the c<strong>on</strong>tributi<strong>on</strong> of the treatments to the significant year × treatment interacti<strong>on</strong>. It can<br />
be seen that treatments in which weed c<strong>on</strong>trol, fertilisati<strong>on</strong>, genotype or all the factors<br />
were minimum were chiefly resp<strong>on</strong>sible for the interacti<strong>on</strong>. Based <strong>on</strong> the mean yield<br />
resp<strong>on</strong>se and the stability, the yield stability (YS) parameters indicated that Treatments<br />
2, 3, 5 and 7 were the most favourable (Table 2).<br />
When the effects of the treatments were investigated in diverse envir<strong>on</strong>ments by<br />
means of linear regressi<strong>on</strong> analysis (Fig. 3), the greatest difference in mean yield<br />
resp<strong>on</strong>se was observed between the treatments in which all the factors were minimum<br />
or optimum. The yield difference between the two treatments (expressed as the distance<br />
between the straight lines) increased with the envir<strong>on</strong>mental mean. The figure shows<br />
that the slight effect of tillage depth could <strong>on</strong>ly be detected for a low envir<strong>on</strong>mental<br />
mean. When fertilisati<strong>on</strong> was at a minimum the magnitude of the yield reducti<strong>on</strong> was<br />
c<strong>on</strong>siderable and increased with the value of the envir<strong>on</strong>mental mean. The magnitude<br />
and trend of yield reducti<strong>on</strong> was similar when the genotype was at a minimum. There<br />
was also a similar tendency for the plant density, but the yield reducti<strong>on</strong> was smaller in<br />
this case, particular in the case of a high envir<strong>on</strong>mental mean.<br />
24<br />
F<br />
E<br />
E<br />
D<br />
C<br />
B<br />
A
Table 2. Variance and regressi<strong>on</strong> parameters of the stability of the producti<strong>on</strong> factors<br />
in the l<strong>on</strong>g-<strong>term</strong> experiment (1960-2001)<br />
Stability<br />
parameters<br />
Coefficient of<br />
1 2 3 4 5 6 7<br />
variati<strong>on</strong> (CV%)<br />
Ecovalence<br />
45.7 19.5 21.9 34.8 24.5 25.1 36.6<br />
(W 2 )<br />
Stability variance<br />
117.7 51.0 53.5 219.2 57.1 163.2 325.9<br />
(σ 2 )<br />
Yield stability<br />
3.22 0.94 1.02 6.68 1.15 4.77 10.32<br />
(YS) **<br />
Correlati<strong>on</strong><br />
+ +<br />
+<br />
+<br />
coefficient (r)<br />
Regressi<strong>on</strong><br />
0.796 0.947 0.962 0.713 0.919 0.690 0.899<br />
coefficient (b)<br />
Mean yield<br />
0.500 1.145 1.236 0.860 1.020 0.580 1.659<br />
resp<strong>on</strong>se t ha -1<br />
2.094F 8.585A 8.315B 5.205E 6.355D 4.979E 7.014C<br />
Treatments *<br />
* Treatments: 1. Minimum values of all factors, 2. Optimum values of all factors, 3.<br />
Minimum depth of soil cultivati<strong>on</strong>, 4. Minimum level of fertilisati<strong>on</strong>, 5. Minimum level<br />
of plant density, 6. Minimum for variety, 7. Minimum weed c<strong>on</strong>trol.<br />
** +: variants selected by the STABLE programme.<br />
Table 3. AMMI Analysis for the l<strong>on</strong>g-<strong>term</strong> experiment (1960-2001)<br />
Source df SS MS F-value<br />
Total 1175 8540<br />
Treatments 6 5054 842.3 1969.7 ***<br />
Envir<strong>on</strong>ments 41 2091 51.0 76.1 ***<br />
Block 126 84 0.67<br />
Interacti<strong>on</strong>s 246 988 4.0 9.39 ***<br />
IPCA1 46 467 10.2 23.8 ***<br />
IPCA2 44 238 5.4 12.7 ***<br />
Residuals 156 282 1.8 4.23 ***<br />
Error 756 323 0.43<br />
Compared with the other producti<strong>on</strong> factors the weed c<strong>on</strong>trol minimum exhibited the<br />
opposite envir<strong>on</strong>mental dependence, indicating that weed infestati<strong>on</strong> had the greatest<br />
yield-reducing effect at low envir<strong>on</strong>mental means and the smallest effect at an<br />
envir<strong>on</strong>mental index of 10 t ha –1 . C<strong>on</strong>sequently, it can be stated that Treatments 2, 3 and<br />
5 have average stability (regressi<strong>on</strong> coefficients of around 1), while the approximately b<br />
= 0.5 value recorded for Treatments 1 and 6 and the b
Maize grain yield t ha -1<br />
Maize grain yield t ha -1<br />
Maize grain yield t ha -1<br />
18<br />
16<br />
14<br />
12<br />
10<br />
18<br />
16<br />
14<br />
12<br />
10<br />
8<br />
6<br />
4<br />
2<br />
0<br />
8<br />
6<br />
4<br />
2<br />
0<br />
18<br />
16<br />
14<br />
12<br />
10<br />
8<br />
6<br />
4<br />
2<br />
0<br />
Optimum values<br />
of all factors<br />
Minimum<br />
values of all<br />
factors<br />
0 2 4 6 8 10 12<br />
Optimum values<br />
of all factors<br />
0 2 4 6 8 10 12<br />
Optimum values<br />
of all factors<br />
Minimum level<br />
of fertilisati<strong>on</strong><br />
Minimum for<br />
genotype<br />
0 2 4 6 8 10 12<br />
Envir<strong>on</strong>mental index t ha -1<br />
Figure 3. Yield resp<strong>on</strong>se of maize for minimum levels of producti<strong>on</strong> factors vs.<br />
optimum level of all factors in comparisi<strong>on</strong> to the envir<strong>on</strong>mental index in the l<strong>on</strong>g<strong>term</strong><br />
experiment (1960-2001)<br />
26<br />
18<br />
16<br />
14<br />
12<br />
10<br />
8<br />
6<br />
4<br />
2<br />
0<br />
18<br />
16<br />
14<br />
12<br />
10<br />
8<br />
6<br />
4<br />
2<br />
0<br />
18<br />
16<br />
14<br />
12<br />
10<br />
8<br />
6<br />
4<br />
2<br />
0<br />
Optimum values<br />
of all factors<br />
Minimum depth of<br />
soil cultivati<strong>on</strong><br />
0 2 4 6 8 10 12<br />
Optimum values of<br />
all factors<br />
Minimum plant<br />
density<br />
0 2 4 6 8 10 12<br />
Optimum values<br />
of all factors<br />
Minimum weed<br />
c<strong>on</strong>trol<br />
0 2 4 6 8 10 12<br />
Envir<strong>on</strong>mental index t ha -1
The b = 1.7 value recorded for Treatment 7 can be explained partly by the lower extent<br />
of weed interference in a favourable envir<strong>on</strong>ment.<br />
The main effects of year and treatments and the interacti<strong>on</strong>s were also examined<br />
using AMMI analysis, which combines ANOVA and principal comp<strong>on</strong>ent analysis<br />
(Table 3). The first and sec<strong>on</strong>d principal comp<strong>on</strong>ents (PCA1, PCA2) explained 71.4%<br />
of the interacti<strong>on</strong> SS. The results of the AMMI analysis are illustrated in Figure 4, with<br />
the yield average <strong>on</strong> the X axis and the value of the first principal comp<strong>on</strong>ent <strong>on</strong> the Y<br />
axis, for seven treatments in 42 envir<strong>on</strong>ments (years). The higher the value of the<br />
principal comp<strong>on</strong>ent, the greater the c<strong>on</strong>tributi<strong>on</strong> of the treatment to the interacti<strong>on</strong>, i.e.<br />
the lower the yield stability. It can be seen that Treatments 7, 6, 1 and 4 made the<br />
greatest c<strong>on</strong>tributi<strong>on</strong>s to the interacti<strong>on</strong>, while Treatments 2, 3 and 5 had the greatest<br />
yield stability.<br />
First principal comp<strong>on</strong>ent scores<br />
3<br />
2<br />
1<br />
0<br />
-1<br />
-2<br />
G1<br />
G7<br />
.<br />
E40<br />
E27<br />
E33 .<br />
E32<br />
.<br />
E26<br />
E42 E3 7<br />
E22 .<br />
E38 G3 E30<br />
E17 E35 E23 .<br />
.<br />
E19 E16<br />
. E39 E10 E41 G2<br />
E34 E14 .<br />
. . . . .<br />
. . . . . G5 E36<br />
. . . . . . E21 . . . . .<br />
. . .<br />
. . . E28 . . . . . . . .<br />
E18<br />
E31<br />
.<br />
E1 E20<br />
.<br />
E4E13<br />
E8<br />
.<br />
E24<br />
E15<br />
.<br />
. E1 2<br />
E9<br />
E11<br />
E5 E7<br />
E25<br />
E6<br />
.<br />
.<br />
E2<br />
E29<br />
E3<br />
.<br />
.<br />
G4<br />
G6<br />
2 4<br />
6<br />
8<br />
Mean yields t ha -1<br />
Figure 4. Plot of the mean yields and first principal comp<strong>on</strong>ent scores of crop<br />
producti<strong>on</strong> factors in 42 envir<strong>on</strong>ments<br />
ACKNOWLEDGEMENTS<br />
This research was supported by a grant from the Nati<strong>on</strong>al Committee for Technological<br />
Development (OMFB-00895/2005).<br />
27<br />
10<br />
12
REFERENCES<br />
Berzsenyi Z., Győrffy B. (1995): Effect of various crop producti<strong>on</strong> factors <strong>on</strong> the yield<br />
and yield stability of maize (Különböző növény<strong>term</strong>esztési tényezők hatása a<br />
kukorica <strong>term</strong>ésére és <strong>term</strong>ésstabilitására). Növény<strong>term</strong>elés, 44: 507-517.<br />
Crossa, J. (1990): Statistical analyses of multilocati<strong>on</strong> trials. Advances in Agr<strong>on</strong>omy,<br />
44: 55-85.<br />
Finlay, K.W., Wilkins<strong>on</strong>, G.N. (1963): The analysis of adaptati<strong>on</strong> in a plant breeding<br />
programme. Aust. J. Agric. Res., 14: 742-754.<br />
Győrffy B (1969): Effect of crop producti<strong>on</strong> factors <strong>on</strong> maize yields, Experiment code:<br />
Komplex I (Különböző növény<strong>term</strong>esztési tényezők hatása a kukorica <strong>term</strong>ésére,<br />
Komplex I). In: I’só I. (ed.), Maize Producti<strong>on</strong> <strong>Experiments</strong> 1965–1968<br />
(Kukorica<strong>term</strong>esztési kísérletek 1965-1968). Akadémiai Kiadó, Budapest.<br />
Kang, M.S. (1995): Simultaneous selecti<strong>on</strong> for yield and stability in crop performance<br />
trials: c<strong>on</strong>sequences for growers. Agr<strong>on</strong>omy Journal, 85: 754-757.<br />
Sváb J. (1981): Biometrical Methods in Research (Biometriai módszerek a kutatásban).<br />
Mezőgazdasági Kiadó, Budapest.<br />
Tollenaar, M., Lee, E.A. (2002): Yield potential, yield stability and stress tolerance in<br />
maize. Field Crops Research, 75: 161-169.<br />
28
THE IMPACT OF PLANT BREEDING ON SEED OIL CONTENT AND<br />
QUALITY IN EVENING PRIMROSE CROPS<br />
ABSTRACT<br />
Andrew F. Fieldsend<br />
University of Debrecen Centre of Agricultural Sciences<br />
4032 Debrecen, Böszörményi ut 138., Hungary<br />
In recent decades, evening primrose (Oenothera spp.) has been grown as an agricultural<br />
crop in several parts of the world including western and central Europe, North America<br />
and New Zealand. Its value is derived from its seed oil which c<strong>on</strong>tains unusually high<br />
levels of gamma linolenic acid (GLA), an uncomm<strong>on</strong> fatty acid which is used in<br />
pharmaceuticals and nutriti<strong>on</strong>al supplements. Whilst such improvements have led to a<br />
reducti<strong>on</strong> in the seed price, increases in seed oil c<strong>on</strong>tent and quality (i.e. percentage of<br />
GLA) can lead to further reducti<strong>on</strong>s in the cost of the end product. The breeding<br />
programme of Scotia Pharmaceuticals Ltd, UK was effective in achieving such<br />
increases. For example, in spring-sown crops in New Zealand, seed oil c<strong>on</strong>tent<br />
increased from around 25% in older cultivars grown in the period 1990-1992 to around<br />
28% in the cultivar “Rigel” grown in the period 1992-1999. Furthermore, GLA c<strong>on</strong>tent<br />
increased by almost 50%, from around 9.5% in older cultivars to more than 13% in<br />
“Rigel”. It has been estimated that these improvements through plant breeding have led<br />
to an 18% reducti<strong>on</strong> in product costs.<br />
Keywords: Evening primrose; Oenothera spp; breeding; oil c<strong>on</strong>tent; oil quality<br />
INTRODUCTION<br />
In recent decades, evening primrose (Oenothera spp.) has been grown as an agricultural<br />
crop in several parts of the world including western and central Europe, North America<br />
and New Zealand. Its value is derived from its seed oil which c<strong>on</strong>tains unusually high<br />
levels of gamma linolenic acid (GLA), an uncomm<strong>on</strong> fatty acid which is used in<br />
pharmaceuticals and nutriti<strong>on</strong>al supplements (Horrobin, 1992). Although found in some<br />
other plant and fungal oils, in evening primrose oil GLA is thought to occur in its most<br />
bioactive state, as part of a triacylglycerol called Enotherol® (Horrobin, 1994). A brief<br />
botanical descripti<strong>on</strong> of evening primrose was given by Fieldsend (1996a).<br />
Early commercial crops of evening primrose were grown from seed stocks available<br />
through the horticultural trade (Fieldsend, 1996a). One such “traditi<strong>on</strong>al” stock, grown<br />
in New Zealand, was simply known as “421”. This and similar lines were unsuitable for<br />
agriculture in many ways, for example in having seed capsules which split as they<br />
matured, meaning that a significant proporti<strong>on</strong> of seed was lost prior to harvest. This<br />
factor simply compounded the problem that the potential seed yield of evening primrose<br />
was already much lower than that of other combinable crops (Nix, 1995).<br />
UK-based Agricultural Holdings, owned by the Balint family, was an important early<br />
seed producer, sourcing much of its seed from Hungary. Its subsidiary, which in time<br />
became Scotia Pharmaceuticals Ltd, a leading manufacturer of products derived from<br />
evening primrose oil, set up an evening primrose breeding programme in the mid-<br />
1970s. Whilst development of cultivars with n<strong>on</strong>-splitting capsules was an important<br />
29
eeding objective, as higher seed yields would lead to a reducti<strong>on</strong> in the price per t<strong>on</strong>ne<br />
of seed, Scotia recognised that of equal importance was an increase in the oil (for<br />
nutriti<strong>on</strong>al supplements) and GLA (for pharmaceuticals) c<strong>on</strong>tents of the seed. The<br />
difference is that whilst the former are normally c<strong>on</strong>sumed by users as a fixed volume<br />
of oil per day, for the latter the daily dose of GLA is the important criteri<strong>on</strong>.<br />
Evening primrose breeding is made difficult by its unusual genetic system (Cleland,<br />
1972). The formati<strong>on</strong> of chromosome rings at meiosis, ordered chromosome movement<br />
and the presence of lethal genes limit segregati<strong>on</strong> (and therefore scope for selecting<br />
improved lines) in the generati<strong>on</strong>s following the initial cross. Variable establishment,<br />
coupled with seed dormancy in many genotypes, and the l<strong>on</strong>g growing seas<strong>on</strong> of the<br />
overwintered crop in temperate climates (approximately 14 m<strong>on</strong>ths) also present<br />
practical problems (Fieldsend and Moris<strong>on</strong>, 2000a). For unimproved lines, field sowing<br />
would be an unreliable method of trialling, and selecti<strong>on</strong> of lines to trial would need to<br />
be made before the previous generati<strong>on</strong> of breeding material had been harvested.<br />
This paper describes the methods developed by Scotia to breed evening primrose and<br />
the improvements in oil c<strong>on</strong>tent and quality of commercial crops in New Zealand which<br />
have resulted from the use of cultivars developed by the programme.<br />
MATERIALS AND METHODS<br />
The Scotia breeding programme began with the collecti<strong>on</strong>, from many sources, of over<br />
2000 individual “accessi<strong>on</strong>s” which were numbered in a single series from X1 upwards.<br />
Each accessi<strong>on</strong> was evaluated in a single spring-transplanted row of 21 plants. Seeds<br />
were sown in potting compost in 750 mm diameter pots in January and placed in a<br />
refrigerator set at 4ºC for 2-3 weeks to break dormancy, following which the pots were<br />
transferred to a propagator hotbox at a temperature of 27ºC. So<strong>on</strong> after emergence 24<br />
seedlings of each accessi<strong>on</strong> were pricked out into potting compost in modules and kept<br />
in a heated glasshouse with a mean temperature of 13ºC. The rosettes were moved out<br />
of the glasshouse to acclimate during late April and transplanted into 5 m rows with a<br />
0.55 m row spacing in the field in early May. Assessments <strong>on</strong> each row were carried out<br />
during the growing seas<strong>on</strong> and at maturity representative single plants were hand<br />
harvested and dried using a forced-air drier before the seed was removed from the<br />
capsules by hand-threshing. Following cleaning, seed samples were analysed for oil and<br />
GLA c<strong>on</strong>tent using methods described by Fieldsend and Moris<strong>on</strong> (2000b).<br />
Eventually, a crossing programme was established to generate new breeding lines.<br />
Crosses were carried out using plants grown in pots in an insect-proof glasshouse. Prior<br />
to the pollen becoming viable, the anthers were removed from flowers of the plants to<br />
be used as female parents and, 24 hours later, the stigmas were hand pollinated using<br />
anthers from the male parent. Reciprocal crosses were carried out as the progeny of<br />
reciprocal crosses within Oenothera can be genetically completely different from each<br />
other (Cleland, 1972). The seed was hand-harvested and stored during the winter.<br />
Subsequent generati<strong>on</strong>s were evaluated in breeding rows as described above. In extreme<br />
cases, the F1 populati<strong>on</strong> of Oenothera is genetically uniform and c<strong>on</strong>tains <strong>on</strong>e complete<br />
genome from each parent. No genetic segregati<strong>on</strong> takes place in subsequent generati<strong>on</strong>s.<br />
Even in less extreme cases, <strong>on</strong>ly limited segregati<strong>on</strong> occurs. Hence, Scotia adopted a<br />
30
modified pedigree breeding method, entailing a large number of crosses and very small<br />
(21 plant) F2 populati<strong>on</strong>s. Unpromising breeding lines could be discarded after the F2<br />
generati<strong>on</strong> whilst in some cases populati<strong>on</strong>s were uniform and showed enough<br />
commercial promise for seed multiplicati<strong>on</strong> to start.<br />
Clearly, this method does not allow the breeder to evaluate seed yields or the ability of<br />
the breeding line to overwinter. During the 1980s, overwintering ability was assessed <strong>on</strong><br />
autumn-transplanted rows of the breeding lines showing the most promise in F3 springsown<br />
rows. Selecti<strong>on</strong> of lines for trialling was based <strong>on</strong> field assessments from the F1,<br />
F2 and (until mid-August when the choice of lines for trialling had to be made) F3<br />
breeding rows plus the oil and GLA c<strong>on</strong>tent data from the F2 rows. By the 1990s,<br />
however, the establishment ability of the breeding lines had improved to such an extent<br />
that the most commercially promising advanced breeding material could be evaluated in<br />
autumn and spring-sown replicated yield trials (c.f. Fieldsend and Moris<strong>on</strong>, 2000a).<br />
New Zealand is an important source of evening primrose seed, not least because the sixm<strong>on</strong>th<br />
difference in harvest date compared to the northern hemisphere spreads the<br />
demands of harvest, storage and payment for seed more equably throughout the year.<br />
Overwintered crops are normally sown in February and harvested in March of the<br />
following year, whilst spring-sown crops are sown in September or October and<br />
harvested in April. The husbandry of spring-sown crops is described by Fieldsend<br />
(2003). The total area of crops grown under c<strong>on</strong>tract to Scotia in 1990-1997 ranged<br />
between 200 ha and 450 ha, with an approximately equal split between overwintered<br />
and spring-sown crops in the earlier years and a subsequent emphasis <strong>on</strong> spring<br />
producti<strong>on</strong>. This area would be divided between over 20 farmers, who would normally<br />
use field sizes of between 4 and 16 ha (Fieldsend, 2003). After harvest, a seed sample<br />
from each field was analysed for oil and GLA c<strong>on</strong>tents and the annual means of these<br />
data (not corrected for differences in field size) are used in this paper.<br />
RESULTS AND DISCUSSION<br />
Oenothera is a very diverse genus and most accessi<strong>on</strong>s were immediately rejected as<br />
being unsuitable for commercial seed producti<strong>on</strong> after <strong>on</strong>e year of trials. However, three<br />
accessi<strong>on</strong>s were “off-type” single plants with n<strong>on</strong>-splitting capsules taken from<br />
commercial crops of the “traditi<strong>on</strong>al” stock growing in the UK and Hungary. X444 and<br />
X851 were commercialised as the cultivars “Paul” and “Peter” respectively (similar<br />
accessi<strong>on</strong> X852 was not commercialised) and were awarded Plant Variety Rights (PVR)<br />
in several countries including the UK and New Zealand. The reduced risk of seed loss<br />
from “Paul” and “Peter” resulted in an approximately 50% increase in seed yields over<br />
“421” when grown as overwintered crops in New Zealand in 1990 and 1991 (Fig. 1),<br />
but these cultivars were similar to “421” in every other respect. Seed oil c<strong>on</strong>tents were<br />
in the regi<strong>on</strong> of 26.5% from autumn sowing (Fig. 2) and 25% from spring sowing (Fig.<br />
3) whilst the GLA c<strong>on</strong>tents of the oil were around 9% and 9.5% respectively. Higher oil<br />
c<strong>on</strong>tents and lower GLA c<strong>on</strong>tents from overwintered crops are typical of evening<br />
primrose (Fieldsend, 2000b).<br />
No other accessi<strong>on</strong>s were directly commercialised but many were used in the crossing<br />
programme. For example, the cultivar “Merlin” resulted from the cross (X444 x X390).<br />
This was a high yielding, easy to grow cultivar much liked by farmers which was also<br />
31
awarded PVR in several countries. The very high yield in 1990 (Fig. 1) was obtained<br />
from just 2.6 ha so this result must be treated with cauti<strong>on</strong> but does show its yield<br />
potential. In 1991-1993 “Merlin” outyielded "Paul” and “Peter” whilst its oil and GLA<br />
c<strong>on</strong>tents tended to be higher as well (Figs 2 and 3).<br />
Seed yield (t ha -1 )<br />
2,0<br />
1,6<br />
1,2<br />
0,8<br />
0,4<br />
0,0<br />
1990 1991 1992 1993 1994 1995 1996 1997<br />
Year of harvest<br />
Figure 1: Mean seed yield of crops of evening primrose harvested in New Zealand<br />
from 1990 to 1993. Closed symbols: overwintered; open symbols: spring sown;<br />
circles: 421; triangles up: Paul/Peter; squares: Merlin; triangles down: Rigel.<br />
A series of plants collected from the wild in Ontario, Canada produced seed with high<br />
oil c<strong>on</strong>tents and a very high GLA c<strong>on</strong>tent in the oil (approximately 15%) but were<br />
unsuitable for commercial use. One of these, X945, was crossed with another accessi<strong>on</strong>,<br />
X1021, in 1985 and the resulting breeding line (LQB) also produced high quality oil but<br />
the seed yield was low. In turn, in 1988, LQB was crossed with “Merlin” and the result<br />
was the cultivar “Rigel”. This cultivar successfully combined the n<strong>on</strong>-splitting capsules<br />
of “Paul” with the high oil and GLA c<strong>on</strong>tents of X945 and a high seed yielding ability.<br />
The great potential of the breeding line was c<strong>on</strong>firmed by the laboratory results from the<br />
F3 breeding row and the unusual step was taken of sending seed from the breeding row<br />
(despite this not having been raised in isolati<strong>on</strong> from other lines) to New Zealand for<br />
trialling. A 1000 m 2 plot was sown <strong>on</strong> 21 October 1991 and yielded 1121 kg seed ha -1<br />
when harvested in 1992. The oil and GLA c<strong>on</strong>tents were 28.5% and 13.3% respectively,<br />
substantially higher than cultivars then in commercial producti<strong>on</strong> (Fig. 3).<br />
32
“Rigel” seed multiplicati<strong>on</strong> commenced with a 1000 plant, 200 m 2 spring transplanted<br />
plot grown in the UK in 1992 which yielded 25 kg seed. 14.5 ha of “Rigel” crops were<br />
sown in New Zealand in October 1992 and harvested oil and GLA c<strong>on</strong>tents were again<br />
much higher than “Merlin”. Unfortunately, yield data after 1990 are not available but<br />
Fieldsend (2003) reported yields of up to 1.87 t ha -1 from spring-sown “Rigel” in New<br />
Zealand in 1998 whilst in 1999 some “Rigel” crops yielded more than 2 t ha -1 (Scotia,<br />
commercial data). This cultivar was again awarded PVR and was the sole cultivar to be<br />
grown commercially for Scotia after 1994.<br />
GLA c<strong>on</strong>tent of oil (%)<br />
Seed oil c<strong>on</strong>tent (%)<br />
14<br />
13<br />
12<br />
11<br />
10<br />
9<br />
8<br />
29<br />
28<br />
27<br />
26<br />
25<br />
24<br />
1990 1991 1992 1993 1994 1995 1996 1997<br />
Year of harvest<br />
Figure 2: Seed oil c<strong>on</strong>tent and GLA c<strong>on</strong>tent of oil of overwintered crops of evening<br />
primrose harvested in New Zealand from 1990 to 1997. Cultivars are designated<br />
thus: circles: 421; triangles up: Paul/Peter; squares: Merlin; triangles down: Rigel.<br />
33
Thus, by the late 1990s, oil and GLA c<strong>on</strong>tents of overwintered crops in New Zealand<br />
averaged around 28.5% and 12.5% respectively whilst the equivalent data from springsown<br />
crops were 28% and 13.5%. The impact <strong>on</strong> the GLA c<strong>on</strong>tent of the seed was an<br />
increase from around 2.5% in 1990 to over 3.5% by 1999, with spring-sown crops<br />
normally giving the highest results (Fig. 4). The big difference between sowing dates in<br />
1994 is due to the fact that overwintered crops were almost exclusively “Paul” and<br />
“Merlin” whilst all spring-sown crops were “Rigel”. The Scotia breeding programme<br />
closed in 1999 but “Rigel” c<strong>on</strong>tinues to be commercially grown.<br />
GLA c<strong>on</strong>tent of oil (%)<br />
Seed oil c<strong>on</strong>tent (%)<br />
14<br />
13<br />
12<br />
11<br />
10<br />
9<br />
8<br />
29<br />
28<br />
27<br />
26<br />
25<br />
24<br />
1990 1991 1992 1993 1994 1995 1996 1997<br />
Year of harvest<br />
Figure 3: Seed oil c<strong>on</strong>tent and GLA c<strong>on</strong>tent of oil of spring sown crops of evening<br />
primrose harvested in New Zealand in from 1990 to 1997. Cultivars are designated<br />
thus: circles: 421; triangles up: Paul/Peter; squares: Merlin; triangles down: Rigel.<br />
34
The benefits of the cultivars bred by Scotia can be seen in different ways. For<br />
nutriti<strong>on</strong>al supplements, the daily dose is normally specified as the number of (gelatine)<br />
capsules to be taken but a higher GLA c<strong>on</strong>tent in the oil gives the retailer a marketing<br />
benefit. For pharmaceuticals, where the dose normally much higher and is calculated as<br />
daily GLA intake, the number of capsules taken can be reduced. This has the twin<br />
benefits of improving patient compliance and also, owing to savings al<strong>on</strong>g the entire<br />
manufacturing chain, reducing product costs. It has been estimated (Scotia, commercial<br />
data) that the change to “Rigel” oil led to an 18% reducti<strong>on</strong> in costs.<br />
Seed GLA c<strong>on</strong>tent (%)<br />
4,0<br />
3,5<br />
3,0<br />
2,5<br />
2,0<br />
1990 1991 1992 1993 1994 1995 1996 1997<br />
Year of harvest<br />
Figure 4: Mean seed GLA c<strong>on</strong>tent of crops of evening primrose harvested in New<br />
Zealand from 1990 to 1997. Closed symbols: overwintered; open symbols: spring<br />
sown.<br />
The oil and GLA c<strong>on</strong>tents achieved with “Rigel” are close to the upper limit observed in<br />
the genetic material evaluated by Scotia. However, the high linoleic acid c<strong>on</strong>tent of the<br />
seed oil (approximately 75% in “Merlin” (Fieldsend and Moris<strong>on</strong>, 2000b)) is a substrate<br />
for potentially much higher GLA c<strong>on</strong>tents which could possibly be achieved through<br />
genetic engineering. Borage (Borago officinalis L.) oil, for example, is more complex<br />
and c<strong>on</strong>tains approximately 37% linoleic acid and 21% GLA, i.e. a ratio of less than 2:1<br />
(Senanayake and Shahidi, 2000). de Gyves (2004) dem<strong>on</strong>strated the c<strong>on</strong>cept by using<br />
an Agrobacterium-mediated transformati<strong>on</strong> procedure to deliver a cDNA encoding a 6desaturase<br />
from borage under the c<strong>on</strong>trol of a cauliflower mosaic virus (CaMV) 35S<br />
35
promoter in “Rigel”. Analysis of the transformed plants dem<strong>on</strong>strated an altered profile<br />
of polyunsaturated fatty acids, with an increase in GLA and octadecatetraenoic acid in<br />
leaf tissues when compared with c<strong>on</strong>trol lines. The fatty acid profile of the seed oil was<br />
virtually unaltered, but this is not surprising as CaMV 35S is not a seed-specific<br />
promoter.<br />
Fieldsend (1996b) coined the <strong>term</strong> “factories in the field” to describe the cultivati<strong>on</strong> of<br />
evening primrose as a source of GLA. The farmer is the “factory manager”. Agricultural<br />
research provides him with the correct “equipment” in the form of cultivars adapted to<br />
his growing c<strong>on</strong>diti<strong>on</strong>s, and the know-how, or “operating manual”, in the form of<br />
husbandry guidance. Almost ten years later, the c<strong>on</strong>cept has been adopted as part of the<br />
stakeholder proposal for the “Plants for the Future” technology platform (An<strong>on</strong>, 2005).<br />
In evening primrose, the advances envisaged by the technology programme have<br />
already been achieved in practice.<br />
REFERENCES<br />
An<strong>on</strong>. (2005): Plants for the Future. Stakeholder Proposal for a Strategic Research<br />
Agenda 2025; Part 1: Summary, EPSO, 28 p.<br />
Cleland, R.E. (1972): Oenothera, cytogenetics and evoluti<strong>on</strong>. Academic Press, L<strong>on</strong>d<strong>on</strong>.<br />
Fieldsend, A.F. (1996a): Evening primrose - from garden flower to oilseed crop. The<br />
Horticulturist, 5: (3): 2-5 p.<br />
Fieldsend, A.F. (1996b): Factories in the Field. Green Futures, 2: 20-21 p.<br />
Fieldsend, A.F. (2003): Agr<strong>on</strong>omic factors affecting the yield of crops of evening<br />
primrose (Oenothera spp.) Buletinul USAMV-CN, A-H, 59/2003: 58-63 p. ISSN<br />
1454-2382.<br />
Fieldsend, A.F. and Moris<strong>on</strong>, J.I.L. (2000a): C<strong>on</strong>trasting growth and dry matter<br />
partiti<strong>on</strong>ing in winter and spring evening primrose crops (Oenothera spp.) Field<br />
Crops Research, 68: 9-20 p.<br />
Fieldsend, A.F. and Moris<strong>on</strong>, J.I.L. (2000b): Climatic c<strong>on</strong>diti<strong>on</strong>s during seed growth<br />
significantly influence oil c<strong>on</strong>tent and quality in winter and spring evening<br />
primrose crops (Oenothera spp.) Industrial Crops and Products, 12: 137-147 p.<br />
de Gyves E.M., Sparks C.A., Sayanova O., Lazzeri P., Napier J.A. and J<strong>on</strong>es H.D.<br />
(2004): Genetic manipulati<strong>on</strong> of γ-linolenic acid (GLA) synthesis in a commercial<br />
variety of evening primrose (Oenothera sp.) Plant Biotechnology Journal, 2: (4):<br />
351-357 p.<br />
Horrobin, D. F. (1992): Nutriti<strong>on</strong>al and medical importance of gamma-linolenic acid.<br />
Progress in Lipid Research, 31: 163-194 p.<br />
Horrobin, D.F. (1994): Natural ≠ Safe. Pharmaceutical Technology Europe, December<br />
1994, 2 pp.<br />
Nix, J. (1995): Farm management pocketbook. 26 th editi<strong>on</strong>. Ashford, Kent: Wye<br />
College, University of L<strong>on</strong>d<strong>on</strong><br />
Senanayake, S.P.J.N. and Shahidi, F. (2000): Lipid comp<strong>on</strong>ents of borage (Borago<br />
officinalis L.) seeds and their changes during germinati<strong>on</strong>. Journal of the American<br />
Oil Chemists' Society, 77: (1): 55-61 p.<br />
36
ABSTRACT<br />
SOME LESSONS LEARNED FROM THE NYÍRLUGOS<br />
LONG-TERM FIELD EXPERIMENT<br />
Imre Kádár<br />
Research Institute for Soil Science and Agricultural Chemistry<br />
H-1022, Budapest, Herman Ottó Str. 15, Hungary. E-mail: kadar@rissac.hu<br />
The paper reports results achieved in a 44-year-old field trial set up <strong>on</strong> acid sandy<br />
brown forest soil in the Nyírség regi<strong>on</strong>. Characteristics of the site: pH (KCl) 4.3-4.6,<br />
humus 0.5-0.7%, CEC 3-4 meq/100 g in the ploughed layer. The topsoil was poor in all<br />
five macr<strong>on</strong>utrients (N, P, K, Ca, Mg) and the groundwater depth was 2-3 m. Fertilizers<br />
applied in form of Ca-amm<strong>on</strong>ium nitrate, superphosphate, potassium chloride,<br />
powdered limest<strong>on</strong>e and dolomite. The following c<strong>on</strong>clusi<strong>on</strong>s could be drawn:<br />
1. As the result of the regular N fertilizati<strong>on</strong> the pH (KCl) value in the topsoil dropped<br />
to 3.5 (c<strong>on</strong>trol plots 4.3), while it rose to 6.4 after additi<strong>on</strong> of 1 t/ha/yr CaCO3. As a<br />
functi<strong>on</strong> of liming, the exchangeable Ca 2+ enhanced from 0.13 to 2.18 meq/100 g,<br />
while Al 3+ decreased from 0.68 to 0.40, Fe 2+ from 0.43 to 0.15 meq/100 g; CEC<br />
increased from 3.3 to 3.6-3.8, S-value (sum of bases) from 0.4 to 2.5 meq/100 g, base<br />
saturati<strong>on</strong> (V) from 12% to 69%.<br />
2. The NH4-acetate+EDTA soluble (Lakanen and Erviö 1971) element c<strong>on</strong>tents in<br />
plowlayer also mirrored the changes caused by liming and fertilizati<strong>on</strong>. So, Ca lifted<br />
from 87 to 767, Mg from 18 to 97, Mn from 8 to 36, Sr from 0.4 to 2.7, Co from 0.15<br />
to 0.53, Ni from 0.10 to 0.19 mg/kg.<br />
3. Fertilizer resp<strong>on</strong>ses were time dependent. In the 1 st decade of the experiment N al<strong>on</strong>e<br />
increased the potato and rye yields. In the 2 nd decade, the yield <strong>on</strong> N-plots declined<br />
near to c<strong>on</strong>trol. In the studied last few years, the yields <strong>on</strong> N-plots were negligible,<br />
the soil lost its fertility and became leached and very acid. To maintain or increase<br />
the yields of crops NPKCaMg fertilizati<strong>on</strong> was needed. Fertile plots have a pH (KCl)<br />
5.5-6.0, 120-150 mg/kg amm<strong>on</strong>iumlactate-soluble P2O5 and K2O in plowlayer and<br />
require the applicati<strong>on</strong> of about 150 kg N and 1 t/ha ground dolomite yearly.<br />
Keywords: fertilizati<strong>on</strong>, liming, time dependence, resp<strong>on</strong>ses, l<strong>on</strong>g-<strong>term</strong> field<br />
experiment<br />
INTRODUCTION<br />
There is a nearly universal acceptance of the fact that most l<strong>on</strong>g-<strong>term</strong> ecological<br />
research is inherently interdisciplinary in nature. The successful performance of l<strong>on</strong>g<strong>term</strong><br />
field experiments requires the special recogniti<strong>on</strong> of certain key elements in the<br />
work. The scientific questi<strong>on</strong>s and objectives must be clearly defined and stated because<br />
they must guide research for several generati<strong>on</strong>s of scientists, administrators and<br />
funding agencies.<br />
37
The people involved in the l<strong>on</strong>g-<strong>term</strong> field experiments must possess a shared<br />
philosophy, appropriate training, the acceptance of a team leadership, and mutual trust<br />
and respect. The site at which l<strong>on</strong>g-<strong>term</strong> field experiments are performed must be<br />
representative of the biological and ecological (soil, weather, type of cultivati<strong>on</strong> etc.)<br />
setting. Both the site and research programme must have solid financial support and<br />
c<strong>on</strong>tinuity, and they must have a str<strong>on</strong>g group of people associated with them. Of<br />
course, the actual applicati<strong>on</strong> and implementati<strong>on</strong> of l<strong>on</strong>g-<strong>term</strong> field experiments may<br />
vary from place to place and time to time.<br />
Every human use of the natural ecosystem results in the removal of material and<br />
embodied energy which are later not returned or broken down at the same place. This<br />
means that human uses change natural ecosystems through different forms of matter and<br />
energy transports, involving removal or supply. L<strong>on</strong>g-<strong>term</strong> field experiments are vital in<br />
order to represent cumulative effects and unusual events caused by human use,<br />
agricultural practice, such as cultivati<strong>on</strong>, manuring etc.<br />
L<strong>on</strong>g-<strong>term</strong> field experiments make visible processes and events often invisible in most<br />
short-<strong>term</strong> experiments. As Magnus<strong>on</strong> et al. (1983) stated: "Because we are unable to<br />
directly sense these slow changes and because we are even more limited in our abilities<br />
to interpret cause and effect relati<strong>on</strong>s for these slow changes, processes acting over<br />
decades are hidden and reside in the invisible present." This is the time scale of acid<br />
depositi<strong>on</strong>, the introducti<strong>on</strong> of synthetic chemicals, acidificati<strong>on</strong> of soils, air borne<br />
polluti<strong>on</strong> and climate changes made by man.<br />
Slow phenomena, or accumulative changes in soil properties may be captured <strong>on</strong>ly in<br />
l<strong>on</strong>g-<strong>term</strong> studies. Limited durati<strong>on</strong> might miss important results, or worse, cause the<br />
results to be misinterpreted. <strong>Experiments</strong> in different years yield different results<br />
because of weather changes. L<strong>on</strong>g-<strong>term</strong> experiments are <strong>on</strong>e of the sure ways to<br />
de<strong>term</strong>ine slow processes.<br />
The various experiments at Rothamsted, including both the classic and modern,<br />
symbolize the value of l<strong>on</strong>g-<strong>term</strong> studies in general. The original goals of several of<br />
these experiments have l<strong>on</strong>g since been fulfilled. However, they c<strong>on</strong>tinue to be of value<br />
as dem<strong>on</strong>strati<strong>on</strong>s, and as sources of c<strong>on</strong>tinuing insight into agricultural practice<br />
(Johnst<strong>on</strong> 1989). In Hungary, most l<strong>on</strong>g-<strong>term</strong> field experiments are less than 40 years<br />
old, though this number has been increasing since the late 1980’s, as the regi<strong>on</strong>al<br />
research stati<strong>on</strong>s and institutes were established in the late 1950’s and 1960’s.<br />
MATERIALS AND METHODS<br />
The trial was established in autumn 1962 <strong>on</strong> brown forest soil, acid sand with thin<br />
interstratified layers of colloid and sesquioxide accumulati<strong>on</strong> called “kovárvány”. The<br />
soil has a particle-size distributi<strong>on</strong> in plow-layer as follows: sand over 0.05 mm 70-<br />
85%, loam 0.05-0.002 mm 8-20%, clay under 0.002 mm 3-6%. Clay in colloid<br />
accumulati<strong>on</strong> layers makes up to 10-18%. The saturati<strong>on</strong> percentage was 25-30, pH<br />
(H2O) 5.4, pH (KCl) 4.3, humus 0.5-0.8%, CEC 3-5 meq/100 g. The site is situated<br />
38
about 100 m above sea-level with yearly precipitati<strong>on</strong> of 550-600 mm and sunny hours<br />
1900-2000 h/year. The min/max temperature was about –25 Cº and +35 Cº interval, the<br />
watertable level found at a depth of 2-3 m. The site is extremely drought sensitive.<br />
The trial has 32 treatments x 4 replicati<strong>on</strong> = 128 plots with 5 x 10 = 50 m 2 plot size and<br />
randomized block design. The forms of fertilizers applied were Ca-amm<strong>on</strong>ium nitrate,<br />
superphosphate, muriate of potash, powdered limest<strong>on</strong>e and dolomite. The crop<br />
sequence was potato-rye in the first 10 years, than potato-winter wheat (8 years), than<br />
followed different crops the next 10 years (like white lupine, sunflower, grasses, spring<br />
barley, tobacco). In 1991, the 29 th year of the trial, a triticale m<strong>on</strong>oculture was<br />
established which is now 16 years old. The nutrients applied in the trial are shown in<br />
table 1.<br />
Table 1: Fertilizati<strong>on</strong> and liming applied in the experiment, kg/ha/year (Brown forest<br />
soil, acid sand, Nyírlugos, Nyírség regi<strong>on</strong>)<br />
Nutrient<br />
Applied nutrients, kg/ha/year<br />
levels N P2O5 K2O CaCO3 MgCO3<br />
0 0 0 0 0 0<br />
1 50 60 60 250 140<br />
2 100 120 120 500 280<br />
3 150 180 180 1000 -<br />
Remark: Ca-amm<strong>on</strong>ium-nitrate, muriate of potash, powdered limest<strong>on</strong>e and dolomite<br />
The plant material was analysed after wet ashing with cc.HNO3 + cc.H2O2, for the N<br />
de<strong>term</strong>inati<strong>on</strong> with cc.H2SO4 + cc.H2O2 treatment. The soil samples were analysed<br />
using NH4-acetate + EDTA method (Lakanen and Erviö 1971), basic soil properties<br />
according to Baranyai et al. (1987), adsorptive soil characteristics according to<br />
Bascomb (1964). All the mineral elements were measured using ICP technics. The<br />
composite soil samples per plots were made of 20 subsamples, the composite plant<br />
samples were made of 1 m 2 plant material per plot. Material and methods and the main<br />
results of the experiment were summarized elsewhere (Láng 1963, 1973, 1984; Kádár<br />
és Szemes 1990, 1994; Kádár et al. 1999; Márt<strong>on</strong> 2002 a, b, c).<br />
RESULTS AND DISCUSSIONS<br />
As we can see in table 2 the value of pH (H2O) dropped down from 5.4 (c<strong>on</strong>trol) to 4.6<br />
<strong>on</strong> the N3 plot, while lifted up to 6.8 <strong>on</strong> the limed plot. The pH (KCl) values showed<br />
analoge picture between 3.5 minimum and 6.4 maximum. The hydrolytic acidity<br />
followed the movement of pH within a 4-14 interval. The humus c<strong>on</strong>tent did not change<br />
significantly in the ploughed layer as a functi<strong>on</strong> of fertilizati<strong>on</strong> and liming. The NH4acetate+EDTA<br />
soluble phosphorus and potassium mirrored the P-and K-applicati<strong>on</strong>.<br />
The P-supply of soil turned to a satisfactory, while the K-supply of soil still remained<br />
under a satisfactory level in the topsoil. The downward movement of K-fertilizer<br />
probably enriched partly the subsoil.<br />
39
Table 2: Effect of fertilizati<strong>on</strong> and liming <strong>on</strong> the soil chemical characteristics in 2006,<br />
in the 44 th year of the trial. Ploughed layer.<br />
Treatment<br />
code<br />
pH<br />
(H2O)<br />
pH<br />
(KCl)<br />
Hydrolytic<br />
acidity,y1<br />
Humus<br />
%<br />
NH4-acetate+EDTA<br />
soluble<br />
P2O5 mg/kg K2O mg/kg<br />
C<strong>on</strong>trol 5.4 4.3 7.6 0.6 92 67<br />
N1 5.0 4.2 9.6 0.8 85 43<br />
N2 4.7 3.6 12.6 0.8 105 41<br />
N3 4.6 3.5 13.6 0.7 89 44<br />
N2P1 4.9 3.8 10.6 0.8 135 43<br />
N2P2 4.6 3.9 11.6 0.9 158 50<br />
N2P3 4.7 3.9 11.6 0.8 191 41<br />
N2K1 4.7 3.7 11.4 0.8 101 56<br />
N2K2 4.9 3.8 10.4 0.7 92 78<br />
N2K3 4.8 3.7 11.1 0.8 99 73<br />
N2P2K2 4.8 3.8 11.6 0.8 163 67<br />
N2P2K2Ca3 6.8 6.4 3.6 0.7 225 62<br />
N2P2K2Mg2 6.3 6.0 4.8 0.8 198 69<br />
N2P2K2Ca2Mg2 6.7 6.2 3.8 0.7 220 65<br />
LSD5% 0.3 0.5 2.1 0.3 41 12<br />
Mean 5.2 4.3 9.5 0.8 139 57<br />
Table 3 gives an overview about the effect of some treatments <strong>on</strong> soil chemical<br />
properties in the topsoil as a functi<strong>on</strong> of pH and the total grain yields of triticale during<br />
the last two 4-years periods. The close positive correlati<strong>on</strong>s am<strong>on</strong>g the pH (H2O), pH<br />
(KCl) and hydrolytic acidity values are unambiguous. The exchangeable cati<strong>on</strong>s Al 3+<br />
and Fe 2+ dropped down with increasing pH, while Ca 2+ and Mg 2+ i<strong>on</strong>s lifting up when<br />
Ca or Mg was applied. Exchangeable cati<strong>on</strong>s expressed in % the S-value (sum of bases)<br />
show relative relati<strong>on</strong>ships am<strong>on</strong>g these elements.<br />
On the leached and very acid soil Ca 2+ amounted altogether to 32%, while <strong>on</strong> the limed<br />
<strong>on</strong>e to 88%. In case of Mg 2+ , the N3 plot soil had 8%, while the fertilized with dolomite<br />
<strong>on</strong>e 22% relative weight. The K + made out 22% in the very acid soil, while 6% in the<br />
limed soil. C<strong>on</strong>sidering the adsorpti<strong>on</strong> characteristics, <strong>on</strong>e can state that the cati<strong>on</strong><br />
exchange capacity shows increasing trend. The sum of bases and the % of base<br />
saturati<strong>on</strong> enhanced 5-6 times in the limed soils compared to that of the N3 plot (Table<br />
3).<br />
40
Table 3: Effect of treatments <strong>on</strong> soil chemical properties in ploughed layer in 2006<br />
and <strong>on</strong> the cumulative yield of triticale. 2003-2006.<br />
Measured and<br />
Treatments code<br />
LSD5%<br />
calculated characteristics N3 N2 N1 C<strong>on</strong>trol NPKMg NPKCa<br />
pH(H2O) 4.6 4.7 5.0 5.4 6.3 6.8 0.3<br />
pH (KCl) 3.5 3.6 4.2 4.3 6.0 6.4 0.5<br />
Hidrolytic acidity (y1) 13.6 12.6 9.6 7.6 4.8 3.6 2.1<br />
Exchangeable cati<strong>on</strong>s, meq/100 g<br />
Al 3+ 0.68 0.72 0.67 0.63 0.48 0.40 0.08<br />
Fe 2+ 0.43 0.40 0.32 0.25 0.20 0.15 0.07<br />
Ca 2+ 0.13 0.25 0.41 0.85 1.51 2.18 0.28<br />
Mg 2+ 0.03 0.04 0.08 0.11 0.49 0.05 0.06<br />
Adsorpti<strong>on</strong> characteristics<br />
CEC, meq/100 g 3.3 3.4 3.4 3.4 3.8 3.6 0.4<br />
S-value, meq/100 g 0.4 0.5 0.7 1.2 2.2 2.5 0.3<br />
Base saturati<strong>on</strong>, % 12 15 21 35 58 69 8<br />
Exchangeable cati<strong>on</strong>s as % of sum of bases (S-value)<br />
Ca 2+ 32 50 57 71 68 88 11<br />
Na + 28 22 14 9 5 4 7<br />
K + 22 16 13 12 6 6 7<br />
Mg 2+ 8 8 11 9 22 2 5<br />
NH4-acetate+EDTA soluble, mg/kg<br />
Fe 152 158 124 104 121 111 25<br />
Al 134 146 123 103 100 103 15<br />
Ca 87 117 161 238 456 767 88<br />
Mg 18 19 25 28 97 22 12<br />
Mn 8 16 28 35 26 36 9<br />
Sr 0.40 0.78 1.20 2.22 2.88 2.76 0.56<br />
Co 0.15 0.30 0.47 0.47 0.35 0.53 0.13<br />
Ni 0.10 0.16 0.16 0.20 0.18 0.19 0.04<br />
Grain yield, t/ha/ 4 yrs* 6.1 9.3 8.6 6.1 18.0 18.5 3.2<br />
Grain yield, t/ha/ 4 yrs** 1.9 4.8 5.5 3.9 10.0 10.6 2.6<br />
*Sum of the 1999-2002. years<br />
**Sum of the 2003-2006. years.<br />
The NH4-acetate+EDTA soluble element c<strong>on</strong>tents change also as a functi<strong>on</strong> of the pH<br />
(KCl). The c<strong>on</strong>tent of Fe and Al diminished significantly with the increased pH (KCl)<br />
values while the c<strong>on</strong>tent of Ca, Mg and Mn enhanced. The limed soil was enriched also<br />
41
with microelements Sr, Co and Ni. According to our analysis, the used powdered<br />
limest<strong>on</strong>e had 0.3% Mg, 70 mg/kg Mn, 158 mg/kg Sr, 1.3 mg/kg Ni and 0.2 mg/kg Co.<br />
The used dolomit powder c<strong>on</strong>tained around 24% Ca, 12% Mg, 6-7 mg/kg Mn, 75 mg/kg<br />
Sr and 0.4-0.5 mg/kg Ni. The c<strong>on</strong>tent of Co was under detecti<strong>on</strong> limit of 0.08 mg/kg.<br />
The Fe made out 730 mg/kg in limest<strong>on</strong>e and 150 mg/kg in dolomite, while Al was 410<br />
mg/kg in limest<strong>on</strong>e and 140 mg/kg in dolomite powder. In spite of the Fe and Al input,<br />
the NH4-acetate+EDTA soluble Fe and Al c<strong>on</strong>tent in the ploughed layer diminished. So,<br />
the solubility of these elements lessened, assessed with this method. The Sr is usually<br />
moving in line with Ca both in rocks and soils which can be followed here. The same<br />
time, the very low pool of Mn, Sr, Co and Ni in the extremely acid N3 soil might be a<br />
c<strong>on</strong>sequence of leaching of these elements.<br />
Table 4: Effect of fertilizati<strong>on</strong> and liming <strong>on</strong> the yield of triticale in the 41 st – 44 th<br />
years of the experiment. (The 12 th – 16 th years of the triticale m<strong>on</strong>oculture)<br />
Treatments 2003 2004 2005 2006<br />
code Grain Straw Grain Straw Grain Straw Grain Straw<br />
C<strong>on</strong>trol 1.2 1.8 1.8 1.8 0.3 2.3 0.6 0.7<br />
N1 1.2 1.8 3.6 3.9 0.3 2.7 0.4 0.6<br />
N2 1.1 1.6 3.0 3.5 0.4 2.9 0.3 0.3<br />
N3 0.6 1.0 1.1 1.5 0.1 1.2 0.1 0.2<br />
N2P1 1.1 1.7 4.0 4.7 0.5 4.3 0.3 0.4<br />
N2P2 1.8 2.5 4.6 5.3 0.4 3.2 0.3 0.2<br />
N2P3 1.7 2.1 5.0 5.3 0.3 3.2 0.3 0.4<br />
N2K1 1.1 1.5 2.2 3.0 0.2 2.1 0.2 0.2<br />
N2K2 1.7 2.9 3.2 4.1 0.3 3.0 0.3 0.4<br />
N2K3 1.0 1.6 2.3 2.6 0.3 3.0 0.4 0.3<br />
N2P2K2 1.7 2.4 4.2 4.5 0.5 4.8 0.4 0.6<br />
N2P2K2Ca3 1.8 2.7 5.6 5.8 0.9 9.1 2.1 1.8<br />
N2P2K2Mg2 2.2 2.7 5.7 6.1 0.8 7.0 1.3 2.1<br />
N2P2K2Ca2Mg2 2.1 3.3 6.7 6.7 0.9 8.1 1.9 2.5<br />
LSD5% 0.6 0.8 1.5 1.4 0.2 1.8 0.7 0.6<br />
Mean 1.5 2.1 3.8 4.2 0.4 4.1 0.6 0.8<br />
The yield of triticale was depressed <strong>on</strong> the acidified soil. Fertilizer resp<strong>on</strong>ses were time<br />
dependent (Table 4). In the 1 st decade of the experiment N al<strong>on</strong>e increased the potato<br />
and rye yields. In the 2 nd decade, the yield <strong>on</strong> N-plots declined near to c<strong>on</strong>trol. In the<br />
studied last few years, the yields <strong>on</strong> N-plots were negligible, the soil lost its fertility,<br />
became leached and very acid. To maintain or increase the yields of crops NPKCaMg<br />
fertilizati<strong>on</strong> was needed. Fertile plots have a pH (KCl) 5.5-6.0, 120-150 mg/kg<br />
42
amm<strong>on</strong>iumlactate-soluble P2O5 and K2O in plowlayer and require the applicati<strong>on</strong> of<br />
about 150 kg N and 1 t/ha ground dolomite yearly.<br />
REFERENCES<br />
Baranyai F.- Fekete A.- Kovács I. (1987): A magyarországi talaj tápanyagvizsgálatok<br />
eredményei. Mezőgazdasági Kiadó. Budapest.<br />
Bascomb, C. (1964): Rapid method for the de<strong>term</strong>inati<strong>on</strong> of cati<strong>on</strong> exchange capacity<br />
of calcareous and n<strong>on</strong>calcareous soils. J. Soil Sci. Food Agri. 15:821-823.<br />
Johnst<strong>on</strong>, A.E. (1989): The value of l<strong>on</strong>g-<strong>term</strong> experiments, a pers<strong>on</strong>al view. In: L<strong>on</strong>g<strong>term</strong><br />
studies in ecology; approaches and alternatives. Ed: Linkens, E.G. 175-179.<br />
Springer Verlag. New York. USA.<br />
Kádár I.- Szemes I. (1990): Műtrágyázás és meszezés tartamhatásának vizsgálata<br />
savanyú homoktalaj<strong>on</strong>. Növény<strong>term</strong>elés. 39:147-155.<br />
Kádár I. – Szemes I. (1994): A nyírlugosi tartamkísérlet 30 éve. MTA Talajtani és<br />
Agrokémiai Kutató Intézete. Budapest.<br />
Kádár I. (1999): A hazai homoktalajok műtrágyaigényéről. Agrokémia és Talajtan.<br />
48:217-223.<br />
Lakanen, E. & Erviö, R. (1971): A comparis<strong>on</strong> of eight extractants for the<br />
de<strong>term</strong>inati<strong>on</strong> of plant available micr<strong>on</strong>utrients in soil. Acta Agr. Fennica.<br />
123:223-232.<br />
Láng I. (1963): A homoktalajok műtrágyázásának kérdései. MTA Agrártud. Oszt.<br />
Közl. 22:431-434.<br />
Láng I. (1973): Műtrágyázási tartamkísérletek homoktalajok<strong>on</strong>. Akad. Doktori<br />
Disszertáció. Kézirat. MTA. Budapest.<br />
Láng I. (1984): Homoktalajok <strong>term</strong>őképességének fokozása. Agrokémia és Talajtan.<br />
14:75-87.<br />
Magnus<strong>on</strong>, J.J., Bowser, C.J. & Beckel, A.L. (1983): The invisible present, l<strong>on</strong>g-<strong>term</strong><br />
ecological research <strong>on</strong> lakes. L and S Magazine. College of Letters and Science.<br />
Univ. Wisc<strong>on</strong>sin-Madis<strong>on</strong>. Fall. 3-6.<br />
Márt<strong>on</strong> L. (2002a): A csapadék és a tápanyagellátás és az őszi búza (Triticum<br />
aestivum L) <strong>term</strong>ése közötti kapcsolat. Növény<strong>term</strong>elés. 51:529-542.<br />
Márt<strong>on</strong> L. (2002b): A csapadék és tápanyagellátottság hatásának vizsgálata a tritikale<br />
<strong>term</strong>ésére tartamkísérletben. Növény<strong>term</strong>elés. 51:687-701.<br />
Márt<strong>on</strong> L. (2002c): Az éghajlatingadozás és a N-műtrágyázás hatása a rozs (Secale<br />
cereale L.) <strong>term</strong>ésére. Növény<strong>term</strong>elés. 51:199-210.<br />
43
ABSTRACT<br />
ANALYZE OF BIOMASS PRODUCTIVITY BY TIMESERIES<br />
REMOTESENSING DATA IN REGION OF NYÍRLUGOS<br />
János Tamás – Tibor Bíró – Nikolett Szőllősi<br />
University of Debrecen Agricultural Sciences,<br />
Faculty of Agr<strong>on</strong>omy,<br />
Department of Water and Envir<strong>on</strong>mental Management<br />
The biomass growth can be described in regi<strong>on</strong>al space and time by c<strong>on</strong>tinuous<br />
equati<strong>on</strong>s. Effective calculati<strong>on</strong> of biomass was not available because there was not<br />
large scale resoluti<strong>on</strong> punctual data. We have introduced a math treatment first time in<br />
Hungarian biomass research which is made by dissociati<strong>on</strong> of 6 years time series from<br />
earth satellite multispectral remote sensing data in regi<strong>on</strong> of Nyírlugos. This model<br />
c<strong>on</strong>sists of linear trend, periodic, autoregressive and random comp<strong>on</strong>ent which were<br />
successfully transformed forward. Result of trend analyze filtered out l<strong>on</strong>g <strong>term</strong> effect.<br />
This means that the affects of biomass increase and decrease like wet or drought years<br />
were detached. L<strong>on</strong>g <strong>term</strong> climate effects forecast was difficult but periodic and<br />
autoregressive comp<strong>on</strong>ent in short-<strong>term</strong> and in middle <strong>term</strong> forecast provided more<br />
resp<strong>on</strong>sible values. Finished and practical method is more fitting than foregoing to<br />
de<strong>term</strong>inate biomass potential.<br />
Keywords: biomass, time series, multispectral, remote sensing<br />
INTRODUCTION<br />
According to Csete and Láng sustainable competitiveness is accomplished through<br />
sustainable farming methods, whose key motives are sustainable producti<strong>on</strong>,<br />
adaptability, quality to any extent and favourable investments levels, c<strong>on</strong>sequently this<br />
kind of competitiveness is altogether different from any old practice.<br />
The U.S. Department of Agriculture (USDA) and the Nati<strong>on</strong>al Aer<strong>on</strong>autics and Space<br />
Administrati<strong>on</strong> (NASA) signed a Memorandum of Understanding (MOU) to strengthen<br />
future collaborati<strong>on</strong>. In support of this collaborati<strong>on</strong>, NASA and the USDA Foreign<br />
Agricultural Service (FAS) jointly funded a new project to assimilate NASA's Moderate<br />
Resoluti<strong>on</strong> Imaging Spectroradiometer (MODIS) data and products into an existing<br />
decisi<strong>on</strong> support system (DSS) operated by the Producti<strong>on</strong> Estimates and Crop<br />
Assessment Divisi<strong>on</strong>. It produces objective, timely and regular assessments of global<br />
agricultural producti<strong>on</strong> outlook and the c<strong>on</strong>diti<strong>on</strong>s affecting food security. In m<strong>on</strong>itoring<br />
crop c<strong>on</strong>diti<strong>on</strong>s for a specific regi<strong>on</strong>, remotely sensed vegetati<strong>on</strong> index data are used to<br />
track the evoluti<strong>on</strong> of the growing seas<strong>on</strong> compared to reference l<strong>on</strong>g-<strong>term</strong> mean<br />
c<strong>on</strong>diti<strong>on</strong>s (Tucker, 1985). A global normalized difference vegetati<strong>on</strong> index (NDVI) is<br />
produced from MODIS data, and is referred to as the "c<strong>on</strong>tinuity index" similar to the<br />
existing archive of NOAA-AVHRR derived NDVI . SPOT Vegetati<strong>on</strong> NDVI data and<br />
44
MODIS NDVI data from the Terra and Aqua platforms represent improvements in the<br />
ability to m<strong>on</strong>itor land photosynthetic capacity . The croplands are highly variable both<br />
temporally and spatially. Croplands vary from year to year due to events such as<br />
drought and fallow periods, and they vastly differ across the globe in accordance with<br />
characteristics such as cropping intensity and field size (Anyamba, et al., 1998). To<br />
describe this temporally heterogeneity a global NDVI time-series database, with a<br />
spatial resoluti<strong>on</strong> of 250 meters has been assembled using a 16-day compositing period,<br />
allowing for interannual comparis<strong>on</strong>s of growing seas<strong>on</strong> dynamics (Tamás - Németh,<br />
2005).<br />
MATERIAL AND METHOD<br />
Basic data source is made after dual average by MODIS Terra remote sensing data. This<br />
MODIS NDVI dataset is reprojected and mosaicked to suit the Nyírlúgos regi<strong>on</strong>.<br />
There was made maps in Nyírlugos which is after mean of territorial matrix of 10x10<br />
km with 250 meter rescissi<strong>on</strong>. These data are m<strong>on</strong>thly frequency data of 16 day period<br />
which had respect for in 6 years durati<strong>on</strong>. The analyzed areas are in black square in Fig<br />
1. Bright fields signify higher biomass intensity.<br />
RESULTS AND DISCUSSIONS<br />
The progress of biomass process is a climate dependent periodicity in time so it is<br />
manifest to analyze the observati<strong>on</strong> of time series by time steps methods.<br />
Forecast, operati<strong>on</strong>, simulati<strong>on</strong> are solvable this way. Basic data source c<strong>on</strong>sists of dual<br />
averaging NDVI values counted by MODIS Terra remote sensing data. There was made<br />
maps in Nyírlugos which is originated from means of territorial matrix of 10x10 km<br />
with 250 meter resoluti<strong>on</strong>. These data are m<strong>on</strong>thly frequency covering 16 day periods<br />
which had respected for in 6 years durati<strong>on</strong> (Figure 2.).<br />
45
Figure 1. NDVI time series of Nyírlugos in 2006<br />
(January-December from to left by lines)<br />
46
NDVI<br />
0,8<br />
0,75<br />
0,7<br />
0,65<br />
0,6<br />
0,55<br />
0,5<br />
0,45<br />
0,4<br />
0,35<br />
0,3<br />
1 5 9 13 17 21 25 29 33 37 41 45 49 53 57 61 65 69<br />
Time (m)<br />
Figure 2. The time series of NDVI data sources (2001-2006, n=72)<br />
Regi<strong>on</strong> of Nyírlúgos<br />
Lineáris (Regi<strong>on</strong> of<br />
Nyírlúgos)<br />
The mean of presented time series of NDVI data sources in regi<strong>on</strong> of Nyírlugos is<br />
0.536; standard deviati<strong>on</strong>: 0.136; maximum: 0.759; while minimum: 0.316.<br />
Discrete time steps in time describe phenomen<strong>on</strong> of biomass growth. The comp<strong>on</strong>ents<br />
of time series model were de<strong>term</strong>ined in four steps. The applied theoretical model were<br />
described by K<strong>on</strong>tur et al., (1993), used for another phenomen<strong>on</strong> (groundwater<br />
hydrograph) which was c<strong>on</strong>tinuous in time and space. This adopted time series analysis<br />
based <strong>on</strong> the next formula:<br />
Y = T + P + A + V<br />
(eqv. 1.)<br />
i<br />
i<br />
i<br />
i<br />
i<br />
Where Ti trend, Pi periodic, Ai autoregressive and Vi c<strong>on</strong>tingent comp<strong>on</strong>ent. In additi<strong>on</strong><br />
data sources in Figure 1. were <strong>term</strong>inated by equati<strong>on</strong> 1.<br />
First of all the trend was detached from the series for <strong>term</strong> 2001-2006. Due to this<br />
process the l<strong>on</strong>g <strong>term</strong> effect was eliminated from the data source. This means the effect<br />
of serial of wet and drought years resulted of biomass producti<strong>on</strong> changing was filtered<br />
out.<br />
The detachment of linear trend comp<strong>on</strong>ent:<br />
Ti = d0 + d1* i (eqv. 2.)<br />
i= 1, 2, …., N (timesteps)<br />
47
With proper replacement the element of parameter vectors d0 and d1, independent<br />
variable i=1, 2,…N, dependent variable Y1, Y2,… YN and errors y1, y2,…yN can be<br />
obtained the series without trend.<br />
d<br />
2(<br />
N − 2)<br />
− 3<br />
∗Y<br />
− i ∗Yi<br />
= 3<br />
2(<br />
N + 2)<br />
− 3 N 1<br />
−<br />
3 2<br />
0 +<br />
where d0 = c<strong>on</strong>stant of trend functi<strong>on</strong><br />
d1 = coefficient of trend functi<strong>on</strong><br />
N = number of element<br />
i = i th element<br />
Y = mean of original time series<br />
P i<br />
= a<br />
0<br />
2π<br />
2π<br />
+ acos<br />
∗i<br />
+ bsin<br />
∗i<br />
r r<br />
d<br />
1<br />
N + 1<br />
− ∗Y<br />
+ i ∗Yi<br />
=<br />
2<br />
2<br />
N −1<br />
12<br />
(eqv. 3.)<br />
Calculated result of Equati<strong>on</strong> 2. is the following: Ti=0.5113+0.0007i.<br />
The Pi periodic comp<strong>on</strong>ent of time steps of annual biomass intensity is exists in<br />
temperate climate. As next step the periodic comp<strong>on</strong>ent was detached by using the<br />
model. In that case when the periodic time of periodic comp<strong>on</strong>ent is known, the<br />
de<strong>term</strong>inati<strong>on</strong> of the period amplitude has to be d<strong>on</strong>e <strong>on</strong>ly. We suppose there is a 12<br />
m<strong>on</strong>ths periodic in biomass change. The calculati<strong>on</strong> of <strong>on</strong>e time period is the following:<br />
where:<br />
(eqv. 4.)<br />
a0 = c<strong>on</strong>stant of period<br />
a és b = coefficient of functi<strong>on</strong><br />
The relevant period time is r = 12 m<strong>on</strong>ths. The value of i can be counted in m<strong>on</strong>th. The<br />
a0, a and b parameters were de<strong>term</strong>ined. In this case y1 time step without trend with Pi<br />
periodic comp<strong>on</strong>ent was c<strong>on</strong>verged (Eqv. 5.):<br />
yi = Pi + p (eqv.5.)<br />
The value of pi is: pi = yi - Pi<br />
The de<strong>term</strong>inati<strong>on</strong> “a” and “b” (Eqv. 6.) is needed to count Pi.<br />
N 2 2π<br />
a = yi<br />
cos i<br />
N 12<br />
∑<br />
i=<br />
1<br />
N 2 2π<br />
b = yi<br />
sin i<br />
N 12<br />
48<br />
∑<br />
i=<br />
1<br />
(eqv. 6.)
The a*cos(2π*i/12) periodic comp<strong>on</strong>ent value was 0,000131776, while b*sin(2 π<br />
i*i/12) periodic comp<strong>on</strong>ent value was 2,80205E-05 between 2001-2006. Pi periodic<br />
comp<strong>on</strong>ent and substances are dem<strong>on</strong>strated by Figure 3.<br />
c<br />
0,2<br />
0,15<br />
0,1<br />
0,05<br />
0<br />
-0,05<br />
-0,1<br />
-0,15<br />
-0,2<br />
1 5 9 13 17 21 25 29 33 37 41 45 49 53 57 61 65 69<br />
p ′ ′ + V ′<br />
′ i = c1<br />
∗ pi−1<br />
1<br />
N −1<br />
1<br />
N<br />
N −1<br />
∑<br />
i=<br />
1<br />
′ 1 = r1<br />
=<br />
N<br />
∑<br />
i=<br />
1<br />
p<br />
p<br />
i<br />
2<br />
i<br />
p<br />
i−1<br />
i<br />
a*cos(2π*i/12)<br />
b*sin(2π*i/12)<br />
Figure 3. Time steps of biomass producti<strong>on</strong> periodic comp<strong>on</strong>ent and substances<br />
After the isolati<strong>on</strong> of the periodic comp<strong>on</strong>ent from the data source the autoregressive<br />
and random comp<strong>on</strong>ents are remained.<br />
During the interpretati<strong>on</strong> of the prepared model the autoregressive comp<strong>on</strong>ent expressed<br />
the c<strong>on</strong>necting effect of biomass periods (year by year) which are written in plant<br />
producti<strong>on</strong> practice as year effect while random comp<strong>on</strong>ent de<strong>term</strong>ines the uncertainty<br />
of model.<br />
Then analyzes was c<strong>on</strong>tinued with de<strong>term</strong>inati<strong>on</strong> of pi random comp<strong>on</strong>ent supposing the<br />
trend and periodic comp<strong>on</strong>ent effects are not exist anymore. So there is <strong>on</strong>ly<br />
autocorrelati<strong>on</strong> between time series values. The <strong>on</strong>e step autoregressive sub-model is<br />
the following:<br />
Pi<br />
(eqv. 7.)<br />
where t refers time step and Vi is autoregressive comp<strong>on</strong>ent, the c1is c<strong>on</strong>stant of the<br />
above functi<strong>on</strong>.<br />
c1’equal to autocorrelati<strong>on</strong> factor, which is rj (Eqv.8):<br />
49<br />
(eqv. 8.)
Value of c1’ is 0,274262498 while mean of Vi value is 0,04 the minimum 0,08 the<br />
maximum 0,11. When taking into c<strong>on</strong>siderati<strong>on</strong> the all regularity these values mean the<br />
precisi<strong>on</strong> of the de<strong>term</strong>inati<strong>on</strong> of biomass producti<strong>on</strong>. The random comp<strong>on</strong>ent depends<br />
<strong>on</strong> the climate and cultivati<strong>on</strong> c<strong>on</strong>diti<strong>on</strong> mainly. But <strong>on</strong>ly that part of it which is not<br />
affected by the periodicity and the trend. Higher random factor can be occur anywhere<br />
(e.g. extreme year) but it is more likely <strong>on</strong> irregular increasing interval of biomass-curve<br />
(e.g. inde<strong>term</strong>inate sow c<strong>on</strong>diti<strong>on</strong>s).<br />
SUMMARY<br />
We have introduced a math treatment first time in Hungarian biomass research which is<br />
made by dissociati<strong>on</strong> of 6 years time series from earth satellite multispectral remote<br />
sensing data in regi<strong>on</strong> of Nyírlugos. This model c<strong>on</strong>sists of linear trend, periodic,<br />
autoregressive and random comp<strong>on</strong>ents which were successfully transformed forward.<br />
Result of trend analyze filtered out l<strong>on</strong>g <strong>term</strong> effect. This means that the affects of<br />
biomass increase and decrease like wet or drought years were detached.<br />
With detaching periodic comp<strong>on</strong>ent sinus changing was separated from data source.<br />
Autoregressive analyses quantify statistical rules which resulted from dependence of<br />
sequential years.<br />
Random comp<strong>on</strong>ent dem<strong>on</strong>strated the uncertainty of model. This is the factor which<br />
makes biomass producti<strong>on</strong> characteristics unde<strong>term</strong>ined in an added point or in<br />
investigati<strong>on</strong>al space. This means by taking into c<strong>on</strong>siderati<strong>on</strong> the all well-known<br />
influential factor divergent value can be appeared according to the expected value.<br />
After adjusting model comp<strong>on</strong>ent successfully backward transformati<strong>on</strong> was carried out<br />
which procedure proved the applicability clearly. This c<strong>on</strong>firms the fact that significant<br />
regularity can be assumed in biomass growth in such a dynamic model was set in this<br />
study.<br />
REFERENCES<br />
Anyamba, A., J. Eastman R., Tucker, C. J. (1998). Warm Enso Event of 1997/98: NDVI<br />
Precursors and Drought Pattern Predicti<strong>on</strong> for Southern Africa. Greenbelt, NASA pp. 1-<br />
40.<br />
Csete, L., Láng, I.(2005) A fenntartható agrárgazdaság és vidékfejlesztés. Magyarország<br />
az ezredfordulón.MTA Társadalomkutató Közp<strong>on</strong>t. Marosi-Print Kiadó. Budapest. 1-<br />
313.<br />
K<strong>on</strong>tur I., Koris K., Winter J. (1993). Hidrológiai számítások. Akadémiai Kiadó,<br />
Budapest. pp. 143-184.<br />
Tamás J., Németh T. (szerk.) (2005): Agrárkörnyezetvédelmi indikátorok elmélete és<br />
gyakorlati alkalmazásai. Debreceni Egyetem, Debrecen, p. 138<br />
Tucker, C. J., Vanpraet, C. L., Sharman, M. J , van Ittersum, G. (1985) Satellite remote<br />
sensing of total herbaceous biomass producti<strong>on</strong> in the Senegalese Sahel: 1980–1984,”<br />
Remote Sens. Envir<strong>on</strong>., vol. 17, pp. 233–249,<br />
50
RAINFALL, FERTILIZATION AND LIMING RESPONSE ON TRITICALE<br />
(X Triticosecale W.) YIELD IN THE 44 YEAR OLD NYÍRLUGOS FIELD TRIAL<br />
BETWEEN 1999 AND 2006<br />
Márt<strong>on</strong> László<br />
Research Institute for Soil Science and Agricultural Chemistry of the Hungarian<br />
Academy of Sciences (RISSAC-HAS)<br />
1022 Budapest, Herman O. u. 15. Hungary, e-mail: mart<strong>on</strong>@rissac.hu<br />
ABSTRACT<br />
Precipitati<strong>on</strong> amount, distributi<strong>on</strong> and nitrogen (N)-, phosphorus (P2O5)-, potassium<br />
(K2O)-, calcium (CaO)-, and magnesium (MgO) fertilizati<strong>on</strong> interacti<strong>on</strong> effects were<br />
studied <strong>on</strong> a sandy acidic lessivated brown forest soil; WRB: Haplic Luvisol in the 44<br />
year old Nyírlugos Field Trial (NYFT) in a Hungarian fragile agro-ecosystem in<br />
Nyírség regi<strong>on</strong> (N: 47 0 41’ 60’’ and E: 22 0 2’ 80’’) <strong>on</strong> triticale (X Triticosecale W.)<br />
yield from 1999 to 2006. At the trial set up in 1962, the soil had the following<br />
agrochemical properties: pH (H2O) 5.9, pH (KCl) 4.7, hydrolytic acidity 8.4, hy1 0.3,<br />
humus 0.7%, total N 34 mg kg -1 , amm<strong>on</strong>lactate (AL) soluble-P2O5 43 mg kg -1 , AL-K2O<br />
60 mg kg -1 in the plowed (0-25 cm) layer. From 1980 to 2006, the experiment c<strong>on</strong>sisted<br />
of 32x4=128 plots in randomised block design. The gross plot size was 10x5=50 m 2 .<br />
The average fertilizer rates in kg ha -1 year -1 were nitrogen 75, phosphorus 90 (P2O5),<br />
potassium 90 (K2O), calcium 437.5 (CaCO3) and magnesium 140 (MgCO3). The<br />
groundwater table has had at a depth of 2-3 m below the surface. The main results are as<br />
follows. During drought c<strong>on</strong>diti<strong>on</strong>s the respective yield of the c<strong>on</strong>trol areas was -25%<br />
less than for average years. The applicati<strong>on</strong> of N al<strong>on</strong>e, or of NP and NK treatments, led<br />
to yield reducti<strong>on</strong> of -19.7%, while that of NPK, NPKCa, NPKMg or NPKCaMg caused<br />
an -28.3% drop during these types of years. In the wet years, the yield decreased by -<br />
22.2% <strong>on</strong> the unfertilized soils; in the case of N, NP, or NK nutriti<strong>on</strong> with an -14.1%;<br />
and increased at 13.8% <strong>on</strong> NPK, NPKCa, NPKMg and NPKCaMg treated plots. In the<br />
very wettest year, the yields were dropped -43.1% <strong>on</strong> c<strong>on</strong>trol soils, -39.3% of N, NP, or<br />
NK loadings and -35.8% <strong>on</strong> NPK, NPKCa, NPKMg and NPKCaMg treatments to those<br />
in the average year. The relati<strong>on</strong>ships between rainfall quantitiy during the vegetati<strong>on</strong><br />
period N, P, K, Ca and Mg nutriti<strong>on</strong> and yield were characterised by polynomial<br />
correlati<strong>on</strong>s (c<strong>on</strong>trol: R= 0.7212***, N: R = 0.7410***, NP: R = 0.6452***, NK: R =<br />
0.6998***, NPK: R = 0.5555***, NPKCa: R = 0.5578***, NPKMg: R = 0.4869**,<br />
NPKCaMg: R = 0.4341**). However, the total regressi<strong>on</strong> coefficients ranged from 0.43<br />
to 0.74 in depence <strong>on</strong> the different nutrient applicati<strong>on</strong>. Maximum yields of 5.8-6.0 t .<br />
ha -1 were achieved in the rainfall range of 580-620 mm. At values above and below this<br />
domain of the precipitati<strong>on</strong> the grain yield reduced quadratically. So, it can be stated<br />
that both, drought and excess rainfall c<strong>on</strong>diti<strong>on</strong>s resulted dramatically in significant<br />
negative effects between fertilizati<strong>on</strong> (N, P, K, Ca, Mg) and triticale yield.<br />
Keywords: precipitati<strong>on</strong>, fertilizati<strong>on</strong>, liming, triticale, yield<br />
51
INTRODUCTION<br />
The hazards associated with climate change are depend <strong>on</strong> the interacti<strong>on</strong> of several<br />
systems with many variables (Johnst<strong>on</strong>, 2000). Accummulati<strong>on</strong> of carb<strong>on</strong> dioxide,<br />
methane, water vapor, oz<strong>on</strong>e, nitrous oxide, sulfur hexafluoride, hydrofluorocarb<strong>on</strong>s,<br />
perfluorocarb<strong>on</strong>s, chlorofluorocarb<strong>on</strong>s (build-up of greenhouse gases) in the atmosphere<br />
and trends in their emissi<strong>on</strong>s suggest that we can expect significant envir<strong>on</strong>mental<br />
changes in the 21th century (Cynthia and Ana, 2006; Eric 2006). However, a recent<br />
c<strong>on</strong>sensus has emerged that between the greenhouse gases rising of atmospheric<br />
c<strong>on</strong>centrati<strong>on</strong>s of carb<strong>on</strong> dioxide could become the more dangerous bacause it causing<br />
the global warming (Láng, 2005).<br />
Today, most researchers believe that higher temperature, drought and rainfall<br />
excess caused by climate change will depress crop yields in many places in the coming<br />
decades (Kádár et al., 1999; Jolánkai, 2005). Thus, in the last decades many agricultural<br />
investigati<strong>on</strong>s focused <strong>on</strong> understanding the relati<strong>on</strong> between mean climate change and<br />
crop producti<strong>on</strong> (Runge, 1968; Várallyay, 1992). Changes in weather patterns were<br />
observed thoughout Europe including Hungary as early as 1850. Am<strong>on</strong>g the natural<br />
c<strong>on</strong>sequences of changing weather patterns, years of drought (rainfall deficit) and wet<br />
(rainfall excess) c<strong>on</strong>diti<strong>on</strong>s, resulted in problems am<strong>on</strong>g plant nutriti<strong>on</strong> and field crop<br />
producti<strong>on</strong> (EU, 2003). Triticale (Kádár et al., 1999; Márt<strong>on</strong>, 2002) is a crop of<br />
worldwide importance, limited research exists about the effects of climate change <strong>on</strong><br />
these crop. The crop is sensitive to the prevailing weather c<strong>on</strong>diti<strong>on</strong>s (such as rainfall)<br />
and, for this reas<strong>on</strong>, understanding the effects of anthropogenic climate change <strong>on</strong> it’s<br />
producti<strong>on</strong> is important (Márt<strong>on</strong>, 2006). In additi<strong>on</strong> to growing seas<strong>on</strong> triticale<br />
c<strong>on</strong>diti<strong>on</strong>s (e.g., soil agrochemical properties, fertilizati<strong>on</strong>, liming) affect the growth and<br />
yield of crop (Lobell and Asner, 2003) and cause yield variati<strong>on</strong>s. Understanding the<br />
fertilizati<strong>on</strong>, liming and rainfall effects have been a c<strong>on</strong>tinuous endeavor toward<br />
improving farming technology and management strategy to reduce the negative impacts<br />
of fertilizati<strong>on</strong>, liming and rainfall and to increase crop yield (Kádár and Szemes 1994;<br />
Várallyay 1994; am<strong>on</strong>g others).<br />
Our main objective of this research it was study and clarify the precipitati<strong>on</strong><br />
amount and distributi<strong>on</strong> and nitrogen (N)-, phosphorus (P2O5)-, potassium (K2O)-,<br />
calcium (CaO)-, and magnesium (MgO) fertilizati<strong>on</strong> interacti<strong>on</strong> effects <strong>on</strong> a sandy<br />
acidic lessivated brown forest soil; WRB: Haplic Luvisol in the 44 year old Nyírlugos<br />
field trial in a Hungarian fragile agro-ecosystem in Nyírség regi<strong>on</strong> <strong>on</strong> triticale (X<br />
Triticosecale W.) yield from 1999 to 2006. Furthermore, it was our intent to emphasize<br />
that the net effect of multiple envir<strong>on</strong>mental changes is far more important than the<br />
effect of a single factor <strong>on</strong> the crop.<br />
MATERIALS AND METHODS<br />
The net-influence of rainfall (quantity, distributi<strong>on</strong>) and mineral fertilizati<strong>on</strong> (N, P2O5,<br />
K2O, CaO, MgO) were studied in a l<strong>on</strong>g <strong>term</strong> field experiment established at the<br />
Research Institute for Soil Science and Agricultural Chemistry of the Hungarian<br />
Academy of Sciences Experiment Stati<strong>on</strong> (RISSAC-HAS ET) in Hungary <strong>on</strong> a Haplic<br />
Luvisol (sandy acidic lessivated brown forest soil) with triticale (X Triticosecale W.)<br />
indicator crop under fragile agro-ecological circumstances at Nyírlugos for 8 years from<br />
52
1999 to 2006. The main experiment’s soil agrochemical characteristics in the plowed<br />
(0-25 cm) layer are presented in Table 1. at the experimental set up in 1962 (Láng,<br />
1973). From 1980 to 2006 the experiment c<strong>on</strong>sisted of 32x4=128 plots in randomized<br />
block designs. The gross plot size has been having 10x5=50 m 2 . The experimental<br />
treatments and combinati<strong>on</strong>s are shown in Table 2.<br />
The fertilizers were applied in the form of 25% calcium amm<strong>on</strong>ium nitrate, 18%<br />
superphosphate, 40% potassium chloride, calcium carb<strong>on</strong>ate and magnesium sulphate.<br />
The groundwater table has had at a depth of 2-3 m below the surface. The plant samples<br />
had had taken by manually at the harvest time. Rainfall amounts (deviati<strong>on</strong> in rainfall<br />
from the average over many years: dry year -10 - -20%, drought year -20% over, wet<br />
year +10 - +20%, year with excess rainfall +20% over) and other related data<br />
de<strong>term</strong>ined <strong>on</strong> traditi<strong>on</strong>al Hungarian (Harnos, 1993) and Research Institute for Soil<br />
Science and Agricultural Chemistry of the Hungarian Academy of Sciences (Márt<strong>on</strong>,<br />
2004) standards, and MANOVA (Multivariate Analysis of Variance) by SPSS test<br />
(SPSS Inc., 2000).<br />
Table 1: The main soil agrochemical properties in the plowed (0-25 cm) layer at the<br />
experiment set up. (Brown forest soil, acid sand; WRB: Haplic Luvisol, Nyírség<br />
regi<strong>on</strong> Nyírlugos 1962)<br />
C<strong>on</strong>tent PH HA* Hy1 Humus Total AL**<br />
(H2O) (KCl) nitrogen P2O5 K2O<br />
5.9 4.7 8.4 0.3<br />
% 0.7<br />
mg . kg -1 34 43 60<br />
* Hydrolytic acidity, ** amm<strong>on</strong>iumlactate (AL) soluble<br />
Table 2: Fertilizati<strong>on</strong> and liming treatments in the experiments, kg ha -1 year -1<br />
(Brown forest soil, acid sand; WRB: Haplic Luvisol, Nyírség regi<strong>on</strong> Nyírlugos)<br />
Applied kg ha -1 yr -1<br />
Treatment<br />
Levels N P2O5 K2O CaCO3 MgCO3<br />
0 0 0 0 0 0<br />
1 50 60 60 250 140<br />
2 100 120 120 500 280<br />
3 150 180 180 1000 -<br />
Note: In the form of Ca-amm<strong>on</strong>ium-nitrate, Superphosphate, potassium cloride,<br />
powdered limest<strong>on</strong>e and dolomite.<br />
RESULTS AND DISCUSSION<br />
During drought c<strong>on</strong>diti<strong>on</strong>s, in c<strong>on</strong>formity with Adams et al. (1995), Rosenzweig and<br />
Tubiello (1997) and McMaster (1999) the respective yield of the c<strong>on</strong>trol areas was -<br />
25% less than for average years. The applicati<strong>on</strong> of N al<strong>on</strong>e, or of NP and NK<br />
treatments, led to yield losses of -19.6%, while that of NPK, NPKCa, NPKMg or<br />
53
NPKCaMg caused an -28.3% drop during these types of years. In the wet years, the<br />
yield decreased by -22.2% in the unfertilized plots; in the case of N, NP, or NK<br />
nutriti<strong>on</strong> with an -14.1%; and increased at 13.8% <strong>on</strong> NPK, NPKCa, NPKMg and<br />
NPKCaMg treatments. In the very wettest year, the yields were dropped -43.1% <strong>on</strong><br />
c<strong>on</strong>trol soils, -39.3% of N, NP, or NK loadings and -35.8% <strong>on</strong> NPK, NPKCa, NPKMg<br />
and NPKCaMg treatments to those in the average year, reverse of Asbjorn et al. (2004).<br />
The relati<strong>on</strong>ships between rainfall quantitiy during the vegetati<strong>on</strong> period N, P, K, Ca<br />
and Mg nutriti<strong>on</strong> and yield were characterised by polynomial correlati<strong>on</strong>s (c<strong>on</strong>trol: R=<br />
0.7212***, N: R = 0.7410***, NP: R = 0.6452***, NK: R = 0.6998***, NPK: R =<br />
0.5555***, NPKCa: R = 0.5578***, NPKMg: R = 0.4869**, NPKCaMg: R =<br />
0.4341**). However, the total regressi<strong>on</strong> coefficients ranged from 0.43 to 0.74 in<br />
depence <strong>on</strong> the different nutrient applicati<strong>on</strong>. Maximum yields of 5.8-6.0 t . ha -1 were<br />
achieved in the rainfall range of 580-620 mm. At values above and below this range the<br />
grain yield reduced quadratically.<br />
To sum up we can say climate change will gradually and, at some point, be even<br />
abruptly affects Hungary and Europa agriculture. Warming temperatures and a greather<br />
incidence and intensity of extreme weather events possible lead to significant reducti<strong>on</strong>s<br />
in triticale yield. Expanded ranges of crop agrochemicals and altered transmissi<strong>on</strong><br />
dynamics of different irrigati<strong>on</strong> soluti<strong>on</strong>s might exacarbate these reducti<strong>on</strong>s. Since<br />
farmers’ strategies grow out of experience, they can find that the past will be a less<br />
reliable predictor of the future.<br />
ACKNOWLEDGEMENTS<br />
This research was supported by Hungarian Academy of Sciences, H-Budapest and the<br />
Hungarian and Spanish Intergovernmental S & T Cooperati<strong>on</strong> Project of E-2/04-<br />
OMFB-00112/2005 and Hungarian and Indian Intergovernmental S & T Cooperati<strong>on</strong><br />
Project of IND-3/03/2006.<br />
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agriculture. Climatic Change, Vol. 30, pp. 147-167.<br />
Asbjorn, T., Michelle, T. and Bárd, R. (2004): Climate Change Impacts <strong>on</strong><br />
Agricultural Productivity in Norway. CICERO. Oslo<br />
Cynthia, R. & Ana, I. (2006): Potential impact of climate change <strong>on</strong> world food<br />
supply. Data sets from a major crop modeling study. Socioec<strong>on</strong>omic Data and<br />
Applicati<strong>on</strong>s Center. Columbia University. New York<br />
Eric La F. (2006): Adapting crops for climate change. UBC Botanical Garden and<br />
Centre for Plant Researches.<br />
EU (European Uni<strong>on</strong>). (2003): Drought costs EU farmers euro of 11 billi<strong>on</strong>. European<br />
Report, Brussels<br />
Harnos, Zs. (1993): Weather and weather-yield interacti<strong>on</strong> analysis. (In Hungarian) In:<br />
Aszály 1983 (Szerk.: Baráth Cs-né., Győrffy B., Harnos Zs.). KÉE. Budapest<br />
Johnst<strong>on</strong>, A. E. (2000): Some aspects of nitrogen use efficiency in arable agriculture.<br />
K. Scogs-o. Lantbr. Akad. Tidskr. 139:8.<br />
54
Jolánkai M. (2005): Effect of climate change <strong>on</strong> plant cultivati<strong>on</strong>. (In Hungarian) In:<br />
„AGRO-21” Füzetek. 41. 47–58.<br />
Kádár I., Németh T., Szemes I. (1999): Triticale trágyareakciója a nyírlugosi<br />
tartamkísérletben. Növény<strong>term</strong>elés. 48:647-661.<br />
Kádár I., Szemes I. (1994): A nyírlugosi tartamkísérlet 30 éve. MTA Talajtani és<br />
Agrokémiai Kutató Intézete. Budapest.<br />
Láng I. (1973): Műtrágyázási tartamkísérletek homoktalajok<strong>on</strong>. Akad. Doktori<br />
Disszertáció. Kézirat. MTA. Budapest.<br />
Láng I. (2005): Weather and climate change: change-effect-resp<strong>on</strong>se. (In Hungarian)<br />
In: „AGRO-21” Füzetek. 43. 3–10.<br />
Lobell, D. B., Asner, G. P. (2003): Climate and management c<strong>on</strong>tributi<strong>on</strong>s to recent<br />
trends in U.S. agricultural yield. Science. 299. 1032.<br />
Márt<strong>on</strong> L. (2002): A csapadék és tápanyagellátottság hatásának vizsgálata a tritikale<br />
<strong>term</strong>ésére tartamkísérletben. Növény<strong>term</strong>elés. 51:687-701.<br />
Márt<strong>on</strong> L. (2004): Rainfall and fertilizati<strong>on</strong> effects <strong>on</strong> crops yield in a global climate<br />
change. In: Proc. 4 th Agroenvir<strong>on</strong> Symposium. Role of Multipurpose Agriculture<br />
in Sustaining Global Envir<strong>on</strong>ment-AGROENVIRON 2004 (Udine, 20–24 Oct.,<br />
2004). Part 3. 451–456. DPVTA. Udine<br />
Márt<strong>on</strong> L. (2006): Ecological changes of rainfall and artificial fertilizati<strong>on</strong> <strong>on</strong> crop<br />
yield formati<strong>on</strong>. ESA, Memphis, Tenesse<br />
McMaster, H. J. (1999): The potential impact of global warming <strong>on</strong> Hail Losses to<br />
winter crops in New South Wales. Climatic Change, Vol. 43, No. 2, October, pp.<br />
455-476.<br />
Rosenzweig, C. P. and F. N. Tubiello.( 1997): Impacts of global climate change <strong>on</strong><br />
Mediterranean agriculture: Current methodologies and future directi<strong>on</strong>s: An<br />
introductory essay. Mitigati<strong>on</strong> and Adaptati<strong>on</strong> Strategies for Global Change, Vol.<br />
1, pp. 219-232.<br />
Runge, E. C. (1968): Effect of rainfall and temperature interacti<strong>on</strong> during the growing<br />
seas<strong>on</strong> <strong>on</strong> corn yield. Agr<strong>on</strong>. J, 60:503-507.<br />
SPSS. (2000): SigmaPlot for Windows. Ver. 3.2, Chicago, III.: SPSS, Inc<br />
Várallyay, Gy. (1992): Globális klímaváltozások hatása a talajra. Effect of Global<br />
Climate Change to soil. Magyar Tudomány, 9:1071-1076.<br />
Várallyay Gy. (1994): A nyírlugosi tartamkísérlet talajszelvényeinek leírása és<br />
laborvizs-gálati eredményei. In: A nyírlugosi tartamkísérlet 30 éve. 216-226.<br />
(Kádár I.- Szemes I.) MTA Talajtani és Agrokémiai Kutató Intézete. Budapest.<br />
55
EVALUATION OF THE DATA SET OF THE HUNGARIAN LONG-TERM<br />
K-FERTILIZATION FIELD TRIALS, SET UP BETWEEN 1960 AND 2000<br />
Péter Csathó<br />
Research Institute for Soil Science and Agricultural Chemistry of the Hungarian<br />
Academy of Sciences, H-1022 Budapest, Herman O. út 15, Hungary.<br />
ABSTRACT<br />
Potassium is c<strong>on</strong>sidered to be the third most important macr<strong>on</strong>utrient in Hungary<br />
following nitrogen and phosphorus. The magnitude of resp<strong>on</strong>ses to K applicati<strong>on</strong> is<br />
effected not <strong>on</strong>ly by the available K levels in the soil, but by the crops, too.<br />
Evaluating the results of Hungarian 1- to 10-years-old l<strong>on</strong>g-<strong>term</strong> potassium fertilizati<strong>on</strong><br />
field experiments found in the literature between 1960 and 2000, c<strong>on</strong>clusi<strong>on</strong>s were<br />
made as follows:<br />
The c<strong>on</strong>necti<strong>on</strong> between the AL-K c<strong>on</strong>tent of K-c<strong>on</strong>trol (NP) plots and the<br />
resp<strong>on</strong>ses to K, expressed in relative yields (100Yield in NP plot/Yield in NPK plot,%)<br />
could be described by a Mitscherlich-like equati<strong>on</strong>, modified by Bray (1944): / Y'= 100<br />
(1-10 -cx ) /, where x = AL-K c<strong>on</strong>tent in the K-c<strong>on</strong>trol (NP) plots; Y' = relative yield in<br />
the trials, having "x" AL-K c<strong>on</strong>tents in the K-c<strong>on</strong>trol plots; and c = proporti<strong>on</strong>al<br />
c<strong>on</strong>stant (Mitscherlich's "working factor").<br />
As a result of K applicati<strong>on</strong>, surplusses in maize were higher than in winter wheat<br />
or alfalfa. Average surplusses in maize grain yields varied between 0.0 and 1.9 t/ha, in<br />
alfalfa between 0.0 and 1.6 t/ha and in winter wheat between 0.1 and 0.4 t/ha, resp.<br />
The AL- (amm<strong>on</strong>ium-lactate) (Egner, Riehm and Domingo 1960) soluble K indicated<br />
the natural K- supplying power of the different soils adequately.<br />
Keywords: field trials, potassium, soil texture, Bray-Mitscherlich approach, new<br />
fertiliser recommendati<strong>on</strong> system<br />
INTRODUCTION<br />
In the early 60's, when intensive (NP) K fertilizati<strong>on</strong> had not been introduced, about<br />
half of the arable land in Hungary was poorly or moderately supplied with K. For that<br />
reas<strong>on</strong> the synthesis of the nati<strong>on</strong>al field trial series of the above two crops is especially<br />
important (Corey 1987). Alfalfa is also am<strong>on</strong>g the most important crops produced in the<br />
country. As a first attempt, the data set of K fertilizati<strong>on</strong> trials with winter wheat, maize<br />
and alfalfa has been established.<br />
MATERIALS AND METHODS<br />
A database of the results of 1- to 10-year-old Hungarian potassium field experiments<br />
with maize, winter wheat and alfalfa, found in literature was compiled. A large number<br />
of these trials bel<strong>on</strong>g to the Nati<strong>on</strong>al L<strong>on</strong>g-<strong>term</strong> Field Trial Series with Fertilizers<br />
(OMTK), supervised by the Pann<strong>on</strong> Agricultural University, Keszthely. In all trials,<br />
by-products were removed from the fields. Soil characteristic data of NP (K-c<strong>on</strong>trol)<br />
treatments, grain yield data of NP and also NP+K treatments giving maximum<br />
ec<strong>on</strong>omic yields (about 95% of the maximum yields) were collected. The results were<br />
56
classified according to the soil texture groups. The correlati<strong>on</strong> between soil test (AL-K)<br />
values and resp<strong>on</strong>ses to K, using Bray's relative index method, was investigated. The<br />
AL-method has been widely used in the Scandinavian countries, in the Netherlands, in<br />
Portugal and in some Central European countries, and is still the official soil PK test<br />
method in many of these countries.<br />
RESULTS AND DISCUSSIONS<br />
Evaluating the results of Hungarian 1- to 10-years-old potassium field experiments with<br />
maize, winter wheat, and alfalfa found in literature between 1960 and 2000, c<strong>on</strong>clusi<strong>on</strong>s<br />
were made as follows:<br />
As a result of previous NPK experiment series in Hungary with cereals and row<br />
crops, N-resp<strong>on</strong>ses were highest in both groups, which was followed by the Kresp<strong>on</strong>ses<br />
in row crops and P-resp<strong>on</strong>ses in cereals. N-resp<strong>on</strong>ses were twice as high as<br />
K- or P-resp<strong>on</strong>ses (Várallyay sr 1950, Denke 1974a and 1974b, Debreczeni and<br />
Debreczeni 1994).<br />
Regarding the texture of Hungarian soils, 16% are sands, 10% sandy loams, 43%<br />
loams, 19% clay loams and 7% are clayey soils.<br />
The soils of the experimental fields did not receive previous significant potassium<br />
doses so the results of the trials helped in estimating the original K supplying power of<br />
the soils with different soil texture.<br />
Soils with higher clay c<strong>on</strong>tents provided a better original (native) K-supplying power.<br />
Am<strong>on</strong>g clay minerals, usually smectite was dominant in heavy soils, and illite in light<br />
<strong>on</strong>es.<br />
For estimating the extractable K-c<strong>on</strong>tents of the soils in Hungary, the AL-method<br />
(0.1 mol/L amm<strong>on</strong>ium lactate + 0.4 mol/L acetic acid, pH = 3.7, according to Egnér,<br />
Riehm and Domingo (1960) has been official since the 60's.<br />
On the basis of K field experiments with winter wheat, maize and alfalfa, carried<br />
out between 1960 and 2000, the AL-method indicated the native K-supplying power of<br />
the soils quite accurately (Figures 1 to 3). By this method probably not <strong>on</strong>ly the K<br />
adsorbed and in the soil soluti<strong>on</strong>, but also some parts of fixed K was extracted.<br />
According to the results of Sárdi and Németh (1993), AL- or neutral NH4-acetate extractants dissolve similar amounts of K from the same soil samples, resp.<br />
The c<strong>on</strong>necti<strong>on</strong> between the AL-K c<strong>on</strong>tent of K-c<strong>on</strong>trol (NP) plots and the<br />
resp<strong>on</strong>ses to K, expressed in relative yields (100Yield in NP plot/ Yield in NPK plot,%)<br />
could be described by a Mitscherlich-like equati<strong>on</strong>, modified by Bray (1944): / Y'= 100<br />
(1-10 -cx ) /, where x = AL-K c<strong>on</strong>tent in the K-c<strong>on</strong>trol (NP) plots; Y' = relative yield in<br />
the trials, having "x" AL-K c<strong>on</strong>tents in the K-c<strong>on</strong>trol plots; and c = proporti<strong>on</strong>al<br />
c<strong>on</strong>stant (Mitscherlich's "working factor").<br />
The "c" values - similarly to the findings of Bray (1944) - were different for maize<br />
(0.0083 and for the other crops: alfalfa (0.0109) and for winter wheat (0.0153),<br />
respectively.<br />
The de<strong>term</strong>inati<strong>on</strong> coefficient (R2) for maize and alfalfa trials was higher (0.77*** and<br />
0.80***) than winter wheat trials (0.67***), respectively.<br />
The c<strong>on</strong>necti<strong>on</strong> between the AL-K c<strong>on</strong>tents of K-c<strong>on</strong>trols and surplusses (yield in<br />
NPK-NP, t/ha) for the same crops could be described by a hyperbolic curve.<br />
57
Relative yield (100 NP/NPK), %<br />
Relative yield (100 NP/NPK), %<br />
1.A. Winter wheat 1.B. Winter wheat<br />
Y’ = 100 (1-10 –cx ); n = 59; Y’ = 1/(ax+b); n = 59<br />
c: = 0.0153; r = 0.67<br />
4.0<br />
a = 0.04; b = -0.29; r = 0.72<br />
100<br />
80<br />
60<br />
40<br />
20<br />
0<br />
0 100 200 300 400 500<br />
AL-K2O, mg/kg<br />
Grain increases (NPK-NP), t/ha<br />
3.0<br />
2.0<br />
1.0<br />
0.0<br />
0 100 200 300 400 500<br />
AL-K2O, mg/kg<br />
2..A. Maize 2.B. Maize<br />
Y’ = 100 (1-10 –cx ); n = 86; Y’ = 1/(ax+b); n = 86<br />
c: = 0.0083; r = 0.77 a = 0.01; b = -0.02; r = 0.70<br />
100<br />
4.0<br />
80<br />
60<br />
40<br />
20<br />
0<br />
0 100 200 300 400 500 0 100 200 300 400 500<br />
AL-K2O, mg/kg<br />
AL-K2O, mg/kg<br />
Figures 1 and 2. Relati<strong>on</strong>ship between the AL-K2O c<strong>on</strong>tents of K c<strong>on</strong>trol (NP) plots and<br />
resp<strong>on</strong>ses of winter wheat and corn to K fertilizati<strong>on</strong>, calculated from the database of<br />
Hungarian field trials <strong>on</strong> K fertilizati<strong>on</strong>, 1960–2000, using the Bray-Mitscherlich approach<br />
Resp<strong>on</strong>ses to K fertilizati<strong>on</strong> were eliminated when soil K test values exceeded 160-170<br />
mg/kg for winter wheat, 170-180 mg/kg for alfalfa and 200 mg/kg for maize (Figure 1-<br />
3.).<br />
58<br />
Grain increases (NPK-NP), t/ha<br />
3.0<br />
2.0<br />
1.0<br />
0.0
Relatív <strong>term</strong>és (100 NP/NPK), %<br />
3.A. Alfalfa 3.B. Alfalfa<br />
Y’ = 100 (1-10 –cx ); n = 23; Y’ = 1/(ax+b); n = 23<br />
c: = 0.0109; r = 0.80 a = 0.03; b = 0.94; r = 0.74<br />
100<br />
4.0<br />
80<br />
60<br />
40<br />
20<br />
0<br />
0 100 200 300 400 500<br />
AL-K2O, mg/kg<br />
Terméstöbblet (NPK-NP), t/ha<br />
3.0<br />
2.0<br />
1.0<br />
0.0<br />
0 100 200 300 400 500<br />
AL-K2O, mg/kg<br />
Figure 3. Relati<strong>on</strong>ship between the AL-K 2O c<strong>on</strong>tents of K c<strong>on</strong>trol (NP) plots and resp<strong>on</strong>ses of<br />
alfalfa to K fertilizati<strong>on</strong>, calculated from the database of Hungarian field trials <strong>on</strong> K<br />
fertilizati<strong>on</strong>, 1960–2000, using the Bray-Mitscherlich approach<br />
The AL- (amm<strong>on</strong>ium-lactate) (Egner, Riehm and Domingo 1960) soluble K indicated<br />
the natural K- supplying power of the different soils adequately. With higher soil clay<br />
c<strong>on</strong>tents in the trials, the original K supplying capacity of the soils was usually better.<br />
Maize (and wheat) resp<strong>on</strong>ses to K showed a better correlati<strong>on</strong> with the soil clay<br />
c<strong>on</strong>tents, than with the clay mineral compositi<strong>on</strong>s, resp.<br />
Table 1. The effect of soil texture <strong>on</strong> resp<strong>on</strong>ses to K applicati<strong>on</strong> in Hungarian field trials with<br />
maize, set up between 1960 and 2000<br />
Soil<br />
texture<br />
groups<br />
Number<br />
of trials<br />
(n)<br />
homok h. vályog vályog a. vályog agyag<br />
K A * AL-K 2 O<br />
mg/kg<br />
(NP)<br />
K 2 O dose<br />
kg/ha**<br />
(K-c<strong>on</strong>trol)<br />
Grain yield<br />
<strong>on</strong> NP plots<br />
(NP/NPK)<br />
Relative<br />
yield,%<br />
(NPK-NP)<br />
Surplus,<br />
t/ha<br />
(NPK-NP)<br />
Sands 13 29 80 120 4,27 71 1,91<br />
S. loams 8 36 128 100 5,54 88 0,71<br />
Loams 20 39 159 54 6,17 93 0,43<br />
C. loams 27 45 187 33 6,26 97 0,19<br />
Clay soils 5 55 206 0 7,81 100 0,00<br />
Mean 73 40 155 59 5,90 90 0,61<br />
*Upper limit of plasticity, according to Arany (similar to the US plasticity index).<br />
** K 2 O doses needed for maximum ec<strong>on</strong>omic yields (~ 95% of maximum yield)<br />
59
Table 2. The effect of soil texture <strong>on</strong> resp<strong>on</strong>ses to K applicati<strong>on</strong> in Hungarian field trials with<br />
winter wheat, set up between 1960 and 2000<br />
Soil<br />
texture<br />
groups<br />
Number<br />
of trials<br />
(n)<br />
K A * AL-K 2 O<br />
mg/kg<br />
(NP)<br />
K 2 O dose<br />
kg/ha**<br />
(K-c<strong>on</strong>trol)<br />
Grain yield<br />
<strong>on</strong> NP plots<br />
(NP/NPK)<br />
Relative<br />
yield,%<br />
(NPK-NP)<br />
Surplus,<br />
t/ha<br />
(NPK-NP)<br />
Sands 5 28 75 112 3,08 89 0,41<br />
S. loams 7 35 120 66 4,22 94 0,32<br />
Loams 17 39 160 15 4,15 98 0,06<br />
C. loams 20 46 210 7 3,98 98 0,08<br />
Clay soils 5 56 218 0 4,41 100 0,00<br />
Mean 54 42 171 26 4,02 97 0,13<br />
*Upper limit of plasticity, according to Arany (similar to the US plasticity index).<br />
** K 2 O doses needed for maximum ec<strong>on</strong>omic yields (~ 95% of maximum yield)<br />
Table 3. The effect of soil texture <strong>on</strong> resp<strong>on</strong>ses to K applicati<strong>on</strong> in Hungarian field trials with<br />
alfalfa, set up between 1960 and 2000<br />
Soil<br />
texture<br />
groups<br />
Number<br />
of trials<br />
(n)<br />
K A * AL-K 2 O<br />
mg/kg<br />
(NP)<br />
K 2 O dose<br />
kg/ha**<br />
(K-c<strong>on</strong>trol)<br />
Grain yield<br />
<strong>on</strong> NP plots<br />
(NP/NPK)<br />
Relative<br />
yield,%<br />
(NPK-NP)<br />
Surplus,<br />
t/ha<br />
(NPK-NP)<br />
Sands 2 27 60 120 5.55 78 1.60<br />
S. loams 3 35 163 13 8.42 98 0.26<br />
Loams 2 42 205 46 8.95 90 0.74<br />
C. loams 12 45 172 16 7.74 98 0.18<br />
Clay soils 3 55 150 0 8.50 100 0.00<br />
Mean 22 40 175 20 8,92 96 0,33<br />
*Upper limit of plasticity, according to Arany (similar to the US plasticity index).<br />
** K 2 O doses needed for maximum ec<strong>on</strong>omic yields (~ 95% of maximum yield)<br />
On the basis of these data sets, new AL-K 2 O supply categories were elaborated for<br />
maize, winter wheat and alfalfa, according to their different resp<strong>on</strong>ses to K fertilizati<strong>on</strong>.<br />
The new AL-K 2 O supply categories for clay loam and clay soils are much lower<br />
than those in former fertilizer advisory systems.<br />
CONCLUSIONS<br />
For evaluating the c<strong>on</strong>necti<strong>on</strong> between AL-K2O values and resp<strong>on</strong>ses to K fertilizati<strong>on</strong><br />
in Hungarian field trials with maize, alfalfa and winter wheat, Bray's relative index<br />
method was a useful tool. The difference between the resp<strong>on</strong>ses of maize, winter wheat<br />
and alfalfa to K fertilizati<strong>on</strong> was also indicated in the different "c" values. AL- K values<br />
indicated the K-supply of the different soils adequately.<br />
60
ACKNOWLEDGEMENT<br />
This study was financially supported by the Hungarian Nati<strong>on</strong>al Scientific Research<br />
Fund (OTKA), under grant N.o. T 06511.<br />
REFERENCES<br />
Bray, R. H. (1944) Soil-plant relati<strong>on</strong>s: I. The quantitative relati<strong>on</strong> of exchangeable<br />
potassium to crop yields and to crop resp<strong>on</strong>se to potash additi<strong>on</strong>s. Soil Sci. 58,<br />
305-324.<br />
Corey, R.B. (1987) Soil test procedures: Correlati<strong>on</strong>. In: Soil testing: Sampling,<br />
correlati<strong>on</strong>, calibrati<strong>on</strong>, and interpretati<strong>on</strong> (ed.: Brown, J.R.) 15-22. SSSA Special<br />
Publ. N.O. 21. Madis<strong>on</strong>, Wisc. USA.<br />
Csathó, P. (1994) Database of Hungarian K-fertilizati<strong>on</strong> field trials with maize and<br />
winter wheat, 1960-1990. In: (Eds.: Borin, M. and Sattin, M.) Proc. 3rd ESA<br />
C<strong>on</strong>gress, Abano-Padova, Italy. 464-465.<br />
Denke, J. (1974a) (Ed.) Results of fertilizati<strong>on</strong> researches. 2. Maize. KAE Keszthely,<br />
Hungary. 72 p. (In Hungarian)<br />
Denke, J. (1974b) (Ed.) Results of fertilizati<strong>on</strong> researches. 5. Alfalfa. KAE Keszthely,<br />
Hungary. 48 p. (In Hungarian)<br />
Debreczeni, B. and Debreczeni, K. (eds.). (1994) Fertilizati<strong>on</strong> Researches, 1960-1990.<br />
Akadémiai Kiadó, Budapest. (In Hungarian)<br />
Egner, H., et al. (1960) Untersuchungen über die chemische Bodenanalyse als<br />
Grundlage für die Beurteilung des Nahrstoffzustandes der Böden. II. Kungl.<br />
Lantbrukshögskolans Annaler, 26: 199-215.<br />
Sárdi, K. and Németh, T. (1993) Studies <strong>on</strong> the plant available K-c<strong>on</strong>tent of different<br />
soils at c<strong>on</strong>stant moisture. In "Proc. Symp. <strong>on</strong> Soil Resilience and Sustainable<br />
Land Use" (Ed. Szabolcs I) Agrokémia és Talajtan 42,183-194.<br />
Várallyay, Gy. sr. (1950) <strong>Experiments</strong> and analyses directing fertilizer use. Agrokémia<br />
2. 287-302.<br />
61
LONG-TERM FIELD EXPERIMENTS AS THE BASE OF SITE SPECIFIC<br />
FARMING<br />
KESZTHELY LONG-TERM FIELD EXPERIMENTS<br />
Tamas Kismányoky – Zoltan Toth<br />
University of Pann<strong>on</strong>ia Georgik<strong>on</strong> Faculty of Agriculture<br />
L<strong>on</strong>g-<strong>term</strong> field experiments existed, exist and will exist at least in places, where their<br />
importance is recognised. L<strong>on</strong>g-<strong>term</strong> field experiments are the source of such iformati<strong>on</strong><br />
from which we can learn a lot about the factors that influence soil fertility and its<br />
sustainability. In most cases effects and interacti<strong>on</strong>s can be understood <strong>on</strong>ly from l<strong>on</strong>g<strong>term</strong><br />
data even if different soil and climatical c<strong>on</strong>diti<strong>on</strong>s are compared to each other.<br />
These informati<strong>on</strong> are valuable for farmers, extensi<strong>on</strong> c<strong>on</strong>sultants, decisi<strong>on</strong> makers as<br />
well as scientists either in local or wide scale general c<strong>on</strong>cern.<br />
L<strong>on</strong>g-<strong>term</strong> field experiments can be c<strong>on</strong>sidered such living laboratories that let us<br />
study the physical and biological parameters of the agricultural envir<strong>on</strong>ment reliably,<br />
further give scientists and policy decesi<strong>on</strong> makers basic informati<strong>on</strong> in order to follow<br />
and c<strong>on</strong>trol envir<strong>on</strong>mental mechanisms.<br />
The survey and the register of the European l<strong>on</strong>g-<strong>term</strong> field experiments were<br />
published by Körschens (1994, 1997, 1999, 2000). Valuable publicati<strong>on</strong>s were also<br />
made in this topic by Mazur et. al (1993), Debreczeni B. (1994), Leigh and Johnst<strong>on</strong><br />
(1994), Merbach et. al (1999), Smith et. al (2001), Láng et. al (2002), Debreczeni K and<br />
Körschens, M. (2003).<br />
Most of the l<strong>on</strong>g-<strong>term</strong> field experiments were set up to study the effects and<br />
interacti<strong>on</strong>s of crop rotati<strong>on</strong>, c<strong>on</strong>tinuous cropping, organic and mineral fertilizers,<br />
microelements, liming, soil tillage, water management, plant protecti<strong>on</strong>, nutrient<br />
resp<strong>on</strong>d of new varieties and hi-breds, etc. Similar goals were the reas<strong>on</strong> for the<br />
estabilishment of the l<strong>on</strong>g-<strong>term</strong> field experiments in Keszthely, Hungary.<br />
The l<strong>on</strong>g-<strong>term</strong> field experiments of the World and Hungary are listed in Table 1.,<br />
Table 2. and Table 3.<br />
Table 1. L<strong>on</strong>g-<strong>term</strong> field experiments of the World<br />
(Debreczeni K., Körschens, M. 2003)<br />
C<strong>on</strong>tinent 10-20 20-50 50-100 > 100 year Total<br />
Europe 24 292 81 20 417<br />
Africa 32 37 69<br />
Asia 28 7 35<br />
Australia 7 14 21<br />
North-America 1 41 21 5 68<br />
South-America 9 1 10<br />
World’s total 25 409 161 25 620<br />
62
Table 2. The oldest and most important l<strong>on</strong>g-<strong>term</strong> field experiments of the World<br />
(Körschens et. al., 1994)<br />
Place Country Year of estabilishment<br />
Rothamsted England 1843<br />
Grign<strong>on</strong> French 1875<br />
Illi<strong>on</strong>is USA 1876<br />
Halle/Saale Germany 1878<br />
Columbia USA 1888<br />
Dakota USA 1892<br />
Askov Denmark 1894<br />
Auburn USA 1896<br />
Bad Lauchstädt Germany 1902<br />
Dikopshof Germany 1904<br />
Table 3. The oldest l<strong>on</strong>g-<strong>term</strong> field experiments of Hungary<br />
Place Year of estabilishment<br />
MTA TAKI 1959<br />
Putnok 1963<br />
Kompolt 1961<br />
Szarvas 1989<br />
Kecskemét 1959/60<br />
Gödöllő 1970<br />
Karcag 1977<br />
Debrecen 1978<br />
GKI 1979<br />
Gyöngyös 1994<br />
Westsik 1929<br />
Nyirlugos 1962<br />
OMTK 1967<br />
Keszthely 1963<br />
Mart<strong>on</strong>vásár 1959/61<br />
The crop producti<strong>on</strong> research of Europe and Hungary has several comm<strong>on</strong> aspects<br />
either in the fields of research, or in the structure of the research organisati<strong>on</strong>s or in the<br />
principle of funding research. On the other hand the process of implementati<strong>on</strong> is not<br />
uniform. In the western European countries the crop producti<strong>on</strong> research is made by the<br />
universities and by regi<strong>on</strong>al research institutes. The research institutes are financed by<br />
the administrati<strong>on</strong>, or by private organisati<strong>on</strong>s. A part of the agrotechnical research is<br />
n<strong>on</strong> profit type (eg. plant nutriti<strong>on</strong>, crop rotati<strong>on</strong>, soil management), while others like<br />
breeding or pesticide research are profit oriented <strong>on</strong>es.<br />
Obviously <strong>on</strong> those fields, where the experimental results has a direct return (new<br />
species, or hibred, new pesticide or machine etc.) the financial background of the<br />
research is evident. There is a completely opposit situati<strong>on</strong> in case of those researches,<br />
63
that have higher costs than their direct incomes, since the reliable results can be reached<br />
and published after several replicati<strong>on</strong> and besides this publicati<strong>on</strong>s are available for<br />
every<strong>on</strong>e wihout any compensati<strong>on</strong>, but <strong>on</strong> the other hand they are important for the<br />
society.<br />
The results of the applied agrotechnical research can be transfered to the practice<br />
by extensi<strong>on</strong> effectively, if the research institutes are located in that particular regi<strong>on</strong>.<br />
One of the prominent examples for this is the Nati<strong>on</strong>al L<strong>on</strong>g-<strong>term</strong> Field Experiment<br />
Network.<br />
A scientific school was formed since 1945 in Keszthely, Hungary, based <strong>on</strong> the<br />
successful research activity of several decades in the field of soil fertility and nutrient<br />
management. Dr. Kolbai Károly, Dr. Kemenesy Ernő, Dr. Nyéki Jenő, Dr. Láng Géza,<br />
Dr. Kováts András professors had an unexegerateble role in this development.<br />
At the beginning research was aimed at organic (farmyard) manure treatments and<br />
applicati<strong>on</strong>, delivering useful results for the practice.<br />
Since the 1960-es, when the mineral fertilizer applicati<strong>on</strong> rate increased<br />
dramatically, the research was focused <strong>on</strong> the effective applicati<strong>on</strong> of fertilizers in order<br />
to utilize the genetical potential of the different species and hibreds.<br />
Although mineral fertilizer applicati<strong>on</strong> had been playing the main role in the<br />
maintainance of soil fertility, the new questi<strong>on</strong>s of organic matter and organic manures<br />
had to be studed as well. In this period the research work of the department included the<br />
study of the utilizati<strong>on</strong> of agricultural by products like straw (incorporati<strong>on</strong> or burning)<br />
as well as slurry.<br />
The studies covered the specificati<strong>on</strong>s of technologies according to the different<br />
requirements of the species, varieties and hibreeds (plant nutriti<strong>on</strong>, sowing date, crop<br />
rotati<strong>on</strong>, plant density, tillage methods, etc.) as well as the methods of physical and<br />
chemical improvement of soils.<br />
The researh activity of the department focused mainly <strong>on</strong> the yield improvement<br />
and cost effective cropping methodes of the leader field crops as wheat, maize, barley,<br />
potato, sunflower, rape, sugarbeet, and others (soybeen, fababeen).<br />
This studies and trials were c<strong>on</strong>ducted in small and large plot field experiments <strong>on</strong><br />
30 ha arable land in three different sites (Keszthely, Szentgyörgyvölgy,<br />
Homokszentgyörgy-Mariettapuszta) and also in pot experiments with a well equipped<br />
lab. The three different experimental site had different type of forest soils. Ramann-type<br />
brown forest soil (Keszthely), Pseudogleyic brown forest soil (Szentgyörgyvölgy) and<br />
leached sandy soil (Homokszentgyörgy-Mariettapuszta). The results from this different<br />
study sites can be applied for the most part of the Transdanubian regi<strong>on</strong>, where the<br />
different classes of the forest soils are typical.<br />
The half of the l<strong>on</strong>g-<strong>term</strong> field experiments were set up in 1963-1970 (Láng Géza,<br />
Kemenesy Ernő, Kováts András, Nyéki Jenő, Cseh Ervin, Németh István). This trials<br />
represent high scientific values. From the 2-3-4 decade l<strong>on</strong>g database reliable<br />
c<strong>on</strong>clusi<strong>on</strong>s can be drawn about the alterati<strong>on</strong> of soil fertility as well as about the effects<br />
of the different crop years in these different agroecological regi<strong>on</strong>s.<br />
The great increase in the volumen of mineral fertilizer producti<strong>on</strong> and applicati<strong>on</strong> –<br />
beside the previously started experiments – required a country scale study of the<br />
applicati<strong>on</strong> of fertilizers in order to provide a fertilizati<strong>on</strong> advisory system based <strong>on</strong><br />
uniform basic principles. A board was estabilished for this purpose and worked out the<br />
64
plan of the uniform countrywide field trial network in 1965. In this board the staff of the<br />
Department of Plant Producti<strong>on</strong> in Keszthely played significant role (Láng Géza,<br />
Márt<strong>on</strong>ffy Tamás). Then in 1966 the administrati<strong>on</strong> allocated funds for the<br />
estabilishment of this trial network and field experiments have been set up in 18 sites<br />
operated by 9 research organisati<strong>on</strong>s (institutes and universities). In the next step the<br />
number of the experimental sites rised up to 26, from which 3 (Keszthely, Lábod-<br />
Vadaspuszta, Nagykanizsa) bel<strong>on</strong>ged to the Department of Plant Producti<strong>on</strong> in<br />
Keszthely.<br />
As a recognati<strong>on</strong> of the research work of the department the Ministry of<br />
Agriculture and Food Industry estabilished an operati<strong>on</strong>al board in 1968 in order to<br />
improve and help the new uniform field trial network which was chaired by Dr. Láng<br />
Géza, and the secretary was Dr. Márt<strong>on</strong>ffy Tamás. The expenditures of the experiments<br />
were covered by different funding programs (MÉM MÜFA fund, OMFB, Chemical<br />
Industry Uni<strong>on</strong> SZP-2 program and G-9 program since 1986). The Department of Plant<br />
Producti<strong>on</strong> was in charge of the operati<strong>on</strong> of this new Nati<strong>on</strong>al Field Trial (NFT)<br />
Network until 1985, then it was taken over by the Department of Agrochemistry under<br />
Dr. Debreczeni Béla’s and Dr. Debreczeni Béláné’s leadership. In 2002 the operati<strong>on</strong> of<br />
the network got back to the Department of Plant Producti<strong>on</strong> and Soil Science.<br />
Nowadays the number of the experimental sites of the NFT network has reduced to 9<br />
and the financial sources run out of supplies.<br />
The success of the research activity of the department was recognised by the<br />
following facts (it was c<strong>on</strong>sidered as a school and a scientific workshop).<br />
The Ministry of Agriculture and Food Industry launched different middle-<strong>term</strong><br />
operative research programs in 1970, from which the program of fertilizati<strong>on</strong><br />
(„Research <strong>on</strong> Soil Power Management”) was run by the department with great<br />
experience in this field with academician Láng Géza’s leadership. He was also in charge<br />
of the domestic and internati<strong>on</strong>al public relati<strong>on</strong>s, as well as of the coordinati<strong>on</strong> of the<br />
research in this field by making research plans and comprehensive reports.<br />
In 1971 the ministry tended to rise the research focusing <strong>on</strong> the improvement of<br />
soil fertility as the basic factor of the increase of producti<strong>on</strong> and technological progress<br />
to the priority of the country scale research programs. Dr. Géza Láng was the pers<strong>on</strong> in<br />
charge of the preliminary arrangements as a head of the operating board.<br />
In 1972 the ministry decided to start the country scale research program: K-9<br />
„Improvement of soil fertility…”. The program had several chapters as fertilizati<strong>on</strong>, soil<br />
improvement, soil tillage and soil c<strong>on</strong>servati<strong>on</strong>. The coordinator of the research<br />
program was Dr. Láng Géza professor the head of the operating board and senior<br />
researcher of the Keszthely university of Agriculture. The subprogram of soil fertility<br />
was operated directly also by Keszthely. This program worked until 1980. Since 1981<br />
the subprograms c<strong>on</strong>tinued as independent programs. The soil fertility subprogram was<br />
c<strong>on</strong>tinued by the department under the title of „ SZP-2 Agrochemical research”. Since<br />
1986 it was runed as the project No. 6 of the G-9 Research Program.<br />
The department research activity covered the field of the „Study of soil<br />
acidificati<strong>on</strong> as a result of intensive plant producti<strong>on</strong>” c<strong>on</strong>necting to the soil fertility<br />
research program between 1982-1986. In the frame of the project the reas<strong>on</strong>s for soil<br />
acidificati<strong>on</strong> and the methods of their compensati<strong>on</strong> were studied <strong>on</strong> different soil types<br />
65
and in field and pot experiments. Farm scale data are also were processed for this<br />
purpose.<br />
The next important work (1986-1990) in this field was c<strong>on</strong>nected to the project No.<br />
OTKA-513 called „Factors influencing the utilizati<strong>on</strong> of yield potential of the new<br />
varieties, the dinamics of plant growth affected by the envir<strong>on</strong>mental c<strong>on</strong>diti<strong>on</strong>s in case<br />
of some arable crop species and varieties.” The research was carried out either in l<strong>on</strong>g<strong>term</strong><br />
or in annual field experiments in Keszthely, Hungary.<br />
In the following years the centralized systematic funding of these research projects<br />
by the administrati<strong>on</strong> came to an end and the R+D sources of the instituti<strong>on</strong>s decreased<br />
dramatically. In the following years the funding of l<strong>on</strong>g-<strong>term</strong> field experiments have<br />
been based <strong>on</strong> short-<strong>term</strong> projects (OTKA, FVM, NKFP, GVOP, etc.) and their<br />
existence is quite uncertain nowadays.<br />
Since the ’90-es the l<strong>on</strong>g-<strong>term</strong> field experiments have been run by the today’s<br />
generati<strong>on</strong> of researchers. The main l<strong>on</strong>g-<strong>term</strong> field experiments of the department are<br />
as follows:<br />
• Comparative study of organic and mineral fertilizer applicati<strong>on</strong> (1960-63)<br />
• Minimum tillage experiment in wheat-maize diculture (1972)<br />
• Mineral and organic fertilizati<strong>on</strong> in different crop rotati<strong>on</strong>s (1963)<br />
• Nitrogen fertilizati<strong>on</strong> experiment with the study of residual effect (1965 Keszthely,<br />
1965 Szentgyörgyvölgy)<br />
• Phosphorus fertilizati<strong>on</strong> experiment with the study of residual effect (1963<br />
Keszthely, 1965 Szentgyörgyvölgy)<br />
• Potassium fertilizati<strong>on</strong> experiment with the study of residual effect (1963<br />
Keszthely, 1965 Szentgyörgyvölgy)<br />
• Mineral fertilizati<strong>on</strong> in c<strong>on</strong>tinuous maize cropping experiment (1969)<br />
• <str<strong>on</strong>g>Internati<strong>on</strong>al</str<strong>on</strong>g> organic and mineral nitrogen fertilizati<strong>on</strong> experiment (1983)<br />
• Nati<strong>on</strong>al L<strong>on</strong>g-<strong>term</strong> Field Experiment (OMTK) Network trial (1967-69)<br />
The results of the l<strong>on</strong>g-<strong>term</strong> field experiments are published in several journals,<br />
c<strong>on</strong>ference proceedings and books. The aim of our field research is to develop such<br />
systems of fertilizati<strong>on</strong> and agrotechnics that resulted in the greatest yields in given<br />
agroecological c<strong>on</strong>diti<strong>on</strong> without envir<strong>on</strong>mental hazards.<br />
During the history of the agricultural research the aim of the trials was always to<br />
clarify the limiting factors. While in the ’60-es farmers had to be persuaded about the<br />
effects of mineral fertilizer applicati<strong>on</strong> now beside the clarificati<strong>on</strong> of particular<br />
questi<strong>on</strong>s the integrated polyfactorial and ecosystem oriented approach of research is<br />
the typical.<br />
SUMMARY<br />
The importance and necessity of l<strong>on</strong>g-<strong>term</strong> field experiments:<br />
• L<strong>on</strong>g-<strong>term</strong> effects can <strong>on</strong>ly be studed after several decades reliably.<br />
• The agrotechnical development (chemical, biological, etc.) of the last decades can<br />
be measured by l<strong>on</strong>g-<strong>term</strong> data and the plans for the future should also be based <strong>on</strong><br />
this data base.<br />
• Nutrient balances can be estimated reliably.<br />
66
• The effect of climate change can be estimated by the comparis<strong>on</strong> of the l<strong>on</strong>g-<strong>term</strong><br />
data of the different study sites.<br />
• L<strong>on</strong>g-<strong>term</strong> database is the background of the computer models.<br />
• L<strong>on</strong>g-<strong>term</strong> field experiments are the references of the the sustainable development<br />
of agriculture and envir<strong>on</strong>ment.<br />
REFERNCES<br />
Debreczeni, B., Debreczeni, Bné. (1994) Trágyázási kutatások 1960-1990. Akadémiai<br />
Kiadó Budapest, p. 411.<br />
Debreczeni, K., Körschens, M. (2003) L<strong>on</strong>g-Term Field <strong>Experiments</strong> of the World<br />
Archives of Agr<strong>on</strong>omy and Soil Science, October 2003, Vol. 49, pp. 465-483.<br />
Körschens, M. (2000) IOSDV <str<strong>on</strong>g>Internati<strong>on</strong>al</str<strong>on</strong>g>e organische<br />
Stickstoffdauerdüngungsversuche, UFZ-Bericht No./15/2000. Leipzig-Halle<br />
GMBH. ISSN 0948-9452, p. 192.<br />
Körschens, M. (1999) Experimentelle Möglichkeiten zur Ableitung optimaler Corg-<br />
Gehalt in Ackerboden., UFZ-Bericht No./9/1999. Beziehungen Zwischenb<br />
organischer Bodensubstanz und bodenmikrobiologischen Prozessen. Leipzig-Halle<br />
GMBH. ISSN 0948-9452, pp. 75-94.<br />
Körschens, M. (1997) Die Wichtigsten Dauerfeldversuche der Welt-Übersicht,<br />
Bedeutung, Ergebnisse. Arch. Acker- Pfl. Boden., Vol. 42, pp. 157-168.<br />
Körschens, M. (1994) Der Statische Düngungsversuch Bad Lauchstädt nach 90 Jahren.<br />
B.G. Teubner Verlagsgesellschaft Stuttgart-Leipzig, p. 179.<br />
Láng, I., Lazányi, J. and Németh, T. (2002) Tartamkísérletek, táj<strong>term</strong>esztés,<br />
vidékfejlesztés. Nemzetközi K<strong>on</strong>ferencia I., Debrecen, Nyírlugos, Nyíregyháza,<br />
Livada Sárköz, Románia 2002. június 6-8. ISBN 963-472-654-20 p. 418.<br />
Leigh, R. A. and Johnst<strong>on</strong>, A. E. (1994) L<strong>on</strong>g-<strong>term</strong> <strong>Experiments</strong> in Agricultural and<br />
Ecological Sciences. Proceedings of a c<strong>on</strong>ference to celebrate the 150 th<br />
Anniversary of rothamsted Experimental Stati<strong>on</strong>, held at Rothamsted, 14-17 July<br />
1993, p. 428.<br />
Mazur, K., Filipek, J. and Mazur, B. (1993) Proceedings of the <str<strong>on</strong>g>Internati<strong>on</strong>al</str<strong>on</strong>g><br />
Symposium ’L<strong>on</strong>g-<strong>term</strong> static fertilizer experiments’ June 15-18, 1993. Warszawa-<br />
Kraków, Poland. 253.<br />
Merbach, W., Schmidt, L., Garz, J., Stumpe, H. und Schliephake, W. (1999) Die<br />
Dauerdüngungsversuche in Halle-ein Überblick. UFZ-Bericht No.24/1999<br />
Dauerdüngungsversuche als Grundlage für nachhaltige Landnutzung und<br />
Quantifizierung v<strong>on</strong> Stoffkreisläufen, ISSN 0948-9452.<br />
Smith, P., Fall<strong>on</strong>, P., Smith, J. U. and Powls<strong>on</strong>, D. S. (2001) GCTE Global Change<br />
and Terrestrial Ecosystems. Report No 7. 2 nd edn. Soil Organic Matter Network<br />
(SOMNET): Model and Experomental Metadata. Published by the CGTE Focus 3<br />
Office Wallingford, UK, p. 223.<br />
67
LONG-TERM EFFECT OF ORGANIC AND MINERAL FERTILIZATION ON<br />
DIFFERENT SOIL-FERTILITY PARAMETERS<br />
ABSTRACT<br />
Sándor Hoffmann and Katalin Berecz<br />
University of Pann<strong>on</strong>ia, Georgik<strong>on</strong> Faculty of Agricultural Sciences<br />
Deák F. str. 16, H-8360 Keszthely/Hungary<br />
Eleven variati<strong>on</strong>s of farmyard manuring (FYM) and mineral fertilizati<strong>on</strong> treatments of a<br />
l<strong>on</strong>g-<strong>term</strong> field trial have been selected for the present investigati<strong>on</strong>s. Attributes of<br />
changes in soil fertility were investigated. FYM and equivalent NPK fertilizati<strong>on</strong><br />
resulted in similar pHH2O values, while FYM increased TOC c<strong>on</strong>tent of the soil by 10%<br />
as compared to equivalent NPK fertilizer. FYM increased K, while equivalent doses of<br />
NPK fertilizer increased P c<strong>on</strong>tent of soil to a higher degree, as compared to nil plot.<br />
Yearly 14 t ha -1 FYM or equivalent NPK fertilizer kept soil P and K c<strong>on</strong>tents at the<br />
range of “medium-well-supplied”. However, not even yearly 21 t ha -1 of FYM resulted<br />
in maximum yields in 1998-2006. 35; 70 and 105 t FYM ha -1 5 year -1 increased grain<br />
yields to a lesser extent (by 0.66, 1.44 and 0.85 t ha -1 , respectively), as compared to<br />
equivalent NPK doses, which was mainly due to the better utilizati<strong>on</strong> of mineral<br />
fertilizer. After harvesting all the plots c<strong>on</strong>tained relatively high amounts of nitrate in<br />
the 0-300 cm soil layer. Nitrate c<strong>on</strong>tent of the n<strong>on</strong>-treated plot was c<strong>on</strong>siderably high as<br />
well. Despite the low yield resp<strong>on</strong>se to FYM-treatments, less nitrate was found in the<br />
soil, than with mineral fertilizati<strong>on</strong>.<br />
INTRODUCTION<br />
Field experiments are essential means for studying the changes in soil fertility as<br />
affected by different fertilizati<strong>on</strong> and farming systems. With changing ec<strong>on</strong>omical and<br />
social c<strong>on</strong>diti<strong>on</strong>s and farming systems, also the judgement of the role of farmyard<br />
manuring (FYM) kept changing. The effect of FYM <strong>on</strong> soil fertility is very different.<br />
However, the nutrient c<strong>on</strong>tent of FYM have the most direct and highest effect, which<br />
can be compared to the effect of equivalent NPK fertilizer doses. This effectiveness is<br />
influenced by various factors and it is generally lower, than in case of fertilizers with<br />
the same active ingredients and equivalent doses. Nevertheless, this tendency may turn<br />
with the increase of clay c<strong>on</strong>tent in the soil (Sarkadi, 1993). The NPK fertilizer<br />
equivalency of FYM can be de<strong>term</strong>ined not <strong>on</strong>ly in experiments comparing the effect of<br />
FYM and NPK fertilizers of equivalent doses, but also in combined polyfactorial<br />
experiments (Baldock and Musgrave, 1980; Bouldin et al., 1984), and the results are<br />
nearly similar (Hoffmann, 1999). Many authors detected that the nutrient effect of FYM<br />
can be fully substituted for fertilizati<strong>on</strong> (Stumpe et al., 2000), others founded that the<br />
effect of fertilizati<strong>on</strong> is higher except for the case of very heavy soils (Árendás and<br />
Csathó, 1994). On the other hand, the sec<strong>on</strong>dary effect of FYM, namely the increase in<br />
general soil fertility is very important and it manifests itself in the structure, organic<br />
matter compositi<strong>on</strong> and microbial activity of soil, and this effect can not be substituted<br />
for mineral N-fertilizati<strong>on</strong> in many instances (Albert, 2001).<br />
68
MATERIALS AND METHODS<br />
An organic and mineral fertilizati<strong>on</strong> field experiment was established with a randomized<br />
block design, four replicati<strong>on</strong>s, and two crop rotati<strong>on</strong>s <strong>on</strong> Keszthely in 1963. The initial<br />
soil (Eutric Cambisol) parameters of the l<strong>on</strong>g-<strong>term</strong> experiment were as follows: (KA =<br />
37; pHH2O = 7.3, TOC = 0.87; AL-P2O5 = 27; AL-K2O = 135 mg kg -1 ). The plot size was<br />
98 m 2 . In the present study, <strong>on</strong>ly the results of crop rotati<strong>on</strong> potato - maize - maize -<br />
winter wheat - winter wheat and 11 of the 15 treatments of the experiment (35, 70, 105 t<br />
ha -1 FYM or mineral fertilizer with equivalent NPK doses or FYM+NPK combinati<strong>on</strong>s<br />
as shown in Table 1) are discussed. In case of simple, double and triple doses, FYM was<br />
distributed before sowing of the potato for 5 years, timed half-and-half to the potato for<br />
5 years, and to the maize (in the autumn of the previous year) in the 3 rd year,<br />
respectively. Grain yields in cereal units [CU] (potato tuber yields were multiplied by<br />
0.3.) in 1998-2006, pHH2O, organic carb<strong>on</strong> (TOC), P, K and mineral N-c<strong>on</strong>tent of the 0-<br />
300 cm soil layer (in increments of 30 cm, 4 sample per plot), as attributes of changes in<br />
soil fertility were evaluated.<br />
Composite soil samples were taken from the 0-20 cm soil layer after harvest, 10<br />
samples for each plot. P and K c<strong>on</strong>tent of the soil was de<strong>term</strong>ined applying amm<strong>on</strong>ium<br />
lactate extractant, which is used in Hungary for routine soil tests (Egner-Rhiem-<br />
Domingo, 1960). TOC c<strong>on</strong>tents in the ploughed soil layer were measured by the<br />
classical methods of Turin. Mineral-N was extracted with 1% KCl then NH4-N and<br />
NO3-N were analysed by steam distillati<strong>on</strong> and titrati<strong>on</strong> procedure.<br />
Treatment<br />
No.<br />
Table 1. Selected treatments of the experiment<br />
Codes of treatments kg NPK ha -1 yr -1<br />
1. C<strong>on</strong>trol -<br />
2. 1 FYM N44P38K49<br />
3. 2 FYM N88P76K98<br />
4. 3 FYM N132P114K147<br />
5. 1 eqv N44P38K49<br />
6. 2 eqv N88P76K98<br />
7. 3 eqv N132P114K147<br />
8. 1 FYM + 3 eqv N176P152K196<br />
9. 4 eqv N176P152K196<br />
10. 1 FYM + NPK N172P110K181<br />
11. 1 eqv + NPK N172P110K181<br />
Legends: 1 FYM= 35 t ha -1 farmyard manure in 5 years<br />
1 eqv = mineral NPK equivalent to 35 t ha -1 FYM in 5 years<br />
NPK = N640P360K660 in 5 years<br />
69
RESULTS AND DISCUSSION<br />
Soil pHH20 in the c<strong>on</strong>trol plot was 7.02, which is 0.3 pHH2O lesser than the initial value<br />
(7.3). Very similar pH values were measured with the different treatments and the<br />
different fertilizer type or doses practically did not change the soil acidity. On an<br />
average of the doses, 40 years l<strong>on</strong>g-<strong>term</strong> FYM and equivalent NPK treatments resulted<br />
in pHH20 values of 6.94 and 6.99, respectively (Figure 1). Not even the treatments with<br />
the highest NPK c<strong>on</strong>tent caused lower pH values.<br />
pH H2O<br />
7,6<br />
7,4<br />
7,2<br />
7<br />
6,8<br />
6,6<br />
6,4<br />
6,2<br />
6<br />
5,8<br />
5,6<br />
5,4<br />
5,2<br />
5<br />
c<strong>on</strong>trol<br />
ab<br />
1FYM<br />
ab<br />
1eqv<br />
ab<br />
2FYM<br />
ab<br />
2eqv<br />
a<br />
3FYM<br />
ab<br />
3eqv<br />
ab<br />
1FYM+3eqv<br />
ab<br />
Figure 1: pHH2O values of soil<br />
4eqv<br />
b<br />
1FYM+NPK<br />
ab<br />
1eqv+NPK<br />
TOC c<strong>on</strong>tent of soil was evaluated in two sampling dates, in the 20 th and 40 th years of<br />
the field experiment. The type of the fertilizer influenced the organic matter c<strong>on</strong>tent of<br />
the soil definitely (Figure 2) FYM treatments al<strong>on</strong>e increased TOC by 17% <strong>on</strong> the<br />
average, while equivalent NPK doses <strong>on</strong>ly by 7%, as compared to the c<strong>on</strong>trol plot. A<br />
clear- cut c<strong>on</strong>clusi<strong>on</strong> about the effect of combined treatments <strong>on</strong> the TOC c<strong>on</strong>tent can<br />
not be drawn. No definite change in the TOC c<strong>on</strong>tent could be detected as an affect of<br />
20- or 40-years organic and/or mineral fertilizati<strong>on</strong>.<br />
70<br />
ab
TOC %<br />
1,2<br />
1<br />
0,8<br />
0,6<br />
0,4<br />
0,2<br />
0<br />
bc b c c<br />
bc bc<br />
ab c<br />
bcab<br />
ab<br />
ab ab<br />
ab<br />
ab ab<br />
a aab<br />
a<br />
a<br />
C<strong>on</strong>t<br />
1FYM<br />
2FYM<br />
3FYM<br />
1 eqv<br />
2 eqv<br />
3 eqv<br />
1FYM+3eqv<br />
4 eqv<br />
Figure 2: TOC c<strong>on</strong>tent of soil<br />
1FYM+NPK<br />
1eqv+NPK<br />
Figures 3 and 4 show data about the l<strong>on</strong>g-<strong>term</strong> impact of organic or mineral<br />
fertilizati<strong>on</strong> <strong>on</strong> the phosphorus and potassium c<strong>on</strong>tent. FYM and equivalent NPK<br />
influenced soil P and K c<strong>on</strong>tent differently. While mineral NPK fertilizati<strong>on</strong> resulted in<br />
22 mg kg -1 more P2O5 in the soil than FYM, K2O c<strong>on</strong>tent of the soil in the FYM treated<br />
plots was 26 mg kg -1 higher than in case of mineral fertilizati<strong>on</strong>.<br />
In spite of the differences between the effect of the organic and mineral fertilizers,<br />
it can be stated, that yearly14 t ha -1 FYM (70 t ha -1 5 yr -1 ) or equivalent PK fertilizati<strong>on</strong><br />
can keep soil P and K c<strong>on</strong>tents at the range of “medium-well-supplied”, taking into<br />
c<strong>on</strong>siderati<strong>on</strong> the absolute soil P and K levels.<br />
P 2O 5<br />
mg kg<br />
250<br />
-1<br />
200<br />
150<br />
100<br />
50<br />
0<br />
C<strong>on</strong>t<br />
1FYM<br />
2FYM<br />
3FYM<br />
1 eqv<br />
2 eqv<br />
3 eqv<br />
1FYM+3eqv<br />
4 eqv<br />
Figure 3: AL-P2O5 c<strong>on</strong>tent of soil<br />
71<br />
1FYM+NPK<br />
1eqv+NPK
K 2O mg kg -1<br />
300<br />
250<br />
200<br />
150<br />
100<br />
50<br />
0<br />
C<strong>on</strong>t<br />
1FYM<br />
2FYM<br />
3FYM<br />
1 eqv<br />
2 eqv<br />
3 eqv<br />
1FYM+3eqv<br />
4 eqv<br />
1FYM+NPK<br />
Figure 4: AL-K2O c<strong>on</strong>tent of soil<br />
1eqv+NPK<br />
Figure 5 shows the crop yields expressed in cereal units and in the average of 1998-<br />
2006.<br />
t ha<br />
8<br />
7<br />
6<br />
5<br />
4<br />
3<br />
2<br />
1<br />
0<br />
-1<br />
c<strong>on</strong>trol<br />
a<br />
1FYM<br />
b<br />
2FYM<br />
b<br />
3FYM<br />
c<br />
Crop yields 1998-2006<br />
1eqv<br />
c<br />
2eqv<br />
d<br />
3eqv<br />
de<br />
1FYM+3eqv<br />
de<br />
4eqv<br />
de<br />
1FYM+NPK<br />
Figure 5: Crop yields (CU) t ha -1 1998-2006<br />
e<br />
1eqv+NPK<br />
Despite the relative high soil P and K levels, neither 14 t ha -1 yr -1 nor 21 t ha -1 yr -1<br />
FYM resulted in maximum yields. The organic fertilizer doses, (35; 70 and 105 t<br />
FYM ha -1 5 year -1 ) increased grain yields to a lesser extent (by 0.66, 1.44 and 0.85 t ha -<br />
1 , respectively), as compared to equivalent NPK doses. The average crop yield<br />
amounted to 5.103 t ha -1 with FYM, while to 6.154 t ha -1 with equivalent NPK. The<br />
increase was 28% as compared to the c<strong>on</strong>trol, which was mainly due to the better<br />
utilizati<strong>on</strong> of fertilizer.<br />
72<br />
e
The much better performance of the mineral nutrient form can be the result of the better<br />
utilizati<strong>on</strong>, especially that of N.<br />
The higher TOC c<strong>on</strong>tents as a result of the FYM treatments suggest, that higher N<br />
quantities were incorporated into the humic substances from the organic fertilizer,<br />
which resulted in a lesser N supply and lower crop yields. The highest yields could be<br />
achieved with the combined treatments with high NPK c<strong>on</strong>tent<br />
NO3-N (mg/kg)<br />
0 20<br />
0<br />
30<br />
60<br />
90<br />
120<br />
150<br />
180<br />
210<br />
240<br />
270<br />
300<br />
C<strong>on</strong>trol<br />
NO3-N (mg/kg)<br />
1 FYM<br />
NO3-N (mg/kg)<br />
1 eqv FYM<br />
NO3 – N mgkg -1<br />
NO3-N (mg/kg)<br />
2 FYM<br />
NO3-N (mg/kg)<br />
2 eqv FYM<br />
NO3-N (mg/kg)<br />
Figure 6 :Nitrate-N c<strong>on</strong>tent of soil (0-300cm)mgkg -1<br />
After harvest all the plots c<strong>on</strong>tained relatively high amounts of nitrate in the 0-300 cm<br />
soil layer (Figure 6). Nitrate c<strong>on</strong>tent of the n<strong>on</strong>-treated plot was c<strong>on</strong>siderably high as<br />
well. Despite the low yield resp<strong>on</strong>se to FYM-treatments, less nitrate was found in the<br />
soil, than with mineral fertilizati<strong>on</strong>. The lower nitrate reserves in the soil with FYM<br />
treatments, in parallel with the lower crop yields, also c<strong>on</strong>firms our suggesti<strong>on</strong> that<br />
higher N quantities were incorporated into the humic substances from the organic than<br />
from the mineral fertilizers.<br />
The amount of amm<strong>on</strong>ium was higher, but it did not show any correlati<strong>on</strong> with<br />
fertilizati<strong>on</strong>.<br />
73<br />
3 FYM<br />
NO3-N (mg/kg)<br />
3 eqv FYM<br />
NO3-N (mg/kg)<br />
4 eqv FYM
REFERENCES<br />
Albert, E. (2001): Effect of l<strong>on</strong>g-<strong>term</strong> different mineral and organic fertilizati<strong>on</strong> <strong>on</strong><br />
yields, humus c<strong>on</strong>tent, netN-mineralizati<strong>on</strong> and N-balance. Arch. Acker-Pfl.<br />
Boden., Vol. 46,. 187-213 p.<br />
Árendás T., Csathó P. (1994): Az<strong>on</strong>os NPK – hatóanyagú szerves- és műtrágyázás<br />
hatása a talajtulajd<strong>on</strong>ságok függvényében Agrokémia és Talajtan 43: 399-408 p.<br />
Baldock, J.O.,Musgrave, R.B. (1980): Manure and mineral fertilizer effect in<br />
c<strong>on</strong>tinous and rotati<strong>on</strong>al crop sequences in Central New York. Agr<strong>on</strong>. J. 72, 511-<br />
518 p.<br />
Bouldin, D.R., Klauser, S.D., Ried; W.S. (1984): Use of nitrogen from manure.<br />
Nitrogen in crop producti<strong>on</strong>, Madis<strong>on</strong> Wisc<strong>on</strong>sin USA.. 221-243 p.<br />
Egner, H., Rhiem, H., Domingo. W.R. (1960): Untersuchungen über die chemische<br />
Bodenanalyse als Grundlage für die Beurteilung des Nährstoffzustandes der<br />
Boden. II. K. Lantbr. Hogsk. Ann. 26:199 p.<br />
Hoffmann, S. (1999): Az istállótrágya N-egyenértékének megállapítása nem<br />
hatóanyagaz<strong>on</strong>osságra alapozott tartamkísérletekben. „Növény<strong>term</strong>esztés és<br />
környezetvédelem”. Szerk.: Ruzsányi-Pepó. MTA Agrártudományok Osztály<br />
Kiadványa. Budapest, 97-101 p.<br />
Sarkadi, J. (1993): Szerves- és műtrágyák tápelemtartalmának érvényesülése<br />
tartamkísérletekben. I. Nitrogénforgalom. Agrokémia és Talajtan 42: 293-308 p.<br />
Stumpe, H. et al. (2000): Effect of humus c<strong>on</strong>tent, farmyard manuring, and mineral-N<br />
fertilizati<strong>on</strong> <strong>on</strong> yields and soil properties in a l<strong>on</strong>g-<strong>term</strong> trial. J. Plant Nutr. Soil.<br />
Sci. 163: 657-662 p.<br />
74
GREENHOUSE GAS EMISSION IN LONG-TERM TILLAGE AND<br />
ROTATION EXPERIMENTS<br />
1 Anita Gál, 1 Péter Hegymegi, 1 Erika Michéli, 2 T<strong>on</strong>y J. Vyn<br />
1 Szent István University, Soil Science and Agrochemistry Department, Gödöllő, HU<br />
2 Purdue University, Department of Agr<strong>on</strong>omy, West Lafayette, Indiana, USA<br />
ABSTRACT<br />
Greenhouse gases such as carb<strong>on</strong>-dioxide (CO2), and nitrous oxide (N2O) are str<strong>on</strong>gly<br />
linked to global warming. Soil organic matter is influenced by tillage and rotati<strong>on</strong>, and<br />
through its decompositi<strong>on</strong> and accumulati<strong>on</strong>, soils have a major effect <strong>on</strong> the gas<br />
compositi<strong>on</strong> of the atmosphere. Tillage systems vary substantially in the intensity and<br />
depth of soil disturbance, in crop residue placement, and in the resulting depths of soil<br />
carb<strong>on</strong> gains or losses. The actual effect of different tillage practices <strong>on</strong> soil carb<strong>on</strong><br />
storage is highly dependent <strong>on</strong> the crops produced in the field. Several studies have<br />
been c<strong>on</strong>ducted to de<strong>term</strong>ine the quantity and distributi<strong>on</strong> of organic carb<strong>on</strong> and total<br />
nitrogen under different tillage and rotati<strong>on</strong> systems in l<strong>on</strong>g-<strong>term</strong> studies but less is<br />
known about the relati<strong>on</strong>ship of OC and N stored in the soil and the emissi<strong>on</strong> of CO2,<br />
and N2O from the soil over the crop producti<strong>on</strong> cycle. In our research we have<br />
compared the l<strong>on</strong>g-<strong>term</strong> effects of c<strong>on</strong>servati<strong>on</strong> (no-till) and c<strong>on</strong>venti<strong>on</strong>al (chisel plow<br />
and moldboard plow) tillage systems <strong>on</strong> soil CO2, and N2O emissi<strong>on</strong>, in c<strong>on</strong>tinuous corn<br />
and soybean-corn rotati<strong>on</strong>. Our results show that both tillage and rotati<strong>on</strong> systems<br />
influence significantly gas emissi<strong>on</strong>. Carb<strong>on</strong>-dioxide and nitrous-oxide emissi<strong>on</strong><br />
declines in the growing seas<strong>on</strong> with time. Nitrous-oxide emissi<strong>on</strong> increases after<br />
applicati<strong>on</strong> of nitrogen fertilizer and after heavy rain events. Corn m<strong>on</strong>oculture gives<br />
the highest emissi<strong>on</strong> of carb<strong>on</strong>-dioxide, and also for nitrous-oxide, rotati<strong>on</strong>s including<br />
corn or corn m<strong>on</strong>oculture have the highest emissi<strong>on</strong>. As for tillage treatments, chisel<br />
plowing results in the highest emissi<strong>on</strong> of nitrous-oxide.<br />
Keywords: organic matter, carb<strong>on</strong> sequestrati<strong>on</strong>, no-tillage, gas emissi<strong>on</strong>, rotati<strong>on</strong><br />
INTRODUCTION<br />
Greenhouse gases such as carb<strong>on</strong>-dioxide (CO2), and nitrous oxide (N2O) are str<strong>on</strong>gly<br />
linked to global warming because of their effects <strong>on</strong> radiati<strong>on</strong> (Sparks, 2003). Organic<br />
matter is a major soil comp<strong>on</strong>ent which is influenced by tillage and rotati<strong>on</strong>, and<br />
through its decompositi<strong>on</strong> and accumulati<strong>on</strong>, soils have a major effect <strong>on</strong> the gas<br />
compositi<strong>on</strong> of the atmosphere (Eswaran et al., 1993, Kimble et al., 1990).<br />
Tillage systems vary substantially in the intensity and depth of soil disturbance, in<br />
crop residue placement, and in the resulting depths of soil carb<strong>on</strong> gains or losses. In notill,<br />
which is the ultimate c<strong>on</strong>servati<strong>on</strong> tillage system, soil disturbance is limited to the<br />
opening of a small slot for seed and fertilizer placement. In c<strong>on</strong>trast, moldboard plowing<br />
inverts the soil and most surface residue to a depth between 20-25 cm (Phillips et al.,<br />
1980). Chisel plowing loosens soil to similar depths but does not invert the soil profile,<br />
so it distributes residues more evenly and at shallower depths than moldboard plow.<br />
After several years, soils which have underg<strong>on</strong>e no-till have a higher carb<strong>on</strong><br />
75
c<strong>on</strong>centrati<strong>on</strong> in the surface layer than moldboard plowed or chiseled plots. On the other<br />
hand, carb<strong>on</strong> levels at lower depths are generally similar in both systems, or higher<br />
under moldboard plow (Angers et al. 1997, Gal, 2005).<br />
The actual effect of different tillage practices <strong>on</strong> soil carb<strong>on</strong> storage is highly<br />
dependent <strong>on</strong> the crops produced in the field. Crop rotati<strong>on</strong> c<strong>on</strong>tributes to increased<br />
yield (Mitchell et al., 1991) and nitrogen c<strong>on</strong>tributi<strong>on</strong> by legumes can be very<br />
important. Crop rotati<strong>on</strong> is known to increase soil microbial activity, which has the<br />
direct effect of breaking down organic matter by the microbes. Residue quantity is the<br />
main factor in de<strong>term</strong>ining soil organic carb<strong>on</strong>, for example lower soil organic carb<strong>on</strong><br />
level corresp<strong>on</strong>ds with sequences including soybean, but organic carb<strong>on</strong> increases as<br />
soybean is not present or corn is present in a sequence according to Studdert et al.<br />
(2000). However, the effect of residue amount is c<strong>on</strong>diti<strong>on</strong>ed by several other factors.<br />
Several studies have been c<strong>on</strong>ducted to de<strong>term</strong>ine the quantity and distributi<strong>on</strong> of<br />
organic carb<strong>on</strong> and total nitrogen under different tillage and rotati<strong>on</strong> systems in l<strong>on</strong>g<strong>term</strong><br />
studies (Kern and Johns<strong>on</strong>, 1993; Angers et al., 1997; VandenBygaart et al., 2002).<br />
Gal et al. (2007) have found <strong>on</strong> the same experimental plots that while no-till clearly<br />
resulted in more organic carb<strong>on</strong> and nitrogen accumulati<strong>on</strong> in the surface 15 cm than<br />
moldboard plow, the relative no-till advantage declined sharply witd depth. Indeed,<br />
moldboard plow resulted in more organic carb<strong>on</strong> and total nitrogen int he 30-50 cm<br />
depth interval. However, less is known about the relati<strong>on</strong>ship of OC and N stored in the<br />
soil and the emissi<strong>on</strong> of CO2, and N2O from the soil over the crop producti<strong>on</strong> cycle. In<br />
order to understand the effect of different tillage and rotati<strong>on</strong> systems <strong>on</strong> carb<strong>on</strong><br />
sequestrati<strong>on</strong> - which is the l<strong>on</strong>g-<strong>term</strong> storage of carb<strong>on</strong> in soil and can help to stop the<br />
increase of greenhouse gases in the atmosphere - we have to examine these processes<br />
more in details. Extended knowledge of the factors influencing soil capacity to be a net<br />
sink or source of greenhouse gases in the atmosphere is <strong>on</strong>e of the most relevant and<br />
urgent needs in agricultural policy making and also in extensi<strong>on</strong>.<br />
The objective of the research was to compare the l<strong>on</strong>g-<strong>term</strong> effects of c<strong>on</strong>servati<strong>on</strong><br />
(no-till) and c<strong>on</strong>venti<strong>on</strong>al (chisel plow and moldboard plow) tillage systems <strong>on</strong> soil<br />
CO2, and N2O emissi<strong>on</strong>, in c<strong>on</strong>tinuous corn and soybean-corn rotati<strong>on</strong>.<br />
MATERIALS AND METHODS<br />
Research area<br />
The sampling site chosen for the study is located at Purdue University (West Lafayette,<br />
IN, USA), at the Agr<strong>on</strong>omy Center for Research and Educati<strong>on</strong>. The soil is a poorly<br />
drained Chalmers silty clay loam (Typic Haplaquoll according to the US Tax<strong>on</strong>omy)<br />
dark, with approximately 4 % organic matter in the surface 30 cm, developed under<br />
prairie vegetati<strong>on</strong> (Vyn et al., 2000). The L<strong>on</strong>g-Term tillage experiment has been<br />
c<strong>on</strong>ducted for 30 years. Moldboard plow, chisel plow and no-till tillage treatments were<br />
compared in c<strong>on</strong>tinuous corn and soybean-corn rotati<strong>on</strong>.<br />
The experiment is a randomized complete block in a split-plot (small plots) design<br />
with treatments replicated four times. Crop rotati<strong>on</strong>s are the main units, and the tillage<br />
treatments the subunits. In the course of this experiment, starter N (36 kg N ha -1 as 34-0-<br />
0) and side-dressed N (222 kg N ha -1 as 28% urea amm<strong>on</strong>ium nitrate, UAN) were<br />
applied.<br />
76
Carb<strong>on</strong>-dioxide and nitrous-oxide sampling<br />
Carb<strong>on</strong>-dioxide and nitrous-oxide fluxes were measured by the vented flux chamber<br />
method. In each measurement plot, duplicate anchors measuring 73.7 X 35.4 X 12.0 cm<br />
capped with U-shaped channels (1.8 cm wide by 1.9 cm deep) welded to the outer-edge<br />
were driven about 10 cm into the soil. Within each plot, anchors were placed 10 m<br />
apart and a carpenter’s level was used to ensure that the anchors were level with the soil<br />
surface. Gas fluxes were measured by placing chambers that measured 75.8 X 38 X 13<br />
cm over the anchors to cover the soil surface. To ensure that there was no gas exchange<br />
between chamber and atmosphere during sampling, the U-shaped channel of the anchor<br />
was filled with water to form an air-tight seal before sampling began. Samples were<br />
then collected from the chambers through a rubber septum at regular intervals of 0, 5,<br />
10, and 15 min after deployment using a 20 mL polypropylene syringe and pressurized<br />
into 12 mL pre-evacuated vials. Gas sampling was d<strong>on</strong>e 14 times <strong>on</strong>t he following days:<br />
June 15, 19, 20, 22, 27, 29; July 6, 10, 14, 21, 26; and August 4, 9, 16. CO2, and N2O<br />
c<strong>on</strong>centrati<strong>on</strong> of the gas samples was de<strong>term</strong>ined using Varian CP 3800 Gas<br />
Chromatograph.<br />
Soil temperature and soil moisture data<br />
Soil moisture c<strong>on</strong>tent (%) and soil temperature (°C) measurement was d<strong>on</strong>e together<br />
with gas sampling <strong>on</strong> the same sampling points. Soil moisture was de<strong>term</strong>ined with a<br />
TRIME-TDR equipment at depth of 12 cm. Soil temperature was measured with a<br />
thermometer at 10 cm depth close to the gas samling chambers. Daily precipitati<strong>on</strong><br />
(mm) and average daily air temperature (°C) data were compiled from the<br />
meteorological stati<strong>on</strong> of the experimental farm (http://agmetx.agry.purdue.edu).<br />
Statistical analysis<br />
Statistical analysis was performed with SAS Versi<strong>on</strong> 8.2 program (SAS Institute, Inc.,<br />
2002). Multivariate linear regressi<strong>on</strong> analysis was carried out to de<strong>term</strong>ine the effect of<br />
soil moisture c<strong>on</strong>tent, soil temperature and day of sampling (after applicati<strong>on</strong> of<br />
nitrogen fertilizer), besides the effect of tillage and rotati<strong>on</strong> treatments for carb<strong>on</strong>dioxide<br />
and nitrous-oxide emissi<strong>on</strong>. Predicted values of gas emissi<strong>on</strong> were calculated<br />
using the regressi<strong>on</strong> equati<strong>on</strong>s.<br />
RESULTS AND DISCUSSION<br />
Carb<strong>on</strong>-dioxide emissi<strong>on</strong><br />
Carb<strong>on</strong>-dioxide emissi<strong>on</strong> was significantly (P=0,05) influenced by soil temperature and<br />
sampling day. Carb<strong>on</strong>-dioxide emissi<strong>on</strong> was the highest in no-till , although the<br />
emissi<strong>on</strong> was higher than 6000 mg/m 2 /hr in all treatments; am<strong>on</strong>g crop rotati<strong>on</strong>s corn<br />
m<strong>on</strong>oculture showed the highest emissi<strong>on</strong> values. Figure 1 shows the predicted values<br />
of CO2 emissi<strong>on</strong> influenced by soil temperature and sampling day.<br />
77
CO 2 flux (mg/m 2 /hr)<br />
CO 2 emissi<strong>on</strong> (mg/m 2 /hr) BC-CH+NT vs BC-PL vs CC-CH vs CC-NT vs CC-PL<br />
12000<br />
10000<br />
8000<br />
6000<br />
4000<br />
2000<br />
60<br />
50<br />
40<br />
30<br />
Day<br />
20<br />
Day vs Temp vs Pred BC CH+NT CO 2<br />
Day vs Temp vs Pred BC PL CO 2<br />
Day vs Temp vs Pred CC CH CO 2<br />
Day vs Temp vs Pred CC NT CO 2<br />
Day vs Temp vs Pred CC PL CO 2<br />
10<br />
20<br />
22<br />
24<br />
26<br />
0<br />
Soil temperature ( o C)<br />
Figure 1 Predicted values of CO2 emissi<strong>on</strong> in soybean-corn rotati<strong>on</strong> and corn<br />
m<strong>on</strong>oculture<br />
Regressi<strong>on</strong> equati<strong>on</strong>s (R 2 = 0,330973):<br />
BC CH+NT CO2 = 3016,09 + 179,75 Temp – 1,78 Temp * Day<br />
BC PL CO2 = 1996,81 + 179,75 Temp – 1,78 Temp * Day<br />
CC CH CO2 = 4674,25 + 179,75 Temp – 2,92 Temp * Day<br />
CC NT CO2 = 5177,25 + 179,75 Temp – 2,92 Temp * Day<br />
CC PL CO2 = 5347,96 + 179,75 Temp – 2,92 Temp * Day<br />
78<br />
28<br />
30
Nitrous-oxide emissi<strong>on</strong><br />
Nitrous-oxide emissi<strong>on</strong> was influenced significantly (P=0,05) by soil temperature and<br />
soil moisture c<strong>on</strong>tent. Am<strong>on</strong>g the three tillage treatments chisel plowing resulted in the<br />
highest nitrous-oxide emissi<strong>on</strong>, and there was no significant difference between<br />
moldboard plowing and no-till, so these latter treatments were c<strong>on</strong>sidered as <strong>on</strong>e group<br />
<strong>on</strong> the figures. Am<strong>on</strong>g crop rotati<strong>on</strong>s, corn m<strong>on</strong>oculture had higher values than<br />
soybean-corn rotati<strong>on</strong>, however these differences were not significant. Figure 2 shows<br />
the predicted and actual values of N2O emissi<strong>on</strong> influenced by soil temperature and soil<br />
moisture c<strong>on</strong>tent in soybean-corn rotati<strong>on</strong> and in corn m<strong>on</strong>oculture.<br />
N 2 O flux (mg/m 2 /hr)<br />
70<br />
60<br />
50<br />
40<br />
30<br />
20<br />
10<br />
0<br />
-10<br />
20<br />
22<br />
24<br />
26<br />
28<br />
30<br />
Soil temperature ( o C)<br />
N 2 O emissi<strong>on</strong> (mg/m 2 /hr) CH vs PL+NT<br />
80<br />
70<br />
60<br />
50<br />
Soil moisture (%)<br />
40<br />
30<br />
20<br />
Pred Moist vs Pred Temp vs CH Pred<br />
Pred Moist vs Pred Temp vs PL-NT Pred<br />
CH Moist vs CH Temp vs CH N2O<br />
PL-NT Moist vs PL-NT Temp vs PL-NT N2O<br />
Figure 2. Actual and predicted values of N2O emissi<strong>on</strong> in soybean-corn rotati<strong>on</strong> and<br />
in corn m<strong>on</strong>oculture (effect of tillage treatments)<br />
Regressi<strong>on</strong> equati<strong>on</strong>s (R 2 = 0,282134):<br />
79
CH N2O = - 8,856 + 0,701 * Moist – 0,012 Moist * Temp<br />
PL + NT N2O = - 8,856 + 0,607 * Moist – 0,012 Moist * Temp<br />
Although there was no significant difference between crop rotati<strong>on</strong>s, Figure 3. shows<br />
the predicted values in soybean-corn rotati<strong>on</strong> and in corn m<strong>on</strong>oculture.<br />
N 2 O flux (mg/m 2 /hr)<br />
40<br />
30<br />
20<br />
10<br />
0<br />
Soil temperature ( o C)<br />
-10<br />
20<br />
2224262830<br />
N 2 O emissi<strong>on</strong> (mg/m 2 /hr) CH-BC vs CH-CC vs PL+NT-BC vs PL+NT-CC<br />
80<br />
70<br />
60<br />
50<br />
Soil moisture (%)<br />
40<br />
Moist vs Temp vs CH-BC Pred N2O<br />
Moist vs Temp vs CH-CC Pred N2O<br />
Moist vs Temp vs PL+NT-BC Pred N2O<br />
Moist vs Temp vs PL+NT-CC Pred N2O<br />
Figure 3. Predicted values of N2O emissi<strong>on</strong> in soybean-corn rotati<strong>on</strong> and in corn<br />
m<strong>on</strong>oculture (effect of tillage and rotati<strong>on</strong>)<br />
Regressi<strong>on</strong> equati<strong>on</strong>s (R 2 = 0,287105):<br />
80<br />
30<br />
2
CH BC N2O = - 8,839 + 0,6904 * Moist – 0,0125 Moist * Temp<br />
CH CC N2O = - 8,839 + 0,6904 * Moist – 0,0114 Moist * Temp<br />
PL + NT BC N2O = - 8,839 + 0,5961 * Moist – 0,0125 Moist * Temp<br />
PL + NT CC N2O = - 8,839 + 0,5961 * Moist – 0,0114 Moist * Temp<br />
Carb<strong>on</strong>-dioxide and nitrous-oxide emissi<strong>on</strong> declined further in the growing seas<strong>on</strong>,<br />
mainly, because the amount of nutrients necessary for microbial activity<br />
(undecomposed crop residues and nitrogen fertilizer) has declined in the soil with time.<br />
Nitrous-oxide emissi<strong>on</strong> increased after applicati<strong>on</strong> of nitrogen fertilizer and after heavy<br />
rain events (approximately 30-40 mm). Corn m<strong>on</strong>oculture seems to give the highest<br />
emissi<strong>on</strong> of carb<strong>on</strong>-dioxide, and also for nitrous-oxide, rotati<strong>on</strong>s including corn or corn<br />
m<strong>on</strong>oculture have the highest emissi<strong>on</strong>. As for tillage treatments, chisel plowing results<br />
in the highest emissi<strong>on</strong> of nitrous-oxide.<br />
Based <strong>on</strong> the results of our study from 2002-2004 we c<strong>on</strong>cluded that tillage effects<br />
are more important in organic carb<strong>on</strong> and nitrogen storage than crop rotati<strong>on</strong> effects.<br />
We have found, that despite the 30 % higher total crop residue returned in c<strong>on</strong>tinuous<br />
corn, the soybean-corn and c<strong>on</strong>tinuous corn rotati<strong>on</strong>s resulted in similar organic carb<strong>on</strong><br />
and nitrogen levels. The greater CO2 and N2O emissi<strong>on</strong> in c<strong>on</strong>tinuous corn relative to<br />
soybean-corn rotati<strong>on</strong> explains the similar organic carb<strong>on</strong> and nitrogen storage under<br />
the two crop sequences.<br />
REFERENCES<br />
Angers, D.A., M.A. Bolinder, M.R. Carter, E.G. Gregorich, C.F. Drury, B.C.<br />
Liang, P.R. Vor<strong>on</strong>ey, R.R. Simard, R.G. D<strong>on</strong>ald, R.P. Beyaert, and J. Martel.<br />
1997. Impact of tillage practices <strong>on</strong> organic carb<strong>on</strong> and nitrogen storage in cool,<br />
humid soils of eastern Canada. Soil and Tillage Research 41:191-201.<br />
Eswaran, H., E. Van Den Berg, and P. Reich. 1993. Organic Carb<strong>on</strong> in Soils of the<br />
World. Soil Science Society of America Journal 57:192-194.<br />
Gál, A. 2005. Depth dependency of soil carb<strong>on</strong> changes with l<strong>on</strong>g-<strong>term</strong> tillage and<br />
rotati<strong>on</strong> systems. MSc thesis, Purdue University<br />
Gál A., T. J. Vyn, E. Michéli, E. J. Kladivko, William W. McFee. 2007. Soil carb<strong>on</strong><br />
and nitrogen accumulati<strong>on</strong> with l<strong>on</strong>g-<strong>term</strong> no-till versus moldboard plowing<br />
overestimated with tilled-z<strong>on</strong>e sampling depths. Soil and Tillage Research. (in<br />
press, doi 10.1016 j.still.2007.02.007)<br />
Kimble, J., T. Cook, and H. Eswaran. 1990. Organic matter in soils of the tropics .<br />
p.250-258. In Proc. Symp. Characterizati<strong>on</strong> and role of organic matter in different<br />
soils. Int. C<strong>on</strong>gr. Soil sci. 14 th, Kyoto, Japan. 12-18 Aug. 1990. ISSS,<br />
Wageningen, the Netherlands.<br />
Kern, J.S., and M.G. Johns<strong>on</strong>. 1993. C<strong>on</strong>servati<strong>on</strong> tillage impacts <strong>on</strong> nati<strong>on</strong>al soil and<br />
atmospheric carb<strong>on</strong> levels. Soil Science Society of America Journal 57:200-210.<br />
Mitchell, C.C., R.L. Wes<strong>term</strong>an, J.R. Brown, and T.R. Peck. 1991. Overview of<br />
l<strong>on</strong>g-<strong>term</strong> agr<strong>on</strong>omic research. Agr<strong>on</strong>omy Journal 83:24.<br />
Om<strong>on</strong>ode, R.A., Gál, A., Stott, D.E., Abney, T.S., Vyn, T.J. Short-<strong>term</strong> versus<br />
c<strong>on</strong>tinuous chisel and no-till effects <strong>on</strong> soil carb<strong>on</strong> and nitrogen. Soil Science<br />
Society of Agr<strong>on</strong>omy Journal 70:419-425.<br />
81
Phillips, R.E., R.L. Blevins, G.W. Thomas, W.W. Frye, and S.H. Phillips. 1980. Notillage<br />
agriculture. Science 208:1108-1113.<br />
Sparks, D.L. 2003. Chemistry of soil organic matter, p. 75-112 Envir<strong>on</strong>mental Soil<br />
Chemistry, Sec<strong>on</strong>d ed. Academic Press (Elsevier Science).<br />
Studdert, G.A., and H.E. Echeverria. 2000. Crop rotati<strong>on</strong>s and nitrogen fertilizati<strong>on</strong><br />
to manage soil organic carb<strong>on</strong> dynamics. Soil Science Society of America Journal<br />
64:1496-1503.<br />
VandenBygaart, A.J., X.M. Yang, B.D. Kay, and J.D. Aspinall. 2002. Variability in<br />
carb<strong>on</strong> sequestrati<strong>on</strong> potential in no-till soil landscapes of southern Ontario. Soil<br />
and Tillage Research 65:231-241.<br />
ACKNOWLEDGEMENT<br />
Anita Gál received a Hungarian State Eötvös Scholarship in 2006 for the durati<strong>on</strong> of the<br />
study.<br />
82
INFLUENCES OF AMELIORATIVE FERTILIZATION ON YIELD<br />
AND NUTRITIONAL STATUS OF MAIZE<br />
Kovačević Vlado , Josipovic Marko 2 , Stojić Biserka 3<br />
1 University J. J. Strossmayer, Faculty of Agriculture HR-31000 Osijek, Croatia<br />
2 Agricultural Institute, Juzno predgradje 17, HR-31000 Osiejk, Croatia<br />
3 Petrokemija d.d. Fertilizer Factory HR-44320 Kutina, Croatia<br />
ABSTRACT<br />
Suboptimal levels of plant available nutrients are oft limiting factor of field crop yields<br />
under envir<strong>on</strong>mental c<strong>on</strong>diti<strong>on</strong>s, especially in middle and north-western part of Croatia.<br />
Farmers mainly have not basic informati<strong>on</strong> about their soils (for example, their<br />
agrochemical characteristics). In frame of educati<strong>on</strong> of farmers in Djulovac (Bjelovar-<br />
Bilogora County), Pakrac (Pozega-Slav<strong>on</strong>ia County) and Okucani (Brod-Posavina<br />
County) municipalities during March of 2003 total 108 soil samples (0-30 cm depth)<br />
were taken form unfertilized fields (total 279 ha) and soil testing was made (soil pH,<br />
plant available phosphorus and potassium according AL-method). About 60% of tested<br />
soil samples are acid ( pH in KCl less than 5). Also, about 40% of soil samples are poor<br />
or moderate supplied with phosphorus (P), while potassium (K) status is better because<br />
<strong>on</strong>ly 13% samples are in this range of K availability. Based <strong>on</strong> soil test, choice of plots<br />
for fertilizati<strong>on</strong> experiment was made. Aim of this study was testing of ameliorative<br />
fertilizati<strong>on</strong> (spring 2003) with P and K in form of NPK fertilizer <strong>on</strong> yield and<br />
nutriti<strong>on</strong>al status of maize (the growing seas<strong>on</strong>s 2003-2005). In the sec<strong>on</strong>d and third<br />
year of testing <strong>on</strong>ly standard fertilizati<strong>on</strong> of the experiment was applied. Maize<br />
resp<strong>on</strong>ded to applied fertilizati<strong>on</strong> by moderate yield increases up to 10% <strong>on</strong>ly. We<br />
presume that Al-method is not suitable soluti<strong>on</strong> for predicti<strong>on</strong> of plant available P status<br />
in this soil. Ameliorative fertilizati<strong>on</strong> resulted by significant influences <strong>on</strong> increased<br />
plant available P, K and Fe status in the soil, while differences of the other tested<br />
elements (S, Ca, Mg, Mn, Zn, Cu and Cd) were in level of statistical error. The applied<br />
fertilizati<strong>on</strong> had the most influences <strong>on</strong> maize nutriti<strong>on</strong>al status in the first year of<br />
testing (2003) because significant differences for P, K, Mn (increasing trend) Ca, Mg,<br />
Mn and Zn (decreasing trend) were found. In the sec<strong>on</strong>d year (2004) n<strong>on</strong>-significant<br />
differences were found, while in the third year (2005) significant differences were found<br />
for P, K and Mg <strong>on</strong>ly. It is important that cadmium status in maize plant was<br />
independent <strong>on</strong> applied fertilizati<strong>on</strong> because there are examples in literature that<br />
applicati<strong>on</strong> of the high P rates is mainly in c<strong>on</strong>necti<strong>on</strong> with increases of Cd<br />
c<strong>on</strong>centrati<strong>on</strong>s in plant. Ameliorative fertilizati<strong>on</strong> had mainly low influences <strong>on</strong> maize<br />
grain compositi<strong>on</strong> because <strong>on</strong>ly in the third year of testing significant differences for K,<br />
Mg (increases) and Zn (decrease) status were found. Cd c<strong>on</strong>centrati<strong>on</strong>s in maize grain<br />
were under detectable range of the applied method (
humanity organizati<strong>on</strong>, in cooperati<strong>on</strong> with Croatian Co-operative Associati<strong>on</strong> and<br />
Croatian Agricultural Extensi<strong>on</strong> Service, in frame of ECRA project – revitalizati<strong>on</strong> of<br />
war affected communities- which are supported by USAID agency, began educati<strong>on</strong> of<br />
farmers in Djulovac (Bjelovar-Bilogora County), Pakrac (Pozega-Slav<strong>on</strong>ia County) and<br />
Okucani (Brod-Posavina County) municipalities. During March of 2003 total 108 soil<br />
samples (0-30 cm depth) were taken form unfertilized fields (total 279 ha) and soil<br />
testing was made (soil pH, plant available phosphorus and potassium according ALmethod).<br />
About 60% of tested soil samples (total 65) were acid ( pH in KCl less than 5)<br />
and liming could be useful. Also, about 40% (total 42) of soil samples were poor or<br />
moderate supplied with phosphorus, while potassium status is better because <strong>on</strong>ly 13%<br />
samples (total 14) were in this range of K availability. These results were in detail<br />
elaborated in the previous study (Kovacevic et al., 2005). Soil test was criteri<strong>on</strong> for<br />
choice of the plots for field experiments with increased fertilizati<strong>on</strong> with P and K<br />
including Korenicani experiment in Bjelovar-Bilogora County. Grain yields of maize in<br />
this experiment for three-year period were elaborated in detail by the previous study<br />
(Kovacevic et al., 2006). In this study, influences of the fertilizati<strong>on</strong> <strong>on</strong> leaf and grain<br />
compositi<strong>on</strong> were shown.<br />
MATERIAL AND METHODS<br />
The field experiment: Four fertilizati<strong>on</strong> treatments by applicati<strong>on</strong> of NPK 7:20:30<br />
fertilizer (the product of Petrokemija Fertilizer Factory Kutina, Croatia: a = 0 = standard<br />
fertilizati<strong>on</strong>, b = a+1250, c = a+2500 and d = a+3750 kg ha -1 ) were applied <strong>on</strong><br />
Korenicani soil (municipality Djulovac, Bjelovar-Bilogora County). Different amounts<br />
of nitrogen (N) for the a-d treatments were equalized by additi<strong>on</strong> of CAN (calcium<br />
amm<strong>on</strong>ium nitrate) in the amounts 974, 649, 325 and 0 kg ha-1, for the a, b, c and d,<br />
respectively. Only standard fertilizati<strong>on</strong> of the experiment for the 2004 and the 2005<br />
growing seas<strong>on</strong>s were applied. The experiment was c<strong>on</strong>ducted in four replicates by<br />
randomized block design. Gross of the basic plot was 92,4 m 2 (11.0 m x 8.4 m). Maize<br />
hybrids OsSK552 (the growing seas<strong>on</strong>s 2003 and 2004) and OsSK499 (the growing<br />
seas<strong>on</strong> 2005) were sown by pneumatic sowing machine in <strong>term</strong>s end of April/beginning<br />
May. Planned (theoretical) plant densities (TPD: plants ha -1 ) were 51948, 54 946 and<br />
57143, for 2003, 2004 and 2005, respectively. Maize was harvested manually (four<br />
rows from each plot). Grain yiels were calculated <strong>on</strong> 14% grain moisture basis and plant<br />
densities as follows: 90% TPD (2003), 70% TPD (2004) and 95% TPD (2005),<br />
respectively.<br />
Sampling and chemical analyses: Soil samples were taken after maize harvest at end<br />
October 2005. The samples were taken by auger to 30 cm of depth (15 individual<br />
sampling in the mean sample). The ear-leaf of maize was taken at beginning of silking<br />
(middle of July; 25 leaves in the mean sample). Soil, leaf and grain sampling were made<br />
from three replicates of the trial.<br />
Nutriti<strong>on</strong>al status of soil (mobile fracti<strong>on</strong> of the elements) was predicted by<br />
extracti<strong>on</strong>s with NH4Acetate + EDTA (pH 4.65) according Lakanen and Ervio (1971)<br />
and their de<strong>term</strong>inati<strong>on</strong> by the ICP-AES procedure. For general soil test AL-extracti<strong>on</strong><br />
was used (Egner et al., 1960). Soil reacti<strong>on</strong> and organic matter were de<strong>term</strong>ined<br />
according to ISO (1994, 1998). The total amounts of the elements in the leaf and grain<br />
84
samples were measured by the ICP-AES technique after their microwave digesti<strong>on</strong><br />
using c<strong>on</strong>centrated HNO3+H2O2. The soil and plant analyses were d<strong>on</strong>e in the<br />
laboratory of the Research Institute for Soil Science and Agricultural Chemistry<br />
(RISSAC), Budapest, Hungary.<br />
The soil characteristics: By soil test (end of March 2003) acid reacti<strong>on</strong> (pH in 1nKCl =<br />
4.98), low organic matter c<strong>on</strong>tents (1.86%) and moderate levels of mobile phosphorus<br />
and potassium (5.20 mg P2O5 and 8.13 mg K2O 100 g of soil -1 according the ALmethod)<br />
were found.<br />
RESULTS AND DISCUSSION<br />
In general, in spite of relative low status of plant available P were found by the<br />
preliminary soil test, resp<strong>on</strong>se of maize to ameliorative fertilizati<strong>on</strong> was low because<br />
yield were increased up to 10% <strong>on</strong>ly (Table 1). We presume that Al-method is not<br />
suitable soluti<strong>on</strong> for predicti<strong>on</strong> of plant available P status in this soil. Ameliorative<br />
fertilizati<strong>on</strong> resulted by significant influences <strong>on</strong> increased plant available phosphorus,<br />
potassium and ir<strong>on</strong> status <strong>on</strong>ly, while differences of the other tested elements was in<br />
level of statistical error. Applied fertilizati<strong>on</strong> influenced <strong>on</strong> moderate increases of soil<br />
reacti<strong>on</strong> (pH value), while humus c<strong>on</strong>tents were similar for all fertilizati<strong>on</strong> treatments.<br />
Also, values of plant available P and K which were de<strong>term</strong>ined by two extracti<strong>on</strong><br />
methods are similar (Table 2).<br />
Table 1. Realized plant density and grain yield of maize as affected by ameliorative<br />
fertilizati<strong>on</strong> - the experiment Korenicani (Kovacevic et al., 2006)<br />
a)<br />
b)<br />
c)<br />
d)<br />
Fertilizati<strong>on</strong>* Realized plant density (RPD in %TPD=theoretical plant<br />
(17. 04. 2003.) density) and grain yield of maize**<br />
kg ha -1 Maize hybrid OsSK 552 Hybrid OsSK 499<br />
2003 2004 2005<br />
RPD Yield RPD Yield RPD Yield<br />
NPK CAN % t ha -1 % t ha -1 % t ha -1<br />
0 974 87.3 12.38 73.7 10.12 98.9 11.78<br />
1250 649 85.9 13.33 72.4 10.87 94.0 11.92<br />
2500 325 89.4 13.52 70.1 11.17 94.6 11.88<br />
3750 0 84.6 13.41 72.7 10.82 96.1 11.89<br />
Mean 86.8 13.16 72.2 10.75 95.9 11.87<br />
TPD (plants ha -1 ) 51948 54946 57243<br />
LSD 5% 0.54 0.17 n.s.<br />
* for 2004 and 2005 standard fertilizati<strong>on</strong> of the a-d treatments; **yield calculati<strong>on</strong>s <strong>on</strong><br />
14% moisture basis and TPD 90% (2003), 70% (2004) and 95% (2005), respectively.<br />
85
NPK<br />
7:20:30<br />
Table 2. Influences of fertilizati<strong>on</strong> (spring 2003) <strong>on</strong> soil characteristics<br />
The experiment Korenicani: 0-30 cm of soil depth<br />
(sampling 20 October 2005)<br />
NH4Acetate-EDTA extracti<strong>on</strong> (pH 4.65): c<strong>on</strong>centrati<strong>on</strong>s in mg kg -1 of soil<br />
kg ha -1 P2O5 K2O Ca Mg S Fe Mn Zn Cu Cd<br />
a) 0<br />
b) 1250<br />
c) 2500<br />
d) 3750<br />
64.7<br />
62.7<br />
154.3<br />
155.0<br />
100.4<br />
109.7<br />
147.0<br />
153.3<br />
1835<br />
1764<br />
2100<br />
2067<br />
299<br />
298<br />
365<br />
357<br />
7.9<br />
7.8<br />
9.2<br />
8.6<br />
448<br />
421<br />
597<br />
590<br />
261<br />
245<br />
269<br />
267<br />
2.68<br />
2.65<br />
3.19<br />
3.23<br />
3.59<br />
4.49<br />
4.39<br />
4.21<br />
0.077<br />
0.073<br />
0.089<br />
0.091<br />
LSD5% 74.9 34.0 ns ns ns 78 ns ns ns ns<br />
Mean 109.2 127.6 1942 330 8.4 2.94 261 514 4.17 0.082<br />
General agrochemical soil test<br />
pH AL-method: mg g -1 %<br />
a) 0<br />
b) 1250<br />
c) 2500<br />
d) 3750<br />
H2O KCl P2O5 K2O Humus<br />
6.45<br />
6.46<br />
6.87<br />
6.82<br />
5.12<br />
5.12<br />
5.64<br />
5.53<br />
84.7<br />
96.3<br />
164.7<br />
172.0<br />
85.0<br />
99.3<br />
149.7<br />
176.7<br />
1.94<br />
1.92<br />
1.99<br />
1.94<br />
LSD5% 0.23 0.36 28.6 24.0 n.s.<br />
Mean 6.65 5.35 129.4 127.7 1.95<br />
The applied fertilizati<strong>on</strong> had the most influences <strong>on</strong> maize nutriti<strong>on</strong>al status in the first<br />
year of testing (2003) because significant differences for P, K, Mn (increasing trend)<br />
Ca, Mg, Mn and Zn as well (decreasing trend) were found. In the sec<strong>on</strong>d year (2004)<br />
n<strong>on</strong>-significant differences were found, while in the third year (2005) significant<br />
differences were found for P, K and Mg <strong>on</strong>ly (Table 3). It is important that cadmium<br />
status in maize plant was independent <strong>on</strong> applied fertilizati<strong>on</strong> because there are<br />
examples in literature that applicati<strong>on</strong> of the high P rates are mainly in c<strong>on</strong>nceti<strong>on</strong> with<br />
increases of Cd c<strong>on</strong>centrati<strong>on</strong>s in plant (Bergmann, 1992; Mengel and Kirkby, 2001).<br />
Ameliorative fertilizati<strong>on</strong> had mainly low influences <strong>on</strong> maize grain compositi<strong>on</strong><br />
because <strong>on</strong>ly in the third year of testing significant differences for K, Mg (increases)<br />
and Zn (decrease) status were found. Cd c<strong>on</strong>centrati<strong>on</strong>s in maize grain were under<br />
detectable range of the applied analytical procedure (Table 4).<br />
L<strong>on</strong>caric et al. (2005) applied 500 and 1000 kg ha -1 P2O5 and K2O al<strong>on</strong>e and in<br />
their combinati<strong>on</strong> <strong>on</strong> standard fertilizati<strong>on</strong>. Although low levels of P and K were found<br />
by the soil test (AL-method), maize yields were similar for applied treatments in both<br />
years. For this reas<strong>on</strong>, in questi<strong>on</strong> is scientific applicati<strong>on</strong> of AL-method in<br />
interpretati<strong>on</strong> of soil nutriti<strong>on</strong>al status for this and similar soil types. Probably acid<br />
reacti<strong>on</strong> of soil could be resp<strong>on</strong>sible for low influences of ameliorative fertilizati<strong>on</strong> <strong>on</strong><br />
maize yield in our study and in the study L<strong>on</strong>caric et al. (2005).<br />
Also, with aim of more precise predicti<strong>on</strong> of soil nutriti<strong>on</strong>al status it is need<br />
inclusi<strong>on</strong> soil testing by applicati<strong>on</strong> of different extracti<strong>on</strong> methods.<br />
86
Table 3. Influences of ameliorative fertilizati<strong>on</strong> <strong>on</strong> nutriti<strong>on</strong>al status of maize<br />
(compositi<strong>on</strong> of the ear-leaf at silking stage)<br />
Korenicani field experiment: Influences of fertilizati<strong>on</strong> in spring 2003 (NPK 7:20:30 <strong>on</strong><br />
standard fertilizati<strong>on</strong>) <strong>on</strong> nutriti<strong>on</strong>al status of maize<br />
NPK C<strong>on</strong>centrati<strong>on</strong>s in dry matter of ear-leaf of maize at silking stage<br />
7:20:30 Percent (%) mg kg -1<br />
kg ha -1 P K Ca Mg S Fe Mn Zn Cu Cd<br />
0<br />
1250<br />
2500<br />
3750<br />
The growing seas<strong>on</strong> 2003<br />
0.29<br />
0.31<br />
0.30<br />
0.32<br />
2.14<br />
2.55<br />
2.50<br />
2.50<br />
0.71<br />
0.65<br />
0.63<br />
0.60<br />
0.44<br />
0.30<br />
0.25<br />
0.24<br />
0.19<br />
0.20<br />
0.18<br />
0.19<br />
122<br />
127<br />
129<br />
135<br />
83<br />
108<br />
105<br />
110<br />
66.4<br />
63.7<br />
56.4<br />
56.4<br />
16.5<br />
17.4<br />
15.3<br />
17.0<br />
0.08<br />
0.08<br />
0.09<br />
0.11<br />
LSD5% 0.02 0.21 0.09 0.03 n.s. n.s. 18 10.1 n.s. n.s.<br />
Mean 0.30 0.24 0.65 0.31 0.19 128 101 60.7 16.5 0.09<br />
0<br />
1250<br />
2500<br />
3750<br />
The growing seas<strong>on</strong> 2004<br />
0.38<br />
0.38<br />
0.39<br />
0.38<br />
2.29<br />
2.25<br />
2.37<br />
2.29<br />
0.75<br />
0.72<br />
0.73<br />
0.72<br />
0.22<br />
0.22<br />
0.21<br />
0.22<br />
0.21<br />
0.21<br />
0.21<br />
0.20<br />
190<br />
190<br />
196<br />
202<br />
81<br />
77<br />
87<br />
84<br />
55.3<br />
58.8<br />
60.5<br />
57.3<br />
17.6<br />
18.7<br />
19.0<br />
17.5<br />
0.06<br />
0.06<br />
0.07<br />
0.07<br />
LSD5% n.s. n.s. n.s. n.s. n.s. n.s. n.s. n.s. n.s. n.s.<br />
Mean 0.38 0.23 0.73 0.22 0.21 194 82 58.0 18.2 0.07<br />
0<br />
1250<br />
2500<br />
3750<br />
The growing seas<strong>on</strong> 2005<br />
0.36<br />
0.38<br />
0.41<br />
0.41<br />
2.06<br />
2.19<br />
2.23<br />
2.47<br />
0.66<br />
0.63<br />
0.66<br />
0.65<br />
0.27<br />
0.23<br />
0.23<br />
0.22<br />
0.21<br />
0.21<br />
0.22<br />
0.21<br />
155<br />
191<br />
189<br />
171<br />
97<br />
100<br />
85<br />
96<br />
42.7<br />
46.5<br />
49.6<br />
48.3<br />
15.0<br />
16.1<br />
16.8<br />
16.1<br />
0.02<br />
0.04<br />
0.03<br />
0.02<br />
LSD5% 0.03 0.30 n.s. 0.04 n.s. n.s. n.s. n.s. n.s. n.s.<br />
Mean 0.39 2.18 0.65 0.24 0.21 177 94 46.8 16.0 0.03<br />
In general, based <strong>on</strong> our field experiment experiences in the last decade period, liming<br />
was more influencing factor of field crops yield compared to ameliorative fertilizati<strong>on</strong><br />
with P and K. For example, Kovacevic et al. (2006) applied carbocalk (waste of sugar<br />
factory, 39% CaO) in five rates up to 90 t/ha. Liming with carbokalk resulted with<br />
c<strong>on</strong>siderable increases of field crop yields (maize-maize-sunflower-barley rotati<strong>on</strong> )as<br />
follows: up to 50% and 36% (maize for 2001 and 2002, respectively), up to 49%<br />
(sunflower 2003) and up to 30% (barley 2004). In general, by applicati<strong>on</strong> of carbocalk<br />
in level of 90 t/ha, yields drastically decreased mainly to level of c<strong>on</strong>trol as affected by<br />
overliming. Using 15 t ha -1 of carbocalk <strong>on</strong> Djakovo pseudogley (the growing seas<strong>on</strong><br />
1993) yields of maize hybrids (total 10) were increased depended <strong>on</strong> hybrids in range<br />
from 6% to 45% (Banaj and Kovacevic, 2000). Liming with carbocalk (30 t ha -1 ) was<br />
especially useful for maize under drought stress of the 2000 growing seas<strong>on</strong> <strong>on</strong> Slatina<br />
pseudogley because yield was doubled compared to the c<strong>on</strong>trol. At the same time,<br />
c<strong>on</strong>tributi<strong>on</strong> of female-sterile plants was decreased from 42% to 7% <strong>on</strong>ly (Antunovic et<br />
al., 2001).<br />
87
Table 4. Influences of ameliorative fertilizati<strong>on</strong> <strong>on</strong> nutriti<strong>on</strong>al status of maize<br />
(compositi<strong>on</strong> of grain at maturity stage for the growing seas<strong>on</strong>s 2004 and 2005)<br />
Korenicani experiment: influences of NPK fertilizati<strong>on</strong> <strong>on</strong> grain compositi<strong>on</strong> of maize<br />
NPK C<strong>on</strong>centrati<strong>on</strong>s in dry matter of grain at maturity stage*<br />
7:20:30 Percent (%) mg kg -1<br />
kg ha -1 P K Ca Mg S Fe Mn Zn Cu<br />
The growing seas<strong>on</strong> 2004<br />
0 0.307 3.63 0.005 0.099 0.098 21.9 4.60 21.2 1.52<br />
3750 0.295 3.61 0.007 0.092 0.084 18.4 4.43 19.5 1.31<br />
LSD5% n.s. n.s. n.s. n.s. n.s. n.s. n.s. n.s. n.s.<br />
Mean 0.301 3.62 0.006 0.095 0.091 20.1 4.52 20.3 1.42<br />
The growing seas<strong>on</strong> 2005<br />
0 311 3.51 0.002 0.087 0.087 22.7 4.81 30.3 1.66<br />
1250 308 3.52 0.002 0.086 0.088 23.1 5.04 22.7 1.69<br />
2500 319 3.83 0.002 0.089 0.083 25.6 5.09 18.0 1.71<br />
3750 312 3.74 0.002 0.096 0.086 22.1 4.97 18.2 1.70<br />
LSD5% n.s. 0.12 n.s. 0.004 n.s. n.s. n.s. 9.1 n.s.<br />
LSD1% n.s. 0.18 0.006 n.s. n.s. n.s. n.s.<br />
Mean 0.312 3.65 0.002 0.089 0.086 23.4 4.98 22.3 1.69<br />
Cd c<strong>on</strong>centrati<strong>on</strong>s in grain were under detectable range Cd (
Egner, H., Riehm, H., Domingo, W.R. (1960): Untersuchungen uber die chemische<br />
Bodenanalyse als Grundlage fur die Beurteilung des Nahrstoffzustandes der Boden<br />
II. Chemische Extracti<strong>on</strong>smethoden zu Phosphor- und Kaliumbestimmung. K.<br />
Lantbr. Hogsk. Annlr. W.R. 1960, 26, 199-215.<br />
ISO (1994): Soil quality. De<strong>term</strong>inati<strong>on</strong> of pH. ISO 10390:1994(E)<br />
ISO (1998): Soil quality. De<strong>term</strong>inati<strong>on</strong> of organic carb<strong>on</strong> by sulfochromic oxidati<strong>on</strong>.<br />
ISO 14235:1998(E).<br />
Kovacevic V., Banaj D., Kovacevic J., Lalic A., Jurkovic Z., Krizmanic M. (2006):<br />
Influences of liming <strong>on</strong> maize, sunflower and barley. Cereal Res. Comm. 34 (1):<br />
553–556.<br />
Kovačević V., L<strong>on</strong>čarić Z., Rastija M. (2005): Analize tla i gnojidba u funkciji<br />
povećanja prinosa kukuruza. U: Zbornik radova, XL. Znanstveni skup hrvatskih<br />
agr<strong>on</strong>oma s međunarodnim sudjelovanjem (Kovačević V. i Jovanovac S<strong>on</strong>ja, ur.),<br />
Opatija, 15-18. veljače 2005. str. 457-458.<br />
Kovačević V., Stojić Biserka, Rastija Mirta, Bukvić Gordana, Antunović Manda<br />
(2006): Reakcija kukuruza na melioracijsku gnojidbu fosforom i kalijem. In:<br />
Proceednings, 41 Croatian & 1st <str<strong>on</strong>g>Internati<strong>on</strong>al</str<strong>on</strong>g> Symposium <strong>on</strong> Agriculture, (S.<br />
Jovanovac and V. Kovacevic Eds.), 13-17. February 2006, Opatija, Croatia,<br />
Faculty of Agriculture, University J. J. Strossmayer in Osijek, p. 423-424.<br />
L<strong>on</strong>čarić Z., Kovačević V., Šeput Miranda, Šimić B. and Stojic Biserka (2005):<br />
Influences of fertilizati<strong>on</strong> <strong>on</strong> yield and nutriti<strong>on</strong>al status of maize. Cereal Research<br />
Communicati<strong>on</strong>s 33 (1):259-262.<br />
Mengel K., Kirkby E. A. (2001): Principles of plant nutriti<strong>on</strong>. Kluwer Academic<br />
Publishers Dordrecht/Bost<strong>on</strong>/L<strong>on</strong>d<strong>on</strong>.<br />
89
YIELD AND QUALITY OF WINTER WHEAT GENOTYPES<br />
(Triticum aestivum L.)<br />
Desimir Knezevic 1 , Veselinka Zecevic 2 , Danica Micanovic 2 , Nevena Djukic 3 ,<br />
Aleksandar Paunovic 4 , Milomirka Madic 4 , Ivica Djalovic 4<br />
1 Faculty of Agriculture, Zubin Potok, University of Pristina, Serbia,<br />
2 Center for Small Grains Kragujevac, S. Kovacevica 31, 34000 Kragujevac, Serbia<br />
3 Faculty of Natural Science, Department of Biology, University of Kragujevac, Serbia,<br />
4 Agricultural Faculty, Cacak, Cara Dusana 32, 32000, Cacak, Yugoslavia<br />
ABSTRACT<br />
Variability of yield and quality parameters were studied in 11 wheat lines and cultivars.<br />
Wheat cultivars were grown in the experimental field in 5 replicati<strong>on</strong>. part of The<br />
differences in average values for all the studied parameters am<strong>on</strong>g investigated cultivars<br />
were de<strong>term</strong>ined. High variability of investigated traits was established. The high<br />
variability for analyzed yield and quality comp<strong>on</strong>ents were.<br />
Keywords: wheat, yield, quality, protein sedimentati<strong>on</strong>, gluten<br />
INTRODUCTION<br />
Increasing of yield in wheat has been an important task of breeders. The breeders in the<br />
last six decades have been investigated impact of genetic, physiological, biochemical,<br />
ecological parameters for increases in yield and quality improvement. Numerous data of<br />
investigati<strong>on</strong> indicated that levels of water, fertilizer, pesticide and herbicide applicati<strong>on</strong><br />
have influence to impovement of wheat yield. In successful breeding program wheat<br />
breeders has produced a series of cultivars with improved resistance to pests and<br />
diseases, wider adaptati<strong>on</strong> to adverse soil and climatic c<strong>on</strong>diti<strong>on</strong>s and better quality<br />
(Bedo et al., 2005). Genetic yield potential may be defined as the yield of a cultivar<br />
grown in envir<strong>on</strong>ments to which it is adapted, with nutrients and water n<strong>on</strong>-limiting,<br />
and with pests, diseases, weeds, lodging and other stresses effectively c<strong>on</strong>trolled. The<br />
genetic yield potential of a wheat cultivar may be dependent <strong>on</strong> favorable c<strong>on</strong>diti<strong>on</strong>s<br />
and good agr<strong>on</strong>omy for its expressi<strong>on</strong>. The ideal cultivar for high grain yield or for any<br />
other desirable traits need to express genetic potential in different envir<strong>on</strong>ment with low<br />
value of variance in different envir<strong>on</strong>mental factors of growing (Joshi et al. 2002). The<br />
grain yield of wheat is variable traits, which depends <strong>on</strong> numerous yield comp<strong>on</strong>ents<br />
and envir<strong>on</strong>mental factors (Kraljevic-Balalic et al., 2001). The characters as height of<br />
plants, length of spike, number of spikelets per spike are in positive correlati<strong>on</strong> with<br />
grain yield (Zecevic, 2005). The yield variability is less studied than yield itself.<br />
The wheat grain and its product are very important in human nutriti<strong>on</strong>. The great<br />
nutriti<strong>on</strong>al importance of wheat have stimilated the investigati<strong>on</strong> of quality parameters.<br />
During the recent few decades significant advanced have be made in the biochemical,<br />
genetic and molecular biology studies of protein comp<strong>on</strong>ents. Proteins play major role<br />
in de<strong>term</strong>inati<strong>on</strong> of quality of grain, flour, dough and bread. A number of genes<br />
encoding proteins were isolated and sequenced.<br />
90
To resolve genetics of grain quality in wheat the main investigati<strong>on</strong>s were focused <strong>on</strong><br />
protein sedimentati<strong>on</strong> volume, protein and gluten c<strong>on</strong>tents, grain hardness and<br />
farinograph characteristics, are genetically c<strong>on</strong>trolled. A combinati<strong>on</strong> of those<br />
parameters and the flour protein c<strong>on</strong>tent can be used as a base for estimati<strong>on</strong> of the<br />
technological quality of selecti<strong>on</strong> materials. For improving of wheat grain quality is<br />
especially important to identify the biotype polymorphisms with numerous favorable<br />
traits and gene d<strong>on</strong>ors c<strong>on</strong>trolling high protein c<strong>on</strong>tent with high grain yield and create<br />
high quality cultivars (Bebyakin, 1985, Knezevic et al., 2006).<br />
The aim of this paper is study of variability of yield, height of plants, thousand<br />
grain mass, hectoliter mass and technological quality parameters in genetically<br />
divergent wheat genotypes.<br />
MATERIALS AND METHODS<br />
The variability of yield and and qualyty parameters (thousand grain mass, hectoliter<br />
mass, protein c<strong>on</strong>tents, protein sedimentati<strong>on</strong> volume, wet and dry gluten c<strong>on</strong>tents,<br />
water absorpti<strong>on</strong>) were investigated in 11 genetically divergent wheat lines and cultivars<br />
(KG-35265-1/94, KG-52, KG-3625/97, KG-3089/97, KG-3062/1-93, KG-4, KG-<br />
3617/98, KG-35171/94, KG-3419-3/97, Pobeda and Partizanka). The experiment was<br />
performed in randomized block design in five replicati<strong>on</strong> <strong>on</strong> the experimental field of<br />
Center for Small Grains, Kragujevac, Serbia. The seeds of cultivars were sown <strong>on</strong> basic<br />
plot 5 m 2 . The technological quality of 11 winter wheat cultivars was estimated by<br />
standardized laboratory methods. Standard milling was performed with a Buller type<br />
MLU-202 experimental mill (Buller, Uzwill, Switzerland). Sedimentati<strong>on</strong> value was<br />
analyzed by Zeleny modified in which 2 or 5 g of wheat could be evaluated<br />
satisfactorily. Rheological quality were analyzed with micro-Brabender farinograph<br />
using 10 g flour sample.<br />
RESULTS AND DISCUSSIONS<br />
The analysis of yield and technological quality parameters showed differences am<strong>on</strong>g<br />
cultivars. Sedimentati<strong>on</strong> volume varied between 39 ml (KG-3089/97) and 68 ml<br />
(Partizanka). According to sedimentati<strong>on</strong> volume wheat lines which had sedimentati<strong>on</strong><br />
volume higher than 40 ml are bel<strong>on</strong>ging into the 1 st quality group. Wheat lines which<br />
had less than 40 ml sedimentati<strong>on</strong> volume are clasified into 2 nd quality group (Table 1).<br />
Sedimentati<strong>on</strong> volumes of analyzed cultivars have been different.<br />
High sedimentati<strong>on</strong> protein values indicate high protein c<strong>on</strong>tent and good gluten quality<br />
(Menkovska et al., 1995). This was established in wheat line KG-3419-3/97. Protein<br />
sedimentati<strong>on</strong> volume, an important quality comp<strong>on</strong>ent, has a positive correlati<strong>on</strong> with<br />
other quality parameters such as gluten quality and loaf volume (Zuoji et al., 1989).<br />
High values of protein sedimentati<strong>on</strong> volume mainly depends from protein<br />
c<strong>on</strong>tents as well from amount of gliadin and glutenin and gliadin/glutenin ratio,<br />
respectively. Glutenin c<strong>on</strong>tent is directly related to protein c<strong>on</strong>tents and therefore<br />
influences sedimentati<strong>on</strong> volume (Knezevic et al., 2000). Protein c<strong>on</strong>tents varied from<br />
11.90% (KG-35265-1/94) to 17.21% (KG-3419-3/97) Table 1. Glutenin c<strong>on</strong>tent had a<br />
positive genetic correlati<strong>on</strong> with technological quality of flour, sedimentati<strong>on</strong> volume<br />
91
whereas gliadin had a negative correlati<strong>on</strong> (Sasek et al., 1987; Pogna et al., 1990;<br />
Menkovska et al., 2002).<br />
Table1. Technological qualityparameters of Kragujevac's winter wheat lines<br />
Cultivar Sv (ml) Wg % Dg % Wac Qsg Qn Pc<br />
KG-35265-1/94 63 43.23 16.55 62.4 A1 91.2 11.90<br />
KG-52 58 38.00 13.28 58.4 A2 89.5 14.02<br />
KG-3625/97 42 37.26 11.25 63.4 C1 40.2 14.36<br />
KG-3089/97 39 39.31 14.45 63.4 A2 76.7 15.22<br />
KG-3062/1-93 47 37.95 12.06 62.6 B1 56.6 14.08<br />
KG-4 40 38.89 12.52 64.0 B2 46.7 15.10<br />
KG-3617/98 41 43.68 13.59 63.8 B1 55.4 15.22<br />
KG-35171/94 50 39.26 13.34 62.4 B1 66.4 15.45<br />
KG-3419-3/97 62 47.84 17.28 66.2 A2 71.6 17.21<br />
Pobeda 56 41.21 13.51 62.8 A2 72.8 14.02<br />
Partizanka 68 45.91 14.59 64.2 A2 80.0 13.15<br />
Sv=Sedimentati<strong>on</strong> value (ml); Wg=Wet gluten %; Dg=Dry gluten %; Wac=Water<br />
absorpti<strong>on</strong> capacity ; Qsg=Quality subgroup;Qn= Quality number; Pc=Protein c<strong>on</strong>tents<br />
In this study different value of gluten c<strong>on</strong>tent in wheat cultivars were established. In<br />
average the highest gluten c<strong>on</strong>tent 47.84% (KG-3419-3/97), and the lowest 37.26% in<br />
the KG-3625/97 line. The ratio of variati<strong>on</strong> of gluten c<strong>on</strong>tents is in agreement with data<br />
(Pavlovic et al., 1994) which found that gluten c<strong>on</strong>tent varies from 16 to 52% (crude)<br />
and 5-20% (dry). Grain c<strong>on</strong>sistency, temperature and moisture c<strong>on</strong>diti<strong>on</strong>s during<br />
maturati<strong>on</strong> have influence to gluten quality. High temperature and low moisture<br />
produce str<strong>on</strong>g gluten with less extensibility and the opposite c<strong>on</strong>diti<strong>on</strong>s produce weak<br />
glutens and extensible doughs (Kodanev, 1976). Gluten is formed in the early stages of<br />
ripening (milk) but its quality changes during maturity to the final characteristics.<br />
Gluten structure has a great influence <strong>on</strong> flour rheological properties and quality.<br />
Gluten quality depends <strong>on</strong> the ability of dough to keep CO2 from fermentati<strong>on</strong> in gas<br />
cells in order to provide loaf volume and bread crumb.<br />
All the varieties in study bel<strong>on</strong>ged to semi-dwarf wheat class expressing the plant<br />
height from x= 63.4 cm (partizanka) and x = 76.3 cm (kg-52), according to mean<br />
values over studied period (table 2). This semi-dwarf ccultivars have advantages in<br />
relati<strong>on</strong> to dwarf and tall cultivars as a source of desirable genes in the future breeding<br />
programs. The similar results reported Dimitrijevic et al. (1996) that indicated that<br />
genotype had higher influence than envir<strong>on</strong>ment <strong>on</strong> expressi<strong>on</strong> of plant height. the<br />
proporti<strong>on</strong> of total mass to grain yield in mostly modern cultivars is more than 2:1<br />
which indicates excessive c<strong>on</strong>sumpti<strong>on</strong> of nutrients for straw formati<strong>on</strong> instead of grain.<br />
This is <strong>on</strong>e reas<strong>on</strong> why breeders have c<strong>on</strong>cept of decreasing vegetative part of wheat<br />
and realize optimal proporti<strong>on</strong> in order to increase the harvest index to more than 50%<br />
(Zecevic at al., 2005).<br />
92
Table 2. Average values of analyzed traits of Kragujevac's winter wheat lines<br />
Cultivar Plant height Hectoliter TGW Yield<br />
(ml) mass (kg) (g) (kg)<br />
KG-35265-1/94 67.8 82.30 38.28 4265<br />
KG-52 76.3 80.90 40.19 4556<br />
KG-3625/97 71.6 82.50 37.52 4756<br />
KG-3089/97 68.8 80.10 37.52 4321<br />
KG-3062/1-93 64.3 81.30 36.96 3882<br />
KG-4 67.0 81.30 37.29 4327<br />
KG-3617/98 69.0 82.10 38.98 3718<br />
KG-35171/94 64.4 82.10 39.21 3890<br />
KG-3419-3/97 63.5 80.75 36.96 2875<br />
Pobeda 67.0 81.31 39.36 4753<br />
Partizanka 63.4 82.10 41.47 3962<br />
Decreasing of stem height gave higher possibility of efficient utilizati<strong>on</strong> mineral<br />
elements for the head development. Also translocati<strong>on</strong> of mineral elements is more<br />
efficient in semidwarf cultivars from the vegetative organs to grain.<br />
During investigated periods the highest average value of of thousand grain weight had<br />
Partiyanka cultivar (x=41.47) the lowest average value of thousand grain weight<br />
expressed KG-3062/1-93 cultivar (x=30.62) Table 2. Number of grain per spike and<br />
thousand grain weight have str<strong>on</strong>g influence <strong>on</strong> spike productivity. Yield of analyzed<br />
wheat genotypes varied between 2875 kg ha -1 (KG-3419-3/97) and 4756 kg ha -1 (KG-<br />
3625/97) Table 2. Yield is in high relati<strong>on</strong>ships with the number of stems per m 2 a nd the<br />
percentage of fertile spikes, number of kernels per spike, thousand grain weight. The<br />
genetic c<strong>on</strong>trols of stem height and other yield comp<strong>on</strong>ents are very important, but<br />
envir<strong>on</strong>ment has influence <strong>on</strong> expressi<strong>on</strong> of analyzed traits. Generally speaking,<br />
genotypes reacted similarly to envir<strong>on</strong>mental variati<strong>on</strong> for plant height, and number of<br />
spikiest per spike, number of kernels per spike, thousand grain weight, differing in main<br />
effect. On the c<strong>on</strong>trary, genotype reacti<strong>on</strong> for yield comp<strong>on</strong>ents in interacti<strong>on</strong> making<br />
phenotypic expressi<strong>on</strong> for those trait more unpredictable. The remobilizati<strong>on</strong> of reserve<br />
into harvested organs varies am<strong>on</strong>g the cultivars. Yield is very important agroec<strong>on</strong>omic<br />
traits which will in the focus of investigati<strong>on</strong> in the future.<br />
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Small Grains. (eds. S.Quarrie et all) pp. 9-49<br />
Menkovska, M., Knežević, D., Ivanovski, M., Zečević Veselinka (1995): Technological<br />
Quality of Some Maced<strong>on</strong>ian Bread Wheat Varieties: I. Relati<strong>on</strong> With the<br />
Compositi<strong>on</strong> of Gliadin Comp<strong>on</strong>ents. Bulleten of Chemists & Technologists of<br />
Maced<strong>on</strong>ia,14,1,31-34.<br />
Menkovska Mirjana, Knežević, D., Ivanoski, M. (2002): Protein allelic compositi<strong>on</strong>,<br />
dough rheology, and baking characteristics of flour mill streams from wheat<br />
cultivars with known and varied baking qualities. Cereal Chemestry, 79(5), 720-<br />
725.<br />
Pavlovic, M., Kuburovic, M., Zecevic, V. & Knezevic, D. (1994) Parametars of<br />
technolological quality and comp<strong>on</strong>ents of yield of some winter wheat cultivars. /in<br />
Serbian/ Journal for Cereal and Flour Technology 21, 4, 94-96.<br />
Pogna, N.E., Mazza, M., Redaelli, R. & Ng, P.K.W. (1996) Gluten quality and storage<br />
protein compositi<strong>on</strong> of durum wheat lines c<strong>on</strong>taining the Gli-D1/Gli-D3 loci. In:<br />
CW Wrigley ed. Gluten 96, Cereal Chemistry Devisi<strong>on</strong>, Royal Australian<br />
Chemical Institute, North Melbourne, Sidney, pp. 18-22.<br />
Sasek, A., Kubanek, J. & Cerny, J. (1987) Gliadin and glutenin polymorphism of some<br />
cultivars – Populati<strong>on</strong>s of comm<strong>on</strong> wheat (T. aestivum L.). Scientia Agriculturae<br />
Bohemoslovaca 19, 2, 93-99.<br />
Zuoji Lin, Zhou Hidan, Jie Shenghui, Hu Xueyi, Ding Xiaolin, Jin Maoguo (1989)<br />
Correlati<strong>on</strong> between bread making quality and other quality parameters and yield<br />
in winter wheat. Acta Agr<strong>on</strong>. Sin.(rez. angl.). /in Russian/, 15, 2, 151-159.<br />
Zečević Veselinka, Knežević, D., Mićanović Danica, Pavlović, M., Urošević, D.<br />
(2005): the inheritance of plant height in winter wheat (Triticum aestivum L.).<br />
Genetika, 37, 173-179<br />
Zečević Veselinka, Knežević, D., Mićanović Danica, Pavlović, M., Urošević, D.<br />
(2005): The inheritance of plant height in winter wheat (Triticum aestivum L.).<br />
Genetika, 37, 173-179.<br />
94
CROP ROTATION AND GREEN MANURE INFLUENCE ON YIELD AND<br />
WATER USE EFFICIENCY IN WHEAT AND MAIZE FROM CRIS PLAIN<br />
C. Domuţa, Gh. Ciobanu, Maria Şandor, Alina Samuel, Cornelia Ciobanu,<br />
N.C. Sabău, V. Şcheau, Ioana Borza, Cr. Domuţa<br />
University of Oradea, Oradea, Romania<br />
ABSTRACT<br />
The paper is based <strong>on</strong> the results researches obtained during 2004 – 2006 in Oradea. In<br />
the crop rotati<strong>on</strong> of 4 years with clover in comparis<strong>on</strong> with wheat – maize crop rotati<strong>on</strong>,<br />
the yield gains statistically assured were obtained both in the variants with organic<br />
fertilizati<strong>on</strong> and in the variant with organic + chemical fertilizati<strong>on</strong>. The use of the<br />
mixture lupin + oat in this crop rotati<strong>on</strong> de<strong>term</strong>ined to obtain yield gains in comparis<strong>on</strong><br />
with variants with lupin, pure crop as green manure; yield obtained using lupin + oat<br />
were close to the yield from variant with manure, 25 t/ha. Meliorative crop rotati<strong>on</strong> and<br />
fertilizati<strong>on</strong> de<strong>term</strong>ined the improve of the water use efficiency in comparis<strong>on</strong> with the<br />
c<strong>on</strong>trols; water use efficiency from variants with lupin + oat was bigger than water use<br />
efficiency obtained in the variant with lupin, too.<br />
Key words: crop rotati<strong>on</strong>, green manure, yield, wheat, maize.<br />
INTRODUCTION<br />
In the sustainable agriculture system, the crop rotati<strong>on</strong> is the central pivot and organic<br />
fertilizati<strong>on</strong> is <strong>on</strong>e of the most comp<strong>on</strong>ents (Budoi and Penescu, 1998; Gus and al,<br />
1998; Pakurar and Nagy, 2005, Pepo, 2005, Domuţa, 2006). Green manure can<br />
occupied a very important place between the organic fertilizer types but the correct use<br />
are need c<strong>on</strong>sist of the mixture between lupin and oat (rye) and rape. (Domuţa, 1999,<br />
2005). Due the small C/N rapport the use of the lupin in pure crop de<strong>term</strong>ines explosive<br />
microbiological processes and intense humus mineralizati<strong>on</strong> and finally the decrease of<br />
the soil humus reserve. (Eliade and al, 1983, Samuel and al, 2006).<br />
MATHERIAL AND METHOD<br />
The researches were carried out in Oradea, Western Romania, <strong>on</strong> erosi<strong>on</strong>ed preluvosoil<br />
with slope of 8˚. On ploughing land the pH value is of 6.2, humus c<strong>on</strong>tent is of 2.1,<br />
mobile phosphorum is of 34.1 ppm and mobile potassium c<strong>on</strong>tent is of 209.2 ppm.<br />
Structure degree is of 55.8% and field capacity (24.3%) and wilting point (9.1%) have<br />
median value.<br />
Research period was 2004 – 2006 but the experiment is in the sec<strong>on</strong>d cycle of the<br />
researches. Experimental dispozitive includes three factors:<br />
Factor A: crop rotati<strong>on</strong><br />
a1 : wheat – maize<br />
a2 : oat + clover – clover – wheat – maize<br />
Factor C: annual fertilizati<strong>on</strong><br />
c1: N0P<br />
c2: N90P60K60<br />
95<br />
Factor B: organic fertilizati<strong>on</strong><br />
b1 : c<strong>on</strong>trol<br />
b2 : manure, 25 t/ha<br />
b3 : manure, 50 t/ha<br />
b4 : lupin<br />
b5 : lupin + oat
Number of repetiti<strong>on</strong> used: 4. Surface of the experiment plot: 300 m 2 .Total surface of<br />
the experiment: 6000 m 2 .Green manure was produced in 2001, 2002 and 2003 like main<br />
crop. Sowing rate: 200 kg/ha in lupin pure crop and lupin 100 kg/ha and oat 80 kg/ha in<br />
the mixture. The harvesting was made in the flowering stage of the lupin. After<br />
harvesting, the green manure was kept like mulch <strong>on</strong> the soil surface. After 10 days the<br />
plough land of 25 cm depth was made. The maize was cropped in the first year after<br />
organic fertilizati<strong>on</strong>. Water use efficiency was established dividing the wheat and maize<br />
yield to water c<strong>on</strong>sumpti<strong>on</strong>. Wheat and maize water c<strong>on</strong>sumpti<strong>on</strong> was established using<br />
the soil water balance method; balance depth used was 0 – 150 cm.<br />
RESULTS AND DISCUSSION<br />
Influence of crop rotati<strong>on</strong> <strong>on</strong> wheat yield<br />
The average of the yields <strong>on</strong> the period 2004 – 2006 emphasizes bigger yields in the<br />
crop rotati<strong>on</strong> with clover both in the background N0P0 (4944 kg/ha vs. 3944 kg/ha,<br />
25.3%) and in the background N90P60K60 (6092 kg/ha vs. 5562 kg/ha). The relative<br />
differences between yield registered in the researched crop rotati<strong>on</strong> during the years<br />
were of 18.6% (N0P0) and of 12.4% (N90P60K60) in 2004, of 30.0% (N0P0) and of 9.0%<br />
(N90P60K60) in 2005, of 27.3% (N0P0) and of 7.5% (N90P60K60) in 2006 (table 1).<br />
Influence of crop rotati<strong>on</strong> and green manure <strong>on</strong> maize yield<br />
All the organic fertilizati<strong>on</strong> variants de<strong>term</strong>ined to obtain very significant yield gain in<br />
comparis<strong>on</strong> with c<strong>on</strong>trol both in the crop rotati<strong>on</strong> wheat – maize and in the crop rotati<strong>on</strong><br />
oat + clover – clover – wheat – maize. The biggest yields were obtained in the variant<br />
with manure 50 t/ha.<br />
Table 1: Influence of the crop rotati<strong>on</strong> and fertilizati<strong>on</strong> <strong>on</strong> wheat yield,<br />
Oradea 2004 – 2006<br />
Organic fertilizati<strong>on</strong><br />
N0P0<br />
Chemical fertilizati<strong>on</strong><br />
N90P60K60<br />
2004 2005 2006 Average 2004 2005 2006 Average<br />
Wheat - maize<br />
1. C<strong>on</strong>trol 2740 3010 3580 3110 4010 4690 5010 4570<br />
2. Manure 25 t/ha 3890 3850 4320 4020 5320 5720 6190 5740<br />
3. Manure 50 t/ha 5020 4580 5030 4880 6280 6440 6760 6490<br />
4. Lupin 3690 3420 3840 3650 4960 5130 5530 5210<br />
5. Lupin + oat 4080 3810 4290 4060 5620 5700 6080 5800<br />
Average 3884 3734 4212 3944 5238 5536 5914 5562<br />
Oat + clover – clover – wheat – maize<br />
1. C<strong>on</strong>trol 3640 4010 4520 4060 5020 5330 5840 5400<br />
2. Manure 25 t/ha 4760 5030 5570 5120 5980 6090 6460 6180<br />
3. Manure 50 t/ha 5620 5710 6240 5860 6390 6890 7100 6790<br />
4. Lupin 4210 4540 5010 4590 5720 5680 6010 5800<br />
5. Lupin + oat 4800 4990 5480 5090 6320 6170 6390 6290<br />
Average 4606 4856 5364 4944 5886 6032 6360 6092<br />
96
Organic Chemical Chemical fertilizati<strong>on</strong> x Organic fertilizati<strong>on</strong> x<br />
fertilizati<strong>on</strong> fertilizati<strong>on</strong> Organic fertilizati<strong>on</strong> Chemical fertilizati<strong>on</strong><br />
a1 a2 a1 a2 a1 a2 a1 a2<br />
LSD 5% 140 150 160 110 230 210 200 190<br />
LSD 1% 250 290 230 208 310 420 350 360<br />
LSD 0.1% 590 470 310 390 420 730 560 580<br />
In the variant with lupin + oat, an yields bigger than yields obtained in the variants with<br />
lupin pure crop were obtained 410 kg/ha in the crop rotati<strong>on</strong> wheat – maize and 500<br />
kg/ha in the crop rotati<strong>on</strong> oat + clover – clover – wheat – maize. The yield obtained in<br />
the variants with lupin + oat are very closed to the yields obtained in the variant with<br />
manure 25 t/ha.<br />
Chemical fertilizati<strong>on</strong> applied <strong>on</strong> organic background de<strong>term</strong>ined the increase of<br />
the yields with 28.3% in crop rotati<strong>on</strong> wheat – maize and with 23.3% in crop rotati<strong>on</strong><br />
oat + clover – clover – wheat – maize.<br />
Table 2: Maize yield obtained under the influence of fertilizati<strong>on</strong> and crop rotati<strong>on</strong>,<br />
Oradea 2004 – 2006<br />
Chemical fertilizati<strong>on</strong><br />
Organic fertilizati<strong>on</strong><br />
N0P0<br />
N90P60K60<br />
2004 2005 2006 Average 2004 2005 2006 Average<br />
Wheat - maize<br />
1. C<strong>on</strong>trol 5170 5020 4860 5020 6420 6340 6110 6290<br />
2. Manure 25 t/ha 5990 5620 5520 5710 7030 6830 6820 6890<br />
3. Manure 50 t/ha 6710 6100 6130 6310 7620 7320 7200 7380<br />
4. Lupin 6820 6140 6100 6350 7710 7300 7240 7416<br />
5. Lupin + oat 8010 7320 7930 7750 8760 8520 8410 8560<br />
Oat + clover – clover – wheat – maize<br />
1. C<strong>on</strong>trol 5990 5880 5710 5860 7230 6820 7010 7020<br />
2. Manure 25 t/ha 6810 6400 6320 6510 8010 7350 7560 7640<br />
3. Manure 50 t/ha 7520 6820 6860 7067 8780 7810 7900 8163<br />
4. Lupin 7500 6900 6990 7130 8790 7900 7960 8220<br />
5. Lupin + oat 8410 8020 7910 8113 9220 9000 9020 9080<br />
Organic Chemical Chemical fertilizati<strong>on</strong> x Organic fertilizati<strong>on</strong> x<br />
fertilizati<strong>on</strong> fertilizati<strong>on</strong> Organic fertilizati<strong>on</strong> Chemical fertilizati<strong>on</strong><br />
a1 a2 a1 a2 a1 a2 a1 a2<br />
LSD 5% 180 150 120 110 230 210 208 190<br />
LSD1% 310 290 240 208 410 420 310 360<br />
LSD 0.1% 520 470 410 390 710 730 530 580<br />
Both in the crop rotati<strong>on</strong> wheat-maize and in the crop rotati<strong>on</strong> oat + clover-clover-wheat<br />
+ maize, the lowest maize yields values were obtained in the variant with lupin; the<br />
differences in comparis<strong>on</strong> with the c<strong>on</strong>trol were of 13.7% (N0P0K0) and of 9.5%<br />
(N90P60K60) in the crop rotati<strong>on</strong> wheat-maize and of 11.1% (N0P0K0) and of 8.8%<br />
(N90P60K60) in the crop rotati<strong>on</strong> with clover (table 2).<br />
In the variant with lupin + oat, the maize yields registered were bigger than in the<br />
variants with lupin, pure crop; overall <strong>on</strong> the studied period the differences registered<br />
97
were of 600 kg/ha (N90P60K60) and of 490 kg/ha (N90P60K60) in the crop rotati<strong>on</strong> wheatmaize;<br />
in the crop rotati<strong>on</strong> with clover the differences were of 557 kg/ha (N0P0K0) and<br />
of 523 kg/ha (N90P60K60)).<br />
Generally, in the variant with manure 25 t/ha the yield registered were closed to the<br />
yield obtained in the variant with lupin + oat; the biggest yields was registered in the<br />
variant with manure 50 t/ha in all cases.<br />
Influence of the crop rotati<strong>on</strong> and organic fertilizati<strong>on</strong> <strong>on</strong> wheat water c<strong>on</strong>sumpti<strong>on</strong><br />
All the years, in the crop rotati<strong>on</strong> with clover, a bigger quantity of rainfall was stored<br />
during the cold period. The same phenomen<strong>on</strong> was registered in the variant with organic<br />
fertilizati<strong>on</strong> in comparis<strong>on</strong> with the c<strong>on</strong>trol. In these c<strong>on</strong>diti<strong>on</strong>s the values of the wheat<br />
water c<strong>on</strong>sumpti<strong>on</strong> are bigger in the variants from crop rotati<strong>on</strong> with clover and from<br />
organic fertilizati<strong>on</strong>. (Table 3)<br />
Table 3: Influence of the crop rotati<strong>on</strong> and organic fertilizati<strong>on</strong> <strong>on</strong> wheat water<br />
c<strong>on</strong>sumpti<strong>on</strong>, Oradea 2004 – 2006<br />
Variant of organic<br />
Crop rotati<strong>on</strong><br />
fertilizati<strong>on</strong><br />
Wheat - maize Oat + clover – clover – wheat - maize<br />
2004 2005 2006 2004 2005 2006<br />
1. C<strong>on</strong>trol 3670 3820 4250 4020 4210 4520<br />
2. Manure 25 t/ha 3810 3980 4420 4200 4400 4660<br />
3. Manure 50 t/ha 3960 4210 4760 4420 4580 4880<br />
4. Lupin 3680 3850 4310 4100 4290 4530<br />
5. Lupin + oat 3780 3930 4400 4180 4350 4700<br />
Average 3780 3958 4436 4184 4366 4658<br />
Influence of the crop rotati<strong>on</strong> and green manure <strong>on</strong> maize water c<strong>on</strong>sumpti<strong>on</strong><br />
In the variants with green manure and manure bigger quantities of rainfall were stored<br />
during the cold period leading to values of the maize water c<strong>on</strong>sumpti<strong>on</strong> higher than the<br />
c<strong>on</strong>trol. In the variant with lupin + oat the values of the maize’ water c<strong>on</strong>sumpti<strong>on</strong> were<br />
bigger than in maize’ water c<strong>on</strong>sumpti<strong>on</strong> in the variant with lupin pure crop. The<br />
highest value of the maize’ water c<strong>on</strong>sumpti<strong>on</strong> was registered in the variant with<br />
manure 50 t/ha. In the crop rotati<strong>on</strong> with clover bigger quantities of rainfall were stored<br />
in the cold period de<strong>term</strong>ining values of the water c<strong>on</strong>sumpti<strong>on</strong> higher than the values<br />
registered in the wheat-maize crop rotati<strong>on</strong> (table 4).<br />
Table 4: Maize water c<strong>on</strong>sumpti<strong>on</strong> obtained under the influence of the organic<br />
fertilizati<strong>on</strong> and crop rotati<strong>on</strong>, Oradea 2004 – 2006<br />
Variant of organic<br />
fertilizati<strong>on</strong><br />
2004<br />
Wheat - maize<br />
2005<br />
Crop rotati<strong>on</strong><br />
Oat + clover – clover – wheat - maize<br />
2006 2004 2005 2006<br />
1. C<strong>on</strong>trol 4910 4760 5370 5360 5110 5620<br />
2. Manure 25 t/ha 4970 4820 5460 5420 5200 5710<br />
3. Manure 50 t/ha 5100 4830 5510 5510 5320 5790<br />
4. Lupin 5120 4900 5500 5560 5300 5600<br />
5. Lupin + oat 5320 5020 5610 5720 5420 5810<br />
Average 5084 4866 5490 5514 5270 5706<br />
98
Influence of the crop rotati<strong>on</strong> and organic fertilizati<strong>on</strong> <strong>on</strong> water use efficiency in wheat<br />
In average <strong>on</strong> the studied period, in the background N0P0, in the crop rotati<strong>on</strong> oat +<br />
clover – clover – wheat – maize the wheat quantity obtained <strong>on</strong> 1 m 3 was bigger than<br />
the value obtained in wheat – maize crop rotati<strong>on</strong> with 44.3% (1.40 kg/m 3 vs. 0.97<br />
kg/m 3 ). The same situati<strong>on</strong> was registered in the background N90P60K60, (1.38 kg/m 3 vs.<br />
1.25 kg/m 3 ) but the relative difference (10.4%) is smaller.<br />
The green manure represented by mixture lupin and oat de<strong>term</strong>ined to obtain a water<br />
use efficiency bigger than the values obtained in the variant with lupin both in crop<br />
rotati<strong>on</strong> wheat – maize and in the crop rotati<strong>on</strong> oat + clover – clover – wheat – maize;<br />
the same situati<strong>on</strong> was registered in the background N0P0 and N90P60K60. Generally, the<br />
water use efficiency obtained in the variant with lupin + oat like green manure was very<br />
closed to water use efficiency obtained in the variant with manure, 25 t/ha.<br />
Table 5: Influence of the crop rotati<strong>on</strong> and fertilizati<strong>on</strong> <strong>on</strong> water use efficiency in<br />
Variant of organic<br />
fertilizati<strong>on</strong><br />
wheat, Oradea 2004 – 2006<br />
Crop rotati<strong>on</strong><br />
Wheat - maize Oat + clover – clover – wheat – maize<br />
2004 2005 2006 Average 2004 2005 2006 Average<br />
1. C<strong>on</strong>trol 0.75 0.79 0.84<br />
N0P0<br />
0.79 1.25 1.31 1.39 1.32<br />
2. Manure 25 t/ha 1.12 0.97 0.98 0.99 1.42 1.38 1.47 1.42<br />
3. Manure 50 t/ha 1.27 1.09 1.06 1.13 1.45 1.50 1.45 1.47<br />
4. Lupin 1.00 0.89 0.89 0.93 1.40 1.32 1.33 1.35<br />
5. Lupin + oat 1.08 0.97 0.98 1.01 1.51 1.42 1.36 1.43<br />
Average 1.02 0.94 0.97 0.97 1.41 1.39 1.40 1.40<br />
N90P60K60<br />
1. C<strong>on</strong>trol 1.00 1.11 1.11 1.07 1.25 1.27 1.29 1.27<br />
2. Manure 25 t/ha 1.27 1.30 1.33 1.30 1.42 1.38 1.39 1.40<br />
3. Manure 50 t/ha 1.32 1.41 1.39 1.37 1.45 1.50 1.45 1.47<br />
4. Lupin 1.15 1.20 1.22 1.19 1.40 1.32 1.33 1.35<br />
5. Lupin + oat 1.28 1.31 1.29 1.29 1.51 1.42 1.36 1.43<br />
Average 1.20 1.27 1.27 1.25 1.41 1.38 1.36 1.38<br />
Influence of the crop rotati<strong>on</strong> and organic fertilizati<strong>on</strong> <strong>on</strong> water use efficiency in maize<br />
In comparis<strong>on</strong> with the c<strong>on</strong>trol, in the variant with lupin pure crop the lowest values of<br />
the water use efficiency were de<strong>term</strong>ined both in the crop rotati<strong>on</strong> wheat-maize (1.08<br />
kg/m 3 in the N0P0K0 background and 1.36 kg/m 3 in the N90P60K60 background) and in<br />
the crop rotati<strong>on</strong> with clover (1.3 kg/m 3 in the N0P0K0 and 1.45 kg/m 3 in the N90P60K60<br />
background). In the variants with lupin + oat, the water use efficiency was better than in<br />
the variant with lupin pure crop. The values registered in the variants with manure 25<br />
t/ha were the same or a little bigger than the values obtained in the variant with lupin +<br />
oat; the highest value of the water use efficiency was registered in the variant with<br />
manure 50 t/ha (table 6)<br />
99
Table 6: Water use efficiency (kg/m 3 ) in maize under the influence of the organic<br />
fertilizati<strong>on</strong> and crop rotati<strong>on</strong>, Oradea 2004 – 2006<br />
Variant of organic<br />
fertilizati<strong>on</strong><br />
Wheat - maize<br />
Crop rotati<strong>on</strong><br />
Oat + clover – clover – wheat – maize<br />
2004 2005 2006 Average 2004 2005 2006 Average<br />
N0P0<br />
1. C<strong>on</strong>trol 1.05 1.05 0.91 1.00 1.31 1.33 1.14 1.26<br />
2. Manure 25 t/ha 1.12 1.17 0.96 1.08 1.41 1.42 1.25 1.36<br />
3. Manure 50 t/ha 1.32 1.26 1.11 1.23 1.49 1.52 1.31 1.44<br />
4. Lupin 1.33 1.25 1.11 1.23 1.51 1.49 1.32 1.44<br />
5. Lupin + oat 1.51 1.46 1.41 1.46 1.65 1.75 1.50 1.63<br />
Average 1.27 1.24 1.10 1.20 1.47 1.50 1.30 1.42<br />
N90P60K60<br />
1. C<strong>on</strong>trol 1.31 1.33 1.14 1.26 1.35 1.33 1.37 1.35<br />
2. Manure 25 t/ha 1.41 1.42 1.25 1.36 1.48 1.41 1.45 1.45<br />
3. Manure 50 t/ha 1.49 1.52 1.31 1.44 1.59 1.47 1.48 1.51<br />
4. Lupin 1.51 1.49 1.32 1.44 1.58 1.49 1.50 1.52<br />
5. Lupin + oat 1.65 1.70 1.50 1.62 1.61 1.66 1.66 1.64<br />
Average 1.47 1.49 1.53 1.42 1.52 1.47 1.45 1.49<br />
CONCLUSIONS<br />
1. The yields wheat obtained in the crop rotati<strong>on</strong> oat + clover – clover – wheat –<br />
maize were bigger than yields obtained in the wheat – maoze crop rotati<strong>on</strong> both in<br />
N0P0 background and in N90P60K60 background.<br />
2. In the variant with lupin+oat, the maize yield obtained was bigger than the maize<br />
yield obtained in the variant with lupin pure crop both in the wheat-maize crop<br />
rotati<strong>on</strong> and in the crop rotati<strong>on</strong> with clover. The same situati<strong>on</strong> was registered both<br />
in the N0P0K0 background and in the N90P60K60 background.<br />
3. Organic fertilizati<strong>on</strong> (manure 25 t/ha, manure 50 t/ha, lupin, lupin + oat) but<br />
especially organic fertilizati<strong>on</strong> associated with annual background N90P60K60<br />
de<strong>term</strong>ined to obtained very significant yield gains in comparis<strong>on</strong> with c<strong>on</strong>trol.<br />
4. The use of the green manure composed by lupin + oat (applied for maize)<br />
de<strong>term</strong>ined yield gains bigger than green manure composed by lupin like pure crop.<br />
The yields gains were registered in the twice crop rotati<strong>on</strong> studied.<br />
5. In the crop rotati<strong>on</strong> with clover the rainfall storages in the cold period were bigger<br />
than in the crop rotati<strong>on</strong> wheat – maize; the same situati<strong>on</strong>, was registered in the<br />
variants with organic fertilizati<strong>on</strong>. Due the values of wheat and maize water<br />
c<strong>on</strong>sumpti<strong>on</strong> increased.<br />
6. Both in wheat and maize the water use efficiency values obtained in the crop<br />
rotati<strong>on</strong> with clover were bigger than in the crop rotati<strong>on</strong> wheat – maize; the values<br />
registered in the background N90P60K60 were bigger than the values obtained in the<br />
background N0P0. In the variant with lupin + oat like green manure the water use<br />
efficincy had bigger values than in the variant with lupin pure crop.<br />
7. The researches emphasizes the importance of the crop rotati<strong>on</strong> in wheat and maize<br />
and recomand to use of the green manure like mixture beetween lupin and oat and<br />
not like lupin pure crop.<br />
100
REFERENCES<br />
1. Budoi Gh., Penescu A. (1996) – Agrotehnica. Ed. Ceres, 427 – 432 p.<br />
2. Domuţa C., (1999) – Ameliorarea fertilităţii solurilor erodate pe terenurile în pantă<br />
din vestul ţării. Cereale şi plante tehnice nr. 7.<br />
3. Domuţa C. (2005) – Agrotehnica terenurilor în pantă din nord – vestul României.<br />
Ed. Universităţii din Oradea, 96 – 117 p.<br />
4. Domuţa C. (2006) – Agrotehnica diferenţiată. Ed. Universităţii din Oradea, 377 –<br />
442 p.<br />
5. Eliade Gh., Ghinea L., Ştefanic Gh., (1983) – Bazele biologice ale fertilităţii<br />
solului. Ed. Ceres, 127 – 130 p.<br />
6. Guş P., Lăzureanu A., Săndoiu D., Jităreanu G., Stancu I. (1998) – Agrotehnica.<br />
Ed. Risoprint, 496 – 499 p.<br />
7. Pakurar M., Nagy I. (2005) - Effect of different tillage treatments <strong>on</strong> the yield of<br />
maize. <str<strong>on</strong>g>Internati<strong>on</strong>al</str<strong>on</strong>g> Symposium A.R.D.S. Oradea, 7 – 8 July, 158 – 165 p.<br />
8. Pepo P. (2005) – New issues in sustainable wheat producti<strong>on</strong>. <str<strong>on</strong>g>Internati<strong>on</strong>al</str<strong>on</strong>g><br />
Symposium A.R.D.S. Oradea, 7 – 8 July, 17 – 28 p.<br />
9. Samuel D., Drăgan – Bularda M., Domuţa C (2006) – The effect of green<br />
manure <strong>on</strong> enzymatic activities in a brown luvic soil. Studia Universitas Babeş –<br />
Bolyai, Biologia, L I, 83 – 93 p.<br />
101
APPLICATION OF A DIGESTATE AS A NUTRIENT SOURCE AND ITS<br />
EFFECT ON SOME SELECTED CROPS AND SOIL PROPERTIES 1<br />
ABSTRACT<br />
Marianna Makádi 1 , Attila Tomócsik 1 , József Lengyel 2 ,<br />
Zsolt Bogdányi 1 , Árpád Márt<strong>on</strong> 1<br />
1 Research Centre of CAS of University of Debrecen<br />
H-4432 Nyíregyháza, Westsik Vilmos u. 4-6.<br />
2 Bátortrade Ltd. H-4300 Nyírbátor, Árpád u. 156/A.<br />
The largest biogas plant of Hungary is operated in the city of Nyírbátor, NE Hungary. A<br />
substantial amount, some 95.000 m 3 of digestate is originating every year, as the<br />
residual material of biogas producti<strong>on</strong>. A study was c<strong>on</strong>ducted <strong>on</strong> the utilizati<strong>on</strong> of this<br />
material as a nutrient source for different crops. In this paper we present the results<br />
obtained with winter wheat and triticale. The use of the digestate has a more<br />
pr<strong>on</strong>ounced effect <strong>on</strong> triticale than <strong>on</strong> winter wheat. On the average, the yield of winter<br />
wheat and triticale were increased by 20.8 % and 51.42 %, respectively, and the grain to<br />
straw ratio was also increased by the treatments with the digestate. As for soil<br />
properties, the treatments caused positive changes in the case of humus c<strong>on</strong>tent, but they<br />
increased the Na c<strong>on</strong>tent of the soil.<br />
Keywords: biogas, digestate, nutrient supply, winter wheat, triticale<br />
INTRODUCTION<br />
According to the regulati<strong>on</strong>s of Law No. XLIII./2000 <strong>on</strong> waste management, by 2014<br />
the amount of biologically degradable waste types disposed in landfills should be<br />
reduced to 35 % of the total amount of the waste landfilled in 1995. A significant<br />
amount of organic waste can be composted and used as natural fertilizer and soil<br />
improving material. The other possible treatment of organic wastes is their utilizati<strong>on</strong> in<br />
biogas plants. The main products of this treatment are biogas (methane) and heat, while<br />
by-product is digestate, which can also be utilized as source of nutrients in agriculture.<br />
This anaerobic treatment is envir<strong>on</strong>mentally friendly because it utilises renewable<br />
energy sources such as plants, and destroys and utilises organic wastes such as offal.<br />
The fermentati<strong>on</strong> residue – the digestate – can substitute for the artificial fertilizers<br />
(Kovács, 2007).<br />
Numerous papers deal with the importance of quality and quantity of basic<br />
materials in biogas producti<strong>on</strong>. But very few papers are available <strong>on</strong> the utilizati<strong>on</strong> of<br />
the digestate as a plant nutrient source. Qi et al. (2005) examined in North China the<br />
effect of fermented waste as organic manure in cucumber and tomato producti<strong>on</strong>. They<br />
have found increasing yield and vitamin C-c<strong>on</strong>tent of about 18 %. Banik and Nandi<br />
(2004) investigated biogas residual slurry manure <strong>on</strong> the yield, protein and mineralc<strong>on</strong>tents<br />
of oyster mushroom. They have found positive effects.<br />
Besides the examinati<strong>on</strong> of the effect of the digestate <strong>on</strong> the crop yields of different<br />
plants, it is also very important to study its effects <strong>on</strong> the soil properties.<br />
1 This project is supported by the grant GVOP-3.1.1.-2004-05-0220/3.0.<br />
102
MATERIALS AND METHODS<br />
The experiment was carried out in the Nyírség regi<strong>on</strong>, the NE Hungary. The biogas<br />
plant is a part of a big agricultural system with crop producti<strong>on</strong>, livestock (beef and<br />
diary cattle), slaughterhouse and processing plant. The biogas plant works with<br />
mesophylic and thermophylic systems, the basic materials are livestock manure, plant<br />
crop and offal. The total capacity of thermophylic and mesophylic digesters are 7,600<br />
m 3 and 9,400 m 3 , respectively. The remaining digestate after anaerobic digesti<strong>on</strong> can be<br />
utilized in crop producti<strong>on</strong> as a nutrient supply.<br />
No. of<br />
treatment<br />
Table 1: Applied treatments in the experiments with digestate.<br />
Phenological stage at the time<br />
of treatment<br />
Proporti<strong>on</strong><br />
Quantity of<br />
digestate<br />
Quantity of water<br />
1. C<strong>on</strong>trol 0 0 0<br />
2. after sowing 100% mean 0<br />
3. after sowing 100% +30% 0<br />
4.<br />
5.<br />
6.<br />
7.<br />
after sowing<br />
plants with 6-7 leafs<br />
after sowing<br />
plants with 6-7 leafs<br />
after sowing<br />
plants with 3-4 leafs<br />
plants with 6-7 leafs<br />
after sowing<br />
plants with 3-4 leafs<br />
plants with 6-7 leafs<br />
50%<br />
50%<br />
50%<br />
50%<br />
25%<br />
50%<br />
25%<br />
25%<br />
50%<br />
25%<br />
mean 0<br />
+30% 0<br />
mean 0<br />
+30% 0<br />
8. after sowing 100% 0 +30%<br />
9. after sowing 100% 0 mean<br />
10.<br />
after sowing<br />
plants with 6-7 leafs<br />
50%<br />
50%<br />
0 mean<br />
11.<br />
after sowing<br />
plants with 6-7 leafs<br />
50%<br />
50%<br />
0 +30%<br />
after sowing<br />
25%<br />
12.<br />
plants with 3-4 leafs<br />
50%<br />
0 +30%<br />
plants with 6-7 leafs<br />
25%<br />
after sowing<br />
25%<br />
13.<br />
plants with 3-4 leafs<br />
50%<br />
0 mean<br />
plants with 6-7 leafs<br />
25%<br />
The rates of the digestate to be applied was de<strong>term</strong>ined by its actual N-c<strong>on</strong>tent, because<br />
N is <strong>on</strong>e of the main macr<strong>on</strong>utrients for plants, but it could be a serious envir<strong>on</strong>mental<br />
polluti<strong>on</strong> if leached to the soil water and accumulates in it. The main parameters of the<br />
digestate are the followings: pH 8.025, density 1025.75 kg m 3 , dry matter c<strong>on</strong>tent<br />
1.1824 m/m% (row matter), total-N 0.376 m/m% (row matter), Mg 32.7 mg kg -1 (r.m.),<br />
Na 272.5 mg kg -1 (r.m.).<br />
103
In the experiments (Table 1.) we applied the “mean” quantity of digestate, which means<br />
the quantity of it equals the average N-demand of the crop plants, while the high dose<br />
was the digestate c<strong>on</strong>taines “mean+30%” nitrogen. The digestate c<strong>on</strong>tains much water,<br />
therefore we applied the same quantity of water in the experiment with winter wheat. In<br />
the triticale-experiment we could not use water. No additi<strong>on</strong>al fertilizer was applied.<br />
The sowing time was 15.10.2005. for winter wheat cv. ‘Sixtus’ (Triticum aestivum<br />
ssp. vulgare L.) and 05.11.2005. for triticale cv. ‘Versus’ (X Triticosecale Wittm.). The<br />
winter wheat was harvested <strong>on</strong> 15.07.2006, the triticale <strong>on</strong> 19.08.2006.<br />
We measured the yield of the crop, physical, chemical and biological parameters of the<br />
soils. One-way analysis of variance, Tukey’s test and T-test were used to de<strong>term</strong>ine<br />
treatment effects by SPSS statistical program.<br />
RESULTS AND DISCUSSIONS<br />
1./ Winter wheat: Winter wheat is the main cereal crop in Hungary. It is mainly<br />
cultivated <strong>on</strong> chernozem soils. In the Nyírség regi<strong>on</strong> the main cereal crop is rye, which<br />
produces satisfactory yield <strong>on</strong> sandy soils. Nevertheless, some better sandy soils are<br />
also appropriate for winter wheat producti<strong>on</strong> with the subsequent delivery of nutrients<br />
in the large-scale farming systems (Bocz (ed.), 1992). In our experiment the nutrient<br />
supply of the winter wheat was ensured by the digestate. The doses of digestate were<br />
calculated <strong>on</strong> the N-demand of the crop, based <strong>on</strong> the literature, and the calculated<br />
amounts were applied to the surface of soil and plants in full, or split in two or three<br />
porti<strong>on</strong>s. In the case of good nutrient supply the crop yield of winter wheat is generally<br />
around 6-7 t ha -1 <strong>on</strong> chernozems and 4-5 t ha -1 <strong>on</strong> sands. In our experiments the average<br />
crop yield was 3.72 t ha -1 , which is less than the average <strong>on</strong> sandy soils. The results of<br />
treatments can be seen in Figure 1.<br />
t*ha -1<br />
6<br />
5<br />
4<br />
3<br />
2<br />
1<br />
0<br />
Winter wheat crop yield<br />
1 2 3 4 5 6 7 8 9 10 11 12 13<br />
No. of treatments<br />
Figure 1: Winter wheat crop yield as affected by the treatments of digestate and<br />
water. Treatments are numbered according to Table 1.<br />
We have not found any significant difference am<strong>on</strong>g the treatments <strong>on</strong> the p
average of watered treatments resulted in a 3.35 % yield increase, while the average of<br />
digestate-treatments increased the yield by 20.8 %, compared to the c<strong>on</strong>trol. The<br />
positive effect of digestate benchmarked against water based <strong>on</strong> its nutrient c<strong>on</strong>tent and<br />
its stimulating effect <strong>on</strong> soil microorganisms.<br />
We have to take into account, that if the digestate is applied in full after sowing,<br />
not all the nutrients can be utilized by plants, part of them will move down to lower soil<br />
layers and may pollute soil water. If the digestate is split into two or three porti<strong>on</strong>s at<br />
the time when the plant needs it, the utilizati<strong>on</strong> of the digestate by plants will be more<br />
efficient, the applicati<strong>on</strong> will be envir<strong>on</strong>mentally friendly. Nowadays in Hungary the<br />
yearly amount of the digestate can be utilized <strong>on</strong>ly before sowing (except our<br />
experiments), because the digestate, as a category, is not defined in Hungarian law. The<br />
digestate is handled as sewage sludge. On the basis of our results, and also c<strong>on</strong>sidering<br />
the results from tests <strong>on</strong> other crops, the applicati<strong>on</strong> of the digestate, based <strong>on</strong> Ndemand<br />
of plants, split in two or three porti<strong>on</strong>s is the right way both in ec<strong>on</strong>omic and<br />
envir<strong>on</strong>mental respects. We computed the grain to straw ratio from the averages of the<br />
water and digestate treatments (Table 2).<br />
Table 2: Grain to straw ratio of winter wheat in the experiment.<br />
C<strong>on</strong>trol<br />
Digestate<br />
Water<br />
Mean + Std. Dev.<br />
0,819 + 0,037<br />
0,907 + 0,110<br />
0,857 + 0,075<br />
The digestate treatments increased this ratio, which means that digestate resulted less<br />
straw and more yield in the treated plots.<br />
2./ Triticale: Triticale is the typical crop of marginal soils. It has higher yield than rye<br />
has, that is why triticale displaced rye from the better sandy soils. The average yield of<br />
this plant <strong>on</strong> the field plots of Bátortrade Ltd, with the use of fertilizers, is 2.8-3.1 t ha -1 .<br />
In our small-plot experiment the samples were collected from sampling plots of 1 m 2 in<br />
size, and after threshing the grain and the straw yield was taken. The yield was higher<br />
than the average as a result of the manual harvesting and handling.<br />
For technical reas<strong>on</strong>s, this experiment was <strong>on</strong>ly treated with the digestate. The<br />
results obtained from the treatments 1-7 can be seen in Figure 2.<br />
Table 3: Grain to straw ratio of triticale in the experiment.<br />
C<strong>on</strong>trol<br />
Digestate<br />
Mean + Std.Dev.<br />
0.702+ 0.074<br />
0.820 + 0.079<br />
Comparing these results to those obtained with winter wheat, treatments were more<br />
efficient since we found significant treatment effect (p
compared to the c<strong>on</strong>trol. It is important to c<strong>on</strong>duct experiments with a large variety of<br />
different crop plants, because they resp<strong>on</strong>d differently to the digestate treatment.<br />
g*m 2<br />
450<br />
400<br />
350<br />
300<br />
250<br />
200<br />
150<br />
100<br />
50<br />
0<br />
a<br />
a<br />
b<br />
Triticale crop yield<br />
a<br />
b<br />
a<br />
b<br />
1 2 3 4 5 6 7<br />
No. of treatments<br />
Figure 2: Triticale crop yield as affected by the treatments of digestate. Treatments<br />
are numbered according to Table 1. Signs a,b means significance groups according to<br />
Tukey’s test (p
Table 4: Changes in soil properties under the winter wheat after 2 years of digestate<br />
applicati<strong>on</strong>. Treatments are numbered according to Table 1.<br />
Treatment pH H2O pH KCl y1 Humus %<br />
NO3-N (mg kg -1 P2O5 ) (mg kg -1 )<br />
K2O (mg kg -1 Na<br />
) (mg kg -1 )<br />
Mg<br />
(mg kg -1 )<br />
Before treatment 5,49 4,52 2,34 1,038 33,834 348 221 17,5 78,4<br />
1 5,89 4,94 8,50 0,867 0,171 138 54 12,8 3,8<br />
2 4,80 3,88 11,00 0,696 7,068 85 95 26,1 10,3<br />
3 5,36 4,40 12,75 0,585 9,013 104 102 14,5 9,7<br />
4 6,35 5,81 8,75 0,978 8,868 98 88 18,8 13,8<br />
5 5,06 4,36 11,25 1,018 21,044 109 132 33,4 15,8<br />
6 4,86 3,90 11,25 1,301 5,509 133 76 32,0 10,5<br />
7 4,76 4,04 11,75 0,897 9,922 194 155 29,7 21,0<br />
8 5,64 4,25 8,50 0,897 0,216 173 52 16,0 3,6<br />
9 6,03 4,98 6,50 0,827 0,184 200 54 14,6 4,9<br />
10 5,63 4,72 10,25 1,139 0,195 197 47 17,7 2,3<br />
11 5,65 4,95 10,25 0,887 0,622 170 46 20,9 7,1<br />
12 5,83 4,46 8,25 0,847 0,480 197 63 19,6 7,1<br />
13 5,66 4,39 12,75 0,948 0,329 192 66 20,1 6,9<br />
Table 5: Changes in soil properties under the triticale after 1 year of digestate<br />
applicati<strong>on</strong>. Treatments are numbered according to Table 1.<br />
Treatment pH H2O pH KCl y1 Humus %<br />
NO3-N (mg kg -1 P2O5 ) (mg kg -1 )<br />
K2O (mg kg -1 Na<br />
) (mg kg -1 )<br />
Mg<br />
(mg kg -1 )<br />
Before treatment 5,46 3,88 9,65 0,44 4,688 245 179 11,5 60,3<br />
1 5,42 3,92 12,75 0,68 0,190 242 131 76,8 3,7<br />
2 5,74 4,29 12,25 0,83 1,322 264 209 67,5 13,0<br />
3 5,57 4,18 11,75 0,76 0,232 173 125 89,8 13,4<br />
4 5,71 4,66 12,25 0,84 0,205 253 210 86,3 12,5<br />
5 5,38 4,01 12,75 0,92 1,433 258 155 49,6 12,8<br />
6 5,26 4,04 8,75 0,79 0,342 258 157 49,5 11,8<br />
7 5,39 4,47 12,50 0,50 2,863 205 136 44,5 3,7<br />
NO3-N and Mg-c<strong>on</strong>tent of soils decreased in both treatments, while Na-c<strong>on</strong>tent was<br />
increased. Increased Na-c<strong>on</strong>tent indicates unfavourable changes in the soils. We<br />
indicated this process for the operators, and the decreasing of Na-c<strong>on</strong>tent of the<br />
digestate was successful.<br />
REFERENCES<br />
Banik, S., Nandi, R. (2004): Effect of supplementati<strong>on</strong> of rice straw with biogas<br />
residual slurry manure <strong>on</strong> the yield, protein and mineral c<strong>on</strong>tents of oyster<br />
mushroom. Industrial Crops and Products, 20: 311-319.<br />
Bocz, E. (ed.) (1992): Szántóföldi növény<strong>term</strong>esztés. Mezőgazda Kiadó, Budapest.<br />
Kovács, K. (2007): Biogas in Hungary: the state of the art in 2007, and future<br />
opportunities. Waste, 2: 11-16.<br />
Qi, X., Zhang, S., Wang, Y., Wang, R. (2005): Advantages of the integrated pigbiogas-vegetable<br />
greenhouse system in North China. Ecological Engineering,<br />
24:177-185.<br />
107
ABSTRACT<br />
SOIL TILLAGE SYSTEMS AND NITROGEN FERTILIZATION<br />
FOR WINTER BARLEY AFTER SOYBEAN 1<br />
Stipesevic, Bojan, Jug, Danijel, Stosic, Miro, Zugec, Ivan, Jug, Irena<br />
Faculty of Agriculture Osijek, Trg Sv. Trojstva 3, 31000 Osijek, Croatia<br />
The winter barley crop producti<strong>on</strong> is not adequately researched regarding soil tillage<br />
systems, especially in crop rotati<strong>on</strong> with the soybean, both crops gaining importance as<br />
food for the animals. The research at experimental site Boksic (Croatia), during the<br />
years 2005 and 2006, showed no difference in yields from c<strong>on</strong>venti<strong>on</strong>al tillage, based<br />
<strong>on</strong> ploughing, and reduced tillage, based <strong>on</strong> diskharrowing, in each of six nitrogen<br />
fertilizati<strong>on</strong> levels (0, 30, 60, 90, 120 and 150 kg N/ha). Regarding N fertilizati<strong>on</strong>, yield<br />
increase was not significantly higher after applied 90 kg N/ha.<br />
Keywords: winter barley, soybean, c<strong>on</strong>venti<strong>on</strong>al tillage, reduced tillage, nitrogen<br />
INTRODUCTION<br />
The soil tillage systems for winter barley producti<strong>on</strong> had been rec<strong>on</strong>sidered during the<br />
last decade, especially in the light of the Croatian needs for more affordable and high<br />
quality cattle fodder. This process is a result of worldwide trends and research results<br />
about tillage simplificati<strong>on</strong>s for higher sustainability of the agriculture, in which the<br />
envir<strong>on</strong>ment protecti<strong>on</strong> and decreases of tillage costs is especially emphasised (Karlen<br />
et al., 1994). In the Slav<strong>on</strong>ia, the most agricultural regi<strong>on</strong> of the Republic of Croatia,<br />
various systems of reduced tillage for different crops have been already tested<br />
(Stipesevic et al., 1997, 2000; Zugec et al., 2000; Filipovic et al., 2006; Jug et al., 2006),<br />
with main goals to decrease the costs of producti<strong>on</strong>, maintain agrosphere sustainability<br />
and to preserve high yield (characteristic for this regi<strong>on</strong>) despite the reducti<strong>on</strong> of applied<br />
agritechniques. Al<strong>on</strong>g with the introducti<strong>on</strong> of reduced tillage systems, the awareness<br />
has been raised of different approach toward fertilizati<strong>on</strong>, soil compacti<strong>on</strong>, weed c<strong>on</strong>trol<br />
and other problems c<strong>on</strong>nected with lesser soil agitati<strong>on</strong>. The simplified soil tillage<br />
particularly raised the questi<strong>on</strong> of efficiency of fertilizers, especially nitrogen, in<br />
interacti<strong>on</strong> with the tillage systems.<br />
MATERIALS AND METHODS<br />
This research was c<strong>on</strong>ducted near Boksic in Eastern Croatia, for the winter barley<br />
(Hordeum vulgare L.) in a crop rotati<strong>on</strong> after soybeans (Glycine max L.) for crop<br />
seas<strong>on</strong>s 2004/05-2005/2006. The site's soil type was de<strong>term</strong>ined as a eutric cambisol,<br />
with loamy clay texture, total porosity between 32.2-44.7%, bulk density from 1.30 to<br />
1.70 kg dm -3 , neutral reacti<strong>on</strong> (pH in KCl 6.8), with rather high c<strong>on</strong>tent of humus (4.%),<br />
and with poor fertility (6.6 mg P2O5 and 6.8 mg K2O per 100 g of soil, 2.8 % of CaCO3)<br />
in 0-30 cm depth. The main experimental set-up was a split-plot design in three<br />
1 This research has been sp<strong>on</strong>sored by Croatian Ministry for Agriculture, Forestry and<br />
Water Management (VIP Project V-29-7/04) and family farm "Oto Kovacevic" from<br />
Boksic, Croatia, <strong>on</strong> whose land this experimental site had been established.<br />
108
epetiti<strong>on</strong>s, where the main treatment was soil tillage with two steps: CT=c<strong>on</strong>venti<strong>on</strong>al<br />
tillage (autumn ploughing up to 25 cm depth, spring diskharrowing, followed by<br />
seedbed preparati<strong>on</strong> with rototiller and standard sowing) and DS=autumn diskharrowing<br />
up to 20 cm depth, seedbed preparati<strong>on</strong> with rototiller in spring and standard sowing.<br />
The sub-treatment of the nitrogen fertilizati<strong>on</strong> c<strong>on</strong>sisted of six steps of nitrogen<br />
fertilizati<strong>on</strong>: N1=0, N2=30, N3=60, N4=90, N5=120 and N6=150 kg N ha -1 (see Table<br />
1 for the nitrogen fertilizati<strong>on</strong> distributi<strong>on</strong>), with the same amount of phosphorus (83 kg<br />
P2O5 ha -1 ) and potassium (124 kg K2O ha -1 ) each seas<strong>on</strong>. The phosphorus and potassium<br />
amounts were de<strong>term</strong>ined by soil analyses and planned crop uptake recommendati<strong>on</strong>s.<br />
The basic experimental plot size was 5 m wide and 30 m l<strong>on</strong>g (total area of 150 m 2 ).<br />
The winter barley cultivar "Trenk" was sown, the creati<strong>on</strong> of the Agricultural Institute<br />
Osijek, Croatia, in recommended plant density of 450 plants m -2 , within the optimal<br />
sowing dates (31. October 2004 and 20. October 2005). During the harvest time, plots<br />
were harvested <strong>on</strong>e by <strong>on</strong>e and complete grain mass from each plot was weighted <strong>on</strong><br />
portable electr<strong>on</strong>ic scale, whereas moisture c<strong>on</strong>tent was de<strong>term</strong>ined by "Dickey John<br />
GAC 2000" grain moisture meter, from ten subsamples taken during the harvest and<br />
preserved in the plastic bags. Soil c<strong>on</strong>e penetrologger "Eikelkamp" model SN with c<strong>on</strong>e<br />
diameter 2.00 cm and 60° c<strong>on</strong>e angle was used for recoding soil resilience up to 60 cm<br />
depth in 1 cm steps with 10 measures for each basic experimental plot, read <strong>on</strong>ce in<br />
May 2005 and May 2006. The split-split-plot ANOVA was performed by SAS statistic<br />
package (V 8.02, SAS Institute, Cary, NC, USA, 1999) with Year as the main level,<br />
Tillage as sub-level and added N as sub-sub-level for the winter barley yields, and with<br />
Year as the main level, Tillage as the sub-level and Depth as the sub-sub-level for the<br />
soil resilience. The Fisher protected LSD means comparis<strong>on</strong>s were performed for<br />
P=0.05 significance levels.<br />
Table 1: Fertilizati<strong>on</strong> scheme for the Boksic experimental trial<br />
Fertilizati<strong>on</strong> levels<br />
Autumn fertilizati<strong>on</strong><br />
0PK NPK Urea<br />
1st<br />
Sidedressing<br />
KAN<br />
2nd<br />
Sidedressing<br />
KAN<br />
0:20:30 8:22:33 46% N 27% N 27% N<br />
N1: 0 kg N ha -1 413 - - - -<br />
N2: 30 kg N ha -1 - 375 - - -<br />
N3: 60 kg N ha -1 - 375 - 111 -<br />
N4: 90 kg N ha -1 - 375 - 111 111<br />
N5: 120 kg N ha -1 - 375 - 167 167<br />
N6: 150 kg N ha -1 - 375 65 167 167<br />
RESULTS AND DISCUSSION<br />
1. Soil resilience: Differences between two tillage treatments are presented in the<br />
Figure 1, where DS treatment formed compacted pan shallower than the CT, which was<br />
emphasised by Birkas et al. (2002) as "diskharrowing pan" and "ploughing pan".<br />
Previous deeper soil tillage residual looseness below "diskharrowing pan" for DS<br />
treatment is also clearly visible between 13-16 cm depth. Lower depth of soil<br />
109
disturbance by reduced tillage was clearly notable through statistically significant<br />
differences between two soil tillage treatments at 10-25 cm soil depth. In any case, the<br />
detected soil resilience values did not present limiting factor for normal growth and<br />
development of the winter barley, although some authors noticed that more compacted<br />
0-30 cm layer under reduced soil tillage can c<strong>on</strong>tribute toward lower grain yield of these<br />
treatments (Izumi et al., 2004).<br />
Figure 1: Soil resiliance for the site "Boksic", average for 2005 and 2006<br />
2. Winter barley yield: The rather favourable weather c<strong>on</strong>diti<strong>on</strong> in both years managed<br />
rather high yields for winter barley under both tillage systems with adequate available<br />
nitrogen, with no statistical differences between Tillage treatments for all Nitrogen<br />
treatment levels (Table 2), although DS tended to be slightly higher, especially at lower<br />
Nitrogen levels (N1 through N3). This effect was probably the result of higher<br />
c<strong>on</strong>centrati<strong>on</strong> of incorporated soybean residues in shallower upper soil layers. At the<br />
fertilizati<strong>on</strong> levels higher than 60 kg N ha -1 (N4 through N6), this difference tended to<br />
decrease due to the sufficient nitrogen supply in both Tillage treatments. These results<br />
were in accordance with some authors whose trials included ploughing and c<strong>on</strong>tinuous<br />
diskharrowing, who reported that, under favourable agroecological c<strong>on</strong>diti<strong>on</strong>s, yields<br />
did not differ am<strong>on</strong>g c<strong>on</strong>venti<strong>on</strong>al tillage and other reduced tillage systems (Halvors<strong>on</strong><br />
et al., 1999). However, some other authors with similar soil tillage treatments recorded<br />
110
that usually c<strong>on</strong>venti<strong>on</strong>al tillage had better results than reduced soil tillage systems<br />
(Varvel et al., 1989; Birkas et al., 2002), although some of them pointed out (Varvel et<br />
al., 1989) that different placement and higher amount of nitrogen could alleviate<br />
negative effects of reduced tillage and thus unfavourable soil preparati<strong>on</strong> for cereals<br />
growth at the yield level achieved with the c<strong>on</strong>venti<strong>on</strong>al tillage based <strong>on</strong> ploughing.<br />
Al<strong>on</strong>g with the increase of N amount, the winter barley resp<strong>on</strong>ded with significant<br />
yield growth between N2 to N3 (2.69 to 3.35 t ha -1 , respectively) and between N3 to N4<br />
(from 3.35 to 3.85 t ha -1 ), with better crop reacti<strong>on</strong> between N2 to N3 (for 663 kg ha -1 or<br />
increase of 25% in comparis<strong>on</strong> with the yield at N2 level), than between N3 to N4 (for<br />
498 kg ha -1 or 15% if compared with N3). These yield leaps are corresp<strong>on</strong>ding with the<br />
Mitscherlich's theory of lowering yield's increase, and roughly de<strong>term</strong>ining Nitrogen<br />
levels N3 and N4 as Baule 2 and Baule 3 values of added nutrient, respectively.<br />
Subsequently, nitrogen rates higher than 90 kg N ha -1 did not produce significantly<br />
higher winter barley yields, which is leading toward the c<strong>on</strong>clusi<strong>on</strong> that, for given<br />
agroecological c<strong>on</strong>diti<strong>on</strong>s and observed cultivar of winter barley, there is ineffective to<br />
fertilize with more than 90 kg N -1 in crop rotati<strong>on</strong> after the soybean.<br />
Table 2: Winter barley grain yield (kg ha -1 ) at 13% grain moistur, Boksic site,<br />
Croatia, average for both years ( 2005 and 2006)<br />
N level<br />
CT<br />
(Ploughing)<br />
DS<br />
(Diskharrowing)<br />
Average (N)<br />
N1 (0 kg N ha -1 ) 2380 a † 2571 a 2475 A ‡<br />
N2 (30 kg N ha -1 ) 2625 a 2752 a 2688 A<br />
N3 (60 kg N ha -1 ) 3224 b 3479 b 3351 B<br />
N4 (90 kg N ha -1 ) 3841 c 3858 bc 3849 C<br />
N5 (120 kg N ha -1 ) 3923 c 3928 bc 3925 C<br />
N6 (150 kg N ha -1 ) 4036 c 4107 c 4072 C<br />
average (Tillage) 3338 3449<br />
†<br />
The winter barley yields within the same Tillage level and labeled with the same<br />
lowercase letter are not different at the P=0.01 significance level<br />
‡<br />
The winter barley yields labeled with the same uppercase letter are not different at the<br />
P=0.01 significance level<br />
ACKNOWLEDGEMENTS<br />
This research has been sp<strong>on</strong>sored by Croatian Ministry for Agriculture, Forestry and<br />
Water Management (VIP Project V-29-7/04) and family farm "Oto Kovacevic" from<br />
Boksic, Croatia, <strong>on</strong> whose land this experimental site had been established. Authors<br />
would like to express their deep gratitude to Oto and Pavao Kovacevic for their<br />
c<strong>on</strong>tributi<strong>on</strong> during this project. Gratitude should also been given to the "Arvi" ltd.,<br />
Croatian branch, for their c<strong>on</strong>tributi<strong>on</strong> in fertilizers used for this experiment.<br />
111
REFERENCES<br />
Birkas, M., Szalai, T., Gyuricza, C., Gecse, M., Bordas, K. (2002): Effect of disk<br />
tillage <strong>on</strong> soil. c<strong>on</strong>diti<strong>on</strong>, crop yield and weed infestati<strong>on</strong>. Rostlinna Vyroba, 48(1),<br />
20-26 p.<br />
Filipovic, D, Husnjak, S, Kosutic, S., Gospodaric, Z (2006) Effects of tillage systems<br />
<strong>on</strong> compacti<strong>on</strong> and crop yield of Albic Luvisol in Croatia. J. of Terramechanics, 43<br />
(2): 177-189 p.<br />
Halvors<strong>on</strong>, A.D., Black, A.L., Krupinsky, J.M., Merrill, S.D., Tanaka, D.L. (1999):<br />
Dryland winter wheat resp<strong>on</strong>se to tillage and N within an annual cropping system.<br />
Agr<strong>on</strong>. J. 91 (4), 637-642 p.<br />
Izumi, Y., Uchida, K., Iijima, M. (2004), Crop producti<strong>on</strong> in successive wheatsoybean<br />
rotati<strong>on</strong> with no-tillage practice in relati<strong>on</strong> to the root system<br />
development. Plant Prod Sci 7 (3), 329-336 p.<br />
Jug, D., Stipesevic, B., Zugec, I., Horvat, D., Josipovic, M (2006): Reduced soil<br />
tillage systems for crop rotati<strong>on</strong>s improving nutriti<strong>on</strong>al value of grain crops. Cereal<br />
Res. Comm., 34 (1): 521-524 p.<br />
Karlen, D.L., Wollenhaupt, D.C. Erbach, E.C. Berry, J.B. Swan, N.S. Eash, J.L.<br />
Jordahl. (1994): L<strong>on</strong>g-<strong>term</strong> tillage effects <strong>on</strong> soil quality. Soil Tillage Res 32:<br />
313-327 p.<br />
Stipesevic, B., Zugec, I., Juric, I., Petrac, B. (1997): Possibility of reduced soil tillage<br />
for winter wheat in East-Croatia c<strong>on</strong>diti<strong>on</strong>s. Fragmenta Agr<strong>on</strong>omica, 2B/97, 613-<br />
616 p.<br />
Stipesevic, B., Zugec, I., Josipovic, M. (2000): Investigati<strong>on</strong> of Rati<strong>on</strong>al Soil Tillage<br />
for Maize (Zea mays L.) in Eastern Croatia. ISTRO 15th Proc. Fort Worth, Texas,<br />
USA (CD-ROM)<br />
Varvel, G.E., Havlin, J.L., Peters<strong>on</strong>, T.A. (1989): Nitrogen placement evaluati<strong>on</strong> for<br />
winter wheat in three fallow tillage systems, SSSA J. 53 (1), 288-292 p.<br />
Zugec, I., Stipesevic, B., Kelava, I. (2000): Rati<strong>on</strong>al Soil Tillage for Cereals (Winter<br />
Wheat - Triticum aestivum L. And Spring Barley - Hordeum vulgare L.) in Eastern<br />
Croatia. Proceedings of the 15th <str<strong>on</strong>g>C<strong>on</strong>ference</str<strong>on</strong>g> of the <str<strong>on</strong>g>Internati<strong>on</strong>al</str<strong>on</strong>g> Soil Tillage<br />
Research Organizati<strong>on</strong> (ISTRO), CD-ROM, 2-7 VII 2000., Fort Worth, Texas,<br />
SAD<br />
112
RESEARCHES REGARDING THE INFLUENCE OF THE CROP ROTATION ON<br />
SOME INDICATORS OF THE WHEAT YIELD QUALITY IN THE CONDITIONS<br />
OF THE CRISURILOR PLAIN<br />
Gh. Bandici , C. Domuţa, Ileana Ardelean , Ioana Borza , Cr. Domuţa<br />
University of Oradea, Envir<strong>on</strong>mental Protecti<strong>on</strong> Faculty, Oradea, Romania<br />
ABSTRACT<br />
The paper sustain the importance of the crop rotati<strong>on</strong> <strong>on</strong> quality of the wheat yield and is<br />
based <strong>on</strong> the results carried out during 2003-2006 in a l<strong>on</strong>g <strong>term</strong> trial out placed <strong>on</strong> the<br />
preluvosoil from Oradea in 1990. Both in n<strong>on</strong>irrigated and irrigated c<strong>on</strong>diti<strong>on</strong>s the smallest<br />
values of the protein, wet gluten and dry gluten were obtained in wheat m<strong>on</strong>ocrop; the<br />
values increased in the crop rotati<strong>on</strong> wheat maize and the biggest values were registered in<br />
the crop rotati<strong>on</strong> wheat-maize-soybean.<br />
INTRODUCTION<br />
The quality of the wheat yield is influenced by many factors. Protein acumulati<strong>on</strong> in the<br />
grains is influenced by wheat type, cultivar, climate c<strong>on</strong>diti<strong>on</strong>s, natural fertility of the soil,<br />
nitrogen doses used, irrigati<strong>on</strong> (Muntean L.S. et al, 2003, Domuţa C., 2005, Ardelean<br />
Ileana, 2006). Gluten c<strong>on</strong>tent of the wheat grain is influenced first of all by climate<br />
c<strong>on</strong>diti<strong>on</strong>s (Bîlteanu Gh., and Bîrnaure V., 1979, Bandici Gh., 1997).<br />
The influence of the crop rotati<strong>on</strong> and irrigati<strong>on</strong> <strong>on</strong> the protein and gluten c<strong>on</strong>tent is<br />
presented in the paper.<br />
MATHERIAL AND METHODS<br />
The paper is based <strong>on</strong> the research obtained in the l<strong>on</strong>g <strong>term</strong> trial with crop rotati<strong>on</strong> placed<br />
in 1990 in Oradea <strong>on</strong> preluvosoil. On ploughing depth, the soil is low acid (pH= 6,8), humus<br />
c<strong>on</strong>tent is low (1,75%), phosphorus (22,0 ppm) and potassium (845,4 ppm) have medium<br />
values; macroagregates hydrostability (47,5%) is high and bulk density (1,44 g/cm 3 ) is high,<br />
too. The experiment dispositive includes:<br />
Factor A: crop rotati<strong>on</strong><br />
a1 = wheat, m<strong>on</strong>ocrop<br />
a2 = wheat-maize<br />
a3 = wheat-maize-soybean<br />
Factor B: water regime<br />
b1 = n<strong>on</strong>irrigated<br />
b2 = irrigated<br />
The surface of the experiment parcele = 50 m 2 . Number of repetiti<strong>on</strong> = 4. Place methods =<br />
blocks method. Cultivar used: Dropia<br />
113
In the irrigated variant soil water reserve <strong>on</strong> 0-50 cm was maintained between easily<br />
available water c<strong>on</strong>tent and field capacity de<strong>term</strong>ining the soil moisture fifteen to fifteen<br />
days and using the irrigati<strong>on</strong> when the situati<strong>on</strong> required. (Domuta C., 2005).<br />
Dry gluten and wet gluten were de<strong>term</strong>ined by usually methods.<br />
Gross protein was de<strong>term</strong>ined using the following formula: Nt x 5,7; when Nt = total<br />
nitrogen. The rainfall registered during the vegetati<strong>on</strong> period of the wheat from harvesting<br />
were of 110,7 mm in 2003, 177,6 mm in 2004, 223,0 mm in 2005 and 287,2 mm in 2006.<br />
RESULTS AND DISCUSSION<br />
Crop rotati<strong>on</strong> influence <strong>on</strong> protein c<strong>on</strong>tent of the wheat grains<br />
Both n<strong>on</strong>irrigated and irrigated c<strong>on</strong>diti<strong>on</strong>s, crop rotati<strong>on</strong>s influenced the protein c<strong>on</strong>tent of<br />
the wheat yield. There were specifical situati<strong>on</strong> for every year studied.<br />
Protein c<strong>on</strong>tent of the wheat grains de<strong>term</strong>ined in the wheat-m<strong>on</strong>ocrop in 2003 was of 9,1%<br />
in n<strong>on</strong>irrigated c<strong>on</strong>diti<strong>on</strong>s and of 9,0% in irrigated c<strong>on</strong>diti<strong>on</strong>s. The values de<strong>term</strong>ined in the<br />
wheat-maize crop rotati<strong>on</strong>, 11,0% and 10,9%, were significant statistically bigger than<br />
values from wheat m<strong>on</strong>ocrop. The biggest values of the protein c<strong>on</strong>tent were registered in<br />
the wheat-maize-soybean crop rotati<strong>on</strong>, 13,8% and 13,7%; the differences in comparis<strong>on</strong><br />
with m<strong>on</strong>ocrop, 4,7% both in n<strong>on</strong>irrigated and irrigated c<strong>on</strong>diti<strong>on</strong>s is very significant<br />
statistically.<br />
In the year 2006, the smallest values of the protein c<strong>on</strong>tent were registered in the<br />
m<strong>on</strong>ocrop of wheat, too: 71% in n<strong>on</strong>irrigated and 6,9% in irrigated c<strong>on</strong>diti<strong>on</strong>s. In the wheatmaize<br />
crop rotati<strong>on</strong> the values increased with 45% and 46% and in the wheat-maizesoybean<br />
crop rotati<strong>on</strong> with 73% and 77% respectively.<br />
In average <strong>on</strong> the researched period, the smallest values of the protein c<strong>on</strong>tent of the wheat<br />
grains were registered in m<strong>on</strong>ocrop, 7,98% in n<strong>on</strong>irrigated c<strong>on</strong>diti<strong>on</strong>s and 7,73% in irrigated<br />
c<strong>on</strong>diti<strong>on</strong>s. In the wheat-maize crop rotati<strong>on</strong> the values of the protein c<strong>on</strong>tent (10,7% and<br />
10,45%) increased distingue significant in comparis<strong>on</strong> with m<strong>on</strong>ocrop. The biggest values<br />
of the protein c<strong>on</strong>tent was obtained in the wheat-maize-soybean crop rotati<strong>on</strong>, 13,02% in<br />
n<strong>on</strong>irrigated and 12,93% in irrigated c<strong>on</strong>diti<strong>on</strong>s.<br />
Table 1: Crop rotati<strong>on</strong> and irrigati<strong>on</strong> influence <strong>on</strong> protein c<strong>on</strong>tent of the wheat<br />
grain, Oradea 2003 – 2006<br />
Crop rotati<strong>on</strong><br />
Water regime<br />
N<strong>on</strong>irrigated Irrigated<br />
Protein<br />
% % % %<br />
Average <strong>on</strong><br />
the crop<br />
rotati<strong>on</strong><br />
1. Wheat- m<strong>on</strong>ocrop 7,98 100 7,73 100 7,86 Mt<br />
2. Wheat-maize 10,7 135 10,45 135 10,56 **<br />
3. Wheat-maize-soybean 13,02 164 12,93 167 12,98 ***<br />
Average <strong>on</strong> the water regime 10,57 Mt<br />
100 9,73 -<br />
98,1 -<br />
114
Crop rotati<strong>on</strong> Water regime<br />
Water regime x<br />
Crop rotati<strong>on</strong><br />
Crop rotati<strong>on</strong> x<br />
Water regime<br />
LSD 5% 1,17 0,73 1,4 1,43<br />
LSD 1% 2,16 1,46 2,6 2,73<br />
LSD 0,1% 3,96 2,96 4,8 4,43<br />
Crop rotati<strong>on</strong> influence <strong>on</strong> wet gluten c<strong>on</strong>tent of the wheat grains<br />
Crop rotati<strong>on</strong> influenced very str<strong>on</strong>g the wet gluten c<strong>on</strong>tent of the wheat grain. Every year<br />
the smallest c<strong>on</strong>tents were obtained in wheat m<strong>on</strong>ocrop both n<strong>on</strong>irrigated and irrigated<br />
c<strong>on</strong>diti<strong>on</strong>.<br />
The year 2003 was the year with the biggest drught and values of the wet gluten were<br />
the biggest too.In wheat m<strong>on</strong>ocro, the values of the wet gluten were of 22,6% in<br />
n<strong>on</strong>irrrigated c<strong>on</strong>diti<strong>on</strong>s and of 21,9% in irrigated c<strong>on</strong>diti<strong>on</strong>s. The values registered in the<br />
whet-maize crop rotati<strong>on</strong> (29,9% and 29%) and in the wheat-maize-soybean crop rotati<strong>on</strong><br />
(36,1% and 33,8%) were very significant statistically bigger than the values registered in the<br />
wheat-m<strong>on</strong>ocrop. (table 1).<br />
The values of wet c<strong>on</strong>tent registered in 2004 in whet-m<strong>on</strong>ocrop were of 20,4 % in<br />
n<strong>on</strong>irrigated c<strong>on</strong>diti<strong>on</strong>s and of 19,6% in irrigated c<strong>on</strong>diti<strong>on</strong>s. There were very significant<br />
differences in the wheat-maize and wheat-maize-soybean crop rotati<strong>on</strong>; relative defferences<br />
were of 36% and 61% in n<strong>on</strong>irrigated c<strong>on</strong>diti<strong>on</strong>s and of 38% and of 63% in irrigated<br />
c<strong>on</strong>diti<strong>on</strong>s. This year were registered the biggest values of the wet gluten of the studied<br />
period.<br />
In 2005 in wheat-m<strong>on</strong>ocrop, the c<strong>on</strong>tent of the wet gluten from grains were of 21,3 in<br />
n<strong>on</strong>irrigated c<strong>on</strong>diti<strong>on</strong>s and of 21% in irrigated c<strong>on</strong>diti<strong>on</strong>s. Diferences registered in the<br />
wheat-maize and wheat-maize soybean crop rotati<strong>on</strong> were very significant statistically, 31%<br />
and 61% in n<strong>on</strong>irrigated c<strong>on</strong>diti<strong>on</strong>s, 30% and 57% in irrigated c<strong>on</strong>diti<strong>on</strong>s respectivelly.<br />
In the year 2006, the smallest values of the wet gluten were registered in the wheat<br />
m<strong>on</strong>ocrop, 19,9% in n<strong>on</strong>irrigated c<strong>on</strong>diti<strong>on</strong>s and 19,5% in irrigated c<strong>on</strong>diti<strong>on</strong>s; in the wheatmaize<br />
crop rotati<strong>on</strong> the values increased with 36% and 37% and in the wheat-maizesoybean<br />
crop rotati<strong>on</strong> with 59% and 62%.<br />
The average data of the period 2003-2006 show that the smallest c<strong>on</strong>tent of the grain<br />
wet gluten was registered in m<strong>on</strong>ocrop. In wheat-maize and in wheat-maize-soybean crop<br />
rotati<strong>on</strong> were registered the differences very significant statistically in comparis<strong>on</strong> with<br />
wheat-m<strong>on</strong>ocrop: 34% and 60% in n<strong>on</strong>irrigated c<strong>on</strong>diti<strong>on</strong>s, 34% and 55% in irrigated<br />
c<strong>on</strong>diti<strong>on</strong>s, respectivelly. (table 2).<br />
115
Table 2: Crop rotati<strong>on</strong> and irrigati<strong>on</strong> influence <strong>on</strong> wet gluten c<strong>on</strong>tent of the wheat<br />
grains, Oradea 2003 – 2006<br />
Water regime<br />
Average <strong>on</strong><br />
Crop rotati<strong>on</strong><br />
N<strong>on</strong>irrigated Irrigated the crop<br />
% % % % rotati<strong>on</strong><br />
1. Wheat- m<strong>on</strong>ocrop 21,1 100 20,5 100 20,8 Mt<br />
2. Wheat-maize 28,2 134 27,5 134 27,85 ***<br />
3. Wheat-maize-soybean 33,7 160 32,6 159 33,15 ***<br />
Average <strong>on</strong> the water regime 27,7 Mt<br />
100 26,9 o<br />
96,9 -<br />
Crop rotati<strong>on</strong> Water regime<br />
Water regime x<br />
Crop rotati<strong>on</strong><br />
Crop rotati<strong>on</strong> x<br />
Water regime<br />
LSD 5% 1,43 0,75 1,70 1,63<br />
LSD 1% 2,40 1,45 3,03 2,96<br />
LSD 0,1% 4,46 3,41 5,24 5,05<br />
Crop rotati<strong>on</strong> influence <strong>on</strong> dry gluten c<strong>on</strong>tent of the wheat grains<br />
In 2003 the values of the dry gluten c<strong>on</strong>tent from wheat grain for m<strong>on</strong>ocrop were of 10,8%<br />
in n<strong>on</strong>irrigated and of 10,3% in irrigated c<strong>on</strong>diti<strong>on</strong>s. The differences registered in wheatmaize<br />
crop rotati<strong>on</strong> were significant statistically, 19% in n<strong>on</strong>irrigated c<strong>on</strong>diti<strong>on</strong>s and 17% in<br />
irrigated c<strong>on</strong>diti<strong>on</strong>s. In the wheat-maize-soybean crop rotati<strong>on</strong> were distingue significant:<br />
35% in n<strong>on</strong>irrigated c<strong>on</strong>diti<strong>on</strong>s and 39% in irrigated c<strong>on</strong>diti<strong>on</strong>s. (table 2).<br />
The dry gluten c<strong>on</strong>tent of the wheat grains in 2004 in the m<strong>on</strong>ocrop were of 9,8% in<br />
n<strong>on</strong>irrigated c<strong>on</strong>diti<strong>on</strong>s aof 9,3% in irrigated c<strong>on</strong>diti<strong>on</strong>s. The statistically significant of the<br />
differences vs. wheat-m<strong>on</strong>ocrop registered in the wheat-maize-soybean crop rotati<strong>on</strong> have<br />
similar statistically significant with the differences registered in 2003: significant and<br />
distingue significant; the biggest values, 13,7% in n<strong>on</strong>irrigated c<strong>on</strong>diti<strong>on</strong>s and 13,0% in<br />
irrigated c<strong>on</strong>diti<strong>on</strong>s, were registered in wheat-maize-soybean crop rotati<strong>on</strong>. (table 3).<br />
In 2005, the smallest values of the dry gluten were registered in wheat-m<strong>on</strong>ocrop, too:<br />
10,2% in n<strong>on</strong>irrigated c<strong>on</strong>diti<strong>on</strong>s and 9,4% in irrigated c<strong>on</strong>diti<strong>on</strong>s. A similar situati<strong>on</strong> with<br />
2003 regarding statistically significant of the differences in comparis<strong>on</strong> wheat m<strong>on</strong>ocrop<br />
was registered in 2005, too. The biggest values of the dry gluten, 14,0% in n<strong>on</strong>irrigated<br />
c<strong>on</strong>diti<strong>on</strong>s and 13,3% in irrigated c<strong>on</strong>diti<strong>on</strong>s, were registered in the wheat-maize-soybean.<br />
In the year 2006, the smallest values of the dry gluten were registered in the wheat<br />
m<strong>on</strong>ocrop, 9,5% in n<strong>on</strong>irrigated variant and 9,3% in irrigated variant. In the wheat-maize<br />
crop rotati<strong>on</strong> the values of the dry gluten increased with 23% both irrigated and n<strong>on</strong>irrigated<br />
c<strong>on</strong>diti<strong>on</strong>s and in the wheat-maize-soybean crop rotati<strong>on</strong> with 38% and 39% respectivelly.<br />
In average <strong>on</strong> the studied period, the values of the dry gluten c<strong>on</strong>tent of the grains wheat<br />
from m<strong>on</strong>ocrop were of 10,01% in n<strong>on</strong>irrigated c<strong>on</strong>diti<strong>on</strong>s and of 9,58% in irrigated<br />
116
c<strong>on</strong>diti<strong>on</strong>s. The values, registered in wheat-maize crop rotati<strong>on</strong> were significant statistically<br />
bigger: (12,20% and 11,53%) and in wheat-maize-soybean were registered the biggest<br />
values (13,88% and 13,38%) and differences distingue significant in comparis<strong>on</strong> with<br />
wheat-m<strong>on</strong>ocrop. (table 3).<br />
Table 3: Crop rotati<strong>on</strong> and irrigati<strong>on</strong> influence <strong>on</strong> dry gluten c<strong>on</strong>tent of the wheat<br />
grains, Oradea 2003 – 2006<br />
Water regime<br />
Average <strong>on</strong><br />
Crop rotati<strong>on</strong><br />
N<strong>on</strong>irrigated Irrigated the crop<br />
% % % % rotati<strong>on</strong><br />
1. Wheat- m<strong>on</strong>ocrop 10,01 100 9,58 100 9,80 Mt<br />
2. Wheat-maize 12,20 122 11,53 120 11,87 **<br />
3. Wheat-maize-soybean 13,88 139 13,38 140 13,59 ***<br />
Average <strong>on</strong> the water regime 12,03 Mt<br />
100 11,49 -<br />
95,6 -<br />
Crop rotati<strong>on</strong> Water regime<br />
Water regime x<br />
Crop rotati<strong>on</strong><br />
Crop rotati<strong>on</strong> x<br />
Water regime<br />
LSD 5% 0,91 0,65 1,18 1,14<br />
LSD 1% 1,56 1,16 2,12 1,90<br />
LSD 0,1% 2,49 2,14 3,95 3,48<br />
CONCLUSIONS<br />
The results obtained in a l<strong>on</strong>g <strong>term</strong> trial (1990-2006) emphasized the importance of the crop<br />
rotati<strong>on</strong> in the protein, wet gluten and dry gluten of the yield wheat.<br />
During 2003-2006 both n<strong>on</strong>irrigated and irrigated c<strong>on</strong>diti<strong>on</strong>s the smallest values of the<br />
protein, wet gluten and dry gluten were obtained in wheat m<strong>on</strong>ocrop.<br />
In comparis<strong>on</strong> with wheat m<strong>on</strong>ocrop, in the wheat-maize crop rotati<strong>on</strong> the values of the<br />
protein, wet and dry gluten increased significant statistically. The differences very<br />
significant statistically in comparis<strong>on</strong> with the wheat m<strong>on</strong>ocrop were registered every year<br />
in the wheat-maize-soybean crop rotati<strong>on</strong> in all three parameters of the wheat yield quality<br />
analyzed.<br />
Irrigati<strong>on</strong> de<strong>term</strong>ined to obtain smaller values of the protein, wet and dry gluten in the<br />
wheat grains in comparis<strong>on</strong> with n<strong>on</strong>irrigated variants from all the crop rotati<strong>on</strong>s.<br />
117
REFERENCES<br />
1. Ardelean Ileana (2006) - C<strong>on</strong>tributi<strong>on</strong>s in the known and modificati<strong>on</strong> of the crop<br />
rotati<strong>on</strong> influence <strong>on</strong> quantity and quality of the winter wheat yield croped <strong>on</strong> the acid<br />
soils from North-Western Romania. Thesis degree, USAMV Cluj-Napoca<br />
2. Bandici Gh. (1997) – C<strong>on</strong>tributi<strong>on</strong>s to establishing the influence of forreruner plant<br />
and fertilisati<strong>on</strong> level up<strong>on</strong> dinamics acumulati<strong>on</strong> of wheat biomass cultivated <strong>on</strong> soils<br />
with excessive moisture in centre of North-Western Romania. Thesis degree, USAMV<br />
Cluj-Napoca.<br />
3. Bîlteanu Gh., Bîrnaure V. (1979) – Phytotechny. Publishing house Ceres Bucureşti<br />
4. Domuţa C., 2005 – Irrigati<strong>on</strong> crops. Publishing house University of Oradea<br />
5. Muntean L.S., Roman Gh..V., Borcean I., Axinte I., (2003) – Phytotechny. Ed. I<strong>on</strong><br />
I<strong>on</strong>escu de la Brad Iaşi<br />
118
CRISANA -THE FIRST ROMANIAN WHEAT VARIETY WITH TOLERANCE<br />
TO ALUMINIUM IONS TOXICITY FROM ACID SOILS<br />
Gh. Bunta*, Elena Bucurean**<br />
*Agricultural Research and Development Stati<strong>on</strong> Oradea,<br />
**University of Oradea, Oradea, Romania<br />
ABSTRACT<br />
The paper present the new wheat variety registered in 2005 in Romania, Crisana. This<br />
<strong>on</strong>e was created in Agricultural Research and Development Stati<strong>on</strong> Oradea by<br />
hybridizati<strong>on</strong> and repeated genealogical selecti<strong>on</strong>, especial for the acid soils from<br />
Transilvania and other parts of Romania. The acid soils from hill z<strong>on</strong>es have low pH<br />
and aluminum i<strong>on</strong>s toxicity, but Crisana variety is very tolerant to this. In the same<br />
time, this new variety has very good backing qualities, being superior to the presents<br />
Romanian wheat variety cultivated in the north part of Romania. In present, the variety<br />
is in a multiplicati<strong>on</strong> process in the fields of Agricultural Research and Development<br />
Stati<strong>on</strong> Oradea and it will be cultivated <strong>on</strong> acid soils from west and north of Romania.<br />
Keywords: aluminum toxicity, variety, wheat, quality.<br />
INTRODUCTION<br />
In 1960, Neenan described for the first time the differential reacti<strong>on</strong> of genotypes to<br />
aluminum i<strong>on</strong> toxicity from acid soils. After this finding, the tolerance of plants to<br />
aluminum i<strong>on</strong> toxicity was approach by many researchers. At the beginning, the<br />
problem was studied in genetically point of whew in Brasilia and CIMMYT –Mexico<br />
(Camargo and Felicio, 1988) and after this like a breeding objective (Aniol and<br />
Gustafs<strong>on</strong>, 1990).<br />
In Romania, for the first time the problem was studied in 1987 (Bunta et al.), when<br />
started a breeding program in cooperati<strong>on</strong> with Nati<strong>on</strong>al Institute for Agricultural<br />
Research and Development Fundulea. The study begun with elaborati<strong>on</strong> of an original<br />
method of testing in laboratory c<strong>on</strong>diti<strong>on</strong>s, in hydrop<strong>on</strong>ics soluti<strong>on</strong>s. The researches<br />
c<strong>on</strong>tinued with the study of a large collecti<strong>on</strong> of genotypes to identify some genetically<br />
sources of tolerance to i<strong>on</strong>s toxicity. In the same idea, we collected many local wheat<br />
populati<strong>on</strong>s and old cultivates (Bunta, 1997). After this, we initiated studies regarding<br />
heredity of wheat tolerance to aluminum i<strong>on</strong>s toxicity (Bunta, 1999/a; Bunta, 1999/b).<br />
The new variety Crişana, which is for the moment the single Romanian wheat<br />
variety with high tolerance to i<strong>on</strong>s toxicity, has another qualities too, like a good<br />
backing quality and high productive potential.<br />
The studies regarding the breeding for tolerance to aluminum toxicity proceeding<br />
for triticale in Nati<strong>on</strong>al Institute for Agricultural Research and Development Fundulea<br />
(Ittu and Săulescu, 1988) and for wheat Agricultural Research and Development Stati<strong>on</strong><br />
Albota, too (Voica and Dumitrascu 2002).<br />
MATERIALS AND METHODS<br />
Th Crisana variety was created by genealogical repeted selecti<strong>on</strong> from hybrid<br />
combinati<strong>on</strong> Fundulea 4 / Atlas 66.<br />
119
The romanian Fundulea 4 variety vas utilized like a genitor becouse in 1986 it was the<br />
most productive wheat variety in our country. It was too the most addapted to our z<strong>on</strong>e,<br />
an ideotype for north-west of Romania, an wet and colder geografic areal.<br />
Atlas 66, an old variety from United States of America (South Carolina), has a lot<br />
of defficiences, like any old cultivar: sensibility to diseases, tall, sensitive to falling and<br />
too tardive. Hovewer, this variety have a very high c<strong>on</strong>tent in protein and high tolerance<br />
to aluminum i<strong>on</strong>s toxicity, beeing an etal<strong>on</strong> in all genetical study.<br />
The hibridati<strong>on</strong> Fundulea 4/Atlas 66 was efectuated in 1986 and F1 hybrid vas<br />
tested in 1987. During the period 1988 - 1990 (F2 – F4 generati<strong>on</strong>s) we selected every<br />
year plantes with aluminum tolerance, in soluti<strong>on</strong> with growing c<strong>on</strong>centrati<strong>on</strong> of<br />
aluminum in laboratory and in a testing fild near Sudrigiu, in Beius Depressi<strong>on</strong>.<br />
Beginning with the year 1991 (F5 generati<strong>on</strong>) and finishing with 1996 (hybrid<br />
generati<strong>on</strong> F10) the selecti<strong>on</strong> has agr<strong>on</strong>omical objective, in two experimental fields,<br />
Sânmartin and Oradea. During 1997 – 1998 (D1 and D2), the breeding line Oradea 133<br />
G vas tested in comparative micro-trials, simultaneous in all three fields. In the period<br />
1999 – 2001 we proceded to testing of breeding line 133 G 41113124-S353 in paralel in 2<br />
comparative experiments, in Oradea and Sânmartin. Because of it performances during<br />
of all this years, during 2002 – 2004 the breeding line Oradea 133 was tested in network<br />
of State Institute for Variety Testing and Registrati<strong>on</strong> (S.I.V.T.R.). Finally, in 2005 this<br />
breeding line vas registred like variety with the name Crisana.<br />
Simultaneoussly, we started the multiplicati<strong>on</strong> of seeds, than today the new variety<br />
is cultivated <strong>on</strong> more than 200 ha in Agricultural Research Stati<strong>on</strong> Oradea fields.<br />
To caracterize the genotipes regarding the tolerance to aluminium, we used an original<br />
method (Bunta, 1996) and another two method addapted by proffesor dr. Gallia Butnaru<br />
from the University of Agricultural Sciences and Veterinary Medicine of Banat county<br />
(USAMVB) from Timişoara, the laboratory of genetics.<br />
The qualities analyses were effectuated in the laboratory of Nati<strong>on</strong>al Institute for<br />
Agricultural Researches and Development Fundulea and State Institute for Variety<br />
Testing and Registrati<strong>on</strong> Bucureşti.<br />
RESULTS AND DISCUSSIONS<br />
The actuals Romanian cultivars do not have tolerance to aluminium toxicity or here<br />
tolerance level is reduced (Albota, Fundulea 133, Arieşan, Trivale). The cause c<strong>on</strong>sists<br />
in the fact that the Romanian breeding programs did not fallow this objective.<br />
The breeding program for tolerance to aluminum i<strong>on</strong>s toxicity started at<br />
Agricultural Research and Development Stati<strong>on</strong> Oradea in 1982, like a program<br />
regarding creati<strong>on</strong> of varieties well adapted to local c<strong>on</strong>diti<strong>on</strong> of hill z<strong>on</strong>e from western<br />
Romania.<br />
The first stage look to transfer aluminum tolerance from spring wheat varieties<br />
originated from Brazil and another country in local winter wheat genotypes. In the same<br />
time, we look to associate aluminum tolerance with different another characters, like:<br />
plant height, precocity, spike characteristics, the positi<strong>on</strong> and color of lives, etc.<br />
To can realize the main objective of our program, it was necessary to create a<br />
specific breeding scheme. These c<strong>on</strong>sist in the classic method (recombinati<strong>on</strong> and<br />
selecti<strong>on</strong>), with some adaptati<strong>on</strong>s (fig. 1).<br />
120
In additi<strong>on</strong>, we used recurent selecti<strong>on</strong> to broke some correlati<strong>on</strong>s of aluminum<br />
tolerance with: plant heigh, falling susceptibility, disease sensibility, bad coocking<br />
quality and another indesirable phisiological or morphological characters.<br />
SUDRIGIU SÂNMARTIN ORADEA<br />
IIIIIIIIIIIIIIIIII<br />
IIIIIIIIIIIIIIIIII<br />
OIIIOIIIIOIIIIO<br />
OIIIOIIIIOIIIIO<br />
IIIIIIIIIIIIIIIIIII<br />
IIIIIIIIIIIIIIIIIII<br />
SIVTR<br />
cecking<br />
IIIIIIIIIIIIIIIIIIIIII<br />
XXXXXXXXXX<br />
00000000000000<br />
00000000000000<br />
**************<br />
**************<br />
OIIIOIIIIOIIIIO<br />
OIIIOIIIIOIIIIO<br />
IIIIIIIIIIIIIIIIIIIIII<br />
IIIIIIIIIIIIIIIIIIIIII<br />
IIIIIIIIIIIIIIIIIIIIII<br />
IIIIIIIIIIIIIIIIIIIIII<br />
Study of initial material<br />
Hybridizati<strong>on</strong>s<br />
Hybrids F1<br />
Individual selecti<strong>on</strong> in fild<br />
and laboratory, F2-Fn<br />
Micro comparative trials<br />
( c<strong>on</strong>trol field)<br />
Orientative trials<br />
IIIIIIIIIIIIIII Comparative<br />
IIIIIIIIIIIIIII trials<br />
IIIIIIIIIII Preliminary<br />
multiplicati<strong>on</strong><br />
Seed<br />
multiplicati<strong>on</strong><br />
Figure 1: The original breeding scheme for creati<strong>on</strong> of wheat variety with aluminum<br />
tolerance<br />
Finally, the best breeding lines are promovated to be tested in the State Institute for<br />
Variety Testing and Registrati<strong>on</strong> network. Simultaneously the seed of promissing<br />
breeding lines are multiplicated to reduce the period of extensi<strong>on</strong> in the large fields.<br />
121
Class Genotype<br />
Table 1: Regenerating capacity of some wheat breeding lines<br />
48 ours later exposure to high Al 3+ rates<br />
Root el<strong>on</strong>gati<strong>on</strong> (mm)<br />
10 ppp 20 ppm 40 ppm<br />
Al 3+<br />
Al 3+<br />
Al 3+<br />
1 32 B 822 31,0±1,4 15,2±2,4 4,6±2,5<br />
2 14 I 323 11,1±1,1 12,5±5,0 8,7±1,2<br />
3 81 J 475 12,5±2,8 6,0±2,0 0,4±0,3<br />
4 CRISANA 12,0±1,3 4,9±1,0 0,4±0,3<br />
5 138 C 412 17,3±2,9 0 0<br />
6 5 E 122 15,2±6,4 0 0<br />
7 4 K 464 9,2±1,4 0,4±0,2 0<br />
8 88 I 159 5,2±1,4 0 0<br />
9 21 H 454 4,2±1,4 1,4±1,7 0<br />
10 8 J 1010 4,5±1,5 1,5±0,5 0<br />
11 10 K 487 1,4±1,2 0,4±0,4 0<br />
12 29 D 125 1,8±0,9 0 0<br />
13 19 I 197 1,6±0,3 2,9±0,9 0<br />
14 93 J 222 0,3±0,3 0 0<br />
15 10 F 414 0 0 0<br />
16 32 F 132 0 0 0<br />
17 35 F 133 0 0 0<br />
18 76 J 545 0 0 0<br />
19 128 G 128 0 0 0<br />
20 FUNDULEA 29 0 0 0<br />
Estimati<strong>on</strong><br />
Very tolerant<br />
Tolerant<br />
Susceptible<br />
From a large list of breeding lines created to have a high tolerance to aluminum i<strong>on</strong>s<br />
toxicity, some of them were evidentiated everytime, in any method of testing. Thus,<br />
Oradea 133 line (the futhure variety Crisana) restarted his el<strong>on</strong>gati<strong>on</strong> of roots, even at<br />
40 ppm Al 3+ in soluti<strong>on</strong> (table 1).<br />
Another method are based <strong>on</strong> maximal roots length of plants at 4 ppm Al 3+ , after<br />
10 days of exposure (table 2). The hydrop<strong>on</strong>ic soluti<strong>on</strong> c<strong>on</strong>sist <strong>on</strong>ly in distiled water and<br />
aluminium i<strong>on</strong>s, assured from AlCl 3, withouth any other mineral, at 4.5 pH, the<br />
nutritive elements being assured by reserve elements from seed. This method is an<br />
original <strong>on</strong>e, adapted by us for a large series of breeding individuals in segregating<br />
populati<strong>on</strong>s.<br />
122
Table 2: The root length of some wheat genotypes at 4 ppm Al 3+ c<strong>on</strong>centrati<strong>on</strong><br />
Class Genotype Roots length Differ. Differ. Estimati<strong>on</strong><br />
(mm) % (mm) signiff.<br />
1 Crisana 112,3 154,8 +39,7 *** Tolerant<br />
2 Oradea 30 110,1 151,9 +37,6 ***<br />
3 Oradea 33 93,4 128,8 +20,9 ** Medium<br />
4 Fundulea 472 79,9 110,2 +7,4<br />
tolerant<br />
5 Arieşan 76,0 104,7 +3,5<br />
Experimental average 72,5 100,0 0 - -<br />
6 Fundulea 4 40,4 55,7 -32,1 000<br />
7 Lovrin 34 35,8 49,4 -36,7 000 Susceptible<br />
8 Fundulea 29 32,2 44,4 -40,3 000<br />
LSD5%= 10,4 mm; LSD1%= 14,8 mm; LSD0,1%= 21,4 mm.<br />
By total locati<strong>on</strong>s and years average (table 3), the variety Crisana produced a yield<br />
output of 355 kg/ha comparative to check variety Fundulea 4 (106,1%), respective 610<br />
kg/ha comparative to Apullum (110,5%). Comparative to the two-check average, the<br />
yield output was 482,5 kg/ha (108,3%).<br />
Locati<strong>on</strong><br />
Table 3: The yields of Crişana variety in SIVTR network<br />
2002 2003 2004 Locati<strong>on</strong> averages<br />
Kg/ha %<br />
% check<br />
averages<br />
Kg/ha<br />
Fundulea Apullum<br />
4<br />
Satu Mare 8596 3213 9064 6958 107,6 102,8 105,2<br />
Huedin /Luduş 4785 3785 6848 5139 97,0 134,2 115,6<br />
Galda de Jos 5268 4188 5952 5136 108,5 104,2 106,4<br />
Sibiu 6546 5121 7678 6448 105,8 111,0 108,4<br />
Hărman/Tg.Secuiesc 7083 1572 8036 5564 118,5 112,1 115,3<br />
Rădăuţi 4705 4677 7527 5636 99,3 98,6 99,0<br />
YEARS<br />
AVERAGES<br />
6164 3759 7518 5814 106,1 110,5 108,3<br />
A simple estimati<strong>on</strong> of ec<strong>on</strong>omical efficiency dem<strong>on</strong>strate that, accepting an price of<br />
0,11 dollars/kg, <strong>on</strong>ly in the west of Romania (more than 500.000 ha cultivated with<br />
wheat), the total profit possible to be realized by cultivati<strong>on</strong> with this new variety<br />
exceed 25 milli<strong>on</strong> dollars/year.<br />
123
Table 4: Quality indicators of Crisana variety comparative to check varieties utilized<br />
in SIVTR network<br />
Quality indicators Estim.<br />
Variety<br />
unit. Fundulea 4 Arieşan Apullum Crisana<br />
Falling number sec.<br />
Schrot analise<br />
238 236 272 299<br />
Total proteine % 14,12<br />
Flour analise<br />
13,94 14,53 14,71<br />
Wet gluten % 26,35 26,72 27,50 27,28<br />
Dry gluten % 9,75 9,82 10,67 10,70<br />
Glutenic index - 75,09 75,76 84,21 87,56<br />
Sedimentati<strong>on</strong> index Ml 50,2<br />
Flour graphic<br />
52,5 57,5 60,5<br />
Hidratati<strong>on</strong> capacity % 67,0 65,5 66,1 68,5<br />
Elasticity UV 100 95 92,5 95<br />
El<strong>on</strong>gati<strong>on</strong> UV 60 55 52,5 55<br />
Flour strength - 61,2 64,7 63,2 62,5<br />
Valuable index - 65,1 67,5 68,4 68,0<br />
Estimating groupe - B1-A2 B1-A2 B1-A2 A2-B1<br />
Crişana variety have a high yelding pottential (with a maximum of 9.064 kg/ha at Satu<br />
Mare, in 2004), and a high milling and backing quality, dem<strong>on</strong>strated by the results<br />
showed in table number 4. Quality analizes were dune in SIVTR Bucureşti laboratories<br />
of chemistry <strong>on</strong> wheat sample collected from all the testing locati<strong>on</strong>s. Thus, the results<br />
prove the quality stability of this new variety in different climatically c<strong>on</strong>diti<strong>on</strong>s.<br />
Falling number and total protein is higher even than Apullum, c<strong>on</strong>sidered like a superior<br />
quality variety.<br />
The analises of flour (especially glutenic index) proved the superior quality of<br />
Crisana variety. These index and flour graphic analize were utilized to calculate<br />
valuable index and classiffied Crisana variety, by this index, in the groupe A2-B1 (in the<br />
rate of 3:1). Thus, this variety is a very good <strong>on</strong>e, having the capacity to ameliorate the<br />
flour quality of another variety, by mixture.<br />
In 2006, the new variety Crisana was tested for yield capacity and backing quality<br />
comparative to another 15 <strong>on</strong>es (table 5). Crisana has a good yielding potential and<br />
quality indicators, compatible with the best Hungarian varieties created in Mart<strong>on</strong>vasár<br />
Institute. In aditi<strong>on</strong> to the high yielding pottential, the tolerance to soil acidity and<br />
aluminium i<strong>on</strong>s toxicity, the superior milling and backing quality, another characteristics<br />
are very good in case of Crisana variety: the good or very good resistance to disease,<br />
winter frost and falling<br />
124
Table 5: Producti<strong>on</strong>s and quality results realized by the new wheat variety<br />
Crisana in the dem<strong>on</strong>strative field. Biharia, 2006<br />
Class. Variety<br />
Yield Quality index<br />
kg/ha % Wet Total Zeleny<br />
gluten % protein % index<br />
1. Toborzó 7148 118.4 30 13.9 46<br />
2. Pálma 7003 116.0 27 13.7 51<br />
3. Emese 6861 113.6 30 14.1 52<br />
4. Marsall 6787 112.4 30 14.9 54<br />
5. Romulus 6370 105.5 30 14.5 53<br />
6. Süveges 6348 105.1 31 15.7 59<br />
7. Crisana 6165 102.1 31 14.2 54<br />
8. Capo 6086 100.8 29 14.1 58<br />
9. Palotás 6067 100.5 32 15.5 61<br />
Experimental<br />
average<br />
6038 100.0 29.9 14.6 57.1<br />
10. Kristina 5890 97.5 23 12.5 52<br />
11. Magdaléna 5653 93.6 32 15.7 64<br />
12. Csárdás 5369 88.9 32 16.0 63<br />
13. Magvas 5326 88.2 33 15.4 61<br />
14. Verbunkos 5285 87.5 31 15.4 58<br />
15. Joszef 5189 85.9 31 15.0 73<br />
16. Apache 5059 83.8 27 13.6 54<br />
Acording with all this characteristics, the State Institute for Variety Testing and<br />
Registrati<strong>on</strong> recommended in his Official Catalogue (2005) that the new variety Crisana<br />
to be cultivated in Transilvania, <strong>on</strong> acid soils from hill z<strong>on</strong>e.<br />
REFERENCES<br />
Aniol, A., Gustafs<strong>on</strong>, J. P., (1990) - Genetics of tolerance in agr<strong>on</strong>omic plants. Heavy<br />
metal tolerance in plants: evoluti<strong>on</strong>ary aspects. Ed. A. J. Shaw, Boca Rat<strong>on</strong>, USA,<br />
255-267 p.<br />
Bunta, Gh., Bunta, Anuţa, Negruţiu, I., (1987) – Toleranţa la aciditate şi exces de<br />
i<strong>on</strong>i de aluminiu din sol. Principalele obiective de ameliorare a grâului pentru z<strong>on</strong>a<br />
colinară şi de deal din vestul ţării. Probl. genet. teor. aplic., XIX, (4), 173-188 p.<br />
Bunta, Gh. (1996) - Cercetări privind efectul i<strong>on</strong>ilor de aluminiu asupra lungimii<br />
rădăcinilor câtorva soiuri de grâu. Analele Universitatii Oradea, fasc. Agric., II,<br />
121 - 130 p.<br />
Bunta, Gh., (1997) - Identificarea unor surse genetice de toleranţă la aluminiu în cadrul<br />
populaţiilor locale de grâu. Probl. genet. teor. aplic., XXIX, (1-2), 11-21 p.<br />
Bunta, Gh., (1999/a) - Results regarding the genetic c<strong>on</strong>trol of tolerance to aluminum<br />
i<strong>on</strong> toxicity in wheat. Romanian Agricultural Research, (11-12), 1-8 p.<br />
125
Bunta, Gh., (1999/b) - Ereditatea nucleo-citoplasmatică a toleranţei grâului la<br />
toxicitatea i<strong>on</strong>ilor de aluminiu. Probl. genet. teor. aplic., XXXI, (1-2), 1-12 p.<br />
Camargo, C., E., Felicio, J., C., (1988) – Wheat breeding at the Campinas Agr<strong>on</strong>omic<br />
Institute. Wheat breeding for acid soils; review of Brazilian/CIMMYT<br />
collaborati<strong>on</strong>, 1974 - 1986. Mexico, D., F., CIMMYT, Ed. Kohli, M., M. and<br />
Rajaram, S., 39-49 p.<br />
Ittu,Gh., Săulescu, N., (1988) – Cercetări privind crearea de soiuri de grâu de toamnă<br />
tolerante la toxicitatea produsă de i<strong>on</strong>ii de aluminiu. Probl. Genet. Teor. Aplic.,<br />
XX, 3: 139 – 149 p.<br />
Neenan, M., (1960) – The effects of soil acidity <strong>on</strong> the growth of cereals with particular<br />
reference to the differential reacti<strong>on</strong> of varieties thereto. Plant Soil, (12), 324 -338<br />
p.<br />
Voica, Maria, Dumitraşcu, Navara, (2002) – Toleranţa unor genotipuri de grâu de<br />
toamnă la toxicitatea i<strong>on</strong>ilor de aluminiu. An. ICCPT Fundulea, LXIX : 51 – 63 p.<br />
*** 2005 – Catalogul oficial al soiurilor de plante de cultură din România. Bucureşti,<br />
110 – 111p.<br />
126
EXPERIMENTS WITH PERENNIAL RYE (SECALE CEREANUM) IN HUNGARY,<br />
AT UNIVERSITY OF DEBRECEN<br />
ABSTRACT<br />
Erika Halász – Tamás Sipos<br />
Research Centre of Nyíregyháza, University of Debrecen<br />
Perennial rye as a new culture crop has some problems which make more difficult the<br />
growth technology. The most important from these is that germinati<strong>on</strong> of seeds is compared<br />
to winter rye very few (50-70). We found correlati<strong>on</strong> between the rev and the germinati<strong>on</strong><br />
ability, but it wasn’t any correlati<strong>on</strong> between harvesting in <strong>on</strong>e step and two steps.<br />
In the Research Centre of Nyíregyháza we started a development of a new forage<br />
mixture with perennial rye and perennial papili<strong>on</strong>aceae (alfalfa, bird’s-foot trefoil). The two<br />
plants with different fodder value in the mixture are complementary to each other from the<br />
point of view of dietetics; the mixture produce great mass fodder with digestible fiber and<br />
high quantity of crude protein. In sec<strong>on</strong>d year perennial rye produced 30-35 t/ha green<br />
mass, but the mixed stand yielded 78-85 t/ha green mass. We didn’t find relevant<br />
differences between total green yields or dry-matter products of mixed treatments, but the<br />
proporti<strong>on</strong> of two comp<strong>on</strong>ents differed remarkably.<br />
Keywords: perennial rye, germinati<strong>on</strong>, forage mixture<br />
INTRODUCTION<br />
Perennial rye (Secale cereanum) is a stable hybrid of the winter rye (Secale cereale L.) and<br />
the perennial wild rye (Secale m<strong>on</strong>tanum Guss.). Crossings in the genus Secale were made<br />
already at the end of the XIX century to study crossability, relati<strong>on</strong>ship, influence <strong>on</strong><br />
cytological parameters, etc. (Kotvics, 1963). Later the objectives of plant breeding (e.g.<br />
increasing the winter and drought resistance, developing resistance to biotic and abiotic<br />
factors, increasing the protein c<strong>on</strong>tent) get more importance. Many researchers found it<br />
interesting to make a new perennial rye crop. The aim was to develop a hybrid, which all<br />
over its botanical importance unites the best qualities of winter rye and perennial rye in<br />
form of a perennial, highly productive, highly tolerant crop with good fodder value, which<br />
can be grown also <strong>on</strong> weak habitats. The hybrid was developed in several countries.<br />
Kotvics G. made the first plants in Hungary, which were useful for the plant breeding, in<br />
the 1950’s (Kotvics, 1970.).<br />
Kruppa J. developed the variety “Kriszta” from this material at the Research Centre of<br />
Nyíregyháza (University of Debrecen), which got the state allowance in 1998. Another<br />
perennial rye variety was created from the same origin in Hungary, at Research Institute in<br />
Kompolt. Diploid and tetraploid varieties were made in Germany from similar crossings<br />
(Reimann-Philipp, 1995).<br />
Perennial rye is a new crop of the disadvantageous fields, and it has some problems of<br />
its growth technology: the germinati<strong>on</strong> of the seeds is often not good, about 50-70% (likely<br />
because of their form they are sensitive for trash), the spikes are often fragile, etc. These<br />
problems can be solved <strong>on</strong> the <strong>on</strong>e hand through plant breeding, <strong>on</strong> the other hand through<br />
improvement the growth technology – and this latter way seems to be faster.<br />
There aren’t too much possibilities of utilizati<strong>on</strong> of weak acid sandy soils. In our<br />
opini<strong>on</strong> we can get from the same area <strong>on</strong> these soils more valuable fodder if we produce<br />
perennial rye mixed with perennial papili<strong>on</strong>aceae.<br />
127
MATERIALS AND METHODS<br />
Perennial rye variety of our experiments was ‘Kriszta’. Owing to its high green<br />
productivity, low seed productivity and perennial type it is utilized as a green fodder, as<br />
grazing land or grass-land. This variety can be in culture 3-5 years, after this time the stand<br />
grows thinner and thinner, and get weedy. Its seed-producti<strong>on</strong> is about 500-1000 kg pro<br />
year. The plant is 150-180 cm high, thousand-kernel weight: 12-15 g<br />
It can be sown with 3-3.5 milli<strong>on</strong> seeds in September and in the springtime in March, too.<br />
Its drought resistance is very good, this perennial crop adapts well to the disadvantageous<br />
soil and weather c<strong>on</strong>diti<strong>on</strong>s, so it can be sown also <strong>on</strong> weak habitats, loose sandy soils. It<br />
covers the fields the whole year. Attributable to this, to the fast shooting and excellent<br />
stooling (the number of its shoots can reach the 100) perennial rye is instrumental in<br />
protecting the loose sandy soils from erosi<strong>on</strong> and deflati<strong>on</strong> – it fixes the surface of the land.<br />
Expectedly its role will extend in the game farming too, because it is exceedingly suitable<br />
to winter pasture: it can be grazed also in winter and it tillers fast early in spring.<br />
In <strong>on</strong>e of our experiments we analyzed the relati<strong>on</strong>ship between the germinati<strong>on</strong> and<br />
some harvest and thresh methods: we studied the effect of the set of threshing machine and<br />
combine (rev) <strong>on</strong> germinati<strong>on</strong> ability. We harvested perennial rye in two ways: <strong>on</strong>e-stepharvesting<br />
and two-steps-harvesting (threshing <strong>on</strong>e weak after mowing). Several standards<br />
were threshed by hand. The germinati<strong>on</strong> was carried out accordingly to the cereal standard.<br />
In our other experiment we started the development of a new forage mixture with two<br />
comp<strong>on</strong>ents (lucerne (Medicago sativa L.) or bird’s-foot trefoil (Lotus corniculatus L.) and<br />
perennial rye (Secale cereanum)). According to our previous experiments we established<br />
that for this mixture lucerne is suitable, bird’s-foot trefoil is suppressed by perennial rye.<br />
In a forage mix experiment we prepared 7 different mixes with perennial rye (variety<br />
Kriszta) and alfalfa (variety Klaudia) and we compared the mixtures with two standard<br />
treatments (perennial rye al<strong>on</strong>e in 1,8 and 2,7 milli<strong>on</strong> plant per hectare doses). According to<br />
previous experiments, we choose perennial rye doses of 0.4, 0.7, 0.8, 1.4 milli<strong>on</strong> plant/ha,<br />
and 2.5, 3.2, 5.6 milli<strong>on</strong> alfalfa plant/ha. The small parcels of this experiment were sown <strong>on</strong><br />
acid sandy soil (pHKCl=4,35, KA=28, H%=0,78) in Kisvárda in March of 2002. We mowed<br />
the parcels four times a year and weighed the green and dry mass product. We compared<br />
the products of the comp<strong>on</strong>ents by mowing and the results of the mixed sowing and<br />
separately–rows sowing.<br />
RESULTS<br />
The results of germinating study are shown in Figure 1. In cases of either threshing<br />
methods we found correlati<strong>on</strong> between the rev and the germinati<strong>on</strong> ability. By the plot<br />
combine threshing and also by the single ear thresher we found that the lowest rev<br />
eventuates the best results (82,3 and 92,5 %), which d<strong>on</strong>’t differ significantly from the<br />
c<strong>on</strong>trol threshed by hand (89,0 %). The disadvantage of low rev is that after threshing a lot<br />
of seeds stay in the ears. The difference between the two mechanical methods follows<br />
probably from the length of threshing-time: the ears stay much shorter in the threshing<br />
drum of plot combine than in the single ear thresher. Between the <strong>on</strong>e step and two steps<br />
harvesting methods <strong>on</strong> the score of germinati<strong>on</strong> we didn’t find any significant differences.<br />
In the forage mix experience the productivity was c<strong>on</strong>siderably low in the year of sow,<br />
the green yield suffice for grazing. From the sec<strong>on</strong>d year the relati<strong>on</strong> of perennial rye and<br />
papili<strong>on</strong>aceae changed as follows: in the first growth the perennial rye was presented by 40-<br />
60 % in the green mass, this value later decrease and the fourth growth we could harvest<br />
<strong>on</strong>ly alfalfa. Because of the frequent mowing technology (which adapts to alfalfa) the<br />
forage mix can utilize for three years, then the perennial rye get thinner. After this the area<br />
can utilized as alfalfa seed producer field. In sec<strong>on</strong>d year perennial rye produced 30-35 t/ha<br />
128
green mass (6,8-7 t/ha/year dry mass), but the mixed stand yielded 78-85 t/ha green mass<br />
(15,7-16,5 t/ha/year dry mass – Table 1.).<br />
Germinati<strong>on</strong> %<br />
100<br />
90<br />
80<br />
70<br />
60<br />
50<br />
40<br />
30<br />
20<br />
10<br />
0<br />
<strong>on</strong>e-step-harvesting<br />
two-steps-harvesting<br />
Rev 1/min 1000 1500 1800 1000 1500 1800<br />
Threshed by: Plot combine Single ear thresher Manually<br />
Figure 1: Effects of harvest methods <strong>on</strong> germinati<strong>on</strong> ability of perennial rye<br />
Table 1: Yield in the sec<strong>on</strong>d year of forage mix experiment, Kisvárda 2004.<br />
Treatments<br />
Sowing<br />
method<br />
Product of green yield Product of dry-matter<br />
t/ha<br />
t/ha<br />
P. rye Alfalfa Total P. rye Alfalfa Total<br />
1. P. rye 1,8 Mp/ha 30,93 - 30,93 6,87 - 6,87<br />
2. P. rye 2,7 Mp/ha 33,42 - 33,42 7,07 - 7,07<br />
3.<br />
4.<br />
5.<br />
6.<br />
7.<br />
8.<br />
9.<br />
P. rye 1,4 +<br />
Alfalfa 3,2 Mp/ha<br />
P. rye 1,4 +<br />
Alfalfa 5.6 Mp/ha<br />
P. rye 0,7 +<br />
Alfalfa 3,2 Mp/ha<br />
P. rye 0,7 +<br />
Alfalfa 5,6 Mp/ha<br />
P. rye 0,4 +<br />
Alfalfa 3,2 Mp/ha<br />
P. rye 0,4 +<br />
Alfalfa 5,6 Mp/ha<br />
P. rye 0,8 +<br />
Alfalfa 2,5 Mp/ha<br />
mixed 15,18 69,34 84,52 2,88 13,41 16,30<br />
mixed 17,42 64,28 81,70 3,31 12,73 16,03<br />
mixed 9,07 72,88 81,95 1,74 13,41 15,15<br />
mixed 11,25 72,88 84,13 2,04 14,02 16,07<br />
mixed 11,10 73,57 84,67 2,16 14,20 16,36<br />
mixed 8,00 78,90 86,90 1,57 14,95 16,52<br />
separated<br />
rows<br />
21,99 56,78 78,77 4,45 11,30 15,74<br />
129
We didn’t find relevant differences between total green yield or dry-matter products of<br />
mixed treatments, but the proporti<strong>on</strong> of two comp<strong>on</strong>ents differed remarkably (product of<br />
perennial rye was 17,96 % in the 3. treatment and 9,2 % in the 8. treatment).<br />
Our experiment proved that perennial rye is adapted to mixing with alfalfa. The new<br />
forage mixture is able to produce c<strong>on</strong>siderable green yield from the sec<strong>on</strong>d year either <strong>on</strong><br />
acid sandy soil. The two forage crop gave more yield in the experiment when they mixed<br />
than they sowed in separately rows (which is hardly can be implemented in the practice).<br />
DISCUSSION<br />
To bring this new plant species into use it’s essential to solve the problem of seedproducti<strong>on</strong>.<br />
The primary method of this could be the right set of thrashing parameters of<br />
combine-harvester.<br />
Perennial rye can play a role in forage-mixture, mixed with perennial papili<strong>on</strong>aceae. The<br />
new forage mixture has the follow envir<strong>on</strong>mental and ec<strong>on</strong>omic advantages:<br />
• The two plants with different fodder value in the mixture are complementary to each<br />
other from the point of view of dietetics; the mixture produce great mass fodder with<br />
digestible fiber and high quantity of crude protein.<br />
• Because of the lack of fodder <strong>on</strong> pastures in summer it is necessary to eat hay or apply<br />
additi<strong>on</strong>al feeding at this time. The mixture with perennial rye can produce pasturable<br />
green mass from spring to autumn.<br />
• If the mixture is utilized by grazing, the surplus of green yield in spring and autumn<br />
can be utilized by making hay or silage, which can make use of feeding in winter.<br />
• The influence of this new mixture from papili<strong>on</strong>aceae and gramineae is advantageous<br />
for the soil. Alfalfa has deep taproot, that reduces the compactness of soil and it can<br />
collect nitrogen from the air. The fibrous root system of perennial rye c<strong>on</strong>tributes to<br />
development of favorable structure of surface soil.<br />
• It is advantageous from the point of water balance of soil, that perennial rye and alfalfa<br />
utilize the water from different bed owing to they different root-system and they d<strong>on</strong>’t<br />
enter into competiti<strong>on</strong> with each other.<br />
• The forage mixture can profitably be utilized for three-five years, it depends <strong>on</strong> the<br />
soil and intensity of utilizati<strong>on</strong>, so the expense of planting is divided am<strong>on</strong>g some<br />
years and the maintenance cost is low.<br />
• This mixture can play great part in soil protective agriculture, because the perennial<br />
culture overgrow the surface of the soil during the whole year and it affords possibility<br />
of ecological farming.<br />
• The new forage crop associati<strong>on</strong> makes the sustainable farming possible for a l<strong>on</strong>g<br />
time, which is given financial assistance by the European Uni<strong>on</strong> (Sipos 2006/B.).<br />
130
REFERENCES<br />
Kotvics G. (1963): A Secale cereale L., Secale m<strong>on</strong>tanum Guss. és hibridjeinek<br />
morfológiai, citogenetikai és fejlődés-élettani tulajd<strong>on</strong>ságainak vizsgálata, kandidátusi<br />
értekezés, Gödöllő.<br />
Kotvics G. (1970): Investigati<strong>on</strong>s os increasing the protein c<strong>on</strong>tent of Secale cereale L. In<br />
Ed. Bálint A. : Protein Growth by Plant Breeding 89-98 Akadémiai Kiadó, Budapest.<br />
Reimann-Philipp R. (1995): Breeding Perennial Rye. In. Ed. Jules Janick 0-471-.57343-4<br />
Plant Breeding Reviews, Volume 13.<br />
Sipos T., Kruppa J. (2006/A): A Kriszta évelő rozs – alternatív energianövény a gyenge<br />
<strong>term</strong>ékenységű homoktalajokra. Ős<strong>term</strong>elő, 10. évf. 3. sz.<br />
Sipos T., Halász E. (2006/B): A „Kriszta” évelő rozs fajta szerepe a homoktalajok<br />
védelmében. Előadás, V. Alföldi Tudományos Tájgazdálkodási Napok, Mezőtúr 2006.<br />
Lektorált kiadvány 117. o.<br />
131
MYCOTOXINS CONTAMINATION IN FRESH AND SILAGE<br />
OF ALFALFA (Medicago sativa)<br />
B. Gálik, D. Bíro, M. Juráček, M. Šimko, J. Michálková<br />
Slovak University of Agriculture, Department of Animal Nutriti<strong>on</strong>, Nitra,<br />
Slovak Republic<br />
ABSTRACT<br />
The aim of this work was detecti<strong>on</strong> of mycotoxin c<strong>on</strong>taminati<strong>on</strong> in fresh alfalfa (dry<br />
matter: 228 g.kg -1 ) and in alfalfa silage. Ensiled matter was wilted at six hours. Dry<br />
matter c<strong>on</strong>tent was 277.7 g.kg -1 in silage. C<strong>on</strong>centrati<strong>on</strong>s of mycotoxins<br />
(deoxynivalenol 5/5, fum<strong>on</strong>isin, T-2 toxin, aflatoxin and ochratoxin) were de<strong>term</strong>ined<br />
in fresh forage and silage of alfalfa. This was used immuno-enzymatic analysis by<br />
ELISA Reader (Noack, Slovak Republic) for investigati<strong>on</strong> amount of mycotoxins.<br />
C<strong>on</strong>centrati<strong>on</strong>s of mycotoxins were higher in silage in comparis<strong>on</strong> with c<strong>on</strong>centrati<strong>on</strong>s<br />
of mycotoxins in fresh matter.<br />
The value of deoxynivalenol (DON) was 100 μg.kg -1 in fresh matter and 233.3<br />
μg.kg -1 (average c<strong>on</strong>centrati<strong>on</strong>) after c<strong>on</strong>servati<strong>on</strong>. We found 61.9 μg.kg -1 of fum<strong>on</strong>isin<br />
after c<strong>on</strong>servati<strong>on</strong> and 51.2 μg.kg -1 in fresh matter. The c<strong>on</strong>tent of T-2 toxin was 21.9<br />
μg.kg -1 before ensiling and 88.5 μg.kg -1 in silage of alfalfa (minimum was 72.3 μg.kg -1<br />
and maximum was 99.2 μg.kg -1 ). C<strong>on</strong>tents of mycotoxins were produced by storage<br />
fungi (aflatoxin and ochratoxin) were higher in silage than numbers of mycotoxins in<br />
fresh alfalfa. C<strong>on</strong>centrati<strong>on</strong>s of all detected mycotoxins were lower than critical values.<br />
Keywords: mycotoxins, c<strong>on</strong>taminati<strong>on</strong>, fresh matter, silage, alfalfa<br />
INTRODUCTION<br />
Mycotoxins are toxic sec<strong>on</strong>dary metabolites produced by fungi. There are hundreds of<br />
mycotoxins known, but few have been extensively researched and even few have good<br />
methods of analysis available (Nels<strong>on</strong> et al., 1993, Nedělník, Moravcová, 2006).<br />
Mycotoxins exhibit a variety of biological effects in animals: liver and kidney toxicity,<br />
central nervous system effects and estrogenic effects, to name a few (Whitlow et al.<br />
1998). Worldwide, approximately 25% of crops are affected by mycotoxins annually.<br />
Many different mycotoxins have been found to occur <strong>on</strong> forages either in the field or in<br />
storage as hay or silage (Lancey, 1991). The limiting factor for fungi growth in hay is<br />
moisture, in silage pH (Kalac, Woolford, 1982). At present nearly 400 mycotoxins<br />
produced by a wide spectrum of fungal pathogens are described but the occurrence of<br />
<strong>on</strong>ly the most frequent and harmful <strong>on</strong>es is m<strong>on</strong>itored <strong>on</strong> a regular basis (Lew et al.<br />
2001). Fungi and their mycotoxins identified in feeds (silage) may occur either through<br />
their deleterious effects <strong>on</strong> nutrient and hygienic quality (Tapia et al. 2005).<br />
Aflatoxins are a family of extremely toxic, mutagenic and carcinogenic<br />
compounds produced by Aspergillus flavus and Aspergillus parasiticus (Deiner et al.<br />
1987). Zearalen<strong>on</strong>e is estrogenic metabolite of several species of Fusarium.<br />
Zearalen<strong>on</strong>e has been reported to occur in corn, other grains and silage in many areas<br />
of the world (Hagler et al. 1984). Trichothecenes, T-2 toxin and deoxynivalenol too, are<br />
comm<strong>on</strong>ly found in agricultural commodities (Desjardins et al. 1993). Fum<strong>on</strong>isins has<br />
132
een shown to cause leucoencephalomalacia in horses (Marasas et al. 1988), pulm<strong>on</strong>ary<br />
edema in swine (Harris<strong>on</strong> et al. 1990) and hepatoxicity in rats (Geldebrom et al. 1991).<br />
Unlike foodstuffs, ultimate c<strong>on</strong>centrati<strong>on</strong>s of individual mycotoxins are not for<br />
individual categories of animals and feeds in many European countries. Therefore<br />
detected results were compared to recommended limits published in the United States<br />
of America. The critical c<strong>on</strong>centrati<strong>on</strong> of zearalen<strong>on</strong>e in all feeds and categories of<br />
animals is 0.5 mg.kg 1 , the limiting c<strong>on</strong>centrati<strong>on</strong> of T-2 toxin is the same. The critical<br />
c<strong>on</strong>centrati<strong>on</strong> of fum<strong>on</strong>isin is 5 mg.kg -1 . The limiting c<strong>on</strong>centrati<strong>on</strong> of deoxynivalenol<br />
is specific for cattle 10 mg.kg -1 , the critical amount of deoxynivalenol is 5 mg.kg -1 for<br />
pigs.<br />
The intenti<strong>on</strong> of the present experiment was to establish mycotoxins c<strong>on</strong>taminati<strong>on</strong><br />
in fresh forage and silage of alfalfa.<br />
MATERIAL AND METHODS<br />
Sec<strong>on</strong>d cut alfalfa (Medicago sativa) was harvested at the blooming stage, variety<br />
Palava. After wilting (6 hours), alfalfa was chopper by chopper harvester to 15 mm<br />
particle length. Wilted matter of alfalfa was c<strong>on</strong>served (281.6 g.kg -1 of dry matter)<br />
without additives in silage units (capacity 15 dm 3 ). Silage units were opened after two<br />
m<strong>on</strong>ths. Samples of fresh alfalfa and silage were analyzed for mycotoxin<br />
c<strong>on</strong>taminati<strong>on</strong>. Fresh matter and alfalfa silage were tested <strong>on</strong> a number of mycotoxin in<br />
two replicates for each mycotoxin. The following mycotoxins were m<strong>on</strong>itored:<br />
zearalen<strong>on</strong>e, deoxynivalenol 5/5, fum<strong>on</strong>isin, aflatoxin and ochratoxin. We used<br />
immuno-enzymatic method for de<strong>term</strong>inati<strong>on</strong> amounts of mycotoxins by ELISA reader<br />
(Noack, Slovak Republic) with veratox tests (quantitative test). Principle of the test is<br />
de<strong>term</strong>inati<strong>on</strong> of c<strong>on</strong>centrati<strong>on</strong> in extract of feedstuffs by spectrophotometric method<br />
(650 nm). Before extracti<strong>on</strong>, samples had to been homogenous. Extracti<strong>on</strong> was<br />
performed with distilled water (DON 5/5), 70% soluti<strong>on</strong> of methanol (V/V) for<br />
fum<strong>on</strong>isin, zearelen<strong>on</strong>e and aflatoxin or 50% soluti<strong>on</strong> of methanol (V/V) for ochratoxin<br />
and T-2 toxin.<br />
RESULTS AND DISCUSSION<br />
Many different mycotoxins have been found to occur <strong>on</strong> forages either in the field, or in<br />
storage as hay or silage (Lacey, 1991). Silage c<strong>on</strong>tained higher numbers of mycotoxins<br />
in comparis<strong>on</strong> to amounts of mycotoxins in fresh matter (Table 1). C<strong>on</strong>tent of<br />
deoxynivalenol was range from 200 to 300 μg.kg -1 it was 100 μg.kg -1 in fresh matter.<br />
The highest c<strong>on</strong>centrati<strong>on</strong> of fum<strong>on</strong>isin was 71.9 μg.kg -1 . Fum<strong>on</strong>isin has been shown to<br />
reduce milk producti<strong>on</strong> in dairy cattle at 100 mg.kg -1 (Diaz et al., 2000). We found<br />
similar results of zearalen<strong>on</strong>e like Nedělník and Moravcová (2006), range from 635.8 to<br />
703.1 μg.kg -1 . In Slovak climatic c<strong>on</strong>diti<strong>on</strong>s, mycotoxins (aflatoxin and ochratoxin)<br />
have produced by storage fungi (Leibetseder, 1995). We found aflatoxin with toxic<br />
mutagenic and carcinogenic characteristics (Deiner et al., 1987) range from 10.4 μg.kg -1<br />
to 11.9 μg.kg -1 , lower than reported Kalac and Woolford (1982). The c<strong>on</strong>centrati<strong>on</strong> of<br />
ochratoxin was 12.5 μg.kg -1 in fresh matter, it was range from 8.2 to 14.5 μg.kg -1 in<br />
silage.<br />
133
Table 1: C<strong>on</strong>centrati<strong>on</strong>s of tested mycotoxins in fresh matter and alfalfa silage<br />
Indicators of<br />
DON ZON T-2 FUM AFL OT<br />
c<strong>on</strong>taminati<strong>on</strong> µg.kg -1<br />
fresh<br />
matter<br />
alfalfa<br />
silage<br />
n=2 100 210 21.9 51.2 5.2 12.5<br />
Min. 200 635.8 72.3 52.8 10.4 8.2<br />
Max. 300 703.1 99.2 71.9 11.9 14.5<br />
n=6 Average 233.3 451.4 88.5 61.9 7.4 7.6<br />
DON: deoxynivalenol, ZON: zearalen<strong>on</strong>e, T-2 : T-2 toxin, FUM : fum<strong>on</strong>isin,<br />
AFL : alfatoxins, OT : ochratoxin<br />
REFERENCES<br />
Deiner, U.L., Cole, R.J. et al. 1987. Epidemiology of aflatoxin formati<strong>on</strong> by Aspergillus<br />
flavus. In: Annual Review of Phytopathology, vol. 25, 1987, p. 240-270.<br />
Desjardins, A.E., Hohn, T.M., McCormick, S.P. 1993. Trichothecene biosyntesis in<br />
Fusarium sp.: chemistry, genetics and significance. In: Microbiology Reviews,<br />
vol. 57, 1993, p. 594-604.<br />
Diaz, D.E., Hopkins, L.M., et al. 2000. Effect of fum<strong>on</strong>isin <strong>on</strong> lactating dairy cattle. In:<br />
Journal of Dairy Science, vol. 83, 2000, p. 1171 (abstr.).<br />
Geldebrom, W.C.A, Kreik, N.P.J., et al. 1991. Toxicity and carcinogenicity of the<br />
Fusarium m<strong>on</strong>iliforme metabolite, fum<strong>on</strong>isin B1, in rats. In: Carcinogenesis, vol.,<br />
12, 1991, p. 1247-1251.<br />
Hagler, W.M., J<strong>on</strong>es, F.T., Bowman, D.T. 1984. Simultaneous occurrence of<br />
deoxynivalenol, zearalen<strong>on</strong>e and aflatoxin in 1982 scabby wheat from the<br />
Midwestern United States. In: Applied of Enviromental Microbiology, vol. 47,<br />
1984, p. 151-154.<br />
Harris<strong>on</strong>, L.R., Colvin, B.M., et al. 1990. Plum<strong>on</strong>ary edema and hydrothax in swine<br />
produced by fum<strong>on</strong>isin B1, a toxic metabolite of Fusarium m<strong>on</strong>iliforme. In:<br />
Journal of Veterinary Diagn. Invest., vol., 2, 1990, p. 217-221.<br />
Kalac, P., Woolford, M.K. 1982. A review of some aspects of possible associati<strong>on</strong>s<br />
between the feeding of silage and animal health. In: British Veterinary Journal,<br />
vol. 138, 1982, p. 305-320.<br />
Leibetseder, J. 1995. The European perspective <strong>on</strong> mycotoxins. In: Ly<strong>on</strong>s, T.P.,<br />
Jacques, K.A.: Biotechnology in the Feed Industry, Nottingham University Press:<br />
Notthingam, 1. vyd., 496 s., ISBN 18-97-67656-5, s. 65-75.<br />
Lancey, J. 1991. Natural occurrence of mycotoxins in growing and c<strong>on</strong>served forage<br />
crops. In: J.E. Smith, R.E. Henders<strong>on</strong>: Mycotoxins and Animal Foods. 1991, 1.<br />
edt., Boca Rat<strong>on</strong> Florida: CRC Press, p. 363-379.<br />
Lew, H. et al. 2001. Occurrence of toxigenic funghi and related mycotoxins in plants,<br />
food and feed in Austria. In: Occurrence of toxigenic funghi, Cost Acti<strong>on</strong> 835,<br />
European Commisi<strong>on</strong>, 2001, s. 3-12.<br />
134
Marasas, W.F.O., Kellerman, W.C.A, et al. 1988. Leucoencephalomalacia in a horse<br />
induced by fum<strong>on</strong>isin B1 isolated from Fusarium m<strong>on</strong>iliforme. In: Onderstepoort<br />
Journal of Veterinary Research, vol., 55, 1988, p. 197-203.<br />
Nedělník, J., Moravcová, H. 2006. Mycotoxins and forage crops problems of the<br />
occurrence of mycotoxins in animal feeds. In: 12 th <str<strong>on</strong>g>Internati<strong>on</strong>al</str<strong>on</strong>g> Symposium:<br />
Forage C<strong>on</strong>servati<strong>on</strong>, VFU: Brno, 2006, p. 15-26.<br />
Nels<strong>on</strong>, P.E., Desjardins, A.E., Plattner, R.D. 1993. Fum<strong>on</strong>isins, mycotoxins produced<br />
by Fusarium species : Biology, chemistry and significance. In: Annual Review of<br />
Phytopathologie, vol. 31, 1993, p. 233-249.<br />
Tapia, M.O., Stern, M.D., Soraci, A.L. et al. 2005. Patulin-producing molds in corn<br />
silage and high moisture corn and effects of patulin <strong>on</strong> fermentati<strong>on</strong> by ruminal<br />
microbes in c<strong>on</strong>tinuous culture. In: Animal Feed Science and Technology, vol.<br />
119, 2005, p. 247-258.<br />
Whitlow, L.W., Hagler, W.M., Hopkins, B.A. 1998. Mycotoxins occurrence in farmer<br />
submitted samples of North Carolina feedstuffs 1989-1997. In: Journal of Dairy<br />
Science, vol. 81, 1998, p. 1189.<br />
135
USAGE OF NATURAL ADDITIVES FOR BROILER CHICKENS<br />
Erika Horniaková, Ladislav Bušta<br />
Department of Animal Nutriti<strong>on</strong>, Faculty of Agrobiology and Food Resource,<br />
Slovak University of Agriculture - Nitra, Slovakia<br />
ABSTRACT<br />
The goal of this experiment was an additi<strong>on</strong>al verificati<strong>on</strong> of the natural additive<br />
preparati<strong>on</strong> Clinoptilolit of sedimentary origin with an additi<strong>on</strong> of microelements and<br />
citric acid, the influence of this APC feed mixture <strong>on</strong> feed c<strong>on</strong>sumpti<strong>on</strong>, feed<br />
c<strong>on</strong>versi<strong>on</strong>, live weight and weight gain in broiler chickens.<br />
The trial included broiler chickens of the ROSS 308 type which were divided into a<br />
c<strong>on</strong>trol group (C) and 2 experimental groups (T1 and T2), each c<strong>on</strong>taining 100 chickens<br />
with four repetiti<strong>on</strong>s. The feed mixtures differed in c<strong>on</strong>centrati<strong>on</strong> according to crude<br />
protein and clinoptilolit preparati<strong>on</strong>. The feeding periods were de<strong>term</strong>ined by the total<br />
c<strong>on</strong>sumpti<strong>on</strong> of 15 kg HYD-01/1, alternatively 65 kg HYD-01/2, further 100 kg of<br />
HYD-02 and finally 170 kg of HYD-03. The highest c<strong>on</strong>centrati<strong>on</strong> of crude protein was<br />
offered to be 230 g.kg-1 in the c<strong>on</strong>trol group, a lower value of 215 g.kg-1 in the first<br />
trial group and 200 g.kg-1 in the sec<strong>on</strong>d group as starter food (HYD-01/1). During<br />
fattening the protein c<strong>on</strong>centrati<strong>on</strong>s were slowly decreased to 220.0, 210.0 and 200.0<br />
g.kg-1 in HYD-01/2, 200.0, 190.0 and 180.0 g.kg-1 in HYD-02 and for finishing the<br />
feed mixture HYD-03 c<strong>on</strong>tained 190.0, 180.0 and 170.0 g.kg-1 . In the groups T1 and<br />
T2 the additive clinoptilolit was added <strong>on</strong> a 0.2% c<strong>on</strong>centrati<strong>on</strong> level.<br />
The highest total feed intake per chicken per day was in the C group (101.39 g) as<br />
compared with the experimental groups, which c<strong>on</strong>sumed 93.65 % and 96.92 % of feed<br />
respectively. A similar tendency was found in the intake of dry matter, crude protein<br />
and energy. The live weights were 1956 g, 2137 g and 2001 g in the same order of the<br />
c<strong>on</strong>sidered groups. The differences were statistically significant <strong>on</strong> the level P
minerals (Čuvanová et al. 2006). The usage of zeolits <strong>on</strong> an 0.2% dose al<strong>on</strong>g with<br />
nicarbazin and narazin positively affected daily weight gain (P
weight and weight gain. These parameters were de<strong>term</strong>ined individually in the trial<br />
group and were compared with the parameters de<strong>term</strong>ined in the c<strong>on</strong>trol group.<br />
Statistical evaluati<strong>on</strong>s were carried out with the program<br />
RESULTS AND DISCUSSION<br />
In evaluating the average feed c<strong>on</strong>sumpti<strong>on</strong> for the starter period (HYD-01/1) for <strong>on</strong>e<br />
broiler chicken per day (table 3) feed c<strong>on</strong>sumpti<strong>on</strong> in the c<strong>on</strong>trol group was 24.27 g and<br />
in the experimental groups the observed values were 23.55 g for group T1 and 23.85 g<br />
for group T2. The relative values were 2.97 % less in T1 and 1.73 % less in T2 less<br />
compared to the c<strong>on</strong>trol group. For the prefattening period (HYD-01/2) feed<br />
c<strong>on</strong>sumpti<strong>on</strong> in the c<strong>on</strong>trol group was highest (67.90 g) in comparis<strong>on</strong> with T1 and T2,<br />
which resulted in smaller values to be 6.42 % and 4.14 % less. The same parameters in<br />
the groups were evaluated also for the fattening period (HYD-02) and finishing period<br />
(HYD-03). For the total feed c<strong>on</strong>sumpti<strong>on</strong> for <strong>on</strong>e broiler chicken per day, feed<br />
c<strong>on</strong>sumpti<strong>on</strong> in the c<strong>on</strong>trol group was 101.39 g, 94.95 g in the first trial group and 96.92<br />
g in the sec<strong>on</strong>d trial group. Average feed c<strong>on</strong>sumpti<strong>on</strong> per kg live weight showed the<br />
same tendency within the c<strong>on</strong>sidered groups as the other parameters. Values for average<br />
feed c<strong>on</strong>versi<strong>on</strong> were observed to be highest in the c<strong>on</strong>trol group (1.906 kg), while in<br />
the experimental groups values were lower, in the trial groups T1 (1.840 kg) and T2<br />
(1.873 kg) during the whole experiment.<br />
Table 2: Analytical analysis and nutriti<strong>on</strong>al value of FM<br />
FN group CP MEN Ca P<br />
HYD-01/1<br />
HYD-01/2<br />
HYD-02<br />
HYD-03<br />
C 233.4 12.9 13.7 7.8<br />
T1 217.9 12.8 10.2 5.2<br />
T2 206.2 12.8 10.1 5.2<br />
C 227.9 12.9 12.5 7.1<br />
T1 209.1 12.8 8.9 4.9<br />
T2 198.4 12.8 8.0 4.7<br />
C 207.0 12.8 9.8 7.6<br />
T1 192.6 13.0 8.4 5.3<br />
T2 186.9 13.0 7.7 5.2<br />
C 191.3 13.1 10.2 7.1<br />
T1 187.9 13.0 7.5 5.1<br />
T2 178.3 13.1 7.7 6.4<br />
These data of decreasing feed c<strong>on</strong>sumpti<strong>on</strong> per chicken and day and also per live weight<br />
were c<strong>on</strong>firmed by Gezen et al. (2002), and by Agrar Producti<strong>on</strong> and C<strong>on</strong>sulting (2005,<br />
2003) in their experiments with applicati<strong>on</strong>s of zeolit or APC preparati<strong>on</strong>s as additives<br />
to feed mixtures.<br />
The average live weight (table 4) of broiler chickens was compared with the<br />
c<strong>on</strong>trol group in the individual periods starter, prefattening, fattening and finishing<br />
(0.155, 0.494, 1.001 and 1.825 kg). In the experimental groups a decrease of live weight<br />
was observed <strong>on</strong>ly in the first two periods (starter HYD-01/1 and prefattening HYD-<br />
138
01/2). For the latest periods (fattening and finishing) where we used the HYD-02 and<br />
HYD-03 feed mixtures, the values in live weight increased. Within the values of live<br />
weight significant and highly significant differences were noticed. The smaller c<strong>on</strong>tent<br />
of crude protein is resp<strong>on</strong>sible for the delayed growing up at the beginning of fattening.<br />
The additives with clinoptilolit result in a better utilizati<strong>on</strong> of protein and thus a lower<br />
required supply al<strong>on</strong>g with a higher live weight and weight gain at the end of fattening.<br />
Reducti<strong>on</strong> of nitrogen in the feed mixture resulted in better utilizati<strong>on</strong> and a reducti<strong>on</strong> in<br />
nitric manure.<br />
Table 3: Feed c<strong>on</strong>sumpti<strong>on</strong> and feed c<strong>on</strong>versi<strong>on</strong><br />
FM<br />
Feed c<strong>on</strong>sumpti<strong>on</strong> Feed c<strong>on</strong>versi<strong>on</strong><br />
C T1 T2 C T1 T2<br />
HYD-01/1 24.27 23.55 23.85 0.944 1.017 1.018<br />
HYD-01/2 67.90 63.54 65.09 1.852 1.879 1.953<br />
HYD-02 113.95 109.94 111.49 1.968 1.828 1.842<br />
HYD-03 166.05 154.76 159.59 2.069 1.974 2.005<br />
Total 101.39 94.95 96.92 1.906 1.840 1.873<br />
Table 4: Live weight and weight gain<br />
Period C T1 T2 C T1 T2<br />
starter 0.155 0.144 0.142<br />
prefattening 0.494 0.491 0.475 0.340 0.347 0.333<br />
fattening 1.001 1.033 1.020 0.508 0.542 0.545<br />
finishing 1.825 1.897 1.876 0.824 0.864 0.856<br />
REFERENCES<br />
1. Agrar Producti<strong>on</strong> and C<strong>on</strong>sulting, 2003,2005 available <strong>on</strong> : http://www.apcaustria.com/deutsch/produkte/index.htm<br />
2. Reháková, M., Čuvanová, S., Gavaľová, Z., Rimár, J. 2003: Využitie prírodného<br />
zeolitu typu klinoptilolitu v agrochémii a v poľnohospodárstve. Chem. listy 97,<br />
(2003), s. 260-264,.<br />
3. Čuvanová,S., Reháková, M., Rimár, J., Dzivák, M.2006: Klinoptilolit ako nosič,<br />
regulátor a stabilizátor uvoľňovania živín. Acta M<strong>on</strong>tanistica Slovaca Ročník 11<br />
,(2006) , mimoriadne číslo 2, 285-289<br />
4. Skalická, M., Koreneková, B., Makóová, Z., Naď, P.: Vplyv aditív na vybrané<br />
produkčné ukazovatele hydiny. V. Dni výživy a veterinárnej dietetiky. Zborník z<br />
medzinárodnej vedeckej k<strong>on</strong>ferencie, Hrádok, (2002), 237–239.<br />
5. Gezen S.S, Mustafa, E. 2002: Broyler Rasy<strong>on</strong>lazma Katalin Narasin ve<br />
Nikarbazinin, Zeolit Yle Etkilisimin Besi Perfornabsu Userine Etkileri. Uludag,<br />
Univ. Y. Fac. Vet. Med, 21,(2002), p. 95-101<br />
139
PERSPECTIVES ON ROMANIAN-HUNGARIAN CROSS-BORDER<br />
COOPERATION. THE BIHOR - HAJDÚ-BIHAR EUROREGION MODEL<br />
Gabriela Popoviciu 1 , M.C.Neacşu 2<br />
1 University of Oradea, 2 Academy of Ec<strong>on</strong>omic Studies Bucharest - Romania<br />
ABSTRACT:<br />
The promoti<strong>on</strong> of regi<strong>on</strong>al cooperati<strong>on</strong>, including cross-border initiatives, represents a<br />
new c<strong>on</strong>cept for internati<strong>on</strong>al relati<strong>on</strong>s between two or more neighbouring countries,<br />
which have the same interest in interregi<strong>on</strong>al research and development cooperati<strong>on</strong> in<br />
order to stimulate investments in the private sector and lead to a general improvement<br />
of living standards. On the other hand, the regi<strong>on</strong>al development policy promoted some<br />
measures by local public and central administrati<strong>on</strong>, in partnership with the private and<br />
public sector, and volunteers’ community also, for ensuring an ec<strong>on</strong>omic, dynamic and<br />
sustainable growth through the efficient valorisati<strong>on</strong> of the local and regi<strong>on</strong>al potential<br />
aiming.<br />
This is <strong>on</strong>e of the main goals of cooperati<strong>on</strong> between Romania and Hungary.<br />
Particularly, in this case study we will make notificati<strong>on</strong>s about regi<strong>on</strong>al development<br />
between the two countries, in c<strong>on</strong>necti<strong>on</strong> with their natural resources and their ways of<br />
maintaining them in good quality for future generati<strong>on</strong>s. For this reas<strong>on</strong>, both countries<br />
have identified cross-border cooperati<strong>on</strong> as a priority area and have made further steps<br />
to enhance it.<br />
Key words: Development, cooperati<strong>on</strong>, cross-border, perspectives, regi<strong>on</strong>al<br />
INTRODUCTION<br />
In the past few years, the cross-border relati<strong>on</strong>s and cooperati<strong>on</strong> between states have<br />
started to follow a new tendency. These countries have begun to develop new crossborder<br />
cooperati<strong>on</strong> systems, the new <strong>on</strong>e c<strong>on</strong>sisting of Euroregi<strong>on</strong>s.<br />
In this c<strong>on</strong>text, we carried out a research <strong>on</strong> the topic of Cross-border Cooperati<strong>on</strong><br />
in <strong>on</strong>e of the newest Associati<strong>on</strong> Systems between two neighbouring countries,<br />
Romania and Hungary. We will menti<strong>on</strong> here the Bihor-Hajdú-Bihar Euroregi<strong>on</strong>, which<br />
exists in this c<strong>on</strong>figurati<strong>on</strong> since October 2002.<br />
The Bihor-Hajdú-Bihar Euroregi<strong>on</strong> has its origins in the Carpathian Euroregi<strong>on</strong><br />
(composed by Hungary, Poland, Ukraine, Slovakia and Romania), the first cross-border<br />
cooperati<strong>on</strong> form between Central and Eastern European countries. However, over the<br />
years, this Euroregi<strong>on</strong> has had various problems with transposing the entire countries’<br />
programmes for a comm<strong>on</strong> working. This moment was the starting point for the Bihor-<br />
Hajdú-Bihar Euroregi<strong>on</strong>. It c<strong>on</strong>sisted of two cross-border counties, Bihor (RO) and<br />
Hajdú-Bihar (HU). The official agreement’s act of partnership was signed by the<br />
Bihor’s County Council and Self-government’s Hajdú-Bihar al<strong>on</strong>g with c<strong>on</strong>solidating<br />
cooperati<strong>on</strong> am<strong>on</strong>g authorities, other public or private organisati<strong>on</strong>s and, last but not<br />
least, between the populati<strong>on</strong> from those counties.<br />
140
1. GENERAL APPROACH TO EUROREGIONAL PHENOMENA<br />
The moment worth to be symbolically named “1989” and which marks the beginning of<br />
the communist system wane in Europe, is followed by two opposite tendencies:<br />
fragmentati<strong>on</strong> and integrati<strong>on</strong> (Neguţ S., 2005). On the <strong>on</strong>e hand the territorial<br />
disintegrati<strong>on</strong> is obvious, peaceful (the dismemberment of Czechoslovakia, 1993) or<br />
c<strong>on</strong>flicting (e.g. Yugoslavia, 1991-1992, former USSR, 1991). On the other hand, an<br />
ec<strong>on</strong>omic integrati<strong>on</strong> tendency through successive waves of integrati<strong>on</strong> in European<br />
Uni<strong>on</strong> (2004, 2007) manifested.<br />
One of the dimensi<strong>on</strong>s of integrati<strong>on</strong> was the regi<strong>on</strong>ati<strong>on</strong> or euroregi<strong>on</strong>ati<strong>on</strong>,<br />
which has not entailed a fragmentati<strong>on</strong>, but vicinity rather, through the effort to recede<br />
ec<strong>on</strong>omic, social, cultural discrepancies between territorial units spatially close<br />
together, but which bel<strong>on</strong>ged to different states, in an attempt to make the most of<br />
geographical complementarity.<br />
The euroregi<strong>on</strong>al model implies the associati<strong>on</strong> between local territorial<br />
communities, from two or more neighbouring states, creating a unitary regi<strong>on</strong>,<br />
characterized by some comm<strong>on</strong> particularities and interests. Euroregi<strong>on</strong>s c<strong>on</strong>cretize<br />
geographical solidarities, despite state fr<strong>on</strong>tiers, making new spaces of decentralized<br />
cooperati<strong>on</strong>.<br />
The territories of these close cross-border spaces could be characterized as<br />
territorial units of a relative homogeneity with regard to the natural background of the<br />
two or more states from the respective Euroregi<strong>on</strong>, which are similar from the point of<br />
view of their features, traditi<strong>on</strong>s, activities and preoccupati<strong>on</strong>s.<br />
Thus, the collaborati<strong>on</strong> phenomen<strong>on</strong> within Euroregi<strong>on</strong>s c<strong>on</strong>sists of establishing<br />
some direct relati<strong>on</strong>ships between the regi<strong>on</strong>s and communities <strong>on</strong> both sides of a<br />
state’s fr<strong>on</strong>tier, by virtue of the local authorities’ competences, as they are defined in<br />
every country’s nati<strong>on</strong>al legislati<strong>on</strong>.<br />
The prerequisites for an optimal development of <strong>on</strong>e of these Euroregi<strong>on</strong>s,<br />
c<strong>on</strong>diti<strong>on</strong>s that the two counties hereof (Bihor – Romania and Hajdú-Bihar - Hungary)<br />
seem to have successfully accomplished, are: a minimal ec<strong>on</strong>omic equilibrium, relief,<br />
language and culture unity, similar administrative structure and, why not, the same<br />
historical heritage, given the fact that every state has minorities settled in the<br />
neighbouring state. In fact, euroregi<strong>on</strong>s 1 (Neguţ S., 1999) represent a factor of regi<strong>on</strong>al<br />
stability and security at the fr<strong>on</strong>tier, which aims to outrun a negative historical heritage<br />
in the c<strong>on</strong>text of reinstating an ec<strong>on</strong>omic equilibrium able to promote a high living<br />
standard and an improved quality of life.<br />
1 On the occasi<strong>on</strong> of the meeting from Praga, 1998, c<strong>on</strong>cerning the matter of regi<strong>on</strong>al<br />
cooperati<strong>on</strong> formulas in Central and South-East Europe, dr. Sabathil, as member of the<br />
European Commissi<strong>on</strong> Delegati<strong>on</strong>, emphasized: the most traditi<strong>on</strong>al and efective form<br />
for cooperati<strong>on</strong> between cross-border regi<strong>on</strong>s and (…) a relatively flexible instrument<br />
for collaborati<strong>on</strong> in these fr<strong>on</strong>tier z<strong>on</strong>es.<br />
141
2. THE BIHOR-HAJDÚ-BIHAR EUROREGION– A CROSS-BORDER DEVELOPMENT<br />
MODEL<br />
2.1. Geographical, human and ec<strong>on</strong>omic complementarities<br />
An illustrative example for all we have noted above, which meets all the necessary<br />
c<strong>on</strong>diti<strong>on</strong>s and elements for cross-border cooperati<strong>on</strong>, is the Bihor - Hajdú-Bihar<br />
Euroregi<strong>on</strong>. This is the newest cross-border Euroregi<strong>on</strong>al structure 2 and it is situated in<br />
the North-Western part of Romania, in the Romanian-Hungarian cross-border area (fig.<br />
1).<br />
Fig.1. The Bihor-Hajdú-Bihar Euroregi<strong>on</strong><br />
The Bihor – Hajdú-Bihar<br />
Euroregi<strong>on</strong> overlaps <strong>on</strong> three<br />
essential geographical units, which<br />
are the West Plain, the West Hills<br />
and the Apuseni Mountains (<strong>on</strong> the<br />
Romanian side) while <strong>on</strong>ly the plain<br />
is characteristic to the Hajdú-Bihar<br />
County.<br />
The maximum altitude of the<br />
Euroregi<strong>on</strong> reaches almost 2 000 m<br />
(Cucurbăta Mare Peak, 1 849 m)<br />
while the minimum altitude does not<br />
surpass 100 m (Hortobágyi Regi<strong>on</strong>).<br />
The studied Euroregi<strong>on</strong> also has a plentiful hydrological network (Barcău, Crişul<br />
Repede, Crişul Negru, Crişul Alb, Tisa, Körös, Keleti and Hortobágy) and important<br />
natural resources.<br />
Similar to other cross-border cooperati<strong>on</strong> structures (Ilieş Al., 2004) formed by<br />
including the state fr<strong>on</strong>tier, the Bihor-Hajdú-Bihar Euroregi<strong>on</strong> shares some of their<br />
general characteristics (such as the <strong>on</strong>es regarding organisati<strong>on</strong> and working mode) but<br />
also has a few specific features, which we will present as follows.<br />
The Bihor-Hajdú-Bihar Euroregi<strong>on</strong> represents an open cross-border associati<strong>on</strong>,<br />
which does not possess its own juridical pers<strong>on</strong>ality (art.4 from Euroregi<strong>on</strong> Statute).<br />
Within the cooperati<strong>on</strong> process, the parts have identical rights and obligati<strong>on</strong>s. Through<br />
the present cooperati<strong>on</strong>, the parts have expressed their accord that their acti<strong>on</strong>s be based<br />
<strong>on</strong> mutual trust, spirit of collaborati<strong>on</strong> and good neighbouring relati<strong>on</strong>s (art.5 from<br />
Euroregi<strong>on</strong> Statute).<br />
From a structural point of view, the Bihor-Hajdú-Bihar Euroregi<strong>on</strong> is an<br />
associati<strong>on</strong> of administrative-territorial units of the same rank (Bihor county and Hajdú-<br />
Bihar county), <strong>on</strong> <strong>on</strong>e hand, and other individual or in associati<strong>on</strong> inferior rank units,<br />
(Bihor’s Fr<strong>on</strong>tier Localities Associati<strong>on</strong> and Hajdú-Bihar’s Fr<strong>on</strong>tier Localities<br />
2 Romania and Hungary also participate together to the following euroregi<strong>on</strong>s of the<br />
cross-border type: the Carpathian Euroregi<strong>on</strong> (al<strong>on</strong>gside Poland, Slovakia and Ukraine,<br />
with a 148 095 km² surface and over 15 mil. inhabitants; actually, the Bihor-Hajdú-<br />
Bihar Euroregi<strong>on</strong> was created within this macro-regi<strong>on</strong>) and the Danube-Cris-Mures-<br />
Tisa Euroregi<strong>on</strong> (al<strong>on</strong>gside Serbia; this euroregi<strong>on</strong>al structure extends over 77 243 km²<br />
and has about 6 mil. inhabitants).<br />
142
Associati<strong>on</strong> – fig. 2), <strong>on</strong> the other hand. As we can see, the cooperati<strong>on</strong> within our<br />
Euroregi<strong>on</strong> is extending both horiz<strong>on</strong>tally and vertically 3 .<br />
This structure was organised through bilateral participati<strong>on</strong>, which endows it with a<br />
high rate of functi<strong>on</strong>ality and an increased focus <strong>on</strong> specific problems that derive from<br />
the interfacing positi<strong>on</strong> of the two areas.<br />
The favourable cross-border cooperati<strong>on</strong> premises in the Bihor - Hajdú-Bihar<br />
Euroregi<strong>on</strong> also derive from the fact that, especially during the past few years, the<br />
majority of the ec<strong>on</strong>omic and decisi<strong>on</strong>al factors have accepted the perspective of crossborder<br />
development. The following aspects de<strong>term</strong>ine the complexity of the crossborder<br />
dimensi<strong>on</strong> in our regi<strong>on</strong>:<br />
- Dimensi<strong>on</strong>ally speaking, the Euroregi<strong>on</strong> is a structure of medium value because<br />
of its surface (13 755<br />
km 2 ) and its number<br />
of inhabitants (about<br />
1.1 mil.), which<br />
means that it<br />
represents a substate<br />
structure.<br />
- With regard to<br />
the shape of the<br />
Euroregi<strong>on</strong>, it is<br />
similar to that of a<br />
highly functi<strong>on</strong>al<br />
system, thus ensuring<br />
a balanced<br />
distributi<strong>on</strong> of<br />
localities in the<br />
territory in relati<strong>on</strong> to<br />
the county’s residence<br />
(Oradea and<br />
Debrecen) as well as<br />
in relati<strong>on</strong> to the other<br />
cities.<br />
- The Euroregi<strong>on</strong><br />
represents a crossborder<br />
cooperati<strong>on</strong><br />
structure (Ilieş Al.,<br />
2003) situated in<br />
Crişurilor’s and Tisa’s<br />
Fig.2. The Fr<strong>on</strong>tier Localities Associati<strong>on</strong>s from Bihor -<br />
Hajdú-Bihar Euroregi<strong>on</strong><br />
hydrographical basins, with an in tread morphology of the main units of relief and an<br />
3 Reference is making to the internati<strong>on</strong>al agreement c<strong>on</strong>cerning the partnership<br />
principle extended <strong>on</strong> both a horiz<strong>on</strong>tal and vertical level. This <strong>on</strong>e refers to the<br />
collaborati<strong>on</strong> for mutual support and a good neighbouring policy between nati<strong>on</strong>s,<br />
states, localities, authorities etc.<br />
143
accent <strong>on</strong> complementarity which benefits by a de<strong>term</strong>inative compatibility in the<br />
administrative-territorial system.<br />
- The Euroregi<strong>on</strong> benefits from a communicati<strong>on</strong> network favourable to<br />
complementarity as a basis for ec<strong>on</strong>omic development.<br />
In the area of the Euroregi<strong>on</strong> the infrastructure of 16 “potential” cross-border roads<br />
exists, out of which three are operative in the internati<strong>on</strong>al regime (Sal<strong>on</strong>ta - Mehkerek,<br />
Borş - Artand and Valea lui Mihai - Nyirabrany) and two are meant for occasi<strong>on</strong>al<br />
traffic (Toboliu - Körösnagyhomorog and Săcuieni - Letavertes). Four railroad systems<br />
can be added to these, out of which three are active in the regime of internati<strong>on</strong>al traffic<br />
(Sal<strong>on</strong>ta - Kötegyan, Episcopia Bihor - Biharkeresztes and Valea lui Mihai -<br />
Nyirabrany) (Stamate, G., 1997).<br />
- The Euroregi<strong>on</strong> has a main Romanian-Hungarian ethnic base with a balanced<br />
share of the two ethnic<br />
groups. This ethnic base<br />
is characterised by a<br />
powerful traditi<strong>on</strong> of<br />
cohabitati<strong>on</strong> between the<br />
majority and the local<br />
minorities. Other coinhabiting<br />
nati<strong>on</strong>alities<br />
(fig. 3) can be added, in a<br />
larger or smaller number<br />
<strong>on</strong> both sides of the<br />
fr<strong>on</strong>tier.<br />
- The Euroregi<strong>on</strong> evinces<br />
a higher level of<br />
ec<strong>on</strong>omic development<br />
as compared to the<br />
average in the Eastern<br />
part of Europe.<br />
- Currently, the<br />
Euroregi<strong>on</strong> includes <strong>on</strong>ly<br />
<strong>on</strong>e state fr<strong>on</strong>tier sector<br />
(which has a military<br />
Fig. 3. The ethnic structure in the Bihor – Hajdú-<br />
Bihar Euroregi<strong>on</strong> (the values are in %)<br />
functi<strong>on</strong> de<strong>term</strong>ined by<br />
the NATO membership<br />
of both countries 4 , and a<br />
c<strong>on</strong>trol functi<strong>on</strong> of the people and merchandise fluxes).<br />
- The Romanian-Hungarian border length <strong>on</strong> the Bihor County’s side is of approx.<br />
166 km, out of which approx. 160 km are <strong>on</strong> land and 6 km <strong>on</strong> water. These totalize<br />
about 37% of the entire length of the Romanian-Hungarian border (Stamate G., 1997).<br />
Due to the land support of this fr<strong>on</strong>tier sector, there is an active potential for<br />
4 Starting with 2007, <strong>on</strong>ce with Romania’s adhesi<strong>on</strong> to the EU, this sector has<br />
become<strong>on</strong>e of its internal fr<strong>on</strong>tiers<br />
144
communicati<strong>on</strong> between the two associated territorial systems, which ensures a crossborder<br />
flux similar to the <strong>on</strong>e in other European countries.<br />
- The local administrative organisms of the Euroregi<strong>on</strong> established as priorities<br />
those acti<strong>on</strong>s that refer to the c<strong>on</strong>solidati<strong>on</strong> of administrative-instituti<strong>on</strong>al capacities,<br />
which will ensure their practical role, but also the creati<strong>on</strong> of an informati<strong>on</strong>al<br />
framework regarding the potential of the cooperati<strong>on</strong> between Romanians and<br />
Hungarians.<br />
- On both counties’ territory, there are important natural resources and anthropic<br />
edifices, marked by landscape diversity and a natural complementary background. This<br />
provides a special potential for tourism within the Euroregi<strong>on</strong>. For this purpose, “The<br />
Romanian-Hungarian Corridor for C<strong>on</strong>servati<strong>on</strong> of Biodiversity” was created in the<br />
wetland p<strong>on</strong>ds’ area of Cefa, in the Natural Zoological Reservati<strong>on</strong> from Rădvani<br />
(Romania), as well as in the Körös-Máros Nati<strong>on</strong>al Park (Hungary), locati<strong>on</strong>s that are<br />
actually situated “in mirror positi<strong>on</strong>” from <strong>on</strong>e another.<br />
The variety of<br />
geographical c<strong>on</strong>diti<strong>on</strong>s<br />
is essential for a unitary<br />
development of the<br />
Euroregi<strong>on</strong>, because it<br />
can lead to multiple<br />
possibilities of land<br />
usage. In other words,<br />
the Bihor – Hajdú-Bihar<br />
Euroregi<strong>on</strong> develops<br />
according to its natural<br />
c<strong>on</strong>diti<strong>on</strong>s (industrial<br />
activities in the<br />
mountainous z<strong>on</strong>e,<br />
agricultural activities<br />
and services in the other<br />
areas) (fig. 4).<br />
If we compare the<br />
two counties, we can<br />
see that the industrial<br />
sector still plays the<br />
most important role,<br />
Fig. 4. The populati<strong>on</strong> structure <strong>on</strong> the activities sectors<br />
in Bihor - Hajdú-Bihar Euroregi<strong>on</strong><br />
especially in the ec<strong>on</strong>omy of Bihor County, unlike in the Hajdú - Bihar County, where<br />
the services segment is dominant. At the same time, the general trend is to diminish the<br />
importance of agriculture, in favour of increasing the <strong>on</strong>e of services.<br />
145
2.2. Oradea-Debrecen the next urban cross-border agglomerati<strong>on</strong> 5<br />
Although the historical evoluti<strong>on</strong> of the two cities has been different, the<br />
complementarity of these two future attracti<strong>on</strong> poles is obvious: <strong>on</strong> <strong>on</strong>e hand, Oradea is<br />
menti<strong>on</strong>ed in a 1113 document as a fortress set in a marshy area and it evolves<br />
spatiotemporally under the ruling of its political-administrative functi<strong>on</strong>, and Debrecen<br />
<strong>on</strong> the other hand, of a similar age, renowned ec<strong>on</strong>omical centre.<br />
After 1920, al<strong>on</strong>g with the delimitati<strong>on</strong> of state fr<strong>on</strong>tiers, the two cities suffered a<br />
slight syncope in the evoluti<strong>on</strong> of their interdependence relati<strong>on</strong>s, each of them<br />
bel<strong>on</strong>ging to its own decisi<strong>on</strong>al centre. The functi<strong>on</strong>ality of the urban system revitalized<br />
after 1990 through a cultural c<strong>on</strong>vergence (university exchange programs, both cities<br />
endowed with renowned and very dynamic Universities) as well as a business related<br />
<strong>on</strong>e.<br />
Nowadays the two centres represent the pillars of a future cross-border urban axis<br />
grafted <strong>on</strong> the two metropolitan areas: Oradea (the city of Oradea and the administrative<br />
territory of the adjoining villages – Biharia, Bors, Cetariu, Paleu, Nojorid, Oşorhei,<br />
Sânmartin and Sântandrei), with a populati<strong>on</strong> of over 250 000 inhabitants (out of which<br />
the largest part is located in Oradea – about 87%) and Debrecen (with a populati<strong>on</strong> just<br />
over 200 000 inhabitants), which would spread over about 75 kms <strong>on</strong> the Oradea –<br />
Biharkeresytes – Berettyóújfalú – Derecske – Debrecen axis (Săgeată R. et. al., 2004,<br />
Ilieş Al., 2004).<br />
3. THE JURIDICAL-INSTITUTIONAL BACKGROUND REGARDING REGIONAL<br />
DEVELOPMENT IN THE BIHOR – HAJDÚ-BIHAR EUROREGION. OBJECTIVES<br />
Because Euroregi<strong>on</strong>s represent fr<strong>on</strong>tier cooperati<strong>on</strong> structures, which c<strong>on</strong>tribute not to<br />
regi<strong>on</strong>al isolati<strong>on</strong> but to regi<strong>on</strong>al development (Ghidul U.E., 2004), both states have<br />
adopted the C<strong>on</strong>venti<strong>on</strong> of Madrid (May 21, 1980) regarding cross-border cooperati<strong>on</strong><br />
between territorial communities and authorities. Moreover, they are both part of the<br />
European Charter of Local Self-Government (Strasbourg, Oct. 15, 1985). In order to<br />
render functi<strong>on</strong>al the settlements comprised in these two European acts, the legislative<br />
step made by both countries was the adopti<strong>on</strong> of two nati<strong>on</strong>al laws c<strong>on</strong>cerning local<br />
administrati<strong>on</strong> (Law no. 215/2001 – Romania and Law no. XXI/1996 - Hungary).<br />
With regard to the legislative and statutory aspects, the two laws are compatible, in<br />
the sense that they both designate the authorities, communities and organisms playing a<br />
role in exercising regi<strong>on</strong>al functi<strong>on</strong>s as the local and county’s Council (for the<br />
Romanian side) and the county’s and town hall’s local self-government (for the<br />
Hungarian side). We must menti<strong>on</strong> here that <strong>on</strong>ly the typology of the two laws is<br />
different, as the understanding of the settlement is identical.<br />
In accordance with Article 2, paragraph 1 of the Euroregi<strong>on</strong>’s C<strong>on</strong>stituti<strong>on</strong><br />
Agreement, “the cross-border cooperati<strong>on</strong> is exercised by the qualified territorial<br />
collectivities and authorities, as they are defined in the internal law.”<br />
5 This model is currently in the phase of research project initiated by the “Forum”<br />
Foundati<strong>on</strong> from Oradea, with the participati<strong>on</strong> of the University of Oradea and<br />
University of Debrecen, Thinking the future together. The Debrecen-Oradea Crossborder<br />
Agglomerati<strong>on</strong> (2020).<br />
146
The juridical settlement of the Bihor – Hajdú-Bihar Euroregi<strong>on</strong> was created by a private<br />
law c<strong>on</strong>tract, whose settlements and provisi<strong>on</strong>s are available <strong>on</strong>ly <strong>on</strong> the Euroregi<strong>on</strong>’s<br />
territory. According to the Statute and Agreement of Associati<strong>on</strong> of the two counties,<br />
the main objectives of the partnership between the two territorial-administrative<br />
authorities are:<br />
• Encouraging the plan for unitary development of the rural communities in both areas;<br />
• Encouraging the development of communities both locally and regi<strong>on</strong>ally, based <strong>on</strong><br />
the existing natural and human resources;<br />
• Stimulating the native efforts for community’s development and promoting local<br />
initiatives;<br />
• Reinforcing the partnership and creating a network between different regi<strong>on</strong>al and<br />
local n<strong>on</strong>-governmental organizati<strong>on</strong>s, local administrati<strong>on</strong>s and the business sector;<br />
• Reinforcing the managerial capacities of local and regi<strong>on</strong>al n<strong>on</strong>-governmental<br />
organizati<strong>on</strong>s, as well as those of local administrati<strong>on</strong>s.<br />
4. CONCLUSIONS<br />
The Bihor – Hajdú-Bihar Euroregi<strong>on</strong> has a comm<strong>on</strong> natural and cultural heritage. For<br />
the c<strong>on</strong>servati<strong>on</strong> of those comm<strong>on</strong> values, c<strong>on</strong>crete forms of cross-border cooperati<strong>on</strong><br />
are necessary.<br />
The European project of Euroregi<strong>on</strong>s and cross-border cooperati<strong>on</strong> between<br />
neighbouring communities is built <strong>on</strong> the comm<strong>on</strong> patrim<strong>on</strong>y of European societies, <strong>on</strong><br />
specific nati<strong>on</strong>al principles, values and regulati<strong>on</strong>s, which may be identified in the two<br />
counties reunited into the Bihor – Hajdú-Bihar Euroregi<strong>on</strong>.<br />
The c<strong>on</strong>siderati<strong>on</strong> of all these aspects will open new l<strong>on</strong>g-<strong>term</strong> perspectives for a<br />
sure and durable development of the Bihor – Hajdú-Bihar Euroregi<strong>on</strong> and for the<br />
inclusi<strong>on</strong> of its inhabitants into the world development circuit.<br />
In accordance to all of the above, the Bihor – Hajdú-Bihar Euroregi<strong>on</strong> maintains a<br />
high functi<strong>on</strong>ality, which we believe to be accentuated by the positi<strong>on</strong> of the two<br />
associated counties at the border between two different political entities, with comm<strong>on</strong><br />
collaborati<strong>on</strong> and cooperati<strong>on</strong> premises, which render viable the existence of the studied<br />
Euroregi<strong>on</strong>.<br />
REFERENCES<br />
1. Ilieş, Alexandru (2003), România între milenii. Fr<strong>on</strong>tiere, areale fr<strong>on</strong>taliere şi<br />
cooperare transfr<strong>on</strong>talieră (I),. Universităţii din Oradea Publishing House;<br />
2. Ilieş, Alexandru (2004), România. Euroregiuni, Universităţii din Oradea Publishing<br />
House;<br />
3. Neguţ, Silviu (1999), Les Euroregi<strong>on</strong>s, in „Revue Roumaine de Geographie”,<br />
Academiei Publishing House, Bucharest;.<br />
4. Neguţ, Silviu (2005), Europa între fragmentare şi integrare, in Introducere în<br />
geopolitică, Meteor Press Publishing House, Bucharest, p.161-166.<br />
5. Săgeată, Radu (coord.) (2004), Soluţii de optimizare a organizării administrativteritoriale<br />
a României în perspectiva aderării la Uniunea Europeană, Ars Docendi<br />
Publishing House, Bucharest, p.43.<br />
147
6. Stamate, Grigore (1997), Fr<strong>on</strong>tiera de stat a României, Militară Publishing House,<br />
Bucharest;<br />
7. * * * European Institute of Romania (2004), Ghidul U.E., Aquis comunitar nr. 1<br />
series, 2 nd editi<strong>on</strong>, coord. Tamara Ferluşcă.<br />
8. * * * (1996), Legea nr. XXI din 1996 privind administraţia publică locală a<br />
Ungariei.<br />
9. * * * (2001), Legea nr. 215/2001 privind administraţia publică locală, în M. Of. al<br />
României nr. 204 din 23 aprilie 2001, Bucureşti.<br />
10. * * * Kozp<strong>on</strong>ti Statisztikai Hivatal (2002), Hajdú-Bihar megyei Igazgatósága.<br />
11. * * * Nati<strong>on</strong>al Institute of Statistics (2003), Date statistice după recensământul<br />
populaţiei din 2002.<br />
12. * * * (2002), The Euroregi<strong>on</strong>’s C<strong>on</strong>stituti<strong>on</strong> Accord and The Statute of The<br />
Euroregi<strong>on</strong> Bihor – Hajdú-Bihar.<br />
13. Web sites: www.euroregiuni.ro, www.freeweb.hu/romanul, www.mie.ro<br />
148
EFFECT OF BORON FERTILIZATION ON ANNUAL FLUCTUATION OF B<br />
IN SWEET CHERRY LEAVES AND FRUIT QUALITY<br />
Péter Tamás Nagy 1 - Sándor Thurzó 2 - Ida Kincses 1 - Zoltán Szabó 2 - József Nyéki 2<br />
1 Department of Agricultural Chemistry and Soil Science, University of Debrecen,<br />
Centre of Agricultural Sciences, Faculty of Agr<strong>on</strong>omy<br />
2 Institute for Extensi<strong>on</strong> and Development, University of Debrecen, Centre of<br />
Agricultural Sciences<br />
ABSTRACT<br />
The goal of the study was to examine resp<strong>on</strong>se of sweet cherries (Prunus avium L.) to<br />
bor<strong>on</strong> (B) fertilizati<strong>on</strong>. The experiment was c<strong>on</strong>ducted during 2005–2006 in West<br />
Hungary <strong>on</strong> mature cv. ‘Germersdorfi 3’ grafted <strong>on</strong> Prunus mahaleb rootstock.<br />
Sweet cherry trees planted <strong>on</strong> a calcareous chernozem soil. Trees were foliar-fertilized<br />
with B. Foliar B sprays were performed: (1) in the spring, at the stage of white bud,<br />
beginning of flowering, and (2) repeated 5 weeks after full bloom. In each of spring<br />
spray treatments, B was applied at a rate of 0.15 kg ha −1 . Trees untreated with B served<br />
as a c<strong>on</strong>trol.<br />
The results showed that B fertilizati<strong>on</strong> had effect <strong>on</strong> B c<strong>on</strong>centrati<strong>on</strong> in leaf tissues,<br />
mostly after ripening. Mean fruit weight was slightly increased by B fertilizati<strong>on</strong>. Fruit<br />
sensitivity to cracking was not influenced by B fertilizati<strong>on</strong>. Nevertheless, from our data<br />
it can be c<strong>on</strong>clude that the sensitivity of fruit to cracking is improved when the fruit is<br />
riper, the fruit density and fruit weight are higher. The soluble solids varied between<br />
15.0 and 15.9% according to the treatments. Our results for the m<strong>on</strong>osaccharides<br />
investigated varied between 7.6 and 8.0 as glucose and fructose as well.<br />
It is c<strong>on</strong>cluded that under c<strong>on</strong>diti<strong>on</strong>s of this experiment, B fertilizati<strong>on</strong> can be<br />
recommended in sweet cherry culture to improve fruit quality and their appearance.<br />
Keywords cherry, foliar nutriti<strong>on</strong>, bor<strong>on</strong> fertilizati<strong>on</strong>, fruit quality<br />
INTRODUCTION<br />
However the role of Bor<strong>on</strong> (B) is well-known in the plant nutriti<strong>on</strong>, especially fruit<br />
nutriti<strong>on</strong>, there are very little informati<strong>on</strong> about its applicati<strong>on</strong> and use in Hungarian<br />
orchards.<br />
Bor<strong>on</strong> is an essential micr<strong>on</strong>utrient in plants that is often deficient in most soils<br />
because most of the bor<strong>on</strong> in the soil is adsorbed to clay minerals, hydrous metal oxides,<br />
and organic matter in soils. In additi<strong>on</strong>, bor<strong>on</strong> can be co-precipitated with calcium<br />
carb<strong>on</strong>ate making it unavailable to the roots. Moreover the B uptake is hindered by very<br />
wet or very dry soils, increased leaching and cold soil temperatures.<br />
The major role of B in fruit trees involves fruit set (Faust, 1989). Apple, pear and<br />
cherry flowers are very high in B. The B needed in the flower is transported mainly<br />
from the reserves in the adjacent branches and not from the roots during the<br />
development flower. It is essential for reproducti<strong>on</strong>, aids in the formati<strong>on</strong> of pollen<br />
germinati<strong>on</strong> and pollen tube growth. Bor<strong>on</strong> aids in the metabolism of horm<strong>on</strong>es and in<br />
the translocati<strong>on</strong> of calcium, sugars and growth regulators, required for protein<br />
synthesis. In additi<strong>on</strong> B is important for early growth, flowering and fruit set (Kamali<br />
149
and Childers, 1970), maintains balance between sugar and starch, aids in auxin<br />
regulati<strong>on</strong> and of course it is necessary for cell divisi<strong>on</strong> and differentiati<strong>on</strong>, and root tip<br />
development.<br />
Therefore, the close attenti<strong>on</strong> to B levels is important because both low and high<br />
c<strong>on</strong>centrati<strong>on</strong>s cause poor fruit quality. Low B results in short storage life with the fruit<br />
having a higher susceptibly to storage breakdown and fruit deformities. High B results<br />
in a higher incidence of internal disorders such as watercore and internal breakdown.<br />
MATERIALS AND METHODS<br />
The study was c<strong>on</strong>ducted during 2005–2006 in West Hungary <strong>on</strong> cv. ‘Germersdorfi 3’<br />
grafted <strong>on</strong> Prunus mahaleb rootstock. Trees were planted in the spring of 1999. Trees<br />
spaced 7 x 5 m, and growing in a calcareous chernozem soil at Siófok in West-Hungary.<br />
Orchard was not irrigated in 2005 and 2006. For the purpose of the experiment, 20 trees<br />
were randomly selected from a populati<strong>on</strong> of trees with uniform characteristics. The<br />
applied foliar applicati<strong>on</strong>s are presented in Table 1.<br />
For spraying, Solubor (Disodium Octaborate Tetrahydrate - 20.9% B) was applied.<br />
Table 1: Applied foliar fertilizati<strong>on</strong> system<br />
Applied nutrient Dose (g/L) Time of applicati<strong>on</strong>s Code of<br />
treatment<br />
C<strong>on</strong>trol - - C<br />
B as Na2B8O13·4H2O 0.5<br />
at full bloom<br />
5 weeks after full bloom<br />
1. Soil sampling and preparati<strong>on</strong><br />
Two soil samples were collected from three layers (0-20, 20-40 and 40-60 cm) of each<br />
treatment by using manual soil sampling equipment following the Hungarian sampling<br />
guidelines and according to Nagy et al. (2006). Sampling was performed before<br />
treatments, at the beginning of the vegetati<strong>on</strong> period in March 2005. Sample preparati<strong>on</strong><br />
of the soil samples was performed according to Hungarian guideline (MSZ<br />
20135:1999). The following parameters were measured: pH, KA, c<strong>on</strong>tent of humus and<br />
AL soluble P and K according to Hungarian guidelines.<br />
2. Plant sampling and preparati<strong>on</strong><br />
Plant (leaf) samples were taken, from May to September (14 May, 30 May, 30 June and<br />
10 September) in 2005and 2006. Leaves were taken from all trees according to<br />
internati<strong>on</strong>al c<strong>on</strong>cepti<strong>on</strong> and Hungarian sampling guidelines (Stiles and Reid 1966; MI-<br />
08 0468-81). For leaf analysis 1 g plant sample was ashed in a muffle furnace at 450 °C.<br />
Ash was dissolved in 5 ml of a 1M HCl at room temperature, mixed, and measured by<br />
photometric method (Azomethin-H method).<br />
150<br />
B1<br />
B2
3. Examinati<strong>on</strong>s of fruit quality<br />
For fruit assessments, 50 fruits were examined.<br />
3.1. Fruit cracking (%)<br />
It was calculated as a ratio of the number of cracked fruits and the number of total<br />
fruits.<br />
3.2. Fruit weight (g)<br />
It was measured with a digital analytical scale with 0.1 g punctuality.<br />
3.3. Fruit density (1-9)<br />
This parameter was given a number <strong>on</strong> a subjective scale from 1 to 9 (de<strong>term</strong>ined at<br />
10 trees). Where, 1 means the tree without fruit and 9 means the tree fully covered<br />
fruits. It was established two weeks before harvest.<br />
3.4. Maturity (1-9)<br />
This parameter was given a number <strong>on</strong> a subjective scale from 1 to 9 (de<strong>term</strong>ined at<br />
10 trees). Where, 1 means the unripe fruit and 9 means the totally ripe fruit. It was<br />
established two weeks before harvest.<br />
3.5. Soluble solids and sugars<br />
The sugars (fructose and glucose) and soluble solids (Brix) were studied at harvest<br />
in the sweet cherries (Prunus avium L.) cv. ‘Germersdorfi 3’ in 2005 and 2006. All<br />
fruit samples had been picked at the optimal ripening time. For examinati<strong>on</strong> 12<br />
fruits were performed. Fruits were pressed and the obtained juice was filtered<br />
through (FILTRAK Qual. grade:132) folded filters. Soluble solids and sugars were<br />
de<strong>term</strong>ined in the juice by refractometer (ATAGO PAL series) at 20 ºC.<br />
RESULTS AND DISCUSSIONS<br />
1. Soil analysis<br />
The orchard soil type is calcareous chernozem soil. The upper layer of soil (0-60 cm)<br />
c<strong>on</strong>tained 1.7 % humus, 178 mg/kg and 372 mg/kg AL-soluble P and K. The plasticity<br />
index according to Arany (KA) was 39. According to our results the soil is slightly<br />
alkaline (pH(H2O)=7.65) loamy soil and calcareous in deeper layers.<br />
2. Plant analysis<br />
Bor<strong>on</strong> c<strong>on</strong>centrati<strong>on</strong> was varied between 35 and 65 mg/kg in leaves, during examined<br />
vegetati<strong>on</strong> period. It c<strong>on</strong>tinuously increased during examined period in every treatment<br />
till ripening. Then, in the c<strong>on</strong>trol, it significantly decreased while in B treatments its<br />
amount increased c<strong>on</strong>tinuously (Fig. 1). So although shoot leaves from all treatments<br />
collected in summer had similar bor<strong>on</strong> c<strong>on</strong>centrati<strong>on</strong>s, B was present in different<br />
c<strong>on</strong>tent in leaf tissues late (after ripening) in the seas<strong>on</strong>.<br />
This effect is very important for the next year´s growth. Because, the higher foliar B<br />
c<strong>on</strong>tent plays efficient role in transport processes from the leaves into storage tissues for<br />
the next year´s growth. Moreover, we c<strong>on</strong>clude that leaf B after harvest is translocated<br />
to above-ground storage organs before leaf fall and is then used the following seas<strong>on</strong>.<br />
Based <strong>on</strong> the data of ripening, the leaf B values varied between 59 and 61.5. In all<br />
treatments, these values were higher than the optimal range (Mills and J<strong>on</strong>es 1986;<br />
Papp, 2004). Obtained results can be explained by the soil properties and additi<strong>on</strong>al<br />
bor<strong>on</strong> applicati<strong>on</strong>.<br />
151
B mg/kg in dry matter<br />
70<br />
60<br />
50<br />
40<br />
30<br />
20<br />
10<br />
0<br />
May 14 May 30 June 30 Sept. 10<br />
Sampling time<br />
c<strong>on</strong>trol<br />
Figure 1: Effect of applied foliar treatments <strong>on</strong> annual fluctuati<strong>on</strong> of B in leaves<br />
Obtained results c<strong>on</strong>firmed that earlier findings that timing of B maintenance sprays is<br />
not critical for cherry trees if the trees already c<strong>on</strong>tain adequate amounts of B and do<br />
not show visual evidence of B insufficiency (Peryea, 1994; Peryea et al., 2003).<br />
3. Quality factors and fruit analysis<br />
Some investigated fruit quality factors are represented in Figure 2.<br />
Fruit sensitivity to cracking was influenced differently by B fertilizati<strong>on</strong>. B1 treatment<br />
decreased, while B2 increased the number of cracked fruits.<br />
Mean fruit weight was significantly increased by both B fertilizati<strong>on</strong>s.<br />
Quality factors (relative scale)<br />
20<br />
16<br />
12<br />
8<br />
4<br />
0<br />
LSD 0.05 =1.96<br />
Fruit cracking<br />
(%)<br />
LSD 0.05 =0.83<br />
LSD 0.05 =0.57<br />
B1<br />
B2<br />
C<strong>on</strong>trol<br />
B1<br />
B2<br />
LSD 0.05 =0.33<br />
Fruit weight (g) Fruit density Maturity<br />
Figure 2: Effect of applied foliar treatments <strong>on</strong> some quality factors<br />
152
Fruit density was increased applying foliar bor<strong>on</strong> treatments, but <strong>on</strong>ly the B2 treatment<br />
had significant effect <strong>on</strong> it. Maturity was not effected by bor<strong>on</strong> treatments. The lowest<br />
value was founded in the B1 treatment and there was not statistical difference between<br />
B2 treatment and c<strong>on</strong>trol (Fig. 2).<br />
From our data it can be c<strong>on</strong>clude that the sensitivity of fruit to cracking is<br />
improved when the fruit is riper, the fruit density and fruit weight are higher.<br />
Figure 3 shows the soluble solids varied between 15.0 and 15.9% according to the<br />
treatments. Our results are similar to those reviewed by Kaack et al. (1996) and Predieri<br />
et al. (2004). Our results were pointed out that B applicati<strong>on</strong>s influenced the soluble<br />
solids c<strong>on</strong>tent. Result of B1 treatment may be explained by the lower value of maturity.<br />
Moreover the soluble solids c<strong>on</strong>tent is c<strong>on</strong>necti<strong>on</strong> with maturity and fruit weight as<br />
Blažková et al. (2002) reported.<br />
%<br />
18<br />
16<br />
14<br />
12<br />
10<br />
8<br />
6<br />
4<br />
2<br />
0<br />
LSD0.05=0.52<br />
LSD0.05=0.27<br />
C<strong>on</strong>trol<br />
B1<br />
B2<br />
LSD0.05=0.26<br />
Brix Fructose Glucose<br />
Figure 3: Effect of applied foliar treatments <strong>on</strong> c<strong>on</strong>tents of soluble solids (%) and<br />
sugars (g/100g of fresh weight)<br />
Our results for the m<strong>on</strong>osaccharides investigated are similar to those reviewed by<br />
Wrolstad and Schallenberger (1981) who reported mean values for a large variety of<br />
cherries as glucose 7.78 g/100 g; fructose 7.09 g/100 g and Gardiner et al. (1993) who<br />
reported the c<strong>on</strong>tent of glucose is 7.025 g/100 g and fructose is 6.7 g/100 g<br />
approximately. Our values are also comparable to those obtained by Wills et al. (1987)<br />
for Australian cherries and Dolenc and Štampar (1998) in Slovenia. Sucrose was not<br />
detected in the unprocessed cherries, which is c<strong>on</strong>sistent with the findings of Wrolstad<br />
& Schallenberger (1981) who reported the absence of sucrose in some cultivars (e.g.<br />
cherries) and <strong>on</strong>ly low c<strong>on</strong>centrati<strong>on</strong>s in others. No attempt was made to inactivate<br />
invertase in the unprocessed cherries as it is unlikely that this would be d<strong>on</strong>e in a<br />
commercial situati<strong>on</strong>.<br />
153
Furthermore, our results c<strong>on</strong>firmed those earlier results as the c<strong>on</strong>tent of glucose and<br />
fructose is present in approximately equal amounts in most nectar (Van Handel et al.<br />
(1972)).<br />
It is c<strong>on</strong>cluded that under c<strong>on</strong>diti<strong>on</strong>s of this experiment, B fertilizati<strong>on</strong> can be<br />
recommended in sweet cherry culture to improve fruit quality and their appearance.<br />
REFERENCES<br />
J. Blažková - I. Hlušičková and J. Blažek (2002): Fruit weight, firmness and soluble<br />
solids c<strong>on</strong>tent during ripening of Karešova cv. sweet cherry. Hort. Sci. (Prague),<br />
29, 2002 (3): 92–98<br />
Dolenc K. and F. Štampar (1998): De<strong>term</strong>ining the quality of different cherry<br />
cultivars using the hplc method. Acta Hort. (ISHS) 468:705-712<br />
Faust M. (1989): Physiology of temperate z<strong>on</strong>e fruit trees. Wiley, New York.<br />
Gardiner M. A. - R. Beyer - L. D. Melt<strong>on</strong> (1993): Sugar and anthocyanidin c<strong>on</strong>tent of<br />
two processing-grade sweet cherry cultivars and cherry products. New Zealand<br />
Journal of Crop and Horticultural Science, 1993, Vol. 21: 213-218.<br />
Kaack K. – Spayd S.E. – Drake S.R. (1996): Cherry processing. In: Cherries: Crop<br />
Physiology, Producti<strong>on</strong> and Uses. Eds. By Webster A. D. and Lo<strong>on</strong>ey N. E. CAB<br />
<str<strong>on</strong>g>Internati<strong>on</strong>al</str<strong>on</strong>g>, Wallingford. 471-485 pp.<br />
Kamali A.R. and N.F. Childers (1970): Growth and fruiting of peach in sand culture<br />
as affected by bor<strong>on</strong> and fritted form of trace elements. J. Amer. Soc. Hort. Sci.<br />
95:652–656.<br />
Mills H.A. and J.B. J<strong>on</strong>es (1996): Plant Analysis Handbook II. MicroMacro<br />
Publishing, Inc.<br />
MI-08 0468-81: Plant analyses. Orchards. Sampling, preparati<strong>on</strong> of samples, storing of<br />
samples. Hungarian Standards Instituti<strong>on</strong>. Ministry of Agriculture. Budapest (in<br />
Hungarian)<br />
MSZ 20135:1999: De<strong>term</strong>inati<strong>on</strong> of the soluble nutrient element c<strong>on</strong>tent of the soil.<br />
Hungarian Standards Instituti<strong>on</strong>. Budapest (in Hungarian)<br />
Nagy, P. T. - G<strong>on</strong>da I. - Dremák P. and Holb I. (2006): Study <strong>on</strong> the micr<strong>on</strong>utrient<br />
c<strong>on</strong>tent of soil and leaf o fan organic apple orchard in Eastern Hungary.<br />
<str<strong>on</strong>g>Internati<strong>on</strong>al</str<strong>on</strong>g>Papp J. (2004): Fruit Producti<strong>on</strong> 1st Ed.; Mezőgazda Kiadó:<br />
Budapest, Hungary, 686 pp. (in Hungarian)<br />
Peryea F.J. (1994): Bor<strong>on</strong> nutriti<strong>on</strong> in deciduous tree fruit, p. 95–99. In: A.B. Peters<strong>on</strong><br />
and R.G. Stevens (eds.). Tree fruit nutriti<strong>on</strong>. Good Fruit Grower, Yakima, Wash.<br />
Peryea F.J. - D. Nielsen and G. Neilsen (2003): Bor<strong>on</strong> maintenance sprays for apple:<br />
Early-seas<strong>on</strong> applicati<strong>on</strong>s and tank-mixing with calcium chloride. HortScience<br />
38:542–546.<br />
Predieri, S. - R. Dris and F. Rapparini (2004): Influence of growing c<strong>on</strong>diti<strong>on</strong>s <strong>on</strong><br />
yield and quality of cherry: II. Fruit quality. Food, Agriculture & Envir<strong>on</strong>ment<br />
Vol.2 (1) : 307-309.<br />
Stiles W.C., and Reid W.S. (1966): Orchard nutriti<strong>on</strong> management. Cornell<br />
Cooperative Extensi<strong>on</strong>. Informati<strong>on</strong> Bulletin 219.<br />
Van Handel E. – J. S. Haeger and C. W. Hansen (1972): The sugars of some Florida nectars.<br />
Amer. J. Bot. 59(10): 1030-1032.<br />
154
Wills, R. B. H. - Lim, J. S. K. - Greenfield, H. (1987): Compositi<strong>on</strong> of Australian foods. 40.<br />
Temperate fruits. Food technology in Australia 39: 520-521,530.<br />
Wrolstad, R. E. - Schallenberger (1981): Free sugars and sorbitol in fruits—acompilati<strong>on</strong> from<br />
the literature. Journal of the Associati<strong>on</strong> of Official Analytical Chemists 64: 91-103.<br />
155
DRY MATTER PRODUCTIVITY AND NITROGEN UPTAKE OF PERENNIAL<br />
RYEGRASS INFLUENCED BY NITROGEN AND WATER SUPPLY<br />
ABSTRACT<br />
Imre Vágó – János Kátai – Ida Kincses – Andrea Balla Kovács<br />
University of Debrecen, Faculty of Agriculture<br />
Department of Agricultural Chemistry and Soil Science<br />
H-4015 Debrecen, P.O.Box 36.<br />
Besides the genetically coded productivity of the cropped plants, other factors also<br />
de<strong>term</strong>ine the quality and the quantity of yields. Am<strong>on</strong>g these factors the nitrogen and<br />
water supply is very important. Investigati<strong>on</strong>s were made in greenhouse pot<br />
experiments to reck<strong>on</strong> if these factors influence the effects of each other. The same<br />
nitrogen and water supply has different effect in soil types. That is why vegetati<strong>on</strong> pot<br />
experiment <strong>on</strong> three extremely different soil types of Hungary, near of Debrecen was<br />
carried out.<br />
Perennial ryegrass (Lolium perenne L.) was used as a test plant. The effect of<br />
nitrogen (0, 40, and 80 mg kg -1 ) and water (50 and 75% of the total water capacity)<br />
supply <strong>on</strong> the dry matter producti<strong>on</strong> of ryegrass was examined. The nitrogen<br />
c<strong>on</strong>centrati<strong>on</strong> of the shoot was measured and the nitrogen uptake of plants was<br />
calculated.<br />
The dry matter producti<strong>on</strong> of plants reduced during the experimental period from<br />
4.02 g per pot (first cut) to 1.78 g per pot (third cut). The reas<strong>on</strong> is the nutrient<br />
deficiency of small soil mass in the pots and the ageing of grasses. At cuts according to<br />
the mean of treatments the calcareous chernozem was the most productive, while that of<br />
marshy meadow and brown forest soils was similar to each other. The 80 mg kg -1<br />
nitrogen supply doubled the yield in the brown forest soil and calcareous black soil,<br />
while in marshy meadow soil abounding in organic matters the increase of crop mass<br />
was smaller. On the calcareous chernozem and marshy meadow soil, the water supply<br />
increased the yield but <strong>on</strong> the sand the maximum yield at the 50% water capacity was<br />
detected.<br />
During the three cuts summarized extracted nitrogen amount depending <strong>on</strong> the<br />
soil type (P=0.1%). The calcareous chernozem soil and the marshy meadow soil<br />
supplied similar (nearby 200 mg per pot) amount of nitrogen for the ryegrass. This fact<br />
can be explained that the higher shoot producti<strong>on</strong> of ryegrass <strong>on</strong> calcareous black soil<br />
and the higher nitrogen c<strong>on</strong>tent of ryegrass <strong>on</strong> marshy meadow soil equalise each other.<br />
From the brown forest soil <strong>on</strong>ly 110 mg nitrogen per pot in average was extracted by<br />
plant shoots, because of the smaller plant producti<strong>on</strong> and its lower c<strong>on</strong>centrati<strong>on</strong> of<br />
nitrogen.<br />
Keywords: Nitrogen and water supply, productivity and nitrogen uptake of ryegrass<br />
INTRODUCTION<br />
The productivity of the plants is influenced by a lot of different factors. The most<br />
important of them is the genetic potential, but the envir<strong>on</strong>mental and the technological<br />
factors also influence the quantity and the quality of the yields (Mengel, 1976; Loch and<br />
156
Nosticzius, 1992). Am<strong>on</strong>g these, soil parameters, nitrogen and water supply are the<br />
most decisive <strong>on</strong>es. The nitrogen and the water supply significantly modified the<br />
dynamics of nutrient uptake at the different crops (Borchmann, 1963). It should be<br />
noted, however, that there are significant differences in the plant species’ ability to take<br />
up the nutrients (Eichler et al., 1997; Eichler and Köppen, 2001; Eichler, 2004).<br />
The identical nitrogen and water supply has a different effect <strong>on</strong> the soil types. To<br />
investigate this phenomen<strong>on</strong>, a vegetative pot experiment was set up <strong>on</strong> three extremely<br />
different soil types from the near of Debrecen. The advantage of the pot experiment is<br />
that several factors (e.g. water supply) can be c<strong>on</strong>trolled and the same c<strong>on</strong>diti<strong>on</strong>s can be<br />
ensured for all the treatment combinati<strong>on</strong>s. In additi<strong>on</strong>, the amounts of added nutrients<br />
are exactly known due to the size of the pot and the roots of the plant interwove the<br />
whole pot.<br />
Perennial ryegrass (Lolium perenne L.) was used as a test plant. The advantage of<br />
this plant is that it endures well the c<strong>on</strong>diti<strong>on</strong>s of the greenhouse and it can be cut<br />
several times during the vegetati<strong>on</strong> period, therefore, the dynamics of the processes can<br />
be c<strong>on</strong>cluded too.<br />
In the greenhouse experiment, the effect of different nitrogen and water supply <strong>on</strong><br />
the plant’s dry matter producti<strong>on</strong> and its dynamics was examined. The nitrogen<br />
c<strong>on</strong>centrati<strong>on</strong> of the yielded shoot dry matter was measured and the amount of nitrogen<br />
extracted from the soil by the plant was calculated for the different cuts and for the<br />
whole period of the experiment.<br />
MATERIALS AND METHODS<br />
In the Table 1 are included the basic initial parameters of the three untreated soils used<br />
in the pot experiment. The initial total carb<strong>on</strong> and nitrogen c<strong>on</strong>tent of soils was<br />
measured by Nagy, P. T. (2000). Calculated for air-dry state, 2.5 kg of brown forest<br />
soil, and of chernozem soil and 2.0 kg of marshy meadow soil were put separately into<br />
plastic pots of 2.0 dm 3 volume.<br />
The standard background treatment of each experimental pots was a joint 10 cm 3<br />
soluti<strong>on</strong> of 40 mg kg -1 P2O5 and 40 mg kg -1 K2O, as pro anal. compounds of KH2PO4<br />
and K2SO4 in distilled water.<br />
The treatment combinati<strong>on</strong>s applied <strong>on</strong> the three soils are shown in Table 2. Each of<br />
them was carried out in four replicati<strong>on</strong>s to ensure the possibility of statistical<br />
evaluati<strong>on</strong>s.<br />
The transpirated and evaporated water was supplemented by irrigating the soils<br />
with distilled water. In the treatment plan given water supply level set up was ensured<br />
by daily irrigati<strong>on</strong> of the vegetati<strong>on</strong> pots to a calculated specific weight. At high<br />
vegetative mass, and also <strong>on</strong> the hot days plants were irrigated in the afterno<strong>on</strong> to avoid<br />
the threat of wilting.<br />
Plants were cut three times, first time five weeks after the sowing, the 2 nd and the 3 rd<br />
times in three weeks intervals. The grass shoot yield was first air-dried and after that<br />
dried at 60 ºC until reaching c<strong>on</strong>stant mass in a desiccator. The dry matter producti<strong>on</strong><br />
per pot was measured by analytical balance.<br />
157
Table 1: Parameters of untreated soils<br />
Soil parameter<br />
Soil site<br />
Pallag Látókép Dombos<br />
Soil type Brown forest soil Calcareous Marshy<br />
chernozem meadow soil<br />
Soil texture Sand Loam Clayes loam<br />
KA 28 46 58<br />
pH-H2O 5.38 6.13 7.60<br />
pH-KCl 4.04 5.45 7.27<br />
pH-CaCl2 4.62 5.91 6.98<br />
Hydrolytic acidity y1 8.16 7.85 -<br />
CaCO3 c<strong>on</strong>tent (%) - - 26.7<br />
Salt c<strong>on</strong>tent (%) 0.002 0.039 0.022<br />
Total C c<strong>on</strong>tent (%) 0.34 1.93 7.20<br />
Total N c<strong>on</strong>tent (%) 0.043 0.206 0.473<br />
C/N ratio 7.97 9.35 15.24<br />
Organic C (%) 0.26 0.97 1.75<br />
Organic/total C ratio 76.5 50.3 24.3<br />
0.01 M dm -3 CaCl2 extractable (mg kg -1 )<br />
NO3 - -N 0.51 4.95 3.44<br />
NH4 + -N 0.31 3.67 0.56<br />
Inorganic N 0.82 8.62 4.00<br />
Organic N 0.14 0.59 1.46<br />
Total soluble N 0.96 9.21 5.46<br />
The nitrogen c<strong>on</strong>tent of the dry matter was measured <strong>on</strong> the principle of the Kjeldahl<br />
method with a Kjel-Foss analyzer. From the mass of the shoot and the nitrogen<br />
c<strong>on</strong>centrati<strong>on</strong> of it, the amount of nitrogen extracted from the soil per cutting was<br />
de<strong>term</strong>ined. Finally, the total amount of nitrogen extracted was calculated. The obtained<br />
results were evaluated by a three-variate analysis of variance.<br />
Table 2: Combinati<strong>on</strong>s of treatments<br />
Combinati<strong>on</strong> N treatment<br />
code (mg kg -1 Water supply<br />
soil) (% of water capacity)<br />
1. 0 50<br />
2. 40 50<br />
3. 80 50<br />
4. 0 75<br />
5. 40 75<br />
6. 80 75<br />
In the analysis of variance, F and SD 5 % values were calculated for each factor and for<br />
each double and triple interacti<strong>on</strong>, then the significancy levels were de<strong>term</strong>ined.<br />
158
RESULTS AND DISCUSSIONS<br />
The dry matter producti<strong>on</strong> of the perennial ryegrass reduced with time (main average:<br />
1 st cut: 4.02; 2 nd cut 2.04; 3 rd cut: 1.78 g per pot). The reas<strong>on</strong>s for the reducti<strong>on</strong> of dry<br />
matter producti<strong>on</strong> are the nutrient-deficiency originating from low soil mass and the<br />
ageing of grasses. The nutrient supply of soils heavily influenced the dry matter<br />
producti<strong>on</strong> of plants at all cuts. In the average of treatments, the calcareous chernozem<br />
soil was the most productive (significant at P = 0.1 % level), while the productivity of<br />
the brown forest and the marshy meadow soil was similar (Table 3).<br />
The dry matter producti<strong>on</strong> increase due to the nitrogen treatment varied for the different<br />
soils. This is supported by the fact, that the interacti<strong>on</strong> of factors A*B (N dose*soil<br />
type) is highly significant (mostly P = 0,1 %). The applicati<strong>on</strong> of 80 mg kg -1 nitrogen<br />
doubled the yield <strong>on</strong> the brown forest and the calcareous chernozem soil, while <strong>on</strong> the<br />
marshy meadow soil with high organic matter c<strong>on</strong>tent the increase was also significant,<br />
but smaller (30 %).<br />
Table 3: Dry matter producti<strong>on</strong> of ryegrass (g pot -1 ) Sum of 1 st , 2 nd and 3 rd cuts<br />
W.c. (%) Soil site N supply (mg kg -1 ) (A)<br />
(C) (B) 0 40 80 Average<br />
Pallag 4.38 7.43 10.53 7.44<br />
50 Látókép 6.92 8.75 11.57 9.08<br />
Dombos 5.10 6.60 7.27 6.32<br />
Average 5.47 7.59 9.79 7.62<br />
Pallag 4.40 5.80 6.73 5.64<br />
75 Látókép 7.33 10.82 14.43 10.86<br />
Dombos 6.48 7.70 8.85 7.68<br />
Average 6.07 8.11 10.00 8.06<br />
Pallag 4.39 6.61 8.63 6.54<br />
Average Látókép 7.13 9.79 13.00 9.97<br />
Dombos 5.79 7.15 8.06 7.00<br />
Average 5.77 7.85 9.90 7.84<br />
Table of variance<br />
Factor FG MQ F SD 5%<br />
A 2 102.30 117.6 *** 0.54<br />
B 2 83.18 95.6 *** 0.54<br />
A*B 4 6.68 7.7 *** 0.94<br />
C 1 3.51 4.0 * 0.44<br />
A*C 2 0.26 0.3 0.77<br />
B*C 2 22.88 26.3 *** 0.77<br />
A*B*C 4 5.18 6.0 *** 1.33<br />
Error 51 0.87<br />
*** Significant at P=0.1 % level<br />
* Significant at P=5.0 % level<br />
The higher level of water supply increased the perennial ryegrass dry matter producti<strong>on</strong><br />
at every cut. The significance of the B*C (soil*water capacity) interacti<strong>on</strong> (P=0,1 %)<br />
159
proves that the highest yield can be achieved by 75 % water supply <strong>on</strong> calcareous<br />
chernozem and marshy meadow soil and by 50 % water supply level <strong>on</strong> brown forest<br />
soil. It can also be stated that the effect of nitrogen fertilizati<strong>on</strong> was not modified by the<br />
different water supply. The positive effect of nitrogen treatments was detectable in the<br />
dry matter producti<strong>on</strong> of plants at each cuts (significant at P = 0.1 % level).<br />
The nitrogen c<strong>on</strong>tent of plants is the highest (2.81 %) <strong>on</strong> marshy meadow soil<br />
(P = 0.1 %), while <strong>on</strong> brown forest soil and calcareous chernozem the nitrogen c<strong>on</strong>tent<br />
of ryegrass was lower, 1.34 % and 1.91 %, respectively (Table 4). The tendency is in<br />
accordance with the original total nitrogen c<strong>on</strong>tents of the experiment soils (Table 1).<br />
The higher water supply decreased the nitrogen c<strong>on</strong>tent of plants in almost every<br />
treatment and soil combinati<strong>on</strong> (P = 1.0 % or 5.0 %).<br />
In the mean of the experiment, the amount of nitrogen extracted per pot reduced with<br />
time (1st cut: 87.7; 2 nd cut 42.8 and 3 rd cut: 37.2 mg N per pot). The total amount of<br />
nitrogen extracted by the three cuts is also c<strong>on</strong>siderable with 167.7 mg per pot. The<br />
decrease in the amount of nitrogen extracted can be explained by the gradually<br />
decreasing yield.<br />
Table 4: Nitrogen c<strong>on</strong>tent (%) of the ryegrass 1 st cut<br />
W.c. (%) Soil site N supply (mg kg -1 ) (A)<br />
(C) (B) 0 40 80 Average<br />
Pallag 1.13 1.27 1.83 1.41<br />
50 Látókép 1.38 2.00 2.52 1.97<br />
Dombos 1.85 2.90 3.72 2.83<br />
Average 1.45 2.06 2.69 2.07<br />
Pallag 1.00 1.27 1.52 1.27<br />
75 Látókép 1.35 1.78 2.45 1.86<br />
Dombos 2.10 2.75 3.55 2.80<br />
Average 1.48 1.93 2.51 1.98<br />
Pallag 1.06 1.27 1.67 1.34<br />
Average Látókép 1.36 1.89 2.49 1.91<br />
Dombos 1.97 2.83 3.64 2.81<br />
Average 1.47 2.00 2.60 2.02<br />
Table of variance<br />
Factor FG MQ F SD 5%<br />
A 2 7.72 291.5 *** 0.09<br />
B 2 13.27 501.0 *** 0.09<br />
A*B 4 0.56 21.1 *** 0.16<br />
C 1 0.15 5.71 * 0.08<br />
A*C 2 0.08 2.85 + 0.13<br />
B*C 2 0.02 0.82 0.13<br />
A*B*C 4 0.05 1.99 0.23<br />
Error 51 0.03<br />
*** Significant at P=0.1 % level; * Significant at P=5.0 % level<br />
+ Significant at P=10.0 % level<br />
160
The amount of nitrogen uptake by the plants corresp<strong>on</strong>ds with the increasing nitrogen<br />
supply. The amount of the nitrogen extracted from the soils was 97.2; 159.4 and<br />
246.5 mg N per pot in the average of c<strong>on</strong>trol treatments, at 40 mg kg -1 and at 80 mg kg -1<br />
nitrogen doses, respectively (Table 5).<br />
The amount of nitrogen extracted was dependent <strong>on</strong> the soil (P = 0,1 %). The<br />
calcareous chernozem and the marshy meadow soil supplied the grass with<br />
approximately the same amount of nitrogen, which can be explained by the fact that the<br />
higher grass yield <strong>on</strong> the chernozem soil and the higher nitrogen c<strong>on</strong>tent of plants <strong>on</strong><br />
marshy meadow soil offset each other. However, the ryegrass plants extracted <strong>on</strong>ly 110<br />
mg nitrogen per pot from the brown forest soil.<br />
Table 5: Nitrogen uptake of ryegrass (g pot -1 ) Sum of 1 st , 2 nd and 3 rd cuts<br />
W.c. (%) Soil site N supply (mg kg -1 ) (A)<br />
(C) (B) 0 40 80 Average<br />
Pallag 61.6 113.3 206.2 127.0<br />
50 Látókép 113.8 169.4 276.4 186.5<br />
Dombos 112.8 189.7 248.9 183.8<br />
Average 96.1 157.5 243.8 165.8<br />
Pallag 60.0 95.7 125.0 93.6<br />
75 Látókép 98.7 186.2 330.5 205.1<br />
Dombos 136.4 202.2 292.1 210.2<br />
Average 98.3 161.4 249.2 169.9<br />
Pallag 60.8 104.5 165.6 110.3<br />
Average Látókép 106.3 177.8 303.4 195.8<br />
Dombos 124.6 196.0 270.5 197.0<br />
Average 97.2 159.4 246.5 167.7<br />
Table of variance<br />
Factor FG MQ F SD 5%<br />
A 2 134992.9 295.5 *** 12.40<br />
B 2 59348.8 129.9 *** 12.40<br />
A*B 4 4742.3 10.4 *** 21.48<br />
C 1 265.3 0.6 10.13<br />
A*C 2 14.1 0.0 17.54<br />
B*C 2 6361.1 13.9 *** 17.54<br />
A*B*C 4 3207.4 7.0 *** 30.38<br />
Error 51 344.0<br />
*** Significant at P=0.1 % level<br />
In spite of the fact, that water dose had an effect both <strong>on</strong> yield quantity and the nitrogen<br />
c<strong>on</strong>tent of plants, the level of water supply did not influence the amount of nitrogen<br />
uptake significantly. The reas<strong>on</strong> for this was that while yield increased paralelly with<br />
the water supply, the nitrogen c<strong>on</strong>tent decreased. These two counter-effects balanced<br />
each other in the nitrogen uptake of plants.<br />
161
ACKNOWLEDGEMENTS<br />
The research work was carried out with the financial support of the Hungarian Scientific<br />
Research Fund (OTKA T 032343).<br />
REFERENCES<br />
Borchmann, W. (1963): Über die Abhängigkeit des Mineralstoffgehaltes verschiedener<br />
Futterpflanzen v<strong>on</strong> der Höhe der Wasserversorgung. III. Zeitschrift für<br />
Landwirtsch. Versuchs- und Untersuchungswesen, 9: 173-198. p.<br />
Eichler, B. (2004): Möglichkeiten zur Einflussnahme auf Phosphorkreisläufe für die<br />
Gestaltung nachhaltiger Bodennutzungssysteme. Habilitati<strong>on</strong>sschrift. Rostock.<br />
Eichler, B. – Köppen, D. (2001): Die Nutzung ausgewählter Zwischenfrüchte zur<br />
Verringerung v<strong>on</strong> Phosphatverlusten in der Landwirtschaft. Mitt. Gesellschaft für<br />
Pflanzenbauwiss., 13: 166-167 p.<br />
Eichler, B. – Köppen, D. – Grüner, A. (1997): Nutzung v<strong>on</strong> Phosphorvorräten durch<br />
Zwischenfrüchte im Gefäßversuch. 41. Jahrestagung vom 24.-27. 09. 1997 in Kiel,<br />
Tagungsband S. 309<br />
Loch J. – Nosticzius Á. (1992): Agrokémia és növényvédelmi kémia. Mezőgazda<br />
Kiadó, Budapest<br />
Mengel, K. (1972): Ernährung und Stoffwechsel der Pflanze. VEB Gustav Fischer<br />
Verlag, Jena<br />
Nagy, P.T. (2000): Égetéses elven működő elemanalizátor alkalmazhatósága talaj- és<br />
növényvizsgálatokhoz. Agrokémia és Talajtan, 49: 521-534 p.<br />
162
ABSTRACT<br />
THE EFFECT OF NITROGEN SUPPLY, LIMING AND NICKEL<br />
CONTAMINATION ON THE PRODUCTIVITY OF RYEGRASS<br />
(Lolium perenne L.) IN A GREENHOUSE EXPERIMENT<br />
Marianna Sipos, Imre Vágó<br />
University of Debrecen, Faculty of Agriculture<br />
Department of Agricultural Chemistry and Soil Science<br />
H-4015 Debrecen, P.O.Box 36.<br />
It is well known that the dry matter producti<strong>on</strong> of plants is de<strong>term</strong>ined by the nutrient<br />
and water supply and calcium carb<strong>on</strong>ate c<strong>on</strong>tent of soils and the amount of toxic<br />
materials (besides their genetic potential). However, it is not known how the<br />
anthropogenic polluti<strong>on</strong>s (e. g. the heavy metal i<strong>on</strong> c<strong>on</strong>tent of sewage) modify the effect<br />
of these factors.<br />
For studying this phenomen<strong>on</strong>, experiments were carried out <strong>on</strong> two extremely<br />
different soils (leached calcareous chernozem with good fertility and acidic shifting<br />
sand soil with low buffering capacity and humus c<strong>on</strong>tent). In the pot experiment, the<br />
effect of four factors (soil, nitrogen supply, nickel loading and CaCO3 treatment) <strong>on</strong> the<br />
dry matter producti<strong>on</strong> of perennial ryegrass (Lolium perenne L.) was studied.<br />
In the experiment, the dry matter yield of plants was measured in the different<br />
treatments and treatment combinati<strong>on</strong>s.<br />
On chernozem soil, nitrogen had the greatest effect <strong>on</strong> yield. In the sum of cuts,<br />
lime had a yield-increasing effect, while nickel had no significant effect <strong>on</strong> yielding<br />
capacity.<br />
On shifting sand soil significant differences were found in the effect of treatments<br />
<strong>on</strong> yield: nitrogen and lime, and nickel i<strong>on</strong> have exerted their positive and negative<br />
effect, respectively; under high Ni-loading chlorotic symptoms could be observed<br />
visually <strong>on</strong> plants. The yield-reducing effect of nickel was striking mainly in the<br />
treatments without lime, while in limed variants the yield-depressing effect of Ni<br />
c<strong>on</strong>taminati<strong>on</strong> was more moderate.<br />
Calcium carb<strong>on</strong>ate had a str<strong>on</strong>ger influencing effect <strong>on</strong> dry matter formati<strong>on</strong> than<br />
nitrogen treatments. These results indicate that improving the pH value of acidic soils is<br />
important since in additi<strong>on</strong> to improving yielding-capacity, it also limits the depressing<br />
effect of toxic elements.<br />
Keywords: Nitrogen, calcium carb<strong>on</strong>ate, nickel c<strong>on</strong>taminati<strong>on</strong>, productivity of ryegrass<br />
INTRODUCTION<br />
Nowadays, envir<strong>on</strong>mental c<strong>on</strong>siderati<strong>on</strong>s, study of envir<strong>on</strong>mental polluti<strong>on</strong> and<br />
mitigati<strong>on</strong> of damages are becoming increasingly important. Anthropogenic polluti<strong>on</strong><br />
represents a serious burden for the living and n<strong>on</strong>-living envir<strong>on</strong>ment. It might happen<br />
that our striving to protect the envir<strong>on</strong>ment and reduce the polluti<strong>on</strong> will not be efficient<br />
enough without satisfactory knowledge <strong>on</strong> the sources of danger. A good example of<br />
this is the heavy metal polluti<strong>on</strong> from sewage sludge applicati<strong>on</strong>s, which can limit the<br />
favourable yield-increasing effect of sewage or can even reduce yields.<br />
163
Fertilizers, liming materials and manure have a low nickel c<strong>on</strong>tent, therefore, it is not<br />
probable that they would cause serious nickel c<strong>on</strong>taminati<strong>on</strong> (Sim<strong>on</strong>, 1999).<br />
In a low exterior c<strong>on</strong>centrati<strong>on</strong>, nickel has a favourable effect <strong>on</strong> the germinati<strong>on</strong><br />
of plant germinati<strong>on</strong> and growth. Nickel has <strong>on</strong>ly <strong>on</strong>e physiological functi<strong>on</strong> in plant<br />
life: it has a role in building the urease enzyme and its functi<strong>on</strong>ing. For the functi<strong>on</strong>ing<br />
of the enzyme, 6-8 Ni 2+ i<strong>on</strong>s are necessary per molecule, therefore, it is an essential<br />
element for plants (Láng, 1998).<br />
Luckily, nickel c<strong>on</strong>tent is so low in most soils, that no toxic effect should be<br />
expected: the average Ni 2+ c<strong>on</strong>centrati<strong>on</strong> in n<strong>on</strong>-c<strong>on</strong>taminated soils is between 5 and 50<br />
mg kg -1 (Stefanovits et al., 1999). However, in case of its accumulati<strong>on</strong>, it hinders<br />
growth and can result in chlorosis or necrosis. It influences the uptake of i<strong>on</strong>s with two<br />
positive chargings, primarily that of ir<strong>on</strong> and it hinders the translocati<strong>on</strong> of ir<strong>on</strong> into the<br />
shoot. This has a disadvantageous effect <strong>on</strong> the water management of the plant and<br />
results in an oxidative stress. The human health risks of nickel are significant, it was<br />
proven that nickel accumulating in the human body has a primary carcinogenic effect.<br />
In the 1950s, coating of seeds with heavy metal compounds was a comm<strong>on</strong> practice also<br />
in Hungary. In large-scale companies, coating with NiCl2 soluti<strong>on</strong> was also used am<strong>on</strong>g<br />
others against rust and smut diseases of wheat. The improper use of the remaining<br />
soluti<strong>on</strong>s at the place of coating resulted in punctual c<strong>on</strong>taminati<strong>on</strong>s (Csathó, 1994).<br />
In recent decades, it became a general practice to use sewage for fertilizati<strong>on</strong> after<br />
composting or directly. By the applicati<strong>on</strong> of sewage, valuable nutrients are provided<br />
for soils (Balláné and Filep, 2000).<br />
However, large amounts of heavy metal can enter the soil with the applicati<strong>on</strong> of<br />
sewage, even if the c<strong>on</strong>centrati<strong>on</strong> of toxic elements is under the official limit since high<br />
volumes are applied. Although the amount of toxic elements that can be applied to the<br />
soil is low: As 0,3; Cd 0,02; Cr and Cu 10-10; Hg 0,1; Mn 20; Ni 2; Pb and Zn 20-20<br />
kg ha -1 , the danger of accidental overdosing and the fact that repeating applicati<strong>on</strong>s<br />
result in accumulati<strong>on</strong> of pollutants in the soil should be c<strong>on</strong>sidered.<br />
A significant part of the c<strong>on</strong>taminati<strong>on</strong> in the soil fortunately becomes immobile<br />
and <strong>on</strong>ly a fracti<strong>on</strong> of it enters the plants. The heavy metal binding capacity of the soils<br />
varies greatly. It should also be c<strong>on</strong>sidered that envir<strong>on</strong>mental and producti<strong>on</strong> factors<br />
(e.g. nutrient supply, changes in soil pH, precipitati<strong>on</strong>) can modify the mobility of<br />
nutrient elements (Kincses et al., 2005) and pollutants.<br />
All the above facts indicate that it is important to study the effects and interacti<strong>on</strong>s<br />
of factors influencing nickel-uptake with special regards to plant yields.<br />
MATERIALS AND METHODS<br />
The objective of the experiment was to reveal the interacti<strong>on</strong>s between and direct effects<br />
of nickel c<strong>on</strong>taminati<strong>on</strong> and factors having a favourable effect <strong>on</strong> plant development<br />
(fertilizati<strong>on</strong>, CaCO3 applicati<strong>on</strong>).<br />
For studying the interacti<strong>on</strong> between nickel c<strong>on</strong>taminati<strong>on</strong>, nitrogen supply and<br />
CaCO3 applicati<strong>on</strong>, a pot experiment was set up at the Department of Agricultural<br />
Chemistry in May 2006. The advantage of the pot experiment is that several factors<br />
(e.g. water supply) can be c<strong>on</strong>trolled and the same c<strong>on</strong>diti<strong>on</strong>s can be ensured for all the<br />
treatment combinati<strong>on</strong>s. In additi<strong>on</strong>, the amount of nutrients, pollutants and CaCO3 are<br />
164
exactly known due to the size of the pot and the roots of the plant interwove the whole<br />
pot. Perennial ryegrass (Lolium perenne L.) was used as a test plant. The advantage of<br />
this plant is that it endures well the c<strong>on</strong>diti<strong>on</strong>s of the greenhouse and it can be cut<br />
several times during the vegetati<strong>on</strong> period, therefore, the dynamics of the processes can<br />
be c<strong>on</strong>cluded too.<br />
The effect of four factors (soil, nitrogen supply, calcium carb<strong>on</strong>ate applicati<strong>on</strong> and<br />
nickel loading) <strong>on</strong> plants was studied.<br />
As it is already known, the basic soil characteristics such as soil type, humus<br />
c<strong>on</strong>tent, pH etc. have a major effect <strong>on</strong> plant development. Accordingly, the experiment<br />
was carried out <strong>on</strong> two extremely different soils. One of the soils was calcareous<br />
chernozem with high fertility and buffering capacity from Debrecen-Látókép, while the<br />
other was an acidic sandy soil with weak fertility from Újfehértó.<br />
The basic parameters of the untreated soils used in the greenhouse experiment are<br />
included in Table 1. Calculated for air-dry state, 1.0 kg of each soil was put separately<br />
into the vegetati<strong>on</strong> pots.<br />
Table 1: Parameters of untreated soils<br />
Soil site Debrecen-Látókép Újfehértó<br />
Soil type Calcareous chernozem Shifting sand<br />
Soil texture Loam Sand<br />
Hygroscopicity acc. to Arany (KA) 39 26<br />
pH-H2O 6.05 5.01<br />
pH-KCl 5.41 3.98<br />
pH-CaCl2 5.73 4.19<br />
Hydrolytic acidity (y1) 9.07 7.06<br />
Total C c<strong>on</strong>tent (%) 1.89 0.27<br />
Mass volume (kg dm -3 ) 1.19 1.49<br />
Calculated lime requirement (t ha -1 ) 6.16 3.19<br />
Calculated lime requirement (g kg -1 ) 2.07 0.86<br />
The initial total carb<strong>on</strong> c<strong>on</strong>tent of soils was measured by Nagy, P. T. (2000). The<br />
standard treatment of pots was a joint soluti<strong>on</strong> of 100 mg kg -1 P and 160 mg kg -1 K, in<br />
10 cm 3 KH2PO4 and K2SO4. The combinati<strong>on</strong>s of treatments applied <strong>on</strong> the soils are<br />
shown in Table 2.<br />
Nitrogen and nickel were added to the soils as NH4NO3 and NiSO4 soluti<strong>on</strong>,<br />
respectively. Lime requirement of the soils was calculated according to the method of<br />
Filep (1988) as<br />
CaCO3 (kg ha -1 ) = 17.4 * KA * y1<br />
then was added to the soil as fine powder. Each treatment was carried out in four<br />
replicati<strong>on</strong>s.<br />
The water supply was ensured by daily irrigati<strong>on</strong> to a specific weight. The<br />
transpirated and evaporated water was supplemented by irrigating the chernozem and<br />
sandy soil with distilled water to 75% and 60% of the field water capacity, respectively.<br />
At high vegetative mass, plants were irrigated in the afterno<strong>on</strong> also <strong>on</strong> hot days to avoid<br />
wilting.<br />
165
Table 2: Combinati<strong>on</strong>s of treatments<br />
Number of<br />
combinati<strong>on</strong><br />
N treatment<br />
(mg kg -1 )<br />
Ni treatment<br />
(mg kg -1 )<br />
CaCO3<br />
treatment code (!)<br />
1. 80 0 0<br />
2. 80 50 0<br />
3. 80 100 0<br />
4. 80 0 1<br />
5. 80 50 1<br />
6. 80 100 1<br />
7. 160 0 0<br />
8. 160 50 0<br />
9. 160 100 0<br />
10. 160 0 1<br />
11. 160 50 1<br />
12. 160 100 1<br />
(!) 0 = without CaCO3 1 = CaCO3 dose according to calculated lime requirement<br />
Plants were cut two times, 33 and 61 days after sowing, the grass yield was dried at<br />
60ºC until reaching c<strong>on</strong>stant mass in a desiccator and the dry matter producti<strong>on</strong> per pot<br />
was measured by analytical balance.<br />
RESULTS AND DISCUSSIONS<br />
The obtained results were evaluated by a three-variate analysis of variance <strong>on</strong> the basis<br />
of Sváb’s method (1981). In the analysis of variance, F and SD 5 % values were<br />
calculated for each factor and for each double and triple interacti<strong>on</strong>, then the<br />
significance levels were de<strong>term</strong>ined.<br />
Summarized yields of the two cuts and their statistical evaluati<strong>on</strong> <strong>on</strong> chernozem<br />
and sandy soils are shown in Tables 3 and 4.<br />
On the chernozem soil at Látókép, it can be stated when c<strong>on</strong>sidering the sum of dry<br />
matter producti<strong>on</strong> of two cuts that nitrogen had a significant effect <strong>on</strong> yield (P = 0.1 %).<br />
Nitrogen supply had a different effect in the different lime and nickel combinati<strong>on</strong>s.<br />
This is indicated by the fact that the N*Ni*CaCO3 triple interacti<strong>on</strong> was significant at<br />
P = 5.0 % level, meaning that factors (in our case nitrogen) exert their effect depending<br />
up<strong>on</strong> the other factors.<br />
In combinati<strong>on</strong>s not treated with calcium carb<strong>on</strong>ate, the yield-increasing effect of<br />
nitrogen could be observed at all three nickel dosages, however this effect was<br />
decreasing with increasing nickel dosages. Whereas in limed treatments, the increasing<br />
nickel dosages do not limit the yield-increasing effect of nitrogen.<br />
The yield-reducing effect of Ni is not significant <strong>on</strong> this soil in the sum of the two cuts<br />
as c<strong>on</strong>firmed by the statistical analysis.<br />
166
Table 3: Dry matter producti<strong>on</strong> of perennial ryegrass (g pot -1 )<br />
Látókép chernozem, 1st+2nd cut<br />
Factors Unlimed Limed Average<br />
N=80<br />
Ni=0 7.53 8.02 7.77<br />
Ni=50 7.65 7.55 7.60<br />
Ni=100 7.68 7.68 7.68<br />
Average 7.62 7.75 7.68<br />
N=160<br />
Ni=0 10.20 9.18 9.69<br />
Ni=50 9.93 10.05 9.99<br />
Ni=100 9.67 9.60 9.64<br />
Average 9.93 9.61 9.77<br />
N average<br />
Ni=0 8.86 8.60 8.73<br />
Ni=50 8.79 8.80 8.79<br />
Ni=100 8.67 8.64 8.66<br />
Average 8.78 8.68 8.73<br />
Table of variance<br />
Factor MQ DF F SD5%<br />
N 52.292 1 234.9 *** 0.28<br />
Ni 0.076 2 0.34 0.38<br />
N*Ni 0.273 2 1.22 0.48<br />
CaCO3 0.11 1 0.49 0.28<br />
N*CaCO3 0.63 1 2.83 0.398<br />
Ni*CaCO3 0.86 2 0.39 0.48<br />
N*Ni*CaCO3 0.876 2 3.94 * 0.68<br />
*** Significant at P=0.1 % level * Significant at P=5.0 % level<br />
No c<strong>on</strong>siderable differences were found in dry matter yield of the first and sec<strong>on</strong>d cuts.<br />
This suggests that the amount of nutrients applied to 1 kg chernozem soil and the<br />
natural nutrient-supplying capacity of the soil were satisfactory for plant development.<br />
Regarding the summarized yield of the two cuts <strong>on</strong> shifting sandy soil, it can be stated<br />
that all three treatment factors had a significant effect.<br />
Increasing nitrogen dosages increase yield significantly as proven by the statistical<br />
analysis (P = 1.0 %). However, the increasing effect was different at different lime<br />
dosages. Without lime treatment, higher nitrogen doses resulted in a smaller yield<br />
167
increment, than with higher calcium carb<strong>on</strong>ate dosaging. It has to be noted also, that<br />
lime also had a positive effect <strong>on</strong> yield and it was more significant (P = 0.1 %) than that<br />
of nitrogen.<br />
Table 4: Dry matter producti<strong>on</strong> of perennial ryegrass (g pot -1 )<br />
Újfehértó shifting sand, 1st+2nd cuts<br />
Factors<br />
N=80<br />
Unlimed Limed Average<br />
Ni=0 4.73 5.18 4.95<br />
Ni=50 3.38 4.90 4.14<br />
Ni=100 1.65 4.90 3.28<br />
Average<br />
N=160<br />
3.25 4.99 4.12<br />
Ni=0 5.85 6.63 6.24<br />
Ni=50 4.85 7.05 5.95<br />
Ni=100 0.88 5.80 3.34<br />
Average<br />
N average<br />
3.86 6.49 5.17<br />
Ni=0 5.29 5.90 5.59<br />
Ni=50 4.11 5.97 5.04<br />
Ni=100 1.26 5.35 3.31<br />
Average<br />
Table of variance<br />
3.55 5.74 4.65<br />
Factor MQ DF F SD5%<br />
N 13.335 1 11.77 ** 0.62<br />
Ni 22.811 2 20.14 *** 0.76<br />
N*Ni 3.226 2 2.85 + 1.08<br />
CaCO3 57.422 1 50.67 *** 0.62<br />
N*CaCO3 2.385 1 2.11 0.88<br />
Ni*CaCO3 12.393 2 10.94 *** 1.08<br />
N*Ni*CaCO3 0.491 2 0.43 1.53<br />
*** Significant at P=0.1 % level<br />
** Significant at P=1.0 % level<br />
+ Significant at P=10 % level<br />
The yield-increasing effect of nitrogen had a significant interacti<strong>on</strong> with nickel dosaging<br />
at P = 10 % level. In treatments without nickel, nitrogen had a yield-increasing effect<br />
both in limed and unlimed treatments. In the case of nickel treatment however, the<br />
yield-reducing and yield-increasing effect of nickel and nitrogen, respectively, were<br />
168
more definite in the n<strong>on</strong> limed treatments. Under calcium carb<strong>on</strong>ate treatment, a<br />
significant yield-increment could be observed in all treatment combinati<strong>on</strong>s.<br />
Nickel c<strong>on</strong>taminati<strong>on</strong> had a significant yield-reducing effect in the sum of the two cuts<br />
at P = 0.1 %. In unlimed treatments, a dosage of 100 mg kg -1 Ni had a str<strong>on</strong>ger reducing<br />
effect than the smaller dosage. Data of the table also indicate that in the average of N<br />
treatments, the harmful effect of nickel was str<strong>on</strong>g in the unlimed treatments (yieldreducti<strong>on</strong><br />
from 5.29 g pot -1 to 1.26 g pot -1 ), while in the limed treatments, the effect was<br />
much weaker (yield-reducti<strong>on</strong> from 5.90 g pot -1 to 5.35 g pot -1 ).<br />
C<strong>on</strong>sidering the effect of N treatment, it can be observed that yield-reducti<strong>on</strong> was<br />
smaller both in limed and unlimed treatments when smaller N dosage was applied than<br />
at higher dosages. This is probably due to the fact that plants used more nutrients and<br />
c<strong>on</strong>sequently they took up more Ni for producing higher yields.<br />
CaCO3 treatment had a significant effect <strong>on</strong> yield in all treatment combinati<strong>on</strong>s. It<br />
can be stated that as a result of the favourable soil pH casued by liming, the dry matter<br />
producti<strong>on</strong> per pot increases. The ratio of this increment is higher than that of the yieldincreasing<br />
effect of nitrogen.<br />
By adding lime, the yield-reducing effect of nickel was reduced or even prevented.<br />
This c<strong>on</strong>clusi<strong>on</strong> is supported by the fact that the significance level of Ni*CaCO3<br />
interacti<strong>on</strong> <strong>on</strong> shifting sand was P = 0.1 %.<br />
As a c<strong>on</strong>tinuati<strong>on</strong> of the study, we plan to de<strong>term</strong>ine the c<strong>on</strong>stituti<strong>on</strong> of elements in<br />
the dry matter of perennial ryegrass. This would enable us to de<strong>term</strong>ine how treatments<br />
influenced the uptake of elements by plants. This knowledge can lead to further<br />
c<strong>on</strong>clusi<strong>on</strong>s.<br />
ACKNOWLEDGEMENTS<br />
The research work was carried out by the support of “Tormay Béla” H<strong>on</strong>our Society of<br />
the University of Debrecen, Centre of Agricultural Sciences.<br />
REFERENCES<br />
Balláné Kovács, A. - Filep, T. (2000): A különböző SO4 2- /SO4 2- +Cl - arányú K-, Ca-,<br />
Mg-só adagok hatása az angolperje tápelem összetételére. Agrokémia és Talajtan<br />
49: 3-4 p.<br />
Csathó, P. (1994): A környezet nehézfém-szennyezettsége és az agrár<strong>term</strong>elés. MTA<br />
TAKI, Budapest.<br />
Filep, Gy. (1988): Talajkémia. Akadémiai Kiadó, Budapest<br />
Kincses, S.-né – Nagy, P. T. – Balláné Kovács, A. – Filep, T. (2005): Az NPKtrágyázás<br />
hatása a búza (Triticum aestivum) és a kukorica (Zea mays) Znfelvételére.<br />
Acta Agr<strong>on</strong>omica Óváriensis, 47: 239-252 p.<br />
Láng, F. (1998): Növényélettan. ELTE Eötvös Kiadó, Budapest<br />
Nagy, P.T. (2000): Égetéses elven működő elemanalizátor alkalmazhatósága talaj- és<br />
növényvizsgálatokhoz. Agrokémia és Talajtan, 49: 521-534 p.<br />
Sim<strong>on</strong>, L. (1999): Talajszennyeződés, talajtisztítás. Környezetgazdálkodási Intézet<br />
Környezet – és Természetvédelmi Szakkönyvtár és Információs Közp<strong>on</strong>t,<br />
Budapest<br />
Sváb J. (1981): Biometriai módszerek a kutatásban. Mezőgazdasági Kiadó, Budapest<br />
169
TOXIGENIC FUNGI AND MYCOTOXINS IN GRAINS AND BAKERY<br />
PRODUCTS FROM ROMANIA<br />
Carmen Puia, Viorel Florian, Alexandra Suciu, Augusta Lujerdean*, Dana Pusta*<br />
Department of Plant Protecti<strong>on</strong>, *Department of Chemistry, University of Agricultural<br />
Sciences and Veterinary Medicine, Cluj Napoca, Romania<br />
ABSTRACT<br />
Our researches want to establish the mycoflora and the natural occurrence of<br />
mycotoxins in grains for industrializati<strong>on</strong> and fodder, bread and bakery products in our<br />
country. We intend to compare these levels with the internati<strong>on</strong>al <strong>on</strong>es and to change<br />
the legal level of mycotoxins admitted by our laws and to align these levels with the<br />
European legislati<strong>on</strong>. The authors reported the identificati<strong>on</strong> of mycoflora in 130<br />
samples of grains, bread, biscuits and cereal flakes from different districts of Romania.<br />
Moulds evaluati<strong>on</strong> was de<strong>term</strong>ined using c<strong>on</strong>venti<strong>on</strong>al methods as blotting test and<br />
Ulster test. The predominant genera were those of toxigenic fungi: Penicillium,<br />
Aspergillus and Fusarium.<br />
Our studies were focused <strong>on</strong> finding rapid methods for screening of aflatoxins B1,<br />
(AB1), aflatoxin B2 (AB2), aflatoxin G1 (AG1), aflatoxin G2 (AG2) and ochratoxin A.<br />
Mycotoxins were extracted in chloroform, separated <strong>on</strong> silicagel thin-layer<br />
chromatography plates and quantificated using densitometric analysis. There were<br />
analyzed 39 cereal samples (wheat and maize) from different Romanian districts.<br />
The c<strong>on</strong>taminati<strong>on</strong> range was: aflatoxin B1: 1.7 - 5.7 µg/kg, aflatoxin B2: 0.02 -2.8<br />
µg/kg, aflatoxin G1: 1.1 – 5.7 µg/kg, aflatoxin G2: 0.12 – 1.8 µg/kg, total aflatoxins: 1.2<br />
– 10.8 µg/kg and ochratoxin A: 4.4 - 30.0 µg/kg. The higher number of c<strong>on</strong>taminati<strong>on</strong><br />
rate was in the case of wheat samples. The mycotoxin analyses reveal to us a high and<br />
dangerous level of ochratoxin A and total aflatoxins. Our food and fodder are dangerous<br />
and our legislati<strong>on</strong> is way too permissive. We have to align our legislati<strong>on</strong> with the<br />
European <strong>on</strong>e to take care about our grains.<br />
Key words: mycoflora, toxigenic fungi, grains, bakery products, mycotoxins<br />
INTRODUCTION<br />
Man has l<strong>on</strong>g benefited from the nutriti<strong>on</strong>al properties of certain microscopic fungi,<br />
such as ferments and yeasts used in making cheese, beer, bread, vinegar and yogurt. But<br />
some moulds of fungal origin are potent pois<strong>on</strong>s, especially the mycotoxin family that<br />
has been the subject of increased attenti<strong>on</strong> over the past 15 years.<br />
Most fungi produce a class of chemical compounds called sec<strong>on</strong>dary metabolites.<br />
These compounds have a wide range of biological activities including antibiotic<br />
(antibacterial and antifungal), acute and chr<strong>on</strong>ic toxicities (plant, animal, and humans),<br />
horm<strong>on</strong>es and growth regulati<strong>on</strong> (plants and animals). Several species of fungi that<br />
col<strong>on</strong>ize grains produce mycotoxins, however not all isolates of a toxigenic species<br />
produce mycotoxins and isolates capable of producing a mycotoxin do not always<br />
synthesize the toxin (Mills, 1990).<br />
Our researches, want to establish the mycoflora and the natural occurrence of<br />
mycotoxins in grains for industrializati<strong>on</strong> and fodder in our country. We intend to<br />
170
compare these levels with the European <strong>on</strong>es and to change the legal level of<br />
mycotoxins admitted by our laws and to align these levels with the European<br />
legislati<strong>on</strong>.<br />
MATERIALS AND METHODS<br />
Different random samples of grains (wheat, corn, Triticale and rye), bread and bakery<br />
products (biscuits, cereal flakes) were collected from 14 districts of Romania. Each<br />
sample (1 kilo) was put in sterilized bag and in laboratory was divided into 2 parts. One<br />
part was used for mycological analysis and the sec<strong>on</strong>d <strong>on</strong>e was used for mycotoxin<br />
analysis. The mycological analysis was made in 5 replicati<strong>on</strong>s, by using the blotting<br />
paper method (Raicu Cristina and Doina Baciu, 1978) and the Ulster method <strong>on</strong><br />
agarized plates (Hulea Ana and all, 1982). We used 10 grains from each sample in 3 or<br />
5 replicati<strong>on</strong>s. In this study was employed a Czapek - Dox agar medium with<br />
Streptomycin as bacteriostatic agent, added after sterilizati<strong>on</strong> (Hulea Ana, 1969). The<br />
identificati<strong>on</strong> of fungi was according to Raicu Cristina and Doina Baciu (1978) and<br />
Domsch and Gams (1972).<br />
Our studies were focused <strong>on</strong> finding rapid methods for screening of aflatoxins B1,<br />
(AB1), aflatoxin B2 (AB2), aflatoxin G1 (AG1), aflatoxin G2 (AG2) and ochratoxin A.<br />
Mycotoxins were extracted in chloroform, separated <strong>on</strong> silicagel thin-layer<br />
chromatography plates and were quantified using densitometric analysis (Braicu and all,<br />
2005). We analyzed samples from different Romanian regi<strong>on</strong>s.<br />
The limits of detecti<strong>on</strong> of this method <strong>on</strong> TLC plate were 0.25 μg/kg for AG1, AG2,<br />
AB1, AB2, respectively 3�μg/kg for ochratoxin A and the limits of quantificati<strong>on</strong> from<br />
samples were 0.83 μg/kg AG1, AG2, AB1, AB2, 10 μg/kg for ochratoxin A<br />
RESULTS AND DISCUSSIONS<br />
The degree of infestati<strong>on</strong> with fungi as presented in Table 1 is very high, above 50%, in<br />
all types of samples collected from the country.<br />
Table 1. The degree of infestati<strong>on</strong> with fungi in grains and bakery products<br />
Product<br />
Analised<br />
samples<br />
(nr)<br />
Number of samples infected with fungi<br />
Potential toxigenic fungi (%)<br />
Total<br />
no (%)<br />
Penicillium Aspergillus Fusariu<br />
sp.<br />
sp. m sp.<br />
Wheat 130 78 / 60.0% 38.46 38.46 76.92<br />
Maize 60 35 / 58.3% 85.71 77.14 68.57<br />
Rye 6 5 / 83.3% 40.0 60.0 60.0<br />
Triticale 6 5 / 83.3% 80.0 100 100<br />
Bread 6 6 / 100% 100 83.3 33.3<br />
Biscuits 6 4 / 66.7% 25.0 25.0 50.0<br />
Flakes 6 3 / 50.0% 100 33.3. 100<br />
Total 220 135 / 61.4% 56.29 53.33 74.07<br />
171
In wheat 60% of samples are infected, in corn 58.3% of samples but the most alarming<br />
issue is that the fungal infecti<strong>on</strong> is present in bread (100%), biscuits and cereal flakes.<br />
The results indicates that in the analyzed samples the percent of potential toxigenic<br />
fungi is high too, we notice Penicillium from 25% to 100% (bread) infected sample,<br />
Aspergillus from 33.3% to 100% (Triticale) infected samples and Fusarium from 33.3%<br />
(bread) to 100% (Triticale and flakes) infected samples.<br />
Analyzing the wheat samples by blotting test and Ulster test, we’ve de<strong>term</strong>ined<br />
fungi related to 12 genera (Table 2) which have developed <strong>on</strong> bread grains: Penicillium,<br />
Aspergillus, Fusarium, Alternaria, Cladosporium, Helminthosporium, Trichotecium,<br />
Stachybotrys, Septoria, Epicoccum, Acrem<strong>on</strong>iella and Rhizopus. On grains we can<br />
observe fungi from field, typical storage mycoflora and in<strong>term</strong>ediate mycoflora (Hulea<br />
Ana and all, 1982; Boariu Carmen, 1988). We can notice higher values especially for<br />
toxigenic fungi as Penicillium, Aspergillus and Fusarium.<br />
Table 2. The occurrence of fungi in the analyzed samples of bread grains<br />
Frequency of fungi de<strong>term</strong>ined by B.p.t. and Ulster test<br />
Fungi<br />
0% under 20% 20-70% above 70%<br />
B.p.t U.t. B.p.t. U.t. B.p.t. U.t. B.p.t U.t.<br />
Penicillium sp. 40,0 15.38 33.3 34.62 26.7 38.46 - 11.5<br />
4<br />
Aspergillus sp. 43.3 15.38 26.7 19.24 26.7 50.0 3.3 15.3<br />
8<br />
A. ochraceus 96.7 84.62 - 7.69 3.3 7.69 - -<br />
A. niger 76.7 53.84 6.7 19.23 13.3 23.08 3.3 3.85<br />
A. nidulans 100 84.62 - 7.69 - 7.69 - -<br />
A. flavus 90.0 61.54 - 19.23 10.0 15.38 - 3.85<br />
A. glaucus 96.7 84.62 - 15.38 3.3 - - -<br />
A. versicolor 100 76.93 - 15.38 - 7.69 - -<br />
A. fumigatus 93.4 92.3 3.3 3.85 3.3 - - 3.85<br />
Fusarium sp. 10.0 26.92 26.7 30.77 53.3 34.62 6.67 7.69<br />
Alternaria sp. 20.0 26.92 13.3 19.23 43.3 38.46 23.3 15.3<br />
9<br />
Cladosporium sp 56.7 38.46 26.6 30.77 16.7 30.77 - -<br />
Trichotecium roseum 93.3 88.77 - 7.69 6.7 11.54 - -<br />
Stachybotrys sp. 96.7 96.15 3.3 - - 3.85 - -<br />
Rhizopus sp. 76.7 61.54 6.7 23.8 13.3 15.38 3.3 -<br />
Septoria sp. 90.0 100 10.0 - - - - -<br />
Helminthosporium sp. 96.7 100 3.3 - - - - -<br />
Acrem<strong>on</strong>iella atra 96.7 96.15 3.3 3.85 - - - -<br />
Epicoccum purpurasc. 96.7 100 3.3 - - - - -<br />
172
Table 3. Natural occurrence of mycotoxins in cereals analyzed by TLC coupled with<br />
densitometry from different Romanian districts<br />
Samples Dist.<br />
No of No. of Ochrat.<br />
examin. c<strong>on</strong>tamin. A<br />
samples samples µg/kg<br />
AB1<br />
µg/kg<br />
AB2<br />
µg/kg<br />
AG1<br />
µg/kg<br />
AG2<br />
µg/kg<br />
Total<br />
Aflat.<br />
µg/kg<br />
Wheat BC 1 1 0 5.3 5.5 10.8<br />
BN 2 2 0 4.3 1.4 0<br />
5.7<br />
6.5 5.0 1.5<br />
6.5<br />
BV 3 2 0<br />
4.2 1.0 5.2<br />
30.0<br />
2.0 1.8 3.8<br />
0<br />
0 1.2 1.2<br />
BH 1 -<br />
HD 3 2 0 5.7 2.6<br />
8.3<br />
3.9 0.9<br />
4.8<br />
CJ 2 -<br />
MM 4 1 0 0.1 0.5 0.6<br />
GR 2 -<br />
SB 1 1 0 5.6 2.7 8.3<br />
SM 1 1 0 4.0 4.0<br />
SJ 1 -<br />
Maize BC 2 2 0 1.7 0<br />
1.7<br />
3.9 1.4<br />
5.3<br />
BV 1 -<br />
MM 1 -<br />
SB 1 1 0 3.3 3.3<br />
HD 2 1 4.4 1.7 0.6 2.3<br />
GR 2 1 0 - 0.02 0.12 0.14<br />
SM 1 -<br />
Triticale BV 3 3 0<br />
0.5 1.1<br />
1.6<br />
5.7 0.8 6.5<br />
5.7 0.8 6.5<br />
Rye SB 1 1 25.6 4.6 2.8 7.4<br />
HD 1 - -<br />
Bread CJ 2 -<br />
Biscuits CJ 1 1 0 4.4 1.8 - 6.2<br />
Limits by Commissi<strong>on</strong> of Regulati<strong>on</strong><br />
(EC), No 472/ 2002<br />
3-5
In c<strong>on</strong>clusi<strong>on</strong>, the fungal charge of grains for food and fodder <strong>on</strong> all samples is enough<br />
high to alarm us, indifferent of the method of analyze. We have to corroborate this data<br />
with mycotoxins analyses to know if our food and fodder is dangerous or not.<br />
In the next table (Table 3) we present the natural occurrence of mycotoxins in<br />
grains analyzed by TLC coupled with densitometry.<br />
As shown in Table 3, we analyzed 39 samples from different Romanian districts,<br />
20 samples were found c<strong>on</strong>taminated with mycotoxins. Four samples were<br />
c<strong>on</strong>taminated with ochratoxin A and the c<strong>on</strong>centrati<strong>on</strong> was higher than legal European –<br />
we notice that in rye and a sample of wheat for bakery, the values were very alarming,<br />
over 20 µg/kg. In 11 samples was identified AB1, in 9 samples the am<strong>on</strong>t of this was<br />
higher than norms; c<strong>on</strong>cerning the total c<strong>on</strong>centrati<strong>on</strong> of aflatoxins 10 sample this was<br />
higher than legal European norms. The c<strong>on</strong>centrati<strong>on</strong> of aflatoxins is very dangerous in<br />
biscuits for kids.<br />
The c<strong>on</strong>taminati<strong>on</strong> range was: aflatoxin B1: 1.7 - 5.7 µg/kg, aflatoxin B2: 0.02 -2.8<br />
µg/kg, aflatoxin G1: 1.1 – 5.7 µg/kg, aflatoxin G2: 0.12 – 1.8 µg/kg, total aflatoxins: 1.2<br />
– 10.8 µg/kg and ochratoxin A: 4.4 - 30.0 µg/kg. The higher number of c<strong>on</strong>taminati<strong>on</strong><br />
rate was in the case of wheat samples.<br />
I n c<strong>on</strong>clusi<strong>on</strong>, we can notice that the fungal charge of grains for food and fodder<br />
and in bakery products <strong>on</strong> all samples is enough high to alarm us, no matter of the<br />
method of analyze. The mycotoxin analyses reveal to us a high and dangerous level of<br />
ochratoxin A and total aflatoxins. Our food and fodder are dangerous and our legislati<strong>on</strong><br />
is way too permissive. We have to align our legislati<strong>on</strong> with the European <strong>on</strong>e to take<br />
care about our grains.<br />
REFERENCES<br />
1. Boariu Carmen, 1988, Cercetări privind modificările fizico – chimice şi biologice<br />
care au loc în timpul păstrării seminţelor destinate c<strong>on</strong>sumului., în „C<strong>on</strong>tribuţii ale<br />
cercetării ştiinţifice la dezvoltarea agriculturii din z<strong>on</strong>a centrală a Câmpiei de<br />
Vest”, C.M.D.P.A., 291-318.<br />
2. Braicu Cornelia, Carmen Puia, C. Bele C., E. Bodoki, Carmen Socaciu, 2005,<br />
Optimisati<strong>on</strong> of screening systems to evaluate relevant mycotoxins from cereals<br />
and bread, 4 th <str<strong>on</strong>g>Internati<strong>on</strong>al</str<strong>on</strong>g> Symposium of USAMV Cluj-Napoca, 16:144-149<br />
3. Domsch K.H., W. Gams, 1972, Fungi in agricultural soils, published by L<strong>on</strong>gman.<br />
4. Hulea Ana, 1969, Ghid pentru laboratoarele de micologie şi bacteriologie, Ed.<br />
Agrosilvică, Bucureşti, 304 p.<br />
5. Hulea Ana, Gh. Tasca, C. Beratlief, 1982, Bolile şi dăunătorii produselor agricole<br />
şi hortiviticole după recoltare, Ed. Ceres, Buc., 193 – 210.<br />
6. Mills J.T., 1990, Mycotoxins and toxigenic fungi <strong>on</strong> cereal grains in western<br />
Canada., Can. J. Physiol. Pharmacol. 1990 Jul; 68 (7):982-6.<br />
7. Raicu Cristina, Doina Baciu, 1978, Patologia semintei, Ed. Ceres, Bucuresti, 207 p.<br />
174
CORRELATIONS OF SOCIAL AND ECONOMIC FUNCTIONS IN<br />
COMMUNITIES AROUND HORTOBÁGY<br />
Erzsébet Csengeri<br />
Tessedik Sámuel College Faculty of Agricultural Water and Envir<strong>on</strong>mental<br />
Management Envir<strong>on</strong>mental Management Institute<br />
1-3 Szabadság Sreet 5540 Szarvas<br />
ABSTRACT<br />
The change of ecosystems is mainly influenced by the material handling activity of man.<br />
This is particularly accentuated in an area sensitive in other respects as well, where a<br />
couple of decades ago man still fought hardly for a farming as efficient as possible <strong>on</strong><br />
this extensive area with saline, bad quality soils. A characteristic feature of Hortobágy is<br />
that it represents a well-delimited natural unit from both botanical and zoological and<br />
hydrographic respects, which preserves several cultural features. I will present the<br />
envir<strong>on</strong>ment-modifying activity of man mainly after the industrial revoluti<strong>on</strong> <strong>on</strong> the<br />
example of communities with large agricultural lands.<br />
Due to the social and ec<strong>on</strong>omic changes brought about by the political change the<br />
communities with dominance of agriculture got into more and more unfavourable<br />
c<strong>on</strong>diti<strong>on</strong>s which resulted in depopulati<strong>on</strong> and the disc<strong>on</strong>tinuati<strong>on</strong> of land use forms,<br />
and thus, a new change of envir<strong>on</strong>ment. From a demographic point of view, Hortobágy<br />
can be divided into two main parts, the core village centre where 65% of the populati<strong>on</strong><br />
lives and the farm centres lying in a radius of 3 to 10 km, which used to be centres of<br />
the former Hortobágy State Farm. By now, these communities partly became<br />
depopulated, partly became parts of Hortobágy. All these communities are characterised<br />
by aging of the populati<strong>on</strong> due to the lack of ec<strong>on</strong>omic income. In respect of farming,<br />
the tendency of shifting from the earlier intensive acitivities to extensive <strong>on</strong>es is typical.<br />
Plants are produced <strong>on</strong>ly in areas with good quality soils which fulfills <strong>on</strong>ly the need for<br />
feeds. Mainly the breeding of indigenous breeds is preferred for rehabilitati<strong>on</strong> of the<br />
original state of vegetati<strong>on</strong> <strong>on</strong> the ploughed areas.<br />
Keywords: human activity, land use industrial revoluti<strong>on</strong> ec<strong>on</strong>omic changes<br />
INTRODUCTION<br />
As the human society plays an ever more de<strong>term</strong>ining role in the ecosystem<br />
metabolism, the research of the activities and material handling of man as a key species,<br />
i.e. of the society metabolism necessarily comes to the fr<strong>on</strong>t. In this, the demography,<br />
ec<strong>on</strong>omy, culture, sociology, political and power structure and operati<strong>on</strong> of local<br />
communities are de<strong>term</strong>ining.<br />
My objective is to show the impact of man as a key species <strong>on</strong> ecosystems <strong>on</strong> the<br />
example of the settlements around Hortobágy. In the rural settlements to be presented, I<br />
analyse in the first place the change of the role of agriculture from the point of view of<br />
the demographic changes of the communities (Oláh, 2006).<br />
There have been differences in the ec<strong>on</strong>omic activity of the chosen settlements, both in<br />
the past and present. The studied settlements were Árkus, Hortobágy-halastó, Kónya,<br />
Máta, Szásztelek and the following farmsteads: Borsós, Kungyörgy, Kuntelek,<br />
175
Várostanya, Malomháza, Faluvégháza, Vill<strong>on</strong>gó I and Hortobágy that administratively<br />
unites these settlements.<br />
The listed settlements are all typical agricultural, rural <strong>on</strong>es. The lack of independence<br />
of the settlements shows already by itself that they are not viable <strong>on</strong> their own, neither<br />
from a demographic nor from an ec<strong>on</strong>omic point of view. According to the<br />
classificati<strong>on</strong> of the rural human organizati<strong>on</strong> levels, they corresp<strong>on</strong>d to small clusters<br />
of communities that are more isolated and offer less services compared to the town of<br />
Hortobágy.<br />
MATERIALS AND METHODS<br />
The demographic and ec<strong>on</strong>omic data were collected pers<strong>on</strong>ally in the settlements. In<br />
additi<strong>on</strong> to my own collecti<strong>on</strong>, necessary informati<strong>on</strong> was obtained from the Central<br />
Statistical Office and the T-STAR database. The demographic indices were de<strong>term</strong>ined<br />
using the parameters of the Clocke index indicating the directi<strong>on</strong>s of the possible future<br />
development of the given settlement. The studied settlements, Árkus, Hortobágyhalastó,<br />
Kónya, Máta, Szásztelek, Borsós, Kungyörgy, Kuntelek, Várostanya,<br />
Malomháza, Faluvégháza, Vill<strong>on</strong>gó I and Hortobágy are typically agricultural<br />
settlements, and thus, their study over more years is interesting in view of the ever<br />
decreasing populati<strong>on</strong> holding capacity of the agriculture.<br />
Changes of the farming structure were studied from the 1960s, broken down by<br />
ten-year periods, giving median values. The study targeted the different agricultural<br />
land uses, changes of the ratios of agriculturally used crop lands and livestock. I made<br />
an in-depth interview with Mr. Csaba Göcz who was in charge of agricultural<br />
producti<strong>on</strong> <strong>on</strong> this territory of over 30 000 ha from the late 1960s.<br />
RESULTS AND DISCUSSIONS<br />
The changes of the resident populati<strong>on</strong> closely follow those of the ec<strong>on</strong>omic life. Earlier<br />
in time, in the 1960s and 1970s, the peripheries had higher populati<strong>on</strong>. In the eighties,<br />
the fiscal goods coming from agriculture, especially from animal husbandry, permitted<br />
the increase of the demands of the peasantry. Because of the better infrastructural<br />
c<strong>on</strong>diti<strong>on</strong>s, the populati<strong>on</strong> moved from the farmsteads to Hortobágy, while retaining<br />
their occupati<strong>on</strong>. This is when the total populati<strong>on</strong> of Hortobágy and the neighbouring<br />
farmsteads reached its maximum of nearly 2000 pers<strong>on</strong>s (Figure 1). Following this, a<br />
slowly decreasing trend started in the 1990s, which has lasted to our days.<br />
176
1600<br />
1400<br />
1200<br />
1000<br />
800<br />
600<br />
400<br />
200<br />
0<br />
1960 1970 1980 1990 2000 2004<br />
Figure1: Run of the populati<strong>on</strong> in periphery and core areas<br />
As a result of the depopulati<strong>on</strong> process, the farmsteads have either become part of<br />
Hortobágy (e.g., Borsós) or totally depopulated like e.g. Várostanya, Malomháza,<br />
Faluvégháza or Kuntelek. In Kungyörgy and Vill<strong>on</strong>gó I, not even ten families live,<br />
while in Máta, Kónya, Hortobágy-halastó, Árkus and Szásztelek, there are more than 10<br />
families left (Egyedné, 1999).<br />
The parameters of the Clocke index show the dynamics of the rural populati<strong>on</strong>.<br />
The parameters study the ratios of the following age classes that, in case of Hortobágy,<br />
show the pattern presented in Table 1 in the studied two years.<br />
Table 1: Demographic structure of, village populati<strong>on</strong><br />
perifery<br />
1985 1995 2000 2004<br />
Aging Ratio 0,10 0,30 0,57 0,70<br />
Generati<strong>on</strong>’s rejuvenati<strong>on</strong> Ratio 0,20 0,20 0,20 0,19<br />
Fertility Ratio 2,0 1,3 1,03 1,15<br />
Sex Ratio 95 97 99 101<br />
Dependants Ratio 0,47 0,35 0,3 0,34<br />
As a result, it can be stated that the aging of the populati<strong>on</strong>, which corresp<strong>on</strong>ds to the<br />
nati<strong>on</strong>-wide trend, is even more pr<strong>on</strong>ounced in agricultural-type rural areas like<br />
Hortobágy and the neighbouring farmsteads. The age compositi<strong>on</strong> of the Hortobágy<br />
settlements shows the following pattern. In 1995, 21% of the populati<strong>on</strong> was children,<br />
67% was middle-aged (15-19 years old), while 12% were over 60. On the basis of the<br />
2001 statistical data, it can be seen that a part of the children of the 1995 statistics<br />
moved to the middle-aged group, but this reducti<strong>on</strong> was not compensated by<br />
177<br />
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eproducti<strong>on</strong>, and therefore, children represented <strong>on</strong>ly 19%. In the middle-aged group,<br />
the same trend can be observed, their share is 69%. The populati<strong>on</strong> over 60 years of age<br />
makes 11%. The generati<strong>on</strong> rejuvenati<strong>on</strong> can be c<strong>on</strong>sidered good, although it is<br />
gradually approaching <strong>on</strong>e. The fertility ratio also shows a decreasing trend.<br />
The dependants’ ratio can be c<strong>on</strong>sidered good in the settlement, as the share of the<br />
actively occupied populati<strong>on</strong> is predominant and unemployment does not exceed 10%.<br />
These results show that the previous emigrati<strong>on</strong> process is now complemented by a<br />
slow aging process, which can have unpredictable c<strong>on</strong>sequences in this area, important<br />
from many respects. This process is also well reflected by the changes in the total<br />
populati<strong>on</strong> of the settlement (Table 2).<br />
Table 2:Run of the populati<strong>on</strong><br />
Years Total populati<strong>on</strong>/head<br />
1960 1508<br />
1970 1791<br />
1980 1848<br />
1990 1741<br />
2000 1757<br />
2004 1681<br />
Next, I study if the changes of the ec<strong>on</strong>omic data corroborate the demographic changes.<br />
Here, I also go back to the early years of the industrial revoluti<strong>on</strong>. After the Sec<strong>on</strong>d<br />
World War, there several ideas were proposed regarding the better exploitati<strong>on</strong> of the<br />
Hortobágy puszta. However, all proposals c<strong>on</strong>curred in that they c<strong>on</strong>sidered animal<br />
husbandry as <strong>on</strong>e of the possible main directi<strong>on</strong>s. It is the principal branch of agriculture<br />
today as well. The required feeds were produced in the ploughed areas of the<br />
Hortobágy.<br />
In view of the soil characteristics of Hortobágy (grassland clay, sodic, chernozem<br />
loam, sandy loam, alluvial and sandy soils) the plant cultivati<strong>on</strong> c<strong>on</strong>sists mainly of the<br />
cultivati<strong>on</strong> of corn and sugar beet satisfying the winter feed demand of livestock,<br />
especially sheep.<br />
In the 1960s, farming started <strong>on</strong> an area of 30 351 ha, whereof, in corresp<strong>on</strong>dence with<br />
the planned agricultural profile, the largest share was given to meadows and pastures<br />
that occupied approximately twice as much area as the plough-lands at the time.<br />
This area would seem big enough to entirely satisfy the feeding needs of the warmblooded<br />
livestock during all the summer and winter period. The real situati<strong>on</strong> is much<br />
worse. In fact, great part of the meadows and pastures are barren from the beginning of<br />
June, they can be used <strong>on</strong>ly as sheep pastures. Areas with such characteristics make<br />
65% of the farmed area of Hortobágy.<br />
The lucerne growing <strong>on</strong> 22% of the study area also serves the feeding needs of<br />
animal husbandry. Plants as poppy, tobacco or soybean were grown <strong>on</strong> a further 5% of<br />
the farmed area, which did not produce adequate results. Autumn wheat was generally<br />
cultivated <strong>on</strong> the sodic or highly sodic soils with the lowest producti<strong>on</strong> capacity, using<br />
intensive wheat sorts and significant quantities of fertilizers (Table 3).<br />
178
Table 3: Land use/ha<br />
1960 1970 1980 1990 2000<br />
Plough-land 5379 5610 5350 3215 2802<br />
Garden 3 4 7 7 7<br />
Vineyard - - - - 0,36<br />
Orchard - - - - 3,61<br />
Meadow<br />
Pasture-land<br />
16179 10221 10100 12821 13006<br />
Reed 551 570 1345 1869 2183<br />
Forest 875 825 820 814 796<br />
Fish-p<strong>on</strong>d 7150 6920 6431 5011 4070<br />
Total bearing surface 30137 24150 24053 23737 22870<br />
Another result of the sixties was the systematic forestati<strong>on</strong> programme started <strong>on</strong> the<br />
lowest-lying parts of the pastures.<br />
The land use trends have lasted throughout the years, although the usable crop land<br />
has shrunk, partly because of their exclusi<strong>on</strong> from cultivati<strong>on</strong> and partly because of land<br />
appropriati<strong>on</strong>s by the Hortobágy Nati<strong>on</strong>al Park founded in 1973.<br />
The village of Hortobágy has its own administrative area since 1 January 1966 (a<br />
total of 28 458 ha), previously it bel<strong>on</strong>ged to Balmazújváros. Of the 28 458 ha, the<br />
surface used by the core areas and excluded from cultivati<strong>on</strong> has been c<strong>on</strong>tinuously<br />
increasing, i.e. the crop land under agricultural cultivati<strong>on</strong> has been diminishing.<br />
The ploughing land use was the most widespread in the 1960s and 1970s because<br />
of rice growing <strong>on</strong> ploughed areas, which <strong>term</strong>inated in 1980. Since that time, the<br />
ploughed lands have been c<strong>on</strong>tinuously shrinking, while the grasslands and – in the<br />
recultivated wetlands – the reedy areas have been growing. The use of the fishp<strong>on</strong>ds<br />
established in the early 20 th century has been c<strong>on</strong>stantly decreasing. The Hortobágy<br />
Nati<strong>on</strong>al Park, established in 1973, has been c<strong>on</strong>tinuously increasing the protected areas<br />
and the area in its property has also grown. By 2000, over 80% of the crop land under<br />
the jurisdicti<strong>on</strong> of the village of Hortobágy was transferred under the administrati<strong>on</strong> of<br />
the Nati<strong>on</strong>al Park, these areas are mainly used by the Hortobágy N<strong>on</strong>-Profit Company,<br />
while a smaller part of them is leased by agricultural entrepreneurs (Szeifert, 1969).<br />
Of the animal husbandry of the 1960s and 1970s, sheep breeding reached<br />
excepti<strong>on</strong>al ec<strong>on</strong>omic levels resulting in significant exports, and thus, incomes, but the<br />
fisheries and poultry farming show similar characteristics, whereof the highest was the<br />
share of the guinea-fowl and duck farming. The cattle and horse breeding dealt <strong>on</strong>ly<br />
with issues related to the rehabilitati<strong>on</strong> of the genetic pool at the time (Table 4).<br />
179
Table 4: Livestock/pieces,<br />
1960 1970 1980 1990 2000<br />
Cattle 2172 2615 2980 2955 2681<br />
Pig 7489 7820 7905 5015 1870<br />
Horse, 630 715 608 520 385<br />
Sheep 31838 33200 29020 14835 6485<br />
Poultry 59949 450300 690100 71500 20891<br />
Bee 48 69 36 55 46<br />
The largest share in the exports and thus, the highest incomes came from fish exports,<br />
however, interestingly, the statistical evaluati<strong>on</strong>s do not provide numerical data for<br />
these stocks.<br />
During the decentralized privatizati<strong>on</strong> of the State Farm, the large-scale intensive<br />
animal farms were c<strong>on</strong>stantly privatized or liquidated between 1988 and 1992, which<br />
resulted mainly in the reducti<strong>on</strong> of the cattle and sheep stocks. In this area, pig breeding<br />
had traditi<strong>on</strong>s mainly in household farms, but it has decreased to a minimum by today.<br />
The poultry stock c<strong>on</strong>sisted of waterfowl (geese, ducks) and guinea-fowl. Between 1960<br />
and 1980, the State Farm raised large quantities of so-called meat geese, p<strong>on</strong>d-reared<br />
broiler ducks and broiler guinea-fowl according to the market demand of the time. By<br />
2000, the poultry stock has reduced to a minimum. It is typical that in the village of<br />
Hortobágy, 100% of the cattle is kept industrially, while 30% of the horses, 65% of the<br />
sheep and 70% of the pigs can be found in small-scale, privately owned farms.<br />
As a result of the survey of the animal husbandry sector of the agriculture, it can be<br />
stated that, in accord with the Hortobágy soil c<strong>on</strong>diti<strong>on</strong>s and the productivity of the<br />
natural vegetati<strong>on</strong> evolved here, rearing cattle that is grazed in the open air all the year<br />
round and sheep is more dominant even to this day, as well as the fish culture, d<strong>on</strong>e<br />
within the frame of partly natural p<strong>on</strong>d culture.<br />
REFERENCES<br />
Clocke P. J. (1997): An index of rurality for England and Wales. Regi<strong>on</strong>al Studies 11:<br />
31-46<br />
Egyedné Kertész I. (1999): A helyi önkormányzatok hatáskörébe tartozó adók<br />
elemzése és ellenőrzése a bevételek alakulása folyamatában Hortobágy Község<br />
Önkormányzatának 1991-1998. évi gazdálkodásán és adóztatási gyakorlatán<br />
keresztül. Államigazgatási Főiskola, Budapest 12-13<br />
KSH (2004): Egyéni agrárgazdaságok és népsűrűségük. Észak-Alföld. KSH<br />
Házinyomda. Debrecen, Szolnok, Nyíregyháza 31-45<br />
Oláh J.(2006): Környezetgazdálkodás. TSF MVKFK, Szarvas<br />
Szeifert I.(1969): Hortobágyi Állami Gazdaság. Hajdú-Bihar Megyei Lapkiadó<br />
Vállalat, Debrecen<br />
180
RIDING TOURISM IN THE NORTHERN GREAT PLAIN REGION<br />
(HUNGARY): STATUS AND POTENTIAL<br />
Réka Incze and Gábor Hevessy<br />
University of Debrecen, Center of Agricultural Sciences<br />
4032 Debrecen, Böszörményi Str. 138<br />
ABSTRACT<br />
Only a small part (
assessed the different c<strong>on</strong>diti<strong>on</strong>s of the Great Plain regi<strong>on</strong> inserted into a Hungarian<br />
regi<strong>on</strong>al comparis<strong>on</strong> system.<br />
Nowadays, tourism is <strong>on</strong>e of the most developing industrial branches; additi<strong>on</strong>ally, it<br />
results the highest incomes from export in the world (UNO, 2002). Tourism offers for<br />
127 milli<strong>on</strong> people possibility of work, with its share of 6 % of the world’s GDP, i.e.<br />
each fifteenth pers<strong>on</strong> works in the field of tourism. If we include also workplaces<br />
c<strong>on</strong>nected indirectly to tourism, then we get 260 milli<strong>on</strong> workplaces, i.e. approx. 10 %<br />
of GDP (World Travel and Tourism Council, 2004). Incomes originated from tourism<br />
achieved the value of 622 milli<strong>on</strong> USD in 2004 exceeding by 18.7 % the incomes of<br />
2003. The above menti<strong>on</strong>ed data do not c<strong>on</strong>tain inland travels, the number of which<br />
may be the sextuple of internati<strong>on</strong>al travels.<br />
Tourism also plays an important role in the ec<strong>on</strong>omy of Hungary. The value of the<br />
foreigners’ c<strong>on</strong>sumpti<strong>on</strong> visiting Hungary was 822 billi<strong>on</strong> HUF in 2004, within which<br />
the value of tourist c<strong>on</strong>sumpti<strong>on</strong> was 596 billi<strong>on</strong> HUF. Moreover, the Hungarian<br />
populati<strong>on</strong> spent 385 billi<strong>on</strong> HUF <strong>on</strong> tourist services. Summing these, the value of<br />
touristical incomes achieved 981 billi<strong>on</strong> HUF. The total GDP of touristical branches is 5<br />
% of the whole Hungarian GDP. Taking into account also multiplier impacts 8.5 % of<br />
the Hungarian GDP originates from tourism. The number of employees employed<br />
directly in the field of tourism was 398,000 pers<strong>on</strong>s in 2004, which is 8.9% of the total<br />
employment. Taking into account multiplier impacts the menti<strong>on</strong>ed proporti<strong>on</strong> increases<br />
to 12.5 %, which means that each 8 th workplace has been produced by tourism.<br />
Table 1.: touristically qualified riding establishments and services in Hungary<br />
Number of horseshoes<br />
5 4 3 2 1 Other* Total<br />
Qualified establishments (pcs) 15 29 32 29 27 26 158<br />
Accommodati<strong>on</strong> (pcs) 831 702 572 445 880 1022 4452<br />
Roofed riding ground (pcs) 9 13 8 1 2 7 40<br />
Own horse number (pcs) 1635 778 757 574 471 1997 6212<br />
Accommodati<strong>on</strong> for foreign horses<br />
(pcs) 232 359 377 166 221 649 2004<br />
* E.g. presentati<strong>on</strong> sites, horse-breeding, riding educati<strong>on</strong> establishments and<br />
establishments rendering services of nomadic character.<br />
The most popular destinati<strong>on</strong>s are Budapest, lake Balat<strong>on</strong> and spa resorts. The average<br />
capacity utilisati<strong>on</strong> of commercial accommodati<strong>on</strong>s was 39.5% in 2005. The best<br />
utilisati<strong>on</strong>, i.e. 48.2 % average utilisati<strong>on</strong> was achieved by hotels. The average<br />
utilisati<strong>on</strong> of guest-houses was 22.7 %. The demand for higher quality accommodati<strong>on</strong>s<br />
has increased, while the demand has decreased for lower quality services. The income<br />
of commercial accommodati<strong>on</strong> was 112.3 billi<strong>on</strong> HUF and the income of hospitality<br />
was 53.6 billi<strong>on</strong> HUF and the income originated from other sources was 48.5 billi<strong>on</strong><br />
HUF in 2005. The total income was 214.4 billi<strong>on</strong> HUF, from which the share of hotels<br />
was the highest (187.1 billi<strong>on</strong> HUF). Anyway, the share of other commercial<br />
accommodati<strong>on</strong>s was <strong>on</strong>ly 27.3 billi<strong>on</strong> HUF. Accommodati<strong>on</strong>s are used almost by<br />
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visitors bel<strong>on</strong>ging to all types of tourism. Henceforth we menti<strong>on</strong> <strong>on</strong>ly the status of the<br />
riding tourism. The riding tourism includes all branches of „riding and coach driving<br />
seas<strong>on</strong>ably or systematically pursued by tourists in their spare time, for hobby, for<br />
maintaining their own health <strong>on</strong> their own at their own costs” (Fehér and Kóródi 2007).<br />
According to the publicati<strong>on</strong> prepared by Magyar Turizmus Zrt. and Hungarian<br />
Tourists’ Associati<strong>on</strong> (2006a) the touristically qualified riding establishments and<br />
services presented in Table 1. can be found in Hungary.<br />
Riding services can be divided into 3 groups according to their frequency (Fehér and<br />
Kóródi 2007):<br />
• frequently, generally offered services (e.g.: riding, riding educati<strong>on</strong>, lease keeping,<br />
riding tour, coach driving, camping, accommodati<strong>on</strong> and catering)<br />
• often, but not generally services offered (e.g.: cross-country riding, show jumping,<br />
dressage, competiti<strong>on</strong>, presentati<strong>on</strong>s, hunting and riding)<br />
• rarely offered services (e.g.: visiting horse-breeding establishments, military)<br />
Within the riding tourism active and passive services can be distinguished. Active<br />
services are e.g. riding, riding educati<strong>on</strong>, western riding, coach driving and other<br />
equestrian sports. Passive services are mainly presentati<strong>on</strong>, e.g. presentati<strong>on</strong> of races,<br />
equestrian archery, coach driving presentati<strong>on</strong>, five-in-hand and equestrian acrobatics.<br />
Hungarian riders’ characteristics, like horse-herders’ attracti<strong>on</strong>s and five-in-hand<br />
mean a special attracti<strong>on</strong> both for Hungarian and foreign tourists. Horse-herders used to<br />
guard herds of horses in the open air all the year in the plain. Nowadays horses are<br />
rarely kept in such a way. Horse-herders maintain the traditi<strong>on</strong> c<strong>on</strong>cerning their tools<br />
and clothing. They use special horse-accessories, like saddles without girths and shortstock<br />
ornamental l<strong>on</strong>g whip. The „five-.in-hand”, „Koch-five-in-hand” or “Hungarian<br />
post” are daring equestrian attracti<strong>on</strong>s: the horse-herder is driving five or more horses<br />
standing <strong>on</strong> the back of the last two horses.<br />
Several groups of interest, e.g. hobby riders and their compani<strong>on</strong>s, pers<strong>on</strong>s<br />
pursuing and organising equestrian sports, horse-breeders, resp. participants of riders’<br />
performances, riding tours and camping are touched by the riding tourism. Furthermore,<br />
groups rendering services c<strong>on</strong>nected with equestrian sports, e.g. pers<strong>on</strong>s or enterprises<br />
following trades c<strong>on</strong>nected with horses. People bel<strong>on</strong>ging to different groups of interest<br />
can be basically squaded into two big groups (Incze and Hevessy 2007). The first<br />
groups is formed by pers<strong>on</strong>s touched rarely or as complementary activity by riding<br />
services. The sec<strong>on</strong>d group includes pers<strong>on</strong>s being engaged with horses as main<br />
activity.<br />
Nowadays riding tourism informati<strong>on</strong> is easily accessible in Hungary. Actual<br />
riding services are published by different types of media, such as periodicals, radio, TV,<br />
advertisement newspapers and Internet. The Internet is probably the most tailor-made,<br />
because it offers the most flexible searching possibilities.<br />
Enterprises rendering different riding services are co-ordinated by the Hungarian<br />
N<strong>on</strong>-Profit Associati<strong>on</strong> of Riding Tourism (MLTKSZ). Aptitude test of horses as well<br />
as the management of the horseshoe qualifying system bel<strong>on</strong>gs to the tasks of the<br />
MLTKSZ. The “horseshoe” indicati<strong>on</strong> promotes the orientati<strong>on</strong> c<strong>on</strong>cerning the quality<br />
of different services. The indicati<strong>on</strong> of the quality extends from <strong>on</strong>e to five horseshoes<br />
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3
depending <strong>on</strong> the character and level of the service (it is similar to the c<strong>on</strong>venti<strong>on</strong>al star<br />
qualifying system of hotels).<br />
Hungary has generally good natural c<strong>on</strong>diti<strong>on</strong>s for equestrian programmes (Incze<br />
and Hevessy 2007). The riding tourism in the Northern Great Plain can not be easily<br />
assessed <strong>on</strong> the basis of up-to-date data. There are 9 touristical regi<strong>on</strong>s in Hungary,<br />
which are used by Magyar Turizmus Zrt in general in its inland communicati<strong>on</strong>. The<br />
Regi<strong>on</strong>al Tourist Service Committees also operate in accordance with this system.<br />
However, <strong>on</strong>ly 5 regi<strong>on</strong>s are used in the external (i.e. internati<strong>on</strong>al) communicati<strong>on</strong>. As<br />
a result of this cumulati<strong>on</strong>, the Northern Great Plain, the Lake Tisza and the Southern<br />
Great Plain are called comm<strong>on</strong>ly as Plain and Lake Tisza cumulated regi<strong>on</strong> (Fig. 1.). It<br />
is a questi<strong>on</strong>, why some recent Hungarian publicati<strong>on</strong> has also <strong>on</strong>ly 5 regi<strong>on</strong>s?<br />
Fig. 1.: Northern Great Plain and divisi<strong>on</strong> of Hungary into 5 regi<strong>on</strong>s<br />
(Source: www.itth<strong>on</strong>.hu).<br />
The horse stock of the Northern Great Plain is nati<strong>on</strong>ally important. The number of<br />
horses is 7 031 in county Hajdú-Bihar (Lóska 2001), 4 091 in county Jász- Nagykun-<br />
Szolnok and 7 047 in county Szabolcs-Szatmár-Bereg. The total is 18 169 horses,<br />
which is approximately 20 % of Hungarian horse stock.<br />
Several riding tourism services are present in the Northern Great Plain: e.g. riding,<br />
hunter riding, dressage, military, voltage, cross-country riding, coach driving,<br />
competiti<strong>on</strong>s, riding educati<strong>on</strong>, visiting of horse-breeding establishments, equestrian<br />
shows. Additi<strong>on</strong>ally, there is catering and accommodati<strong>on</strong>. In some places holiday<br />
check and credit card can be also used.<br />
The riding tourism c<strong>on</strong>diti<strong>on</strong>s of the Northern Great Plain are presented according<br />
to the elements of SWOT analyses, i.e. Strengths, Weaknesses, Opportunities and<br />
Threats (Lisztes 2007, Lóska 2001, Borsy 1998). The micro-relief c<strong>on</strong>diti<strong>on</strong>s of the<br />
Northern Great Plain are varied and so, both advantageous and disadvantageous<br />
c<strong>on</strong>diti<strong>on</strong>s for riding tourism can be found. Extending steppes, sandy terrain, landscape<br />
having mosaic character and gently sloping terrain are advantageous; while the plain<br />
character, the cracked structure soil are disadvantageous. The narrowing c<strong>on</strong>tinuous<br />
terrains and strictly protected areas may mean danger, i.e. decreasing facilities. Anyway,<br />
the relief is suitable for a lot of equestrian programs and it makes possible to elaborate<br />
further paths for riding and coach tours.<br />
184<br />
4
C<strong>on</strong>cerning the weather l<strong>on</strong>g autumn and spring, hard winters and very hot summers are<br />
characteristic. The weather should be taken into account during the organizati<strong>on</strong> of the<br />
programs, e.g.: in summer, equestrian programs can be organized in early hours of the<br />
morning or late the evening. Regarding surface waters, the network of rivers is rich,<br />
here should be menti<strong>on</strong> especially the Tisza. The presence of water may also cause<br />
problems, e.g. in the case of marshes and bogs, not to menti<strong>on</strong> floods. The combinati<strong>on</strong><br />
of water and riding tourism or perhaps organisati<strong>on</strong> of combined programmes may offer<br />
very interesting possibilities. The regi<strong>on</strong> has a very rich fauna and flora, which may be<br />
attractive for naturalist tourists. Near communities, cultivated lands are often<br />
predominant; these, of course, do not attract visitors interested for natural curiosities,<br />
but these places are easily accessible. The (riding)tourism may disturb the natural<br />
envir<strong>on</strong>ment but undesirable impacts can be avoided with an appropriate organizati<strong>on</strong>.<br />
The Hortobágy Nati<strong>on</strong>al Park has a positive attitude towards riding tourism within<br />
reas<strong>on</strong>able limits (Lisztes 2007, Varga 2007). The horse-breeding establishment of<br />
Máta, being <strong>on</strong> the territory of the Nati<strong>on</strong>al Park at issue, plans to renew the stage and to<br />
use more efficiently the roofed riding grounds. There are also weak points in the<br />
collaborati<strong>on</strong> of the nati<strong>on</strong>al park and touristically qualified riding establishments, e.g.<br />
defective informati<strong>on</strong> flow. The collaborati<strong>on</strong> with the nati<strong>on</strong>al parks could offer further<br />
possibilities, e.g. riding system of nature protecti<strong>on</strong> guards.<br />
Summarized, tourism plays an important role both in Hungarian and in world<br />
ec<strong>on</strong>omy. C<strong>on</strong>cerning, riding tourism Hungary has advantageous c<strong>on</strong>diti<strong>on</strong>s and within<br />
the country the Northern Great Plain is especially suitable for this activity. Besides the<br />
above menti<strong>on</strong>ed favorable c<strong>on</strong>diti<strong>on</strong>s and riding establishments, the riding tour path<br />
“Puszta” is a special attracti<strong>on</strong>, it is listed am<strong>on</strong>g the best of the world (Harpers<br />
magazine). The horse stock, touristical riding establishments, as well as natural and<br />
ec<strong>on</strong>omic factors offer a good possibility for further developments.<br />
METHODS<br />
Sec<strong>on</strong>dary data are obtained from documentati<strong>on</strong>, while primary data are obtained by<br />
carrying out own estimati<strong>on</strong>s, analyses and a case-study. The subject of the case-study<br />
is the Debrecen Riding Academy. In the case-study, the documentati<strong>on</strong> was completed<br />
by interviews and field research (from September 2006 until January 2007). We have<br />
c<strong>on</strong>sidered the case-study important because through the Debrecen Riding Academy we<br />
could present noti<strong>on</strong>s, types and possibilities of services by means of actual and<br />
c<strong>on</strong>crete examples. Furthermore, our objective was also to present strengths and<br />
weaknesses, opportunities and threats trough a c<strong>on</strong>crete situati<strong>on</strong>. The data c<strong>on</strong>cerning<br />
the Riding Academy are the data from 2005 and 2006 (there was no significant<br />
difference between these 2 years). Our analyses and comparis<strong>on</strong>s have been based <strong>on</strong><br />
the data of Magyar Turizmus Zrt. and Hungarian Riding Tourism N<strong>on</strong>-Profit<br />
Associati<strong>on</strong>, and they were carried out by means of the software Microsoft Excel. In<br />
comparis<strong>on</strong>s, the c<strong>on</strong>diti<strong>on</strong>s of the Plain and Lake Tisza cumulated regi<strong>on</strong> was<br />
compared with those of the Transdanubia, Balat<strong>on</strong>, Budapest and its surroundings, as<br />
well as with Eger-Tokaj Mountains regi<strong>on</strong>. Furthermore, we have assessed the<br />
relati<strong>on</strong>ship between the Northern and Southern Great Plain within the Plain and Lake<br />
Tisza cumulated regi<strong>on</strong>.<br />
185<br />
5
RESULTS AND CONCLUSIONS<br />
There are no data in literature c<strong>on</strong>cerning the total income originating of riding tourism.<br />
We have made an estimati<strong>on</strong> about the incomes of riding establishments having also<br />
accommodati<strong>on</strong>s. The total number of nights passed in Hungary by tourists was 19 334<br />
750 in 2005. Taking into account the 4452 accommodati<strong>on</strong>s shown in Table 1., and the<br />
fact that the majority of establishments having accommodati<strong>on</strong>s bel<strong>on</strong>gs to the category<br />
of guest-houses (the average use of which was 22.7%), the number of nights passed by<br />
tourists is 368 870, which is 1.9 % of the total number of nights passed by tourists. The<br />
guest-houses achieved an income of 18.8 billi<strong>on</strong> HUF in 2005 with their average use of<br />
22.7 %. This means an income of 3.3 billi<strong>on</strong> HUF, which is 1.54 % of the total nati<strong>on</strong>al<br />
income of 214.4 billi<strong>on</strong> HUF (c<strong>on</strong>nected to commercial accommodati<strong>on</strong>s).<br />
We have assessed the situati<strong>on</strong> of the Plain and Lake Tisza cumulated regi<strong>on</strong> in<br />
comparis<strong>on</strong> with other Hungarian regi<strong>on</strong>s and then, within the Plain and Lake Tisza<br />
cumulated regi<strong>on</strong> the role of the Northern Great Plain. C<strong>on</strong>cerning the number of<br />
touristically qualified riding establishments the Plain and Lake Tisza cumulated regi<strong>on</strong><br />
has the first positi<strong>on</strong> am<strong>on</strong>g the 5 regi<strong>on</strong>s (Table 2.). Am<strong>on</strong>g the 55 touristically<br />
qualified riding establishments 20 are in the Northern Great Plain, which is 12.5 % of<br />
the total number (158) of Hungarian establishments. In the Southern Great Plain there<br />
much more, 34 establishments, which is 21.5% of the nati<strong>on</strong>al total number. There is<br />
also a difference between the Northern and Southern Great Plain c<strong>on</strong>cerning the<br />
categories of higher quality and so there are also proporti<strong>on</strong>ally more 5- or 4-horseshoeestablishments<br />
in the Southern Great Plain. At Lake Tisza there is <strong>on</strong>ly <strong>on</strong>e touristically<br />
qualified establishment.<br />
Table 2.: Touristically qualified riding establishments<br />
Number of<br />
horseshoes<br />
5 4 3 2 1 Other Total<br />
Plain and Lake Tisza cumulated regi<strong>on</strong> 5 14 11 7 7 11 55<br />
Transdanubia 7 7 8 8 8 5 43<br />
Balat<strong>on</strong> 0 4 5 1 2 3 15<br />
Budapest and its surroundings 2 3 8 9 3 5 30<br />
Eger-Tokaj Mountains 1 1 0 4 7 2 15<br />
C<strong>on</strong>cerning the number of accommodati<strong>on</strong>s, the status of the Great Plain is shown in<br />
Fig. 2. The Plain and Lake Tisza cumulated regi<strong>on</strong> has the most (1868)<br />
accommodati<strong>on</strong>s. Within this the Northern Great Plain, there are 810, and in the<br />
Southern Great Plain 1032 accommodati<strong>on</strong>s.<br />
186<br />
6
2000<br />
1800<br />
1600<br />
1400<br />
1200<br />
1000<br />
800<br />
600<br />
400<br />
200<br />
0<br />
1868<br />
Plain and<br />
Tisza lake<br />
cumulated<br />
regi<strong>on</strong><br />
Transdanubian<br />
1434<br />
Balat<strong>on</strong><br />
239<br />
602<br />
Budapest and<br />
its<br />
surroundings<br />
Fig. 2.: Number of accommodati<strong>on</strong>s of establishments having accommodati<strong>on</strong>s<br />
The most of roofed riding grounds are also in the Plain and Lake Tisza cumulated<br />
regi<strong>on</strong> (Fig. 3.). 10 out of 17 roofed riding grounds are in the Northern Great Plain and<br />
so this regi<strong>on</strong> even as an independent regi<strong>on</strong> has an important status in nati<strong>on</strong>al<br />
comparis<strong>on</strong> c<strong>on</strong>cerning this aspect. The most of own horses are also in the Plain and<br />
Lake Tisza cumulated regi<strong>on</strong> (Fig. 4.).<br />
18<br />
16<br />
14<br />
12<br />
10<br />
8<br />
6<br />
4<br />
2<br />
0<br />
17<br />
Plain and<br />
Tisza lake<br />
cumulated<br />
regi<strong>on</strong><br />
Transdanubian<br />
13<br />
Balat<strong>on</strong><br />
1<br />
6<br />
Budapest and<br />
its<br />
surroundings<br />
Eger-Tokaj<br />
Mountains<br />
Eger-Tokaj<br />
Mountains<br />
Fig. 3.: Number of roofed riding grounds in each regi<strong>on</strong><br />
187<br />
309<br />
3<br />
7
2500<br />
2000<br />
1500<br />
1000<br />
500<br />
0<br />
2137<br />
Plain and<br />
Tisza lake<br />
cumulated<br />
regi<strong>on</strong><br />
Transdanubian<br />
1908<br />
Balat<strong>on</strong><br />
367<br />
1288<br />
Budapest and<br />
its<br />
surroundings<br />
Fig. 4.: Number of own horses in touristically qualified riding establishments<br />
Out of 2137 own horses 649 are in the area of the Northern Great Plain. In the Southern<br />
Great Plain, there is about the double of this number (1743 horses). In <strong>term</strong>s of assessed<br />
parameters, the most significant difference lies in the number of horses between the two<br />
regi<strong>on</strong>s.<br />
C<strong>on</strong>cerning the number of services rendered by different establishments there is a<br />
balanced competiti<strong>on</strong> am<strong>on</strong>g regi<strong>on</strong>s (Fig. 5.). The Plain and Lake Tisza cumulated<br />
regi<strong>on</strong> is bigger (in <strong>term</strong>s of its territory) than the other regi<strong>on</strong>s. Besides good riding<br />
c<strong>on</strong>diti<strong>on</strong>s, the size can also influence the favorable positi<strong>on</strong> of the regi<strong>on</strong> in<br />
comparis<strong>on</strong>s with other regi<strong>on</strong>s.<br />
13<br />
12,5<br />
12<br />
11,5<br />
11<br />
10,5<br />
10<br />
12,7<br />
Plain and<br />
Tisza lake<br />
cumulated<br />
regi<strong>on</strong><br />
Transdanubian<br />
12,5<br />
Balat<strong>on</strong><br />
12,2<br />
11,1<br />
Budapest and<br />
its<br />
surroundings<br />
Eger-Tokaj<br />
Mountains<br />
Eger-Tokaj<br />
Mountains<br />
Fig. 5.: Number of service types in 5 regi<strong>on</strong>s.<br />
The data c<strong>on</strong>cerning the riding tourism in different types of literature and media mostly<br />
corresp<strong>on</strong>d to the reality, i.e. essential elements are exacts, but details may c<strong>on</strong>tain<br />
errors. For instance Lóska wrote the following things about the Debrecen Riding<br />
Academy in 2001: „Qualificati<strong>on</strong>: 3-horseshoe. The establishment is operated with the<br />
188<br />
512<br />
11,3<br />
8
technical supervisi<strong>on</strong> of the Debrecen University, Center of Agricultural Sciences. The<br />
academy is engaged in dressage and show jumping, there is also a roofed riding ground<br />
and there are 25 horses, which are N<strong>on</strong>ius breeds and Hungarian Sport Horses having a<br />
medium educati<strong>on</strong>.” There is a mistake in the descripti<strong>on</strong>: in the instituti<strong>on</strong>s there have<br />
never been and there are no N<strong>on</strong>ius breeds. Otherwise, the descripti<strong>on</strong>s corresp<strong>on</strong>ds to<br />
the reality of that time. The Riding Academy is still operated with the technical<br />
supervisi<strong>on</strong> of the University of Debrecen. The “<strong>on</strong>ly” 3-horseshoe qualificati<strong>on</strong> can be<br />
attributed to the <strong>on</strong>ly medium educati<strong>on</strong> of the horses and to the relatively new horse<br />
stock in that time.<br />
The horseshoe qualifying system assists the orientati<strong>on</strong> c<strong>on</strong>cerning the quality of<br />
riding establishments. Nowadays the Debrecen Riding Academy bel<strong>on</strong>gs already to the<br />
4-horseshoe category, which is distinguished ranking (the horseshoe qualifying system<br />
is from 1 to 5). Nowadays, the horse stock does not have a medium, but a good<br />
educati<strong>on</strong>. The instituti<strong>on</strong> has not got the 5-horseshoe qualificati<strong>on</strong>, because it does not<br />
have its own accommodati<strong>on</strong>. To obtain the 4-horseshoe qualificati<strong>on</strong> the instituti<strong>on</strong> has<br />
to corresp<strong>on</strong>d to lots of requirements, e.g.: envir<strong>on</strong>mental aspects, prescripti<strong>on</strong>s of<br />
horse-keeping, professi<strong>on</strong>al skill of horses, aspects related to riding services,<br />
professi<strong>on</strong>al and pers<strong>on</strong>al factors. Furthermore, the qualificati<strong>on</strong> indicates that there are<br />
many horses in the instituti<strong>on</strong> and the Riding Academy renders several types of service.<br />
The number, breed, age and educati<strong>on</strong> of horses are very important c<strong>on</strong>cerning the<br />
popularity of an instituti<strong>on</strong>. The instituti<strong>on</strong> is nowadays suitable for keeping 48 horses.<br />
In winter there are 42 to 48 and in summer 38 to 40 horses. The difference can be<br />
attributed to the attractive force of the roofed rising ground (20 x 60 m), i.e. this<br />
establishment offers advantageous c<strong>on</strong>diti<strong>on</strong>s for riding (independent from the weather).<br />
In <strong>term</strong>s of breeds, there are Gidrans, Hungarian and German Sport Horses, Shagya and<br />
P<strong>on</strong>ies. Ten horses are owned by the University of Debrecen, while the others are in<br />
private ownership. Nowadays, the youngest horse is 4, the oldest horse is 15 years old.<br />
The placement of the establishment, horse stock, service types/prices and last but<br />
not least attendants are de<strong>term</strong>ining aspects c<strong>on</strong>cerning the status of the riding<br />
establishment. The Riding Academy is in the town, but at the same time it is in a green<br />
belt, this is a double advantage. The riding establishment has <strong>on</strong>ly 4 regular staff: 2<br />
grooms and 2 instructors. Besides the regularly employed instructors further 1 to 6<br />
instructors work periodically in the instituti<strong>on</strong>. The main 3 services of the instituti<strong>on</strong> are<br />
educati<strong>on</strong> (2200 -2800 HUF per occasi<strong>on</strong>), rental keeping (40-50000 HUF per m<strong>on</strong>th)<br />
and establishment use (1200-2200 HUF per horse per occasi<strong>on</strong>). The number of riding<br />
less<strong>on</strong>s is 20 to 30 less<strong>on</strong>s a week, which relates to both horses and instructors; this<br />
means full house. There is no precise calculati<strong>on</strong> c<strong>on</strong>cerning the number of visitors; a<br />
daily average can be estimated as follows: 36 riders, 30 attending pers<strong>on</strong>s and 20<br />
visitors. On the basis of this estimati<strong>on</strong>, can be said, that the Riding Academy has about<br />
4000 to 5000 visitors per year; 2000 of them arrive from Debrecen (and its<br />
surroundings), a further 2000 of them from the other parts of Hungary and about 300<br />
come from other countries.<br />
Nowadays, a riding establishment has several possibilities to advertise its services.<br />
For instance, the Riding Academy advertises its services in newspapers, <strong>on</strong> the Internet,<br />
in radio and TV. Since there is full house, no marketing development is planned.<br />
189<br />
9
However, material development is probable because in winter there is a higher demand<br />
for rental keeping and establishment use than the instituti<strong>on</strong> could nowadays offer.<br />
Besides the above menti<strong>on</strong>ed development ideas, the elaborati<strong>on</strong> of a “Riders’<br />
path” offers a further, very interesting possibility in the Northern Great Plain. This path<br />
would be based <strong>on</strong> the collaborati<strong>on</strong> of riding establishments; during this path visitors<br />
could get acquainted with the biological characteristics, different riding activities as<br />
well as occupati<strong>on</strong>s c<strong>on</strong>nected with riding.<br />
LITERATURE<br />
Borsy Z. (editor.). 1998. Általános <strong>term</strong>észet földrajz. (General natural geography)<br />
Budapest: Nemzeti Tankönyvkiadó (Nati<strong>on</strong>al Educati<strong>on</strong> Publisher).<br />
ENSZ (2002) World Ec<strong>on</strong>omic Situati<strong>on</strong> and Prospects 2002 (Sales No. E.02.II.C.2),<br />
United Nati<strong>on</strong>s, New York 2002<br />
I.Fehér and M Kóródi. 2007. A vidéki turizmus sajátosságai (Characteristics of the rural<br />
tourism) . Agroinform. January 2007. (extraordinary issue).<br />
R.Incze ands G.Hevessy 2007. Visz<strong>on</strong>yunk a lóhoz (Our relati<strong>on</strong>ship with the horse).<br />
Agroinform. February 2007..<br />
L. Lisztes 2007. Hortobágyi Nemzeti Park: az értéktől az élményig (Natural Park of<br />
Hortobágy: from value to experience). Presentati<strong>on</strong>: Debrecen, 22.01.2007.<br />
J. Lóska 2001. Az Észak-magyarországi és Észak-alföldi régió lovasturizmusának<br />
helyzete és kib<strong>on</strong>takozásának lehetőségei (Status and possibilities of development<br />
c<strong>on</strong>cerning riding tourism of Northern Hungary and Northern Great Plain)<br />
(Regi<strong>on</strong>ális lovas turisztikai <strong>term</strong>ékfejlesztési tervtanulmány és programjavaslat<br />
(Preliminary study and programme proposal for product development regarding<br />
riding tourism). Vanyarc-Sarlóspuszta.<br />
Magyar Turizmus Zrt. 2006a. Lóhát<strong>on</strong> Magyarország<strong>on</strong> (On horseback in Hungary).<br />
Magyar Turizmus Zrt. 2006b. Turizmus Magyarország<strong>on</strong> (Tourism in Hungary) 2005.<br />
World Travel and Tourism Council (WTTC) (2004) Annual Report: Progress &<br />
Priorities 2004/05<br />
Online sources<br />
www.itth<strong>on</strong>.hu<br />
http://www.mth.gov.hu/<br />
www.equi.hu<br />
www.utik<strong>on</strong>yv.hu<br />
http://lovas.lap.hu/<br />
Interviews<br />
Boros Sándor. Instructor, Debreceni Lovasakadémia. Személyes interjú (Riders’<br />
instructor, Debrecen Riding Academy, pers<strong>on</strong>al interview): Debrecen, 2007.01.09.<br />
László Lisztes. Hortobágyi Nemzeti Park. Személyes interjú (Nati<strong>on</strong>al Park of<br />
Hortobágy, pers<strong>on</strong>al interview): Debrecen, 22.01.2007..<br />
Varga Emese. Mátai Ménes. Telef<strong>on</strong>os interjú (Ph<strong>on</strong>e interview): 2007.01.23.<br />
190<br />
10
TISSUE CULTURE METHODS FOR SCREENING RESISTANCE TO<br />
PECTOBACTERIUM CHRYSANTHEMI IN POTATO<br />
Ildikó Hudák 1 , Mária Hevesi 2 , Judit Dobránszki 1 and Katalin Magyar-Tábori 1<br />
1 Research Centre of University of Debrecen, Westsik V. u. 4-6, H-4400 Nyíregyháza,<br />
2 Corvinus University of Budapest, Faculty of Horticultural Science, Villányi út 29-43.,<br />
H-1118 Budapest, Hungary<br />
ABSTRACT<br />
Pectobacterium chrysanthemi (Erwinia chrysanthemi) cause vascular wilts, stunting,<br />
soft rot, spotting of leaves or parenchymal necroses of a wide range of plant species and<br />
cultivars like Allium cepa, Capsicum anuum, Nicotiana tabacum and Solanum<br />
tuberosum. This bacterium affects potatoes in almost every regi<strong>on</strong> in the world. At the<br />
moment the chemical protecti<strong>on</strong> is not solved (Dobránszki and Hevesi, 2002.). Altought<br />
the sensitivity of produced cultivars are different, resistant cultivars are not available for<br />
management of these diseases. The resistance in potato usually comes from wild<br />
Solanum species. In our Research Centre we have bred potato for more decades. Our<br />
breeding work based <strong>on</strong> using of the different wild Solanum species bearing resistance<br />
or tolerance against Pectobacterium species (Hudák et al., 2002.). The aim of our<br />
research is working out new in vitro methods for evaluati<strong>on</strong> of the level of resistance or<br />
tolerance against potato soft rot. We infected in vitro planlets. For evaluati<strong>on</strong> of the<br />
data, disease rating was used. By the help of disease rate we quantified the differences<br />
am<strong>on</strong>g potato cl<strong>on</strong>es. On the basis of our results the in vitro methods may be suitable<br />
for fast test of the breeding materials, cl<strong>on</strong>es or cultivars against Pectobacterium-strains.<br />
Keywords: Pectobacterium chrysanthemi, soft rot, disease rate, in vitro methods<br />
INTRODUCTION<br />
Pectobacterium species causes plant diseases that include blights, cankers, die back,<br />
leaf spots, wilts, discolorati<strong>on</strong> of plant tissues and especially soft rots variously<br />
described as stalk rot, crown rot, stem rot or fruit collapse (Brenner et al., 2005.).<br />
Pectobacterium chrysanthemi (Erwinia chrysanthemi) is <strong>on</strong>e of the phytopathogenic<br />
enterobacteria, which causes soft rot disease in various plants. This bacterium affects<br />
potatoes in almost every regi<strong>on</strong> in the temperate, semitropical and tropical z<strong>on</strong>e of the<br />
world. Its pathogenecity is due to its ability to secret several extracellular enzymes<br />
including pectate lyases, cellulases and proteases. These extracellular enzymes attack<br />
cell walls and membranes of plants leading to plant tissue macerati<strong>on</strong> (Kelman, 1990.).<br />
Because the field experiments would be dangerous and prol<strong>on</strong>ged therefore an in vitro<br />
method should be elaborated. We carried out with Pectobacterium-strain isolated in our<br />
country. We adopted and compared different in vitro infecti<strong>on</strong> methods. We infected in<br />
vitro planlets. We described the symptoms after in vitro infecti<strong>on</strong>.<br />
MATERIALS AND METHODS<br />
We used the bacterial strain isolated by Mária Hevesi in Hungary. In vitro cultured<br />
shoots of potato Boró, Réka, Rachel, 136/92 and 34/85 were used as test plants. The<br />
shoots were propagated <strong>on</strong> horm<strong>on</strong>e-free MS medium. The cultures were illuminated by<br />
191
white light of 105 μMol s -1 m -2 using 16/8 hour light/dark cycles. The temperature was<br />
20-22 C 0 . Three-week-old in vitro potato plants were infected during the experiments.<br />
Pectobacterium chrysanthemi was grown at 26 C 0 <strong>on</strong> Nutrient medium. The bacterial<br />
suspensi<strong>on</strong> c<strong>on</strong>tained 10 8 CPU. The inoculati<strong>on</strong> was made by a forceps immersed in the<br />
suspensi<strong>on</strong>. On c<strong>on</strong>trol treatment we wounded the shoots with a forceps immersed in<br />
sterile distilled water.<br />
The symptoms were evaluated after <strong>on</strong>e week. For evaluati<strong>on</strong> of the data disease<br />
rating were used. The diseased plants were divided into five categories based <strong>on</strong> the<br />
number of the wilted leaves. The disease categories were de<strong>term</strong>ined as follows:<br />
� Symptom-free plantlets<br />
� 1-25 % of leaves wilted<br />
� 26-50 % of leaves wilted<br />
� 51-75 % of leaves wilted<br />
� 76-100 % of leaves wilted<br />
With modificati<strong>on</strong> of this method a special disease rate (DR) was worked out:<br />
Σ [(N1 x 1) + (N2 x 2) + (N3 x 3) + (N4 x 4) + (N5 x 5)]<br />
DR = --------------------------------------------------------------------------<br />
Σ N<br />
N1-5: Number of plantlets in each disease category<br />
N: All obtained plantlets<br />
RESULTS AND DISCUSSION<br />
The results of experiments are summarized:<br />
During the experiments we chose the virulent strains of Pectobacterium chrysanthemy.<br />
We worked out in vitro methods of infecti<strong>on</strong>. We described the symptoms after in<br />
vitro infecti<strong>on</strong>. Disease rate was used for evaluati<strong>on</strong> of the data. With the help of<br />
disease rate we quantified the observable differences (symptoms). We separated three<br />
sensitivity categories:<br />
If 1 < DR < 2 n<strong>on</strong> sensitive<br />
If 2 < DR < 3 middle sensitive<br />
If 3 < DR very sensitive<br />
Symptoms: The symptoms were evaluated after in vitro infecti<strong>on</strong> <strong>on</strong> the seventh day. In<br />
the c<strong>on</strong>trol treatment the plantlets were healthy. No any leaf or stem symptoms could be<br />
observable. The place of wounding could be seen but became dry (Figure 1). In the case<br />
of cl<strong>on</strong>e 34/85 were not any leaf symptoms, at the worst case below or bey<strong>on</strong>d<br />
wounding stem symptoms could be observed. The place of wounding could be seen<br />
(Figure 2). Wilt of the <strong>on</strong>e-<strong>on</strong>e leaf could be observable in the Réka and Rachel. The<br />
place of the wounding could be seen clearly. Stem symptoms were typical. From the<br />
wounding place, the stem rotted off and split in two (Figure 3). Both stem and leaf<br />
192
symptoms could be observable in the Boró and cl<strong>on</strong>e 136/92. Seventy percent of leaves<br />
of the plantlets rotted off and str<strong>on</strong>g stem symptoms were observable <strong>on</strong> the seventh day<br />
after in vitro infecti<strong>on</strong> (Figure 4).<br />
Fig. 1. Symptoms of the c<strong>on</strong>trol treatment after in vitro infecti<strong>on</strong> <strong>on</strong> the seventh day.<br />
Fig. 2. Symptoms of the 34/85 cl<strong>on</strong>e after in vitro infecti<strong>on</strong> <strong>on</strong> the seventh day.<br />
Fig. 3. Symptoms of the Rachel and Réka after in vitro infecti<strong>on</strong> <strong>on</strong> the seventh day.<br />
193
Fig. 4. Symptoms of the Boró and 136/92 cl<strong>on</strong>e after in vitro infecti<strong>on</strong><br />
<strong>on</strong> the seventh day.<br />
We carried out the infecti<strong>on</strong> of the in vitro plantlets and de<strong>term</strong>ined values of DR in<br />
average of three, independent experiments in the case of five cl<strong>on</strong>es (Figure 5).<br />
Values of disease rate against Pectobacterium chrysanthemi:<br />
• 34/85 cl<strong>on</strong>e (DR = 1,2) n<strong>on</strong> sensitive,<br />
• Rachel (DR = 2,3) middle sensitive,<br />
• Réka (DR = 2,5) middle sensitive,<br />
• 136/92 cl<strong>on</strong>e (DR = 3,2) very sensitive,<br />
• Boró (DR = 3,3) very sensitive.<br />
3,5<br />
3<br />
2,5<br />
2<br />
1,5<br />
1<br />
0,5<br />
0<br />
1,2<br />
34/85<br />
Disease rate (DR)<br />
2,3<br />
Rachel<br />
2,5<br />
Réka<br />
3,2<br />
136/92<br />
3,3<br />
Boró<br />
Fig. 5. Values of disease rate against Pectobacterium chrysanthemi<br />
in 34/85, Rachel, Réka, Boró and 136/92 cl<strong>on</strong>es.<br />
It could be c<strong>on</strong>cluded that the in vitro methods are suitable for testing sensitivity of the<br />
cl<strong>on</strong>es and cultivars. In the future we have the intenti<strong>on</strong> to extend over these<br />
194<br />
DR
experiments the other cl<strong>on</strong>es and we want to compare our results with in vivo<br />
experiments.<br />
ACKNOWLEDGEMENTS<br />
This work was supported by GVOP (project N 0 .: GVOP-3.1.1.-2004-05-0041/3.0).<br />
REFERENCES<br />
Brenner, D. J., Krieg, N. R. and Staley, J. T. 2005. Bergey’s manual of Systematic<br />
Bacteriology, Part II. The protobacteria. P. 721-730.<br />
Kelman, A. 1990. Blackleg, bacterial soft rot. In: W.J. Hooker (ed.): Compendium of<br />
Potato Diseases, Part II. Disease in the Presence of Infectious Pathogens. p. 27-29.<br />
Dobránszki, J. and Hevesi, M. 2002. A burg<strong>on</strong>ya fekete szártőrothadása és<br />
baktériumos lágyrothadása. Burg<strong>on</strong>ya<strong>term</strong>esztés. 2002. augusztus. p. 16-19.<br />
Hudák, I., Dobránszki, J. and Hevesi, M. 2002. Hazánkban izolált, burg<strong>on</strong>yát<br />
károsító Erwinia-fajok vizsgálata. "a Magyar Tudomány Napja 2002" alkalmából<br />
rendezett Szabolcs-Szatmár-Bereg Megyei Tudományos K<strong>on</strong>ferencia. 2002.<br />
november 11.<br />
195
EFFECT OF AUXIN ON CALLUS FORMING CAPACITY OF DIFFERENT<br />
PEA GENOTYPES (PISUM SATIVUM L.) 1<br />
ABSTRACT<br />
Katalin Magyar-Tábori, Judit Iszály-Tóth, Judit Dobránszki,<br />
Nóra Mendler-Drienyovszki<br />
Research Centre of Debrecen University,<br />
Nyíregyháza, Westsik V. street 4-6, H-4400 Hungary<br />
Effect of different auxins (IBA, NAA and 2,4-D) were tested in two c<strong>on</strong>centrati<strong>on</strong>s (2.0<br />
and 4.0 mgl -1 ) <strong>on</strong> callus forming capacity of eight genotypes (Graphis, Baccara, 2107,<br />
Hanka, Hunor, Janus, M49060 and Erbi). Leaflets and stem secti<strong>on</strong> without node were<br />
used as explants and they were placed <strong>on</strong> G57 media supplemented with 3 % sucrose,<br />
0.8 % agar-agar and 1.0 mgl -1 BA. Cultures were incubated in dark for 3 weeks at 26 ºC,<br />
after then they were cultured <strong>on</strong> 16 h photoperiod for further 4 weeks. Callus forming<br />
capacity was observed and scored weekly. Our results c<strong>on</strong>firmed that there are<br />
significant differences between genotypes: 2107 breeding line showed the greatest,<br />
while cv. Janus had the least callus forming capacity. The best results were achieved by<br />
IBA in both c<strong>on</strong>centrati<strong>on</strong>s, although the effect of higher level of NAA was similar,<br />
depending <strong>on</strong> genotypes. 2,4-D was not effective especially in high c<strong>on</strong>centrati<strong>on</strong>.<br />
Inducti<strong>on</strong> of callus was more effective <strong>on</strong> leaflets than <strong>on</strong> stem segments.<br />
Keywords: Pisum sativum L., callus, auxin, in vitro, explant type<br />
INTRODUCTION<br />
The pea (Pisum sativum L.) is an important protein and vegetable crop. Improving of<br />
drought tolerance of peas has been <strong>on</strong>e of the major tasks in most breeding programmes<br />
and in vitro methods can help the breeding work. Results of Ezhova et al. (1995)<br />
suggest that mutati<strong>on</strong>s affecting developmental characters may be expressed in tissue<br />
culture. In vitro callus culture can be used to model breeding for heterosis and to predict<br />
productivity and heterosis in pea plants (Rybtsov et al., 1997). However, the applicati<strong>on</strong><br />
of in vitro selecti<strong>on</strong> system can <strong>on</strong>ly be achieved by means of plant cell and tissue<br />
culture where the inducti<strong>on</strong> of somacl<strong>on</strong>al variati<strong>on</strong> frequently is an inherent feature<br />
(Lehminger-Mertens & Jacobsen, 1993). Because of there is a very few informati<strong>on</strong><br />
about callus culture of peas and resp<strong>on</strong>ses of genotypes to in vitro c<strong>on</strong>diti<strong>on</strong>s can be<br />
different (Dobránszki et al., 1999, Vandorne et al., 1995) it is necessary to work out the<br />
callus inducti<strong>on</strong> method, which can be used for the genotypes included in our breeding<br />
programme. Although in the case of tobacco auxin autotrophic callus could be obtained<br />
(Csiszár et al, 2004) and there is horm<strong>on</strong>e-aut<strong>on</strong>omous callus for sugarbeet (Saunders &<br />
Daub, 1984) in the case of pea the presence of auxin was necessary for callus inducti<strong>on</strong><br />
(Gantchev, 1997). In our experiments effect of auxins and explant type <strong>on</strong> callus<br />
development were tested for eight pea genotypes to select the optimal callusing<br />
treatment.<br />
1 This work was supported by GVOP-3.1.1.-2004-05-0041/3.0) project.<br />
196
MATERIALS AND METHODS<br />
Shoot cultures of eight pea genotypes (Graphis, Baccara, 2107, Hanka, Hunor, Janus,<br />
M49060 and Erbi) were maintained in vitro and explants for the experiments were<br />
isolated from micropropagated plants. Two kinds of explants were used: fully<br />
developed leaflets of the upper third of stem or stem segments without node. Isolati<strong>on</strong><br />
was made in a sterile soluti<strong>on</strong> (0.15 mgl -1 citric acid and 0.1 mgl -1 ascorbic acid) to<br />
prevent the withering of explants and the deleterious oxygenati<strong>on</strong> process. Explants<br />
were placed <strong>on</strong> G 57 media (salts and vitamins) (Gamborg et al., 1968) supplemented<br />
with MS-Fe (Murashige & Skoog, 1962), 3.0 % sucrose, 0.8 % agar-agar and 1.0 mgl -1<br />
benzyl-adenine (BA) as cytokinin. Naphthalane-acetic acid (NAA), indolebutyric acid<br />
(IBA) and 2,4-dichlorophenoxyacetic acid (2,4-D) were applied as auxins in two<br />
c<strong>on</strong>centrati<strong>on</strong>s: 2.0 and 4.0 mgl -1 . Cultures were incubated in dark for 3 weeks at 26 ºC,<br />
after then they were cultured <strong>on</strong> 16 h photoperiod for further 4 weeks. Callus forming<br />
capacity was observed and scored weekly as follows:<br />
1- there is no callus <strong>on</strong> the explant<br />
2- some calluses <strong>on</strong> some end of explant<br />
3- there are some calluses <strong>on</strong> both end of explant<br />
4- str<strong>on</strong>g callus forming but the surface of the explant can be seen<br />
5- the callus overgrows the explant.<br />
Statistical analyses were made by variance analyses followed by Tukey test, using by<br />
SPSS 9.0 for Windows programme.<br />
RESULTS AND DISCUSSION<br />
In our experiments callus inducti<strong>on</strong> and development were obtained in all treatments<br />
and for each genotype. The statistical analysis showed significant interacti<strong>on</strong>s between<br />
genotypes, explant type and auxins. In the average of all treatments the IBA was the<br />
best auxin, and the 2,4-D was the least efficient. Effect of NAA in lower c<strong>on</strong>centrati<strong>on</strong><br />
(2.0 mgl -1 ) was similar to the effect of IBA, although by the end of fourth week the<br />
callus forming capacity was significantly lower (table 1.).<br />
Table 1. The main-effect of auxin <strong>on</strong> callus forming capacity (in the average of both<br />
explants and genotypes).<br />
Callus development <strong>on</strong> the end of<br />
1 st 2 nd 3 rd 4 th<br />
Auxin<br />
treatments<br />
mgl -1 week<br />
NAA 2.0 1.69 d 2.52 d 3.40 cd 4.25 cd<br />
NAA 4.0 1.59 c 2.43 c 3.34 c 4.21 c<br />
2,4-D 2.0 1.51 b 2.22 b 2.81 b 3.39 b<br />
2,4-D 4.0 1.34 a 2.01 a 2.63 a 3.21 a<br />
IBA 2.0 1.64 d 2.52 d 3.47 d 4.36 e<br />
IBA 4.0 1.64 d 2.54 d 3.47 d 4.32 de<br />
Means within column signed by the same letter bel<strong>on</strong>g to the same homogeneous group.<br />
197
The genotypes differentiated c<strong>on</strong>sidering the degree of callus development already from<br />
the end of first week. By the end of fourth week the 2107 breeding line showed the<br />
str<strong>on</strong>gest callus forming capacity, while cv. Janus was the weakest. Only cv. Hunor and<br />
Erbi bel<strong>on</strong>gs to the same group, the others differ from each other significantly (table 2.).<br />
Table 2. Effect of genotypes <strong>on</strong> callus inducti<strong>on</strong> and growth in the average of all<br />
treatments.<br />
Callus development <strong>on</strong> the end of<br />
1 st 2 nd 3 rd 4 th<br />
Genotypes<br />
week<br />
Graphis 2.33 g 3.21 f 4.08 g 4.42 f<br />
Baccara 1.88 f 2.69 d 3.51 e 4.30 e<br />
2107 1.85 f 2.84 e 3.76 f 4.65 g<br />
Hanka 1.76 e 2.67 d 3.20 d 4.05 d<br />
Hunor 1.00 a 1.72 a 2.58 a 3.60 b<br />
Janus 1.32 d 1.85 b 2.51 a 2.97 a<br />
M49060 1.23 c 2.12 c 2.87 c 3.77 c<br />
Erbi 1.10 b 1.79 ab 2.78 b 3.63 b<br />
Means within column signed by the same letter bel<strong>on</strong>g to the same homogeneous group.<br />
Similarly to Gantotti and Kartha (1983), who could induce the callus cultures <strong>on</strong> each<br />
part of plant, excluding the root tip, we also obtained callus culture <strong>on</strong> both type of<br />
explants (table 3.). Because of significant interacti<strong>on</strong>s and since we found significant<br />
differences between callus forming capacity of different type of explants, we c<strong>on</strong>tinued<br />
the statistical analysis as <strong>on</strong>e-way ANOVA.<br />
Table 3. Effect of explant type <strong>on</strong> callus forming capacity in the average of all<br />
treatments.<br />
Callus development <strong>on</strong> the end of<br />
1 st 2 nd 3 rd 4 th<br />
Explant type<br />
week<br />
Leaflets 1.55 a 2.49 a 3.35 b 4.21 b<br />
Stem segments 1.58 a 2.25 a 3.00 a 3.67 a<br />
Means within column signed by the same letter did not differ significantly<br />
Callus development was better <strong>on</strong> the leaf explants so we analysed the effect of auxins<br />
<strong>on</strong> callus development <strong>on</strong> leaf explants. Molnár (1993) found 2,4-D to be suitable for<br />
callus inducti<strong>on</strong>, and we also could induce callus development by the use of 2,4-D,<br />
although the degree of callus development was decreased by 2,4-D, especially in higher<br />
c<strong>on</strong>centrati<strong>on</strong>s, excluding cv. Hanka (table 4). Gantchev (1997) also tested different<br />
c<strong>on</strong>centrati<strong>on</strong>s of 2,4-D (2.0-32.0 mgl -1 ) <strong>on</strong> internodal stem segments of pea, and found<br />
198
that callus inducti<strong>on</strong> and development were obtained in the presence of the lowest level<br />
of 2,4-D.<br />
Table 4. Effect of auxins <strong>on</strong> calli developed <strong>on</strong> leaf explants of different genotypes by<br />
the end of fourth week.<br />
Genotypes<br />
Callus development <strong>on</strong> the media supplemented with<br />
NAA 2,4-D IBA<br />
2.0 4.0 2.0 4.0 2.0 4.0<br />
mgl -1<br />
Graphis 5.00 c 5.00 c 3.80 a 4.14 b 5.00 c 5.00 c<br />
Baccara 5.00 c 5.00 d 4.00 b 3.73 a 4.88 d 4.38 c<br />
2107 5.00 c 5.00 c 4.70 b 4.28 a 4.95 c 4.93 c<br />
Hanka 5.00 c 5.00 c 4.07 b 4.00 b 3.73 a 3.75 a<br />
Hunor 3.60 a 4.06 b 3.94 b 3.86 ab 4.37 c 4.97 d<br />
Janus 4.14 c 3.83 c 2.74 b 1.80 a 4.06 c 4.04 c<br />
M49060 3.50 a 4.00 b 3.37 a 3.51 a 4.00 b 4.03 b<br />
Erbi 4.49 b 3.67 a 3.49 a 3.71 a 4.46 b 4.26 b<br />
Means within row signed by the same letter bel<strong>on</strong>g to the same homogeneous group<br />
Presence of NAA and IBA irrespectively of their c<strong>on</strong>centrati<strong>on</strong>s was enough to induce<br />
and maintain an intense callus development of leaf explants of cv. Graphis, 2107 and cv<br />
Janus. In the case of cv. Baccara the best results were obtained <strong>on</strong> media c<strong>on</strong>taining IBA<br />
at 2.0 mgl -1 c<strong>on</strong>centrati<strong>on</strong>, or NAA in both horm<strong>on</strong>e levels. IBA applied in 4.0 mgl -1<br />
was the best treatment for cv. Hunor, while both levels of IBA were favourable for<br />
M49060 and cv. Erbi. In the case of latter cultivars the effect of NAA was the same<br />
advantageous in c<strong>on</strong>centrati<strong>on</strong> of 2.0 mgl -1 or 4.0 mgl -1 for cv. Erbi and M49060,<br />
respectively. Only the cv. Hanka was the genotype, which resp<strong>on</strong>ded to IBA treatments<br />
by the weakest callus development, while inducti<strong>on</strong> of callus development was the most<br />
successful <strong>on</strong> media with NAA. Even if the 2,4-D is generally used for callus inducti<strong>on</strong><br />
in peas (Gantchev, 1997, Molnár, 1993, ) and in other species (Bayliss & Dunn, 1979,<br />
Arora & Singh, 1978, C<strong>on</strong>ger et al., 1982) we found the other auxins (NAA and IBA) to<br />
be more effective to increase callus forming capacity of the majority of pea genotypes<br />
tested in our experiments.<br />
Our results suggested that callus formati<strong>on</strong> can be induced <strong>on</strong> all genotypes and by use<br />
of each auxins, although the genotypes required an optimal horm<strong>on</strong>e c<strong>on</strong>tent in medium<br />
for intensive callus growth. Further research is underway in our laboratory to examine<br />
the effects of different osmotic materials <strong>on</strong> the rate of callus growth.<br />
REFERENCES<br />
Arora, I.K., Singh, R.N. (1978): Growth horm<strong>on</strong>es and in vitro callus formati<strong>on</strong> of<br />
papaya. Scientia Horticulturae, Vol. 8, Iss. 4, 357-361.<br />
199
Bayliss, M. W., Dunn, S. D. M. (1979): Factors affecting callus formati<strong>on</strong> from<br />
embryos of barley (Hordeum vulgare). Plant Science Letters, Vol. 14, Iss. 4, 311-<br />
316.<br />
C<strong>on</strong>ger, B.V., Hilenski, L.L., Lowe, K.W., Carabia, J.V. (1982): Influence of<br />
different auxins at varying c<strong>on</strong>centrati<strong>on</strong>s <strong>on</strong> callus inducti<strong>on</strong>s and growth from<br />
embryo and leaf-explants in gramineae. Envir<strong>on</strong>mental and Experimental Botany,<br />
Vol. 22., Iss. 1, 39-48.<br />
Csiszár, J., Szabó, M., Erdei, L., Márt<strong>on</strong>, L., Horváth, F., Tari, I. (2004): Auxin<br />
autotrophic tobacco callus tissues resist oxidative stress: the importance of<br />
glutathi<strong>on</strong>e S-transferase and glutathi<strong>on</strong>e peroxidase activities in auxin<br />
heterotrophic and autotrophic calli. Journal of plant Physiology, Vol. 161, iss. 6,<br />
691-699.<br />
Dobránszki, J., Takács-Hudák, Á., Magyar-Tábori, K., Ferenczy, A. (1999): Effect<br />
of medium <strong>on</strong> the callus-forming capacity of different potato genotypes. Acta<br />
Agr<strong>on</strong>omica Hungarica, 47 (1), pp. 59-61.<br />
Ezhova, T. A., Bagrova A. M., Gostimski, S. A. (1995): Cell selecti<strong>on</strong> as a possible<br />
reas<strong>on</strong> for the specificity of somacl<strong>on</strong>al variati<strong>on</strong> in pea. Plant Breeding 114 (6):<br />
520-524, dec.<br />
Gamborg, O. L., Miller, R. A., Ojima, K. (1968): Nutrient requirements of<br />
suspensi<strong>on</strong> cultures of soybean root cells. Exp. Cell. Res. 50, 151-157.<br />
Gantchev, L. S. (1997): Initial stages of callus development <strong>on</strong> internodal stem<br />
segments of pea and faba bean in vitro. Biotechnology & Biotechnological<br />
equipment 11 (1-2): A3-A16.<br />
Gantotti, B.V., Kartha, K.K. (1983): Pea. In: Evans, A.D., Sharp, R.W., Ammirato,<br />
P.V., Yamada, Y. (eds): Handbook of Plant Cell Culture. Vol. 4. Techniques and<br />
Applicati<strong>on</strong>s. McMillan Publ. Co., New York, 370-418.<br />
Lehminger-Mertens, R., Jacobsen, H.J. (1993): Regenerati<strong>on</strong> of plants from<br />
protoplast of pea (Pisum sativum L.). Biotechnology in Agriculture and Forestry,<br />
Vol. 22. pp. 97-104. In: Bajaj, Y.P.S. (ed): Plant Protoplasts and genetic<br />
Engineering III: Springer-Verlaug Berlin, Heidelberg.<br />
Molnár, Z. (1993): Importance of the in vitro micropropagati<strong>on</strong> methods in faba beans<br />
(Vicia faba L.) and peas (Pisum sativum L.) breeding. Acta Agr<strong>on</strong>omica<br />
Óváriensis Vol. 35, No. 1., 119-223.<br />
Murashige, T., Skoog, F. (1962): A revised medium for rapid growth and bioassays<br />
with tobacco tissue cultures. Physiol. Plant. 15, 473-479.<br />
Rybtsov S. A., Ezhova T. A., Gostimskii S. A. (1997): Use of in vitro tissue culture to<br />
study heterosis in the pea. Genetika, 33 (11): 1517-1522, Nov.,<br />
Saunders, J.W., Daub, M.E. (1984): Shoot regenerati<strong>on</strong> from horm<strong>on</strong>e-aut<strong>on</strong>omous<br />
callus from shoot cultures of several sugarbeet (Beta vulgaris L.) genotypes. Plant<br />
Science Letters, Vol. 34., Iss. 1-2, 219-223.<br />
Vandorne, L.E., Marshall, G., Kirkwood, R.C. (1995): Somatic embryogenesis in<br />
pea (Pisum sativum L.) – effect of explant, genotype and culture c<strong>on</strong>diti<strong>on</strong>s.<br />
Annals of Applied Biology 126 (1): 169-179.<br />
200
COMBINING ABILITY STUDIES IN A SEVEN- PARENTAL DIALLEL<br />
CROSS OF DIFFERENT PEAS VARIETES<br />
(PISUM SATIVUM L.)<br />
Nóra Mendler-Drienyovszki, Mándi Lajosné, Katalin Magyar-Tábori,<br />
Györgyi Cs<strong>on</strong>tos<br />
Research Centre of University of Debrecen,<br />
Westsik V. u. 4-6. H-4400. Nyíregyháza, Hungary<br />
ABSTRACT<br />
Results of a 7x7 diallel cross c<strong>on</strong>taining parents and F1 without reciprocals are<br />
discussed after Model 1 and Method 2 of Griffing. Affecting three yielding capacity<br />
traits were studied and analized: pod length, pod per plant, seed per pod.<br />
The difference between genotypes in every traits were significant (P=0,01).<br />
Predominance of additive effects in tested features were typical excluding pod pr plant.<br />
The phenotypical performace is grater ofthe parents, which have higher GCA values.<br />
In general the effects of SCA isn't correlate with performance of F1 generati<strong>on</strong>.<br />
INTRODUCTION<br />
Peas have been grown as an important source of animal feed and human food for many<br />
centuries. Over this time, pea has been selected for these uses. Several thousand<br />
varieties exist throughout the world.<br />
The majority part of our most successful pea varieties are more or less near<br />
relati<strong>on</strong>ship with each other. The genetical background of high yield capacity can be the<br />
same because of the crosses of the best pea cultivars seldom result in better new line<br />
than the parents. However it can be supposed that the parents with good combining<br />
ability can produced better lines than the average in a later generati<strong>on</strong>. It's important in<br />
the plan of different cross- combinati<strong>on</strong>s to choose the parents purposefully. In practice<br />
the genotypes which hold a good phenotype values, not to be sure of inheritance their<br />
desired traits, but another not so good varieties would be valuable like a parent in the<br />
diallel cross.<br />
The diallel analysis is a quantitative genetics method in plant breeding where we<br />
can estimate the interacti<strong>on</strong> between enviroment and genes in a populati<strong>on</strong> or the<br />
offsprings from the selected paretnts (Mohay, 1986).<br />
The diallel crosses are used in generally from mating modells, because we can get<br />
the most completly genetic informati<strong>on</strong> about the genotypes.<br />
The base methods are the analysis of Griffing (1956), Hayman and Jinks (1954), which<br />
is based <strong>on</strong> diallel cross.<br />
These diallel methods are popular all over the world, and it can be used<br />
successfully in other species, for example maize (Daniel and Bajtai, 1975, Pepó et al.<br />
1989), sunflower (Füredi and Frank, 1981), wheat (Matuz, 1976, Du et al. 1999), pea<br />
(Csizmadia, 1985, Lőkös, 1985).<br />
201
MATHERIALS AND METHODS<br />
Based <strong>on</strong> observati<strong>on</strong>s of several years we selected ten varieteies from our varieties<br />
collecti<strong>on</strong> for diallel- parent. Seven varieteies got usable results (1:Léda, 2:Margó,<br />
3:Foremos, 4:Presto, 5:Mariza, 6:Early sweet, 7:Debreceni korai velő) since the<br />
crossing was not successful for all combinati<strong>on</strong>. We would like to produced a new early<br />
pea varieteies with high yielding capacity parameters. We made the crosses in 2002, in<br />
several repetiti<strong>on</strong>s depending the number of flowers. We sowed the parent and n<strong>on</strong>-<br />
reciprocal F1 generati<strong>on</strong> beside net in 2003. We made the observati<strong>on</strong>s in more<br />
qualitative and quantitative traits by instructi<strong>on</strong>s of UPOV. In this paper we analysed<br />
the pod length, seed per pod, and pod number in <strong>on</strong>e plan.<br />
Statistical methods: The Model I. Method II of Griffing (1956) used for the combining<br />
ability analysis of F1 data was:<br />
xijk=μ+gi+gj+sij+rij+bk+eijk<br />
where: μ= populati<strong>on</strong> means<br />
gi= GCA effect for parent i,<br />
gj= GCA effect for parent j,<br />
sij= SCA effect for parents i and j,<br />
rij= reciprocal effect<br />
bk= block effect for block k,<br />
eijk= error.<br />
Diallel Analysis and Simulati<strong>on</strong> Software by Burrow and Coors (1993) was used in this<br />
study.<br />
RESULTS<br />
Phenotypic values<br />
Seed per pod: the populati<strong>on</strong> average is 5,34 seed per pod. The parent number 3<br />
(Foremos) has the most seed number (5,73), and the parent number 6 (Debreceni korai<br />
velő) has the least (4,96). If we study these parent's combinati<strong>on</strong>s we can see that the<br />
best combinati<strong>on</strong> is the 3x4. This combinati<strong>on</strong> has the highest seed number (6,45).<br />
Combinati<strong>on</strong> of 4x6 has the least seed number (4,56).<br />
Pod length: the populati<strong>on</strong> average is 5,35 cm. The Foremos has the l<strong>on</strong>gest pod, too<br />
(5,70 cm) and the Margó ( parent number two) has the shortest pod length (4,96 cm).<br />
We can get the best result for the 4x7 combinati<strong>on</strong> (6,25 cm) in our combinati<strong>on</strong> which<br />
is better than the average. The worth combinati<strong>on</strong> is the 6x7 lines (4,20 cm).<br />
The third trait which was analysed the pod per plant. The populati<strong>on</strong> average is 4,857<br />
pods in <strong>on</strong>e plant. Better then the average is the Léda (parent number 1) (5,50), worst<br />
then the average is the Debreceni korai velő (4,27) (parent number 7).<br />
To study the average of the combinati<strong>on</strong>s themselves have seen the most successfully<br />
combinati<strong>on</strong> is the 1x6, the worst is the 2x4 combinati<strong>on</strong> (3 pods).<br />
Effects of combining ability<br />
The different btween genotypes in every traits were significant (P=0,01).<br />
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Table 1: Mean squares of GCA and SCA in every traits<br />
MS (GCA) MS (SCA)<br />
Pod per plant 3,175 3,524<br />
Pod length 1,817 0,852<br />
Seed per pod 1,126 0,527<br />
The GCA values are higher than <strong>on</strong>e in every traits and larger than SCA values, expect<br />
in the trait of pods per plant. The estimated GCA effects associated with each parent are<br />
shown in Table 2.<br />
Table 2: Generally combinig ability<br />
Parents Trait<br />
Pod per plant Pod length Seed per pod<br />
1. 0,771 0,362 -0,001<br />
2. -0,161 -0,489 0,021<br />
3. -0,095 0,415 0,466<br />
4. -0,228 0,027 0,099<br />
5. 0,104 -0,148 -0,068<br />
6. 0,304 -0,354 -0,464<br />
7. -0,695 0,186 -0,052<br />
se [g(i)] 0,260 0,124 0,157<br />
se [g(i)-g(j)] 0,397 0,190 0,240<br />
The values of GCA very similar to each other, because of selected parents were very<br />
similar respecting of yielding- features (excluding Foremos). The values of pod length<br />
of GCA are correlated with the higher valueses of seed per pod of GCA in Margó,<br />
Foremos, Presto, Mariza. The estimated of SCA effects are given in Table 3.<br />
Table 3: Specific combining ability effects<br />
Combinati<strong>on</strong> Traits<br />
Pod per plant Pod length Seed per pod<br />
1x2 0,200 -0,700 -0,495<br />
1x3 -0,533 -0,152 -0,546<br />
1x4 0,600 0,439 0,319<br />
1x5 -0,066 -0,068 0,151<br />
1x6 1,066 0,714 0,423<br />
1x7 -1,266 -0,232 0,148<br />
2x3 1,400 0,546 -0,056<br />
203
2x4 -1,466 -0,535 -0,249<br />
2x5 0,200 0,003 -0,101<br />
2x6 -1,000 0,393 0,270<br />
2x7 0,666 0,292 0,632<br />
3x4 -0,533 -0,514 0,535<br />
3x5 -0,200 0,185 0,537<br />
3x6 0,600 -0,255 -0,094<br />
3x7 -0,733 0,190 -0,375<br />
4x5 1,600 -0,183 -0,126<br />
4x6 -0,600 0,109 -0,417<br />
4x7 0,400 0,685 -0,062<br />
5x6 -1,266 0,018 -0,149<br />
5x7 -0,266 0,044 -0,310<br />
6x7 1,200 -0,979 -0,032<br />
se [s(i,j)] 0,512 0,246 0,311<br />
se [s(i,j)-s(i,k)] 0,794 0,381 0,481<br />
se [s(i,j)-s(k,l)] 0,687 0,330 0,417<br />
In the case of SCA the results were the same: we could not find any very different<br />
results of SCA in either combinati<strong>on</strong>. There are relatively higher values of SCA for pod<br />
per plant in 2x3 and 4x5 combinati<strong>on</strong>, and for pod- length in 1x6 and 4x7, and for seed<br />
number of pod in 2x7 and 3x5 combinati<strong>on</strong>s. The standard deviati<strong>on</strong>s are higher in<br />
number of pod per plant.<br />
DISCUSSION<br />
The parents in the diallel were selected c<strong>on</strong>sciously. Our aim is to produce an earlier<br />
variety with high yielding capacity thus these results just are the part of the whole<br />
experiment, which can help us to breed the new varieteies.<br />
Predominance of additive effects in tested features were typical excluding pod per<br />
plant thus the features can be improved any further in the respective combinati<strong>on</strong>s.<br />
These results are similar to other author's results. The reports <strong>on</strong> inheritance of yield-<br />
related traits in garden pea are variable as apparent from gene acti<strong>on</strong> that was reported<br />
to be additive (Rathore et al., 1995) and epistatic (Narsinghani et al., 1982) for pods per<br />
plant, additive (Singh and Singh, 1991) and n<strong>on</strong> additive (Srivastava and Singh, 1988)<br />
for seeds per pod.<br />
The studied traits, which has higher GCA effects-values has higher phenotypic<br />
values. According to our results the parents of the best phenotypic values combinati<strong>on</strong><br />
are the parents with the best GCA values parents (Table 4.) The best variety was the<br />
Foremos in the pod length and seed per pod traits, and these were accompained the<br />
highest GCA values. In the combinati<strong>on</strong> with the best phenotypic values, the <strong>on</strong>e parent<br />
is the Foremos (3.). Since the Foremos an earlier variety with l<strong>on</strong>g pod length (8-10 cm)<br />
and many seed per pod (8-9). These traits are just in a few combinati<strong>on</strong> have improving<br />
effects. These improving effects aren't so large rate, that we can stabilize the trait in the<br />
204
combinati<strong>on</strong>. We could not obtained that result what was required from the variety.<br />
Table 4: The best phenotypic combinati<strong>on</strong><br />
The combinati<strong>on</strong>s with the best The parents with the best<br />
performance<br />
GCA<br />
Pod per plant 1x6 1<br />
4x5 6<br />
1x4 5<br />
Pod length 4x7 3<br />
1x4 1<br />
3x7 7<br />
Seed per pod 3x4 3<br />
3x5 4<br />
2x7 2<br />
(<strong>on</strong>ly the three highest values are presented)<br />
We can say same c<strong>on</strong>necti<strong>on</strong> about the parents with worth values as menti<strong>on</strong>ed above,<br />
those parents and combinati<strong>on</strong>s got the least seed number and the shortest pod length.<br />
To study the SCA values the results are various (Table 5.). In general the effects of<br />
SCA isn't correlate with performance of F1 hybrid, except the seed per pod, where are<br />
the three combinati<strong>on</strong> give the rank in the performance and SCA values. Singh et al.<br />
(1991) also found c<strong>on</strong>necti<strong>on</strong> between performances and values of SCA.<br />
Table 5. The combinati<strong>on</strong>s with the best performace and SCA, <strong>on</strong>ly the three highest<br />
values are presented in the table.<br />
The combinati<strong>on</strong> with the<br />
best performace<br />
Pod per plant 1x6<br />
4x5<br />
1x4<br />
Pod length 4x7<br />
1x4<br />
3x7<br />
Seed per pod 3x4<br />
3x5<br />
2x7<br />
The combinati<strong>on</strong> with the<br />
best SCA<br />
4x5<br />
2x3<br />
6x7<br />
1x6<br />
4x7<br />
2x3<br />
2x7<br />
3x5<br />
3x4<br />
Further research works are underway to analise the inheritance of plant-heigh and 1000<br />
seed-yield by the use of diallel programe.<br />
In this year the F5 generati<strong>on</strong> will be observed by instructi<strong>on</strong>s of UPOV. We attend to<br />
205
compare the results of statistical analise to the field performance of breeding lines.<br />
REFERENCES<br />
Burrow, M. D.- J. G. Coors (1993): Diallel analysis and simulati<strong>on</strong>. User's guide.<br />
Dep. of Agr<strong>on</strong>. , Univ. of Wisc<strong>on</strong>sin, Madis<strong>on</strong>.<br />
C. G. Du- L. R. Nels<strong>on</strong>- M. E. McDaniel (1999): Diallel Analysis of Gene effects<br />
C<strong>on</strong>diti<strong>on</strong>ing resistance to Stagnospora nodorum (Berk.) in Wheat. Crop. Science,<br />
Vol. 39, May-June. 686-690.<br />
Csizmadia L. (1989): A borsó egyes mennyiségi tulajd<strong>on</strong>ságának öröklődése.<br />
Zöldség<strong>term</strong>esztési Kutató Intézet Bulletinje Kecskemét 22: 29-40.<br />
Daniel L.- Bajtai I.: (1975): Néhány mennyiségi tulajd<strong>on</strong>ság alakulása 11<br />
csemegekukorica beltenyésztett törzs diallél keresztezésben. Növény<strong>term</strong>elés 24:<br />
285-294.<br />
Füredi J.- Frank J. (1981): Napraforgó v<strong>on</strong>alak kombinálódó- képességének<br />
vizsgálata és a kombinációk genetikai elemzése Griffing- módszerrel.<br />
Növény<strong>term</strong>elés 30: 289- 300.<br />
Griffing, B. (1956.): C<strong>on</strong>cept of general and specific combinig ability in relati<strong>on</strong> to<br />
diallel crossig systems. Australian Journal of biological Sciencis, Melbourne 9.<br />
4:463-492.<br />
Hayman, B. I. (1954): The theory and analysis of diallel crosses. Genetics 39: 789-<br />
809.p.<br />
Jinks, J. L. (1954): The analysis of c<strong>on</strong>tinuous variatio in a diallel cross of Nicotiana<br />
rustica varieties. Genetics 39.:767-788.p.<br />
Matuz J. (1976): Heterózis és kombinálódóképesség vizsgálatok őszi búzafajtában.<br />
Doktori értekezés. Gödöllő.<br />
Mohay, J. (1986.): Diallélkeresztezési rendszer. Genetikai kislexik<strong>on</strong>. Natura Kiadó,<br />
Budapest. 40.<br />
Narsinghani, V. G.,- U.S.N. Rao- S.P. Singh. (1982): diallel cross analysis for<br />
quantitative traits in mutant pea types. Indian J. Agric. Sci. 52:364:367.<br />
Pepó P.- Pásztor K.- Palij A. F.- Pepó P. (1989): Kukorica mutánsv<strong>on</strong>alak genetikai<br />
analízie. Növény<strong>term</strong>elés 38: 193-199.<br />
Rathore, P. K.- V. P. Gupta- P. Plaha (1995): Genetics of seed yield and some other<br />
traits in pea using NCIII and triple cross analysis. Indian J. Pulse Res. 8: 119-122.<br />
Singh, M.N.- Singh, R.B. (1991): Genetics of seed number and seed weight in pea.<br />
Indian J. Pulse Res. 4: 165-167.<br />
Srivastava, C.P.- R.B. Singh (1988): genetics analysis of seeds per pod in peas (Pisum<br />
sativum L.) Veg. Sci. 15:38-48.<br />
Tóthné Lőkös K.- Heszky L. (1994): Az általános kombinálódó- képesség, a<br />
fenotípusos érték és az örökölhetőségi értékszám kapcsolata borsófajták diallél<br />
kísérletében. Növény<strong>term</strong>elés, 1994. Tom. 43. No.2: 101-107.<br />
206
SUSTAINABLE LAND-USE BASED ON WESTSIK’S CROP ROTATION<br />
EXPERIMENT<br />
János Lazányi<br />
University of Debrecen Centre for Agricultural Sciences<br />
H-4032 Debrecen Böszörményi út 138.<br />
SUMMARY<br />
Sustainable agriculture and land use seeks soluti<strong>on</strong>s for envir<strong>on</strong>mental, ec<strong>on</strong>omic and<br />
agricultural problems at the same time. The aim is to develop new producti<strong>on</strong> methods<br />
which provide protecti<strong>on</strong> for nature. In any sustainable agricultural system, the<br />
management of organic matter and, more widely, management of natural resources are<br />
based <strong>on</strong> the total self-sufficiency of the farm. The entire cycle of organic matter<br />
producti<strong>on</strong> and decompositi<strong>on</strong> takes place within the farm boundaries. The rate of<br />
metabolism and the organic matter cycle are characteristic features of each farm unit<br />
and define each unit’s activity for a l<strong>on</strong>g time.<br />
The increased demand for food and the change in ecological requirements <strong>on</strong><br />
agricultural producti<strong>on</strong> mean that scientists must revise comm<strong>on</strong>ly used traditi<strong>on</strong>al<br />
principles, in order to develop alternative crop producti<strong>on</strong> practices. In this respect, they<br />
must strive to adopt technologies which serve to maintain soil fertility, and, at the same<br />
time, meet envir<strong>on</strong>mental protecti<strong>on</strong> goals. Westsik’s crop rotati<strong>on</strong> experiment is an<br />
efficient tool which provides a thorough scientific basis for solving this problem.<br />
Westsik’s experiment models various possible methods for managing nutrients (fallow,<br />
straw, farmyard and green manure), and assists in seeking answers to questi<strong>on</strong>s of the<br />
sustainability of agricultural producti<strong>on</strong>.<br />
A study of sustainability of agricultural producti<strong>on</strong> and the c<strong>on</strong>necti<strong>on</strong>s between<br />
agriculture, land use and envir<strong>on</strong>mental quality have been initiated in developed<br />
countries, but the difficulties in less developed countries without adequate food supply<br />
are even more serious. The aim of this study is to examine the potential for<br />
envir<strong>on</strong>mentally-friendly land use in Hungary, and to draw attenti<strong>on</strong> to the importance<br />
of l<strong>on</strong>g-<strong>term</strong> field experiments in the study of agr<strong>on</strong>omic sustainability.<br />
Key words: Sustainable agriculture, land use, crop rotati<strong>on</strong> experiment<br />
INTRODUCTION<br />
Sustainable development is development that meets the needs of the present generati<strong>on</strong><br />
without compromising the ability of future generati<strong>on</strong>s to meet their own needs.<br />
Sustainable development wants to secure higher standards of living now and for future<br />
generati<strong>on</strong>s. An important recommendati<strong>on</strong> of the United Nati<strong>on</strong>s <str<strong>on</strong>g>C<strong>on</strong>ference</str<strong>on</strong>g> <strong>on</strong><br />
Envir<strong>on</strong>ment and Development held in Rio de Janeiro (1992), was that individual<br />
countries should prepare strategies and acti<strong>on</strong> plans to implement their part of the<br />
agreements. Agenda 21 also lays great emphasis <strong>on</strong> the need for all sectors of society to<br />
participate in the formati<strong>on</strong> of effective nati<strong>on</strong>al strategies for sustainable development.<br />
The problem of agriculture is as old as agriculture itself. The core of the problem has<br />
always been soil erosi<strong>on</strong> and loss of soil fertility. Today, when energy input into<br />
agricultural producti<strong>on</strong> has increased, a new aspect has been added to this problem,<br />
207
further exacerbating the old <strong>on</strong>e. Historical and recent experience with agriculture<br />
suggests that human demand are not likely to be achieved without negative effects <strong>on</strong><br />
natural ecosystems, envir<strong>on</strong>mental quality and rural communities. Agriculture in<br />
developing countries moves from traditi<strong>on</strong>al practices relying <strong>on</strong> natural fertility and the<br />
regenerative power of the soil, to practices depending <strong>on</strong> high levels of inorganic<br />
fertilisers, the intensive use of chemicals for pest c<strong>on</strong>trol, and crops produced mainly<br />
under m<strong>on</strong>oculture.<br />
The best known and most remarkable example of c<strong>on</strong>tinuous producti<strong>on</strong> in<br />
Hungary is the Westsik’s crop rotati<strong>on</strong> experiment established in 1929, which is still in<br />
use to study the effects of organic manure treatment, develop models and predict the<br />
likely effects of different cropping systems <strong>on</strong> soil properties and crop yields. In this<br />
respect, Westsik’s crop rotati<strong>on</strong> experiment provides data of immediate value to farmers<br />
c<strong>on</strong>cerning the applicati<strong>on</strong>s of green, straw and farmyard manure. The experiment also<br />
provides a resource of yield, plant and soil data sets for scientific research, whether into<br />
plant and those soil processes which c<strong>on</strong>trol soil fertility, or into the sustainability of<br />
producti<strong>on</strong>. Maintenance of Westsik’s crop rotati<strong>on</strong> experiment can also be used to<br />
illustrate the value of l<strong>on</strong>g-<strong>term</strong> field experiments.<br />
Sustainable agriculture is both a philosophy and a system of farming. It is rooted<br />
in a set of values that reflect an awareness of social and ecological realities. It<br />
emphasises management which works with natural processes to c<strong>on</strong>serve all resources<br />
and minimise waste, as well as envir<strong>on</strong>mental damage, while maintaining or improving<br />
the profitability of producti<strong>on</strong> based <strong>on</strong> nutrient and water cycles, energy flows,<br />
beneficial soil organisms and natural pest c<strong>on</strong>trols.<br />
Sustainable systems also aim at ensuring the well being of rural communities, and<br />
producing food which is nutritious and not c<strong>on</strong>taminated with products which might be<br />
harmful for the ecosystem. A sustainable farming system seeks to reduce or entirely<br />
avoid the use of synthetic fertilisers, pesticides, growth regulators and other agricultural<br />
chemicals. The system relies <strong>on</strong> crop rotati<strong>on</strong>, crop residues, animal manure, legume<br />
crops and green manure as organic matter, c<strong>on</strong>servati<strong>on</strong> tillage and n<strong>on</strong>-chemical pest<br />
c<strong>on</strong>trol, to maintain the fertility of soil and c<strong>on</strong>trol insects, weeds and diseases.<br />
MATERIALS AND METHODS<br />
The crop rotati<strong>on</strong> experiment established by Vilmos Westsik in 1929 offers an excellent<br />
possibility to study soil fertility management in many respects. The experiment,<br />
c<strong>on</strong>sisting of 15 treatments, makes it possible to study sustainable land use under<br />
different applicati<strong>on</strong>s of green, straw and farmyard manure treatments, to study the<br />
ecological impact as well as ec<strong>on</strong>omic aspects of different producti<strong>on</strong> methods. One of<br />
the main practical objects of the Westsik's crop rotati<strong>on</strong> experiment was to measure the<br />
l<strong>on</strong>g-<strong>term</strong> effects of different organic manure and inorganic fertilisers <strong>on</strong> rye and potato<br />
producti<strong>on</strong>. In this way, the 15 treatments can logically be grouped as follows (i) c<strong>on</strong>trol<br />
treatments where no fertilisers were applied, (ii) straw manure treatments, (iii) farmyard<br />
manure treatments, (iv) green manure treatments, (v) sec<strong>on</strong>d crop green manure<br />
treatments (Westsik 1951, 1965).<br />
The F-1 block received no fertilisers and organic material treatment except the rye<br />
and potato roots and straw incorporated into the soil. The fallow in this block was green,<br />
and the plant material produced was ploughed into the soil. The F-2 block represents<br />
208
green manure treatment, where lupine was grown as a main crop and incorporated into<br />
the soil 4-5 weeks after flowering. The phosphorus and potassium fertilisers in this<br />
treatment were applied the previous autumn, before the lupine was sown. The F-3<br />
represents lupine root manure treatment, where lupine was grown for grain and the total<br />
organic material, except for the grain, was incorporated into the soil.<br />
Blocks F-4 - F-7 represent straw manure treatments. In the F-4 block, rye straw<br />
was applied as mulch. In blocks F-6 and F-7, straw manure was fermented without<br />
nitrogen, and in block F-6, with nitrogen additi<strong>on</strong>. The straw manure was incorporated<br />
into the soil 4-6 weeks before the sowing of rye. F-8 is the <strong>on</strong>ly block with 4 main<br />
crops, where lupine grew twice in 4 years; <strong>on</strong>ce as a main crop produced for grain and<br />
<strong>on</strong>ce as a sec<strong>on</strong>d crop produced after rye and before potato, for green manure. In the F-9<br />
block, lupine was grown as a forage crop and harvested 2-3 weeks after flowering.<br />
Blocks F-10 and F-11 represent farmyard manure treatments without and with<br />
supplementary fertilisers, respectively. In block F-12, lupine is grown after a green<br />
forage crop and sown in May. This block is also evaluated with farmyard manure<br />
treatments to measure the comparative effects of the two treatments. Blocks F-13, F-14,<br />
F-15 represent green manure treatments, where lupine is grown as a sec<strong>on</strong>d crop after<br />
rye and before potato. The F-15 block received no supplementary fertilisers. The<br />
difference between blocks F-13 and F-14 can be found in the time of the incorporati<strong>on</strong><br />
of green manure.<br />
RESULTS AND DISCUSSION<br />
Minimum and maximum yields have varied c<strong>on</strong>siderably during the 77 years of the<br />
experiment. Analysing the average rye yields of different treatments, we can c<strong>on</strong>clude<br />
that the poorest results were achieved in 1963, when the average of the various crop<br />
rotati<strong>on</strong>s was 1.23 t<strong>on</strong>/hectare. The highest yields were harvested in 1984, when the<br />
average was 3.15 t<strong>on</strong>/hectare (Figure 1). The difference is 188 %. The average yield of<br />
potato was the lowest in 1972, and the highest, 12 times as much, in 1991 (Figure 1).<br />
From the two-plant species, rye proved to be the more balanced in productivity. Its yield<br />
variati<strong>on</strong>s may be attributed primarily to climatic factors, while, in the case of potato,<br />
the variati<strong>on</strong>s can be closely related to changes in varieties or in the quality of<br />
propagati<strong>on</strong> material. The first improvement in the average yields of potato occurred in<br />
the early 1940's, when a growing demand arose for potatoes as an important foodstuff.<br />
During and immediately after World War II, the average potato yields decreased in both<br />
crop rotati<strong>on</strong> experiments, as well as nati<strong>on</strong>ally. The next increase was achieved in the<br />
sec<strong>on</strong>d half of the 1950's, and was related to the development in potato research and in<br />
seed potato producti<strong>on</strong> at the Research Institute in Nyíregyháza (Figure 2).<br />
209
Rye Yield (t/ha)<br />
Potato yield (t/ha)<br />
5.5<br />
5.0<br />
4.5<br />
4.0<br />
3.5<br />
3.0<br />
2.5<br />
2.0<br />
1.5<br />
1.0<br />
0.5<br />
0.0<br />
50<br />
45<br />
40<br />
35<br />
30<br />
25<br />
20<br />
15<br />
10<br />
5<br />
0<br />
1931<br />
Maximum Minimum Mean<br />
1935<br />
1939<br />
1943<br />
1947<br />
1951<br />
1955<br />
1959<br />
1963<br />
1967<br />
1971<br />
1975<br />
1979<br />
1983<br />
1987<br />
Figure 1: Rye yields in the crop rotati<strong>on</strong> experiment<br />
Maximum Minimum Mean<br />
1991<br />
1995<br />
1999<br />
2003<br />
1931<br />
1935<br />
1939<br />
1943<br />
1947<br />
1951<br />
1955<br />
1959<br />
1963<br />
1967<br />
1971<br />
1975<br />
1979<br />
1983<br />
1987<br />
1991<br />
1995<br />
1999<br />
2003<br />
Figure 2: Potato yield in the crop rotati<strong>on</strong> experiment<br />
210
In Westsik's crop rotati<strong>on</strong> experiments, the average yields of potato increased again in<br />
the late 1980's and early 1990's. This success could be explained by the breeding of the<br />
virus resistant Boro variety and its involvement in the experiments. Similar yield<br />
increases could be noticed during the analysis of Broadbulk's experiments in<br />
Rothamsted, where the yields of wheat were doubled within <strong>on</strong>ly a few years, by<br />
changing the variety and applying pesticides (Johnst<strong>on</strong> 1969, Johnst<strong>on</strong> and Poult<strong>on</strong>,<br />
1980, Garner and Dyke 1969). Comparing Figures 1 and 2, it is difficult to show any<br />
correlati<strong>on</strong> between the yield trends of rye and potato; however, if we express potato<br />
yields as a functi<strong>on</strong> of the average rye yields, a very str<strong>on</strong>g correlati<strong>on</strong> can be revealed,<br />
except for crop rotati<strong>on</strong>s F-2, F-3 and F-9. Average potato yields can be characterised<br />
with the linear regressi<strong>on</strong>: y = 0.79 + 4.21x. The correlati<strong>on</strong> coefficient is r 2 = 0.9198.<br />
This means that there is no significant difference in the effect of various crop rotati<strong>on</strong>s<br />
<strong>on</strong> the yields of rye and potato (Figure 3). C<strong>on</strong>trol treatments, where fertilisers were not<br />
used in additi<strong>on</strong> to the occasi<strong>on</strong>al organic matter management, are <strong>on</strong> the lower part of<br />
the curve. The deviati<strong>on</strong> of F-2, F-3 and F-9 treatments from the regressi<strong>on</strong> line means<br />
that lupine main crop has different effects <strong>on</strong> the average yields of potato and rye,<br />
independently of the fact whether lupine is grown for green fodder, green manure or<br />
seed.<br />
Potato yield (t/ha)<br />
16<br />
14<br />
12<br />
10<br />
8<br />
6<br />
4<br />
2<br />
0<br />
F-1<br />
F-2<br />
Potato Rye<br />
F-3<br />
F-4<br />
F-5<br />
F-6<br />
F-7<br />
F-8<br />
F-9<br />
F-10<br />
F-11<br />
F-12<br />
F-13<br />
F-14<br />
F-15<br />
Figure 3: Rye and potato yield in the crop rotati<strong>on</strong> experiment<br />
Vilmos Westsik, who explained it with the infecti<strong>on</strong> of Fusarium species, which are<br />
able to decrease the yields of both lupine and potato c<strong>on</strong>siderably, described this<br />
phenomen<strong>on</strong> earlier. Furthermore, we have to add that lupine was ploughed under<br />
directly before rye, so it was this culture that benefited from the favourable effect of<br />
211<br />
3.0<br />
2.5<br />
2.0<br />
1.5<br />
1.0<br />
0.5<br />
0.0<br />
Rye yield (t/ha)
organic matter mineralizati<strong>on</strong>. The effects of treatments <strong>on</strong> potato yield were also<br />
expressed as a functi<strong>on</strong> of c<strong>on</strong>trol treatments to reduce variati<strong>on</strong> between years. In this<br />
case, the mean value of F-1, F-7, F-10 and F-15 was calculated for each year and<br />
studied as to how the increasing mean value effects the value of selected treatments.<br />
Agriculture in developed countries has moved progressively from traditi<strong>on</strong>al<br />
practices relying <strong>on</strong> natural fertility and the regenerative power of the soil, to practices<br />
depending <strong>on</strong> high levels of inorganic fertilisers, the intensive use of chemicals for pest<br />
c<strong>on</strong>trol, and crops produced mainly under m<strong>on</strong>oculture (Lockeretz 1988, Carter 1989).<br />
According to Edwards (1987), alternative agriculture is a farming practice avoiding, or<br />
at least minimising, the use of n<strong>on</strong>-renewable producti<strong>on</strong> inputs, such as fertilisers and<br />
pesticides. Lowrance at al. (1986) and Senanayake (1991) defined sustainable<br />
agriculture as agriculture which does not deplete soil or people.<br />
For many years, farmyard manure has been the key element in the maintenance of<br />
soil fertility (Johnst<strong>on</strong> 1987, Keeney 1989). Composted manure increased its value,<br />
stabilised the nutrients and reduced the risk of envir<strong>on</strong>mental polluti<strong>on</strong>. Composting<br />
provides more stable humus, which improves soil aerati<strong>on</strong>, water infiltrati<strong>on</strong> and water<br />
holding capacity. The high temperature of the composting process destroys microorganisms,<br />
the seeds of most weeds, and reduces water c<strong>on</strong>tents and total volume. In<br />
sustainable agriculture, crop and livestock systems should maximise for mutual support.<br />
Crop residue is an excellent feed for animals. Livestock manure can be composted and<br />
returned to the fields as a source of plant nutrient. Various crop and livestock<br />
comp<strong>on</strong>ents can be integrated to minimise the effects of weather-related adversities, as<br />
they can be crucial for the survival of a farm.<br />
In sustainable agriculture, where the use of synthetic chemicals is reduced or<br />
eliminated, the acti<strong>on</strong> of soil micro-organisms becomes a major factor in nutrient<br />
cycling and also plant growth (Lazányi 1996, 1997). Sustainable agricultural cropping<br />
systems should reduce n<strong>on</strong>-farm inputs to decrease envir<strong>on</strong>mental and health hazards<br />
associated with the use of agricultural chemicals and, at the same time, offset rising<br />
producti<strong>on</strong> costs and maintain soil fertility. The reducti<strong>on</strong> of leguminous plants in the<br />
cropping system results not <strong>on</strong>ly from the availability of fertilisers, but also from the use<br />
of fossil energy as a source of power in agriculture. Because of these changes, farmers<br />
are no l<strong>on</strong>ger compelled to use <strong>on</strong>e part of their land for the producti<strong>on</strong> of forage and<br />
grain for their draft animals. In Hungary many farms has eliminated all livestock from<br />
their operati<strong>on</strong>s, moving to cash grain enterprises and making them entirely dependent<br />
up<strong>on</strong> fertilisers and other chemicals. However, when nitrogen fertiliser is expensive or<br />
not available, the producers depend <strong>on</strong> the nitrogen fixed by legumes to maintain the<br />
nitrogen cycle and to improve the fertility of the soil. The quantity of nitrogen fixed<br />
varies greatly, from zero to several hundred kilograms per hectare, according to the soil<br />
type, structure, soil pH and nutrient c<strong>on</strong>tent, temperature, water regimes and<br />
management of the legumes.<br />
There are other benefits from using legumes in the crop rotati<strong>on</strong>, but they are often<br />
disregarded because of the difficulties in quantifying them. The yields of cereals grown<br />
after a leguminous crop or in crop rotati<strong>on</strong> are greater than those grown in m<strong>on</strong>oculture,<br />
regardless of the amount of fertiliser applied. This resp<strong>on</strong>se is often referred to as the<br />
leguminous effect in crop rotati<strong>on</strong>. As additi<strong>on</strong>al nitrogen does not eliminate this yield<br />
difference, most of the resp<strong>on</strong>se must be due to factors other than nitrogen availability.<br />
212
Crop rotati<strong>on</strong> breaks the weed and insect cycles that often predominate in c<strong>on</strong>tinuous<br />
cropping. Crop rotati<strong>on</strong> also enhances soil structure and improves water regime. In this<br />
way, leguminous crops have l<strong>on</strong>g-<strong>term</strong> benefits in crop rotati<strong>on</strong>, resulting in enhanced<br />
soil organic matter c<strong>on</strong>tent, which not <strong>on</strong>ly improves nitrogen availability, but also<br />
improves soil structure, by reducing soil erosi<strong>on</strong> and cultivati<strong>on</strong> costs.<br />
High input agriculture is relatively new in Hungary. Crop rotati<strong>on</strong> was introduced<br />
in the last century, extensive use of chemical fertilisers and pesticides became important<br />
<strong>on</strong>ly in the sec<strong>on</strong>d half of the XX. century. Agricultural producti<strong>on</strong> has become efficient<br />
per unit of human labour and per unit of land. However, when efficiency is measured<br />
against other criteria - capital, fossil fuel, energy, ec<strong>on</strong>omic or social equity - the results<br />
are less clear. The crop rotati<strong>on</strong> experiment established by Vilmos Westsik in 1929<br />
offers an excellent possibility to study soil fertility management. The experiment,<br />
c<strong>on</strong>sisting of 15 treatments, makes it possible to study sustainability of agricultural<br />
producti<strong>on</strong> under different applicati<strong>on</strong>s of green, straw and farmyard manure treatments,<br />
to study the ecological impact as well as ec<strong>on</strong>omic aspects of different producti<strong>on</strong><br />
methods.<br />
REFERENCES<br />
Carter, H. O. (1989): Agricultural sustainability: An overview and research<br />
assessment. Calif. Agric. 16-17.<br />
Edwards, C. A. (1987): The c<strong>on</strong>cept of integrated systems in lower input sustainable<br />
agriculture. American Journal of Alternative Agriculture 2(4):148-152.<br />
Garner, H. V., Dyke, G. V. (1969): The Broadbalk Wheat Experiment. Rothamsted<br />
Exp. Stati<strong>on</strong>. Report for 1969, 2, 26-45.<br />
Johnst<strong>on</strong>, A. E. (1969): Plant nutrients in Broadbalk soils. Rothamsted Experimental<br />
Stati<strong>on</strong> Report for 1968. Part 2, 93-112.<br />
Johnst<strong>on</strong>, A. E. (1987): Effects soil organic matter <strong>on</strong> yields of crops in l<strong>on</strong>g-<strong>term</strong><br />
experiments at Rothamsted and Woburn. INTECOL Bulletin 15, 9-16.<br />
Johnst<strong>on</strong>, A. E., Poult<strong>on</strong>, P. R. (1980): Effects of soil organic matter <strong>on</strong> cereal yields.<br />
Rothamsted Experimental Stati<strong>on</strong>, Report for 1979, Part 1, 234.<br />
Keeney, R. D. (1989): Towards a sustainable agriculture. Need for clarificati<strong>on</strong> of<br />
c<strong>on</strong>cepts and <strong>term</strong>inology. American J. of Alt. Agr. 4: 101-106.<br />
Lazányi, J. (1996): Results of Westsik's crop rotati<strong>on</strong> experiment for sustainable<br />
agriculture. Hungarian Agricultural Research. 1996. Vol. 5. No. 1. 4-8. p.<br />
Lazányi, J. (1997): Linking c<strong>on</strong>venti<strong>on</strong>al and sustainable agriculture. In Filep, Gy. (ed)<br />
Land Use and Soil Management. Debrecen. 68-86 p.<br />
Lockeretz, W. (1988): Open questi<strong>on</strong>s in sustainable agriculture, American Journal of<br />
Alternative Agriculture. 3:174-181.<br />
Lowrance, R., Hendrix, P. F., Odum, E. P. (1986): A hierarchical approach to<br />
sustainable agriculture. American Journal of Alternative Agriculture 1. 169-173.<br />
Senanayake, R. (1991): Sustainable Agriculture: Definiti<strong>on</strong>s and parameters for<br />
measurement. Journal of Sustainable Agriculture. 1:7-28.<br />
Westsik, V. (1951): Homoki vetésforgókkal végzett kísérletek eredményei.<br />
Mezőgazdasági Kiadó, Budapest.<br />
Westsik, V. (1965): Vetésforgó kísérletek homoktalaj<strong>on</strong>. Akadémiai Kiadó, Budapest.<br />
213
THE EFFECT OF LONG-TERM PHOSPHORUS FERTILIZATION ON THE<br />
TRYPTOPHAN AND ZINC CONTENT OF FEED PEAS<br />
Ágnes Elek- Zoltán Győri- Mária Borbély<br />
University of Debrecen, Centre of Agricultural Sciences<br />
Faculty of Agr<strong>on</strong>omy Institute of Food Science, Quality Assurance and Microbiology<br />
ABSTRACT<br />
The quality of food is de<strong>term</strong>ined by the chemical compositi<strong>on</strong>. One of the most<br />
important nutriti<strong>on</strong>al parameter is the protein, and also its amino-acid compositi<strong>on</strong>. Pea<br />
samples from the c<strong>on</strong>trol (0+0+0 kg/ha) and the 20 th treatment of the 1995 OTK research<br />
were analysed from Putnok, Mos<strong>on</strong>magyaróvár, Keszthely, Bicsérd, Hajdúböszörmény<br />
and Iregszemcse research sites. We analysed the tryptophan c<strong>on</strong>tent and its change as a<br />
result of phosphorus and zinc fertilizati<strong>on</strong> of the pea samples. HPLC analysis was<br />
performed at 110° C for 20 hours after alkaline hydrolysis. Our results show that high<br />
rates of phosphorus fertilizers decreased the tryptophan c<strong>on</strong>tent of peas.<br />
Keywords: tryptophan, HPLC, zinc, amino acids, hydrolysis<br />
INTRODUCTION<br />
The quality of food is de<strong>term</strong>ined by the chemical compositi<strong>on</strong>. One of the most<br />
important nutriti<strong>on</strong>al parameter is the protein, and also its amino-acid compositi<strong>on</strong>.<br />
Amino-acid analysis is usually a good challenge for the analytical laboratories, even they<br />
have the suitable instruments for the de<strong>term</strong>inati<strong>on</strong>. Most of the amino-acids of proteins<br />
in food and their raw materials can be analysed by acidic hydrolysis. Tryptophan has a<br />
sensitive indult-group, therefore it degrades in acidic soluti<strong>on</strong>. For HPLC de<strong>term</strong>inati<strong>on</strong><br />
we introduced a sample preparati<strong>on</strong> with alkaline hydrolysis (Ba(OH)2). We analysed the<br />
tryptophan c<strong>on</strong>tent and change of samples from OTK l<strong>on</strong>g-<strong>term</strong> field trials as an effect of<br />
phosphorus and zinc fertilizati<strong>on</strong>.<br />
Correct de<strong>term</strong>inati<strong>on</strong> of tryptophan in food and feed is very important, as this<br />
amino acid is essential for m<strong>on</strong>ogastric animals. On the other hand, there are less data<br />
about tryptophan c<strong>on</strong>tent of feed, than for the other amino-acids. With full knowledge of<br />
the recent research results, tryptohan seems to have even more important role in human<br />
nutriti<strong>on</strong>. Less tryptophan than it is necessary for human promotes the development of<br />
nervous depressi<strong>on</strong>. Its role was proved also in migraine headache. It is well known that<br />
c<strong>on</strong>sumpti<strong>on</strong> of feed with high protein c<strong>on</strong>tent will decrease the tryptophan-level in the<br />
brain, due to a competiti<strong>on</strong> am<strong>on</strong>g different amino acids, regarding their intake.<br />
Triptrophan c<strong>on</strong>tent de<strong>term</strong>inati<strong>on</strong> requires special methods because of its sensitivity<br />
in acidic c<strong>on</strong>diti<strong>on</strong>s in the presence of oxygen. Measurement can be performed by i<strong>on</strong>exchange<br />
chromatography (Moore and Stein, 1963; Dévényi 1971), photometry<br />
(Mathes<strong>on</strong>, 1974; Basha and Roberts, 1977; Votisky 1984).<br />
The highest tryptophan yield was found in the case of short <strong>term</strong> hydrolysis (60 min),<br />
performed with mercaptoethanesulf<strong>on</strong>icacid at high temperature. The most comm<strong>on</strong><br />
method is the HPLC de<strong>term</strong>inati<strong>on</strong> following an alkaline hydrolysis of proteins with<br />
Sodium- or Barium-hydroxide (Delhaye and Landry, 1992). This method was used in our<br />
laboratory to measure the tryptophan c<strong>on</strong>tent of some plant originated food raw materials.<br />
214
The reliability of the method was checked with recovery experiments with L- Tryptophan<br />
(Fluka, Germany), and by the analysis of reference sample with known tryptophan<br />
c<strong>on</strong>tent.<br />
MATERIALS AND METHODS<br />
Pea samples from the c<strong>on</strong>trol (0+0+0 kg/ha) and the 20 th treatment of the 1995 OTK<br />
research were analysed from Putnok, Mos<strong>on</strong>magyaróvár, Keszthely, Bicsérd,<br />
Hajdúböszörmény and Iregszemcse research sites. Crude protein c<strong>on</strong>tent of the analysed<br />
samples varied from 6 to 23 %, according to the research site.<br />
Hydrolysis: For hydrolysis 8,40 g Ba(OH)2x8H2O and 10 ml distilled water was added to<br />
the sample that c<strong>on</strong>tained some 10 mg Nitrogen in a pressure-resistant test tube with<br />
Tefl<strong>on</strong> pipe. The reacti<strong>on</strong> was performed at 110° C for 20 hours. The mixture was<br />
washed to a flask with 30 ml distilled water, and Barium was precipitated with 0,05 M<br />
Phosphoric acid in the form of Barium phosphate. The pH of the filtrate was adjusted to<br />
3,0 with 6N Hydrochloric-acid and diluted to 100 ml with distilled water : methanol<br />
80:20 solvent mixture. The soluti<strong>on</strong> was filtered through 0,45 µm membrane filter<br />
(Millipore, USA) before HPLC analysis.<br />
Chromatography: High performance liquid chromatography was performed by a<br />
Merck-Hitachi AS 4000A equipment. Gradient program was used for separati<strong>on</strong> <strong>on</strong> C18<br />
column (LiChorCart 125x4) with eluent A:0,01mol/l KH2PO4 (Reanal) /Metanol 95:5 and<br />
B: Methanol (Merck). For detecti<strong>on</strong> Merck-Hitachi F1050 fluorescent detector was used<br />
(ex: 280 nm, em: 356 nm).<br />
RESULTS AND DISCUSSIONS<br />
According to the literature, the protein c<strong>on</strong>tent of feed peas is highly dependent <strong>on</strong> the<br />
crop year and the cropping site (Figure 1). Data show that the protein c<strong>on</strong>tent of the<br />
c<strong>on</strong>trol plots varied from 19 to 25 % (Mos<strong>on</strong>magyaróvár and Iregszemcse). The<br />
fertilizati<strong>on</strong> did not increase the protein c<strong>on</strong>tent of pees in Putnok, at the same time, it<br />
decreased in Keszthely. More (in Mos<strong>on</strong>magyaróvár from 19.59 to 23.63%) or less (in<br />
Hajdúböszörmény from 23.4 to 24.93%)increase was found in the other research sites.<br />
215
P mg/kg<br />
fehérje(%)<br />
30<br />
25<br />
20<br />
15<br />
10<br />
5<br />
0<br />
MO BI KE IR PU HB<br />
k<strong>on</strong>troll<br />
20.kezelés<br />
Figure 1: The effect of fertilizati<strong>on</strong> <strong>on</strong> the protein c<strong>on</strong>tent of peas (1995)<br />
7000<br />
6000<br />
5000<br />
4000<br />
3000<br />
2000<br />
1000<br />
0<br />
MO KE PU IR<br />
1 kezelés<br />
20kezelés<br />
Figure 2: The effect of fertilizati<strong>on</strong> <strong>on</strong> the phosphorus c<strong>on</strong>tent of peas (1995)<br />
216
The increased phosphorus rates increased the phosphorus c<strong>on</strong>tent in peas in<br />
Mos<strong>on</strong>magyaróvár, Keszthely, Putnok, and decreased <strong>on</strong>ly in Putnok (Figure 2).<br />
Zn mg/kg<br />
70<br />
60<br />
50<br />
40<br />
30<br />
20<br />
10<br />
0<br />
MO KE PU IR<br />
Figure 3: The effect of fertilizati<strong>on</strong> <strong>on</strong> the zinc c<strong>on</strong>tent of peas (1995)<br />
1 kezelés<br />
20kezelés<br />
As regards zinc c<strong>on</strong>tent both the cropping sites and the effect of fertilizati<strong>on</strong> had<br />
significant differences. The fertilizati<strong>on</strong> treatments decreased the zinc c<strong>on</strong>tent. The<br />
highest effect was found in Putnok, furthermore, the lowest decreasing effect of<br />
fertilizati<strong>on</strong> <strong>on</strong> zinc was found in Mos<strong>on</strong>magyaróvár (Figure 3).<br />
We de<strong>term</strong>ined the tryptophan c<strong>on</strong>tent of pea samples by hydrolysis, in alkaline<br />
media for 20 hours at 110 ºC. We had to take into c<strong>on</strong>siderati<strong>on</strong> the different crude<br />
protein c<strong>on</strong>tent of the samples during the destructuring in barium-hydroxide media, 8,40<br />
g Ba(OH)2x8H2O and 10 ml water was added to 10 mg N c<strong>on</strong>tent. The rate ranged<br />
between 89-98%.<br />
217
Trp (fehérje %-a)<br />
1,2<br />
1,0<br />
0,8<br />
0,6<br />
0,4<br />
0,2<br />
0,0<br />
Mos<strong>on</strong>magyaróvár<br />
Keszthely<br />
Putnok<br />
Iregszemcse<br />
1. kezelés<br />
20. kezelés<br />
SzD 5%<br />
SD5% between treatments MO: 0,310** SD5% between treatments KE: 0,186*<br />
SD5% between treatments PU: 0,198** SD5% between treatments IR: 0,166**<br />
Figure 4 The effect of fertilizati<strong>on</strong> <strong>on</strong> the tryptophan c<strong>on</strong>tent of peas at different<br />
research sites<br />
NPK fertilizati<strong>on</strong> significantly decreased the tryptophan c<strong>on</strong>tent of peas at the four<br />
examined research sites. These results verify the need for testing the relati<strong>on</strong>ship between<br />
phosphorus and zinc in new varieties/hybrids, <strong>on</strong> the <strong>on</strong>e hand by analysing plant samples<br />
from the early stage of the growing year, <strong>on</strong> the other hand by analysing the peas, thus we<br />
can obtain informati<strong>on</strong> <strong>on</strong> the trypthophan c<strong>on</strong>tent as well.<br />
Our earlier results showed that increasing rates of NPK fertilizers decresed the<br />
tryptophan c<strong>on</strong>tent, as the P-Zn i<strong>on</strong> antag<strong>on</strong>ism decreased the intake of Zn.<br />
CONCLUSIONS<br />
1. It was verified with experiments that the de<strong>term</strong>inati<strong>on</strong> of tryptohan can be reliable<br />
performed by alkaline hydrolysis.<br />
2. Data from literature show that the tryptophan c<strong>on</strong>tent of peas can be reliably<br />
performed using the presented method.<br />
3. Increasing phosphorus rates decreased the zinc intake due to the P-Zn antag<strong>on</strong>ism<br />
and led to decreased tryptophan c<strong>on</strong>tent.<br />
4. Our data was included in the database, and the introduced method can be applied to<br />
analyse samples from field trials examining varieties and fertilizati<strong>on</strong> of leguminous<br />
crops as well.<br />
218
REFERENCES<br />
Basha S.M, Roberts R.M (1977) :A simple colorimetric method for the de<strong>term</strong>inati<strong>on</strong> of<br />
tryptophan ,Anal. Biochem. 378-386.<br />
Csapó,J. (2006) (szerk.) : Élelmiszer és takarmányfehérjék minősítése Mezőgazda<br />
Kiadó, Budapest<br />
Debreczeni, B., Debreczeni, B.-né (1994)(szerk): Trágyázási kutatások 1960-1990.<br />
Akadémiai Kiadó, Budapest<br />
Dévényi T., Gergely J.(1971) :Aminosavak, peptidek, fehérjék Budapest: Medicina<br />
Könyvkiadó<br />
Ágnes Elek, Mária Borbely, Zoltan Győri(2007): Tryptophan c<strong>on</strong>tent of some food<br />
raw materials Cereal Research Communicati<strong>on</strong>s, in press.<br />
Izsáki Zoltán: (2006): A N- és P-ellátottság hatása a kukoricaszem (Zea mays L.)<br />
fehérjetartalmára és aminosav-összetételére - Növény<strong>term</strong>elés Vol 55 Nos 3-4<br />
Landry, J; Delhaye,S(1992) : Simplifed Procedure for the De<strong>term</strong>inati<strong>on</strong> of Tryptophan<br />
in Food and Feedstuffs from Barytic Hydrolysis. J. Agric. Food Chem. 40, 776-779.<br />
Mathes<strong>on</strong>, N. A.(1974): The de<strong>term</strong>inati<strong>on</strong> of tryptophan in purified proteins and in<br />
feeding-stuffs. British Journal of Nutriti<strong>on</strong>, 393-400.<br />
Magyar Takarmánykódex FVM értesítő (2003).LIV évfolyam 11.sz. VIII. sz. melléklet<br />
1711-1715.<br />
MSZ ISO 6496:2001<br />
MSZ 6830-4:1981<br />
Stein W.H., Moore M(1963): Chromatographic de<strong>term</strong>inati<strong>on</strong> of amino acids by the use<br />
of automatic recording equipment. In: "Methods in Enzymology", Vol. 6., Ed.<br />
Colowick S. P. and Kaplan N. O., Academic Press, New York, 1963.<br />
Votisky,E. Colorimetria de<strong>term</strong>inati<strong>on</strong> of tryptophan in corn-cob-mix(CCM) (In:<br />
Progress in Tryptophan and Serotomin Research Ed. H.G. Schlossberger,<br />
W.Kochen, B. Linzen and H. Steinhard,W. de Gruyter and Co. Berlin-New York,<br />
115-117<br />
219
THE IMPORTANCE OF CALCIUM AND MAGNESIUM SUPPLY<br />
ON THE ACID BROWN FOREST SOILS IN THE NYIRSÉG REGION 1<br />
Jakab Loch*, János Lazányi**, Péter Tamás Nagy* and Rita Kremper*<br />
* Department of Agricultural Chemistry and Soil Science, University of Debrecen,<br />
Centre of Agricultural Sciences, Faculty of Agr<strong>on</strong>omy<br />
** Department of Rural Development and land Use, University of Debrecen, Centre of<br />
Agricultural Sciences, Faculty of Agroec<strong>on</strong>omics and Rural Development<br />
ABSTRACT<br />
According to soil tests made in l<strong>on</strong>g-<strong>term</strong> field experiments and to the yield data of nine<br />
years it can be c<strong>on</strong>cluded that <strong>on</strong> a certain habitat triticale yield can be duplicate with<br />
appropriate NPK and Ca, Mg supply. NPK fertilizati<strong>on</strong> and especially N treatments<br />
acidify the soil, and increase the Ca and Mg leaching. For maximum yield it is<br />
necessary to add both Ca and Mg systematically bey<strong>on</strong>d the well balanced NPK<br />
porti<strong>on</strong>s. Ca supplement can be well characterised by AL-Ca values.<br />
Keywords: l<strong>on</strong>g-<strong>term</strong> field experiment, brown forest acid sandy sols, fertilizati<strong>on</strong>,<br />
liming, AL-Ca and AL-Mg c<strong>on</strong>tent of soils.<br />
INTRODUCTION<br />
Soils in Nyírség are extremely varied. One characteristic type of sandy soils is the<br />
brown forest soil with thin clay layer, which is usually poor in calcium and magnesium.<br />
The lack of calcium and magnesium depends <strong>on</strong> the extent of leaching. Calcium and<br />
magnesium can be adsorbed relatively in a greater porti<strong>on</strong> <strong>on</strong>ly in clay stripes lying in<br />
deeper z<strong>on</strong>es. Blownsands are poorer in calcium and magnesium even than this.<br />
The fulfilment of ecological aspects and the realizati<strong>on</strong> of envir<strong>on</strong>mentally friendly<br />
plant cultivati<strong>on</strong> require the study of l<strong>on</strong>g-<strong>term</strong> effects of nutriti<strong>on</strong> supply and the<br />
foundati<strong>on</strong> of sustainable producti<strong>on</strong>. A good possibility for this is provided by a l<strong>on</strong>g<strong>term</strong><br />
experiment adjusted in 1962 by Láng István. The yield data of the l<strong>on</strong>g-<strong>term</strong><br />
experiment reported so far (Láng I. 1973, Kádár - Szemes 1994, Kádár – Németh –<br />
Szemes 1999, Márt<strong>on</strong> 2006) unambiguously prove that appropriate yield <strong>on</strong> brown<br />
forest soil with thin clay layer which is str<strong>on</strong>gly leached in the upper layer can be<br />
achieved <strong>on</strong>ly with balancing the pH c<strong>on</strong>diti<strong>on</strong>s and with a well balanced nutrient<br />
supply.<br />
Productivity of sandy soils is de<strong>term</strong>ined by the low humus c<strong>on</strong>tent accompanied<br />
with low nutrient c<strong>on</strong>tent and by the weak water management characteristics. According<br />
to the soil tests and yield analysis of MTA TAKI l<strong>on</strong>g-<strong>term</strong> experiments in Nyírlugos<br />
we already drew attenti<strong>on</strong> to the importance of the certain nutrients additi<strong>on</strong> (Loch et al,<br />
2002, Loch 2003, Loch et al 2005), and to the necessity of calcium supplement. Soil<br />
acidity hinders the effectiveness of nutrients, lack of calcium and magnesium formed as<br />
a c<strong>on</strong>sequence of leaching can limit the yield.<br />
1 Authors are grateful to co-workers of MTA-TAKI for making soil sampling possible<br />
and for providing the yield data of the experiment.<br />
220
MATERIALS AND METHODS<br />
Sandy soils were sampled from the „Nyírlugos” l<strong>on</strong>g-<strong>term</strong> field experiment, which were<br />
established in 1962. The “Nyírlugos” LTFE was set up <strong>on</strong> acidic brown forest soil with<br />
alternating thin layers of clay substance „kovárvány” in the Nyírség regi<strong>on</strong>. The main<br />
characteristics of the experimental soils are: pH(KCl) 4.5, humus 0.7%, CEC 5-10<br />
mgeq/100 g in the ploughed layer. The topsoil was poor in all five macr<strong>on</strong>utrients (N, P,<br />
K, Ca, Mg) and the groundwater depth was 2-3 m (Kádár et al., 1999).<br />
The “Nyírlugos” experiment is a l<strong>on</strong>g-<strong>term</strong> small-plot (50m 2 ) fertilizati<strong>on</strong> and<br />
liming experiment. The experiment c<strong>on</strong>sists of 32 treatments, 4 replicati<strong>on</strong>s altogether<br />
128 plot experiments. The indicator plant is triticale since 1980.<br />
Fertilizati<strong>on</strong> and liming treatments in the experiment were changed several times in<br />
the last few decades but the levels are c<strong>on</strong>stant since 1980 (Table 1.).<br />
Table 1: Treatments of LTFE in “Nyírlugos”, since 1980<br />
Treatments (kg/ha/year)<br />
Levels N P2O5 K2O CaCO3 MgCO3<br />
C<strong>on</strong>trol - - - - -<br />
1 50 60 60 250 140<br />
2 100 120 120 500 280<br />
3 150 180 180 1000 -<br />
Soil samples were collected from 5 different places in each plot after harvest of triticale,<br />
in August 1998 and 2001.<br />
Collected soil samples are handed and prepared as follows: samples were dried and<br />
drilled. Before grinding, samples were cleaned from plant remains and other possible<br />
dirt, and pass a 2 -mm screen. Finally, samples were stored in plastic boxes in dry place<br />
until the examinati<strong>on</strong>.<br />
In our earlier studies, the effects of treatments were described by the 0.01M CaCl2<br />
soluble nutrient c<strong>on</strong>tents measured from soils collected in 1998 (Nagy et al., 2002; Loch<br />
et al 2005). In this study, the l<strong>on</strong>g <strong>term</strong> effect of treatments <strong>on</strong> the average yield (1996-<br />
2004) of triticale (based <strong>on</strong> the data of Márt<strong>on</strong> (2006)) was described by the pH and AL<br />
soluble Ca and Mg values measured from soils collected in 1998 and 2001.<br />
RESULTS AND DISCUSSIONS<br />
1. Effects of treatments <strong>on</strong> the yield and soil characteristics:<br />
Am<strong>on</strong>g N1, N2, N3 treatments <strong>on</strong>ly the N1=50kg/ha increased the yield significantly, N2<br />
= 100kg/ha dose held <strong>on</strong> a level, while N3 =150kg/ha dose caused depressi<strong>on</strong>. We<br />
referred (Nagy et al 2002) previously that in case of this treatment the amount of<br />
inorganic N fracti<strong>on</strong> increased. Amm<strong>on</strong>ium-nitrate applied in excess acidifies the soil,<br />
pH decreases, and Ca and Mg leaching into deeper level intensifies.<br />
In N2P1, N2P2, N2P3 treatments the yield increases, phosphorous doses improve the<br />
utilizati<strong>on</strong> of N, the acidifying effect of amm<strong>on</strong>ium nitrate decreases, which is proved<br />
by pH values also and leaching of Ca and Mg is also smaller.<br />
221
Yields of N2K1, N2K2, N2K3 treatments are less than that of NP treatments, yield<br />
increase presents <strong>on</strong>ly in case of the largest potassium dose. The N2P2K2 treatment<br />
combinati<strong>on</strong>s verify that the supplement of the three elements is necessary, as the yield<br />
increases significantly. Maximum yield can be achieved <strong>on</strong>ly by NPK treatments<br />
completed with calcium carb<strong>on</strong>ate. In these treatments soil pH, AL-Ca and AL-Mg<br />
c<strong>on</strong>tent increase (Figure 1.).<br />
AL soluble Ca (mg/kg)<br />
1200<br />
1000<br />
800<br />
600<br />
400<br />
200<br />
0<br />
K<strong>on</strong>troll<br />
N1<br />
AL-Ca<br />
pH<br />
Yield<br />
N2<br />
N3<br />
N2P1<br />
N2P2<br />
N2P3<br />
N2K1<br />
N2K2<br />
N2K3<br />
N2P2K2<br />
N2P2K2Ca2<br />
N2P2K2Mg2<br />
N2P2K2Ca2Mg2<br />
Figure 1: Effect of treatments <strong>on</strong> triticale yield, the AL-Ca and pH values (0-20 cm)<br />
2. Formati<strong>on</strong> of triticale yield is in accordance with the soil parameters:<br />
AL-Ca values in accordance with the pH values show that in treatments without liming<br />
a great lack of calcium formed as a l<strong>on</strong>g-<strong>term</strong> effect of NPK treatments. Based <strong>on</strong><br />
Balogh (1988) investigati<strong>on</strong> and our own experiences if AL-Ca value in soil is smaller<br />
than 100-150mg/kg with lack of calcium has to be accounted, and pH values under 4.5<br />
are also c<strong>on</strong>sidered as very critical values. According to Iváncsics (1988) if AL-Ca<br />
values in soil is smaller than 120mg/kg the Ca supply level is weak.<br />
As the figures show, in the 0-20 cm soil layer AL-Ca level is under the critical<br />
value in case of N treatments and in the NPK treatments combinati<strong>on</strong>.<br />
222<br />
t<br />
7<br />
6<br />
5<br />
4<br />
3<br />
2<br />
1<br />
0<br />
Yield (t/ha)and pH (0.01M CaCl 2)
AL soluble Ca (mg/kg)<br />
350<br />
300<br />
250<br />
200<br />
150<br />
100<br />
50<br />
0<br />
K<strong>on</strong>troll<br />
N1<br />
AL-Ca<br />
pH<br />
Yield<br />
N2<br />
N3<br />
N2P1<br />
N2P2<br />
N2P3<br />
N2K1<br />
N2K2<br />
N2K3<br />
N2P2K2<br />
N2P2K2Ca2<br />
N2P2K2Mg2<br />
N2P2K2Ca2Mg2<br />
Figure 2: Effect of treatments <strong>on</strong> triticale yield, the AL-Ca and pH values (20-40 cm)<br />
AL soluble Ca (mg/kg)<br />
200<br />
180<br />
160<br />
140<br />
120<br />
100<br />
80<br />
60<br />
40<br />
20<br />
0<br />
AL-Ca<br />
pH<br />
Yield<br />
K<strong>on</strong>troll<br />
N1<br />
N2<br />
N3<br />
N2P1<br />
N2P2<br />
N2P3<br />
N2K1<br />
N2K2<br />
N2K3<br />
N2P2K2<br />
N2P2K2Ca2<br />
N2P2K2Mg2<br />
N2P2K2Ca2Mg2<br />
Figure3: Effect of treatments <strong>on</strong> triticale yield, the AL-Ca and pH values (40-60 cm)<br />
223<br />
6<br />
5<br />
Yield<br />
4<br />
(t/ha) and<br />
3<br />
pH<br />
2<br />
(0.01M<br />
1CaCl<br />
0<br />
6<br />
5<br />
Yield<br />
4<br />
(t/ha) and<br />
3<br />
pH<br />
2<br />
(0.01M<br />
1<br />
CaCl<br />
0<br />
2)<br />
2)
In 20-40 cm layer there is a definite differentiati<strong>on</strong> (formed in the upper layer) caused<br />
by the treatments, the difference in the upper layer decreases according to liming.<br />
Finally in 40-60 cm soil layer beside the str<strong>on</strong>g differentiati<strong>on</strong> AL-Ca values of each<br />
treatments are below 200mg/kg, and in case of certain treatments it is <strong>on</strong>ly 60 mg/kg.<br />
The represented data unambiguously prove the necessity of calcium-carb<strong>on</strong>at additi<strong>on</strong>.<br />
AL-Mg values show similar differentiati<strong>on</strong> to AL-Ca values. These indicate weak and<br />
medium supply in treatments without Ca and Mg. Increase of magnesium c<strong>on</strong>tent relate<br />
to the fact that lime powder applied for liming c<strong>on</strong>tained magnesium also (Table 2).<br />
Table 2: Effects of treatment <strong>on</strong> AL-Mg (mg/kg) values in different layers<br />
Treatments<br />
Depth (cm)<br />
0-20 20-40 40-60 Átlag<br />
C<strong>on</strong>trol 13.2 13.2 13.2 13.2<br />
N1 7.6 8.3 9.4 8.4<br />
N2 8.4 6.0 6.8 7.1<br />
N3 5.3 8.8 10.1 8.1<br />
N2P1 5.2 10.4 8.4 8.0<br />
N2P2 6.9 10.6 10.4 9.3<br />
N2P3 7.7 9.5 8.7 8.6<br />
N2K1 6.4 5.6 13.0 8.3<br />
N2K2 4.1 6.3 6.9 5.8<br />
N2K3 4.5 7.7 7.1 6.5<br />
N2P2K2 5.9 10.7 6.3 7.6<br />
N2P2K2Ca2 8.8 9.1 9.2 9.0<br />
N2P2K2Mg2 22.2 14.4 16.2 17.6<br />
N2P2K2Ca2Mg2 13.9 15.4 24.6 18.0<br />
CONCLUSION<br />
Data of l<strong>on</strong>g-<strong>term</strong> experiment c<strong>on</strong>firm that <strong>on</strong> <strong>on</strong>e of the characteristic soil type in<br />
Nyírség i.e. <strong>on</strong> acidic brown forest soil with thin clay layer triticale can be cultivated<br />
effectively with an adequate nutrient management. The producti<strong>on</strong> can be duplicated<br />
related to the untreated c<strong>on</strong>trol. Nutrient management and adjusting of pH influence<br />
fertility of soils and its maintenance to a great extent.<br />
NPK fertilizati<strong>on</strong> using by itself do not make it possible to reach the maximum<br />
yield. Acidic pH depending <strong>on</strong> the extent of leaching and the lack of calcium are<br />
limiting factors for yield, therefore there is a need for ordinary liming and occasi<strong>on</strong>ally<br />
magnesium additi<strong>on</strong>.<br />
One-sided N fertilizati<strong>on</strong> is especially to be avoided, while N dominance increase<br />
soil acidifying and the leaching of Ca and Mg. Applying N, P, K fertilizati<strong>on</strong> together<br />
adequately improve the utilizati<strong>on</strong> of N. Yields of NP and NPK treatments refer to the<br />
importance of phosphorous and potassium fertilizati<strong>on</strong> <strong>on</strong> a given cropland.<br />
AL-Ca values are good indicators of calcium lack. Data of the experiments c<strong>on</strong>firm<br />
the earlier experiences, values under 100mg/kg limit the yield, Ca and Mg are leached<br />
224
gradually from soils c<strong>on</strong>taining 100-150 mg/kg AL-Ca therefore their additi<strong>on</strong> is<br />
necessary. The pH values under 4.5 can be c<strong>on</strong>sidered as critical values.<br />
Studies of deeper layers prove that the ordinary Ca and Mg supply has effect<br />
primarily <strong>on</strong> surface layers.<br />
REFERENCES<br />
Balogh I. (1988): Nyírségi savanyú homoktalajok <strong>term</strong>ékenységének növelése kalcium<br />
és magnézium visszapótlással. Kandidátusi értekezés. Karcag.<br />
Iváncsics I. (1988): Kutatási eredmények a gyakorlatnak. Tápanyag-gazdálkodás.<br />
Mezőgazdasági és Élelmezési Minisztérium, Agroinform Kiadó, Budapest 88-93.<br />
Kádár I. – Szemes I. (1994): A nyírlugosi tartamkísérlet 30 éve. MTA – TAKI,<br />
Budapest.<br />
Kádár I. – Németh T. – Szemes I. (1999): A nyírlugosi tartamkísérlet 1998 évi<br />
eredményei. Kézirat, MTA-TAKI Budapest.<br />
Láng I. (1973): Műtrágyázási tartamkísérletek homoktalaj<strong>on</strong>. Akadémiai doktori<br />
disszertáció. Budapest.<br />
Loch J. (1999): A hazai talajok magnéziumellátottságának jellemzése és a<br />
magnéziumtrágyázás. Akadémiai doktori értekezés. Debrecen.<br />
Loch J. – Jászberényi I. – Nagy P.T. (2002): A kalcium- és magnéziumpótlás<br />
jelentősége savanyú magnéziumhiányos nyírségi homoktalaj<strong>on</strong>. In:<br />
Tartamkísérletek, Táj<strong>term</strong>esztés, Vidékfejlesztés Nemzetközi K<strong>on</strong>ferencia (Szerk.:<br />
Láng I. – Lazányi J. – Németh T.) I. kötet, 130-135. o.<br />
Loch J. (2003): A magnéziumtrágyázás jelentősége a savanyú homoktalajok<br />
<strong>term</strong>ékenységének növelésében. In: Talajjavítás – talajvédelem. (Szerk.: Győri Z.<br />
– Jávor A.) DE ATC 109-115.<br />
Loch J. – Kiss Sz. – Vágó I. (2005): A 0,01 M kalcium-klorid talaj-kiv<strong>on</strong>ószer<br />
alkalmazásának hazai tapasztalatai In: Fenntartható homoki gazdálkodás a<br />
Nyírségben, (Szerk. Lazányi J.), Kiadó:Westsik V. Nyírségi Tájfejlesztési<br />
Alapítvány. 137-155.<br />
Márt<strong>on</strong> L. (2006): a műtrágyázás, meszezés és a csapadék változék<strong>on</strong>yságának hatása<br />
a tritikále <strong>term</strong>ésére és <strong>term</strong>éskomp<strong>on</strong>enseire. In: Loch J., Lazányi J. (ed.): A<br />
tápanyag-gazdálkodást segítő talajvizsgálati módszerek alkalmazása a Nyírség<br />
homoktalajain. Westsik Vilmos Nyírségi Tájfejlesztési Alapítvány. Nyíregyháza.<br />
51-77.<br />
Nagy P.T. – Jászberényi I. – Loch J. (2002): A trágyázás hatása a 0,01 M kalciumkloridban<br />
oldható nitrogénformák mennyiségére a nyírlúgosi tartamkísérletben. In.:<br />
Tartamkísérletek, Táj<strong>term</strong>esztés, Vidékfejlesztés Nemzetközi K<strong>on</strong>ferencia (Szerk.:<br />
Láng I. – Lazányi J. – Németh T.) I. kötet, 143-148.<br />
225
COMPARATIVE ANALYSIS OF CHEMICAL AND BIOLOGICAL SOIL<br />
EXAMINATION TO DETERMINE THE PLANT AVAILABLE N CONTENT<br />
OF SOIL IN THE NYÍRLUGOS LONG TERM FIELD EXPERIMENTS 1<br />
Péter Tamás Nagy* – János Lazányi** – Jakab Loch*<br />
* Department of Agricultural Chemistry and Soil Science, University of Debrecen,<br />
Centre of Agricultural Sciences, Faculty of Agr<strong>on</strong>omy<br />
** Department of Rural Development and Land Use, University of Debrecen, Centre of<br />
Agricultural Sciences, Faculty of Agroec<strong>on</strong>omics and Rural Development<br />
ABSTRACT<br />
Chemical and biological (incubati<strong>on</strong>) analysis was carried out the acidic sandy soil of<br />
“Nyírlugos” l<strong>on</strong>g <strong>term</strong> field experiment. For chemical analysis 0.01 M CaCl2 extractant<br />
was used. From extractant mineral and organic nitrogen were de<strong>term</strong>ined.<br />
For biological analyses a modified incubati<strong>on</strong> technique was applied to de<strong>term</strong>ine the<br />
easily soluble, above menti<strong>on</strong>ed N fracti<strong>on</strong>s.<br />
The chemical and biological analysis was carried out chosen samples from 1998<br />
and 2001. From the results of chemical analysis it was evident that the amounts of<br />
easily soluble N forms highly depend <strong>on</strong> the applied treatments. All treatments<br />
increased the c<strong>on</strong>tent of both mineral and organic N compared to the c<strong>on</strong>trol. Moreover,<br />
it was found that excess N doses (100-150 kg/ha) resulted significant yield-decrement.<br />
Results of biological analysis were pointed out that logarithmic rate equati<strong>on</strong> was<br />
founded between the amount of mineralized N and time, during incubati<strong>on</strong>. Our results<br />
open the door to establish the potentially mineralizable N-c<strong>on</strong>tents (Npot) and the<br />
c<strong>on</strong>stant of the mineralizati<strong>on</strong> (k) of soils of different treatments. It can be established<br />
that the applied treatments has not remarkably effect <strong>on</strong> the values of Npot, but higher<br />
c<strong>on</strong>stant rate of mineralizati<strong>on</strong> was achieved in the limed treatment group than the<br />
treatments without lime. The lowest mineralizati<strong>on</strong> rate c<strong>on</strong>stant was established in the<br />
c<strong>on</strong>trol.<br />
From the results it was evident that the used modified incubati<strong>on</strong> method is a<br />
reliable tool to plan and implement of professi<strong>on</strong>al N supplement.<br />
Keywords: easily soluble N forms; incubati<strong>on</strong>; l<strong>on</strong>g <strong>term</strong> field experiment;<br />
INTRODUCTION<br />
The balance between supplying sufficient nitrogen for optimal plant performance and<br />
minimizing polluti<strong>on</strong> from excessive nitrogen applicati<strong>on</strong> is delicate and very important.<br />
It is widely know that N misuse can lead to polluti<strong>on</strong> of the envir<strong>on</strong>ment but it effects<br />
<strong>on</strong> soil productivity and yield is well-known. The availability of soil N to the crop is<br />
therefore of great importance in agricultural systems.<br />
The de<strong>term</strong>inati<strong>on</strong> of actual N-supply are very important and obligate for sustainable<br />
1 Authors are grateful to coo-workers of MTA-TAKI for making soil sampling possible<br />
and for providing the yield data of the experiment.<br />
226
agriculture in Hungary, especially sandy soils, which are very sensitive to<br />
envir<strong>on</strong>mental effects and inefficient human treatment (Németh (1996)).<br />
The formati<strong>on</strong> and fate of soluble N-forms and its resp<strong>on</strong>se to organic matter<br />
mineralizati<strong>on</strong> is not obvious yet and often inc<strong>on</strong>sistent despite the intensive research<br />
(Lazányi et al. (2002)).<br />
The aim of this paper is to present data <strong>on</strong> different easily mineralizable N-forms<br />
of a l<strong>on</strong>g <strong>term</strong> field experiment (LTFE) by incubati<strong>on</strong> and chemical method in the<br />
Nyírség regi<strong>on</strong>, in Hungary.<br />
MATERIALS AND METHODS<br />
1. Soils and experimental designs<br />
Sandy soils were sampled from the „Nyírlugos” l<strong>on</strong>g-<strong>term</strong> field experiment, which were<br />
established in 1962. The “Nyírlugos” LTFE was set up <strong>on</strong> acidic brown forest soil with<br />
alternating thin layers of clay substance „kovárvány” in the Nyírség regi<strong>on</strong>. The main<br />
characteristics of the experimental soils are: pH(KCl) 4.5, humus 0.7%, CEC 5-10<br />
mgeq/100 g in the ploughed layer. The topsoil was poor in all five macr<strong>on</strong>utrients (N, P,<br />
K, Ca, Mg) and the groundwater depth was 2-3 m (Kádár et al., 1999).<br />
The “Nyírlugos” experiment is a l<strong>on</strong>g-<strong>term</strong> small-plot (50m 2 ) fertilisati<strong>on</strong> and liming<br />
experiment. The experiment c<strong>on</strong>sists of 32 treatments, 4 replicati<strong>on</strong>s altogether 128 plot<br />
experiments. The indicator plant is triticale since 1980.<br />
Fertilisati<strong>on</strong> and liming treatments in the experiment were changed several times in the<br />
last few decades but the levels are c<strong>on</strong>stant since 1980 (Table 1.).<br />
Table 1: Treatments of LTFE in “Nyírlugos”, since 1980<br />
Treatments (kg/ha/year)<br />
Levels N P2O5 K2O CaCO3 MgCO3<br />
C<strong>on</strong>trol - - - - -<br />
1 50 60 60 250 140<br />
2 100 120 120 500 280<br />
3 150 180 180 1000 -<br />
Only ten (<strong>on</strong>e-sided N- and N-combined) treatments are summarized in this paper.<br />
Chosen treatments can be divided four groups according to applied nutrients and lime<br />
(Table 2.) Soil samples were collected from 5 different places in each plot after harvest<br />
of triticale, in August 1998 and 2001.<br />
Table 2: Chosen treatments and their variati<strong>on</strong>s of “Nyírlugos”<br />
C<strong>on</strong>trol group N group NPK group Limed group<br />
N1 N2P2 N2P2 K2Ca2<br />
C<strong>on</strong>trols N2 N2K2 N2P2 K2 Mg2<br />
N3 N2P2K2 N2P2 K2 Ca2 Mg2<br />
227
2. Sample preparati<strong>on</strong><br />
Collected soil samples are handed and prepared as follows: samples were dried and<br />
drilled. Before grinding, samples were cleaned from plant remains and other possible<br />
dirt, and pass a 2 -mm screen. Finally, samples were stored in plastic boxes in dry place<br />
until the examinati<strong>on</strong> according to Hungarian guideline (MSZ 20135:1999).<br />
3. Procedure of biological and chemical soil analysis<br />
A modified periodical incubati<strong>on</strong> experiment is carried out in the laboratory to<br />
de<strong>term</strong>ine the actual rate of N-mineralizati<strong>on</strong> (Stanford and Smith, 1972; Filep and<br />
Tóthné, 1980; Filep and Ferencz, 1999). We improved c<strong>on</strong>diti<strong>on</strong>s of this modified<br />
incubati<strong>on</strong> apparatus.<br />
The enlarged amount of soil (without quartz sand) is supported in a leaching glass tube.<br />
A thin glass wool pad is placed above and below the soil sample to avoid soil dispersi<strong>on</strong><br />
when the leaching soluti<strong>on</strong> is poured into the tube. It is leached free of mineral N and<br />
then incubated at optimum temperature (35°C) and moisture (WC=75%) for 16 weeks.<br />
100 ml of 0.01 M CaCl2 soluti<strong>on</strong> is used for each N removing operati<strong>on</strong>. So this way the<br />
mineralized and easily soluble organic N of soil was removed periodically. Vacuum is<br />
applied <strong>on</strong>ly to remove excess soluti<strong>on</strong>.<br />
The leachates of incubati<strong>on</strong> and the chemical extracti<strong>on</strong>s are analysed for mineralized N<br />
(sum of nitrate and amm<strong>on</strong>ium nitrogen) and organic N according to the method<br />
described by Houba et al., (1986). The different N forms were analysed by autoanalyser<br />
(SKALAR Segment Flow Analyser - CFA system).<br />
The main advantage of this method that both the 0.01 M CaCl2 soluble inorganic Nform<br />
and the UV digestable organic-N form of soil are de<strong>term</strong>ined simultaneously<br />
(Nagy, 2004).<br />
We used N-free nutrient soluti<strong>on</strong> c<strong>on</strong>taining 0.007 M CaSO4∗2H2O; 0.002 M MgSO4;<br />
0.01 M Ca(H2PO4)2 and 0.015 M K2SO4 for use in replacing nutrients. The leaching<br />
process is repeated after 2., 3., 5., 7., 9., 12., and 16 weeks.<br />
The incubati<strong>on</strong> can be <strong>term</strong>inated when cumulative N mineralizati<strong>on</strong> approaches a<br />
plateau.<br />
RESULTS AND DISCUSSIONS<br />
The incubati<strong>on</strong> was applied for chosen samples 1998 and 2001 as menti<strong>on</strong>ed above. The<br />
Nmin and Norg c<strong>on</strong>tent of soil was measured al<strong>on</strong>g the incubati<strong>on</strong>. The pH, the cumulated<br />
values of Nmin and Norg and the results of chemical extracti<strong>on</strong> are shown in Table 3.<br />
1. Results of chemical examinati<strong>on</strong><br />
The pH of soil of experiment was str<strong>on</strong>gly acid as earlier results (Kádár et al., 1999) and<br />
the result of c<strong>on</strong>trol showed.<br />
Mostly the <strong>on</strong>e sided N and NP, NK, NPK treatments had decreased the pH, due to<br />
the acidic effect of applied fertilizers and their l<strong>on</strong>g <strong>term</strong> using. Note, that <strong>on</strong>ly the<br />
liming treatments could hinder the further acidificati<strong>on</strong> and increased the pH.<br />
The amount of 0.01 M CaCl2 soluble mineral N was between 2.55 and 9.63 mg/kg<br />
228
according to the treatments. Obtained values corresp<strong>on</strong>d to the soil type and ruling soil<br />
c<strong>on</strong>diti<strong>on</strong>s. Results were pointed out that in these circumstances the mineralizati<strong>on</strong><br />
processes were hindered.<br />
The amount of 0.01 M CaCl2 soluble organic N was between 4.08 and 7.25 mg/kg<br />
according to the treatments. This fracti<strong>on</strong> has changed lesser than the above menti<strong>on</strong>ed<br />
mineral fracti<strong>on</strong> according to the treatments.<br />
The obtained results of 0.01 M CaCl2 soluble N forms can be explained the effects<br />
of applied dosage of fertilizers. The easily soluble N c<strong>on</strong>tent of soil (Nmin + Norg) was the<br />
lowest in the c<strong>on</strong>trol and the highest in the N2P2K2Ca2Mg2 treatment. Increasing dose of<br />
N has a negative effect <strong>on</strong> pH, slightly changed the amount of Norg and significantly<br />
increased the amount of Nmin, especially N3 treatment (Table 3).<br />
Table 3: Results of chemical and biological analysis (“Nyírlugos” 1998-2001)<br />
Chemical method Biological method<br />
pH Nmin Norg Nmin + Norg Nmin Norg Nmin + Norg<br />
Treatments (mg/kg)<br />
C<strong>on</strong>trol 4.31 3,20 4,08 7,28 180.72 47.07 227.79<br />
N1 4.22 4,18 4,15 8,33 175.84 54.81 230.65<br />
N2 4.04 3,40 4,98 8,38 157.59 42.77 200.36<br />
N3 3.74 9,63 4,55 14,18 176.37 55.60 231.97<br />
N2P2 4.07 3,31 4,53 7,84 172.34 52.75 225.08<br />
N2K2 3.93 8,05 6,43 14,48 156.91 48.23 205.14<br />
N2P2K2 4.05 2,55 4,75 7,30 157.41 45.81 203.22<br />
N2P2K2Ca2 4.40 6,15 4,83 10,98 190.67 49.24 239.91<br />
N2P2K2Mg2 4.78 3,75 5,63 9,38 190.47 45.38 235.85<br />
N2P2K2Ca2Mg2 5.09 9,10 7,25 16,35 211.29 39.02 250.30<br />
Average 4.26 5.33 5.12 10.45 177.99 49.07 225.03<br />
LSD5% 0.25 1.66 0.64 2.09 10.82 3.26 16.81<br />
The treatments of NPK group resulted an adjusted balance between mineral and organic<br />
N forms and slightly decreased the soil pH compared to the c<strong>on</strong>trol.<br />
The treatments of limed group increased the pH and the Nmin and Norg c<strong>on</strong>tent of soil<br />
compared to the c<strong>on</strong>trol. The amount of sum of N forms was about approximately<br />
twofold in these treatments than the c<strong>on</strong>trol.<br />
2. RESULTS OF INCUBATION<br />
From the cumulated data of incubati<strong>on</strong> the following results can be establish:<br />
Only the treatments of limed group have increased the Nmin c<strong>on</strong>tent of soil compared to<br />
the c<strong>on</strong>trol (Table 3). One-sided N-treatments decreased the c<strong>on</strong>tent of mineralized N<br />
forms of soil compared to the c<strong>on</strong>trol during the incubati<strong>on</strong>. It can be explained with the<br />
acidificati<strong>on</strong> effects of <strong>on</strong>e-sided N fertilizati<strong>on</strong> which has caused unfavourable<br />
c<strong>on</strong>diti<strong>on</strong>s for mineralizati<strong>on</strong> processes.<br />
Poorer mineralizati<strong>on</strong> c<strong>on</strong>diti<strong>on</strong>s were caused applying treatments of NPK group as<br />
229
well. The effect of these treatments can be interpreted by the decreasing soil pH. While<br />
improving soil pH (limed group) the c<strong>on</strong>diti<strong>on</strong>s of mineralizati<strong>on</strong> would be better and<br />
the amount of Nmin was higher than the c<strong>on</strong>trol (Table 3).<br />
The cumulated Norg was varied between 39.02 and 55.60 mg/kg according to<br />
treatments. The lowest value was obtained in N2P2K2Ca2Mg2 treatment. It can be<br />
explained by the better circumstances of mineralizati<strong>on</strong>. Results of applying <strong>on</strong>e sided<br />
N treatments pointed out that the rate of mineralizati<strong>on</strong> was lower compared to the<br />
c<strong>on</strong>trol and especially the limed treatments. Furthermore, it was found that the<br />
treatments of NPK group had not effect <strong>on</strong> the amount of Norg.<br />
The amount of sum of N forms was the highest in the case of treatments of limed group.<br />
Treatments of NPK group have a negative effect <strong>on</strong> the amount of sum of N forms<br />
compared to the c<strong>on</strong>trol. It can be established that increasingly applied N not improved<br />
the rate of mineralizati<strong>on</strong> remarkably.<br />
Furthermore, the 0.01 M CaCl2 soluble N-fracti<strong>on</strong>s are in mainly inorganic forms<br />
under incubati<strong>on</strong> period but the c<strong>on</strong>tent of organic forms also significant (Table 3.).<br />
The relati<strong>on</strong>ship between the Nmin and time could be studied from the periodical data of<br />
incubati<strong>on</strong> (Table 4 and Figure 1).<br />
According to results typical relati<strong>on</strong>ship was established N mineralizati<strong>on</strong> vs. time<br />
(Figure 1). In spite of the different treatments the shapes of the obtained curves were<br />
analogous. Logarithmic rate equati<strong>on</strong> was founded in the case of all treatments.<br />
Difference was established between coefficients of correlati<strong>on</strong>s and intercepts (Table<br />
4.). Differences were explained the above menti<strong>on</strong>ed effects of treatments. Liming<br />
increased, <strong>on</strong>e-sided N treatments decreased the amount of Nmin.<br />
Cumulated Nmin (mg/kg<br />
250<br />
200<br />
150<br />
100<br />
50<br />
0<br />
0 4 8 12 16<br />
Time (weeks)<br />
Figure 1: Relati<strong>on</strong>ship Nmin vs. Time<br />
230<br />
C<strong>on</strong>trol<br />
N1<br />
N2<br />
N3<br />
NPK<br />
Limed
Table 4: Results of regressi<strong>on</strong> analysis<br />
Treatments Regressi<strong>on</strong> equati<strong>on</strong> R value<br />
C<strong>on</strong>trol y = 11,83Ln(x) + 105,02 0.7652<br />
N1 y = 14,097Ln(x) + 93,719 0.7971<br />
N2 y = 12,989Ln(x) + 89,133 0.8198<br />
N3 y = 11,83Ln(x) + 105,02 0.7652<br />
NPK group y = 13,019Ln(x) + 88,636 0.8043<br />
Limed group y = 15,419Ln(x) + 111,97 0.8153<br />
In the 1970s Stanford and co-workers (Stanford and Smith (1972), Stanford (1973),<br />
Stanford et al. (1974)) advanced the c<strong>on</strong>cept of potentially mineralizable N (Npot) and a<br />
related mineralizati<strong>on</strong> rate c<strong>on</strong>stant (k) for use in characterizing soil-available N. Since<br />
then this c<strong>on</strong>cept has been used, modified, and discussed in great detail by numerous<br />
scientist, but they theory has been accepted generally.<br />
This theory assumed that organic nitrogen mineralizati<strong>on</strong> at optimum temperature and<br />
moisture followed first-order kinetics. Accept the above menti<strong>on</strong>ed theory our results<br />
have made possibility to estimate the potentially mineralizable N of the soil and its rate<br />
c<strong>on</strong>stant. Our results have c<strong>on</strong>firmed that the correlati<strong>on</strong> between 1/Nt and 1/t is linear.<br />
The potentially mineralizable N-c<strong>on</strong>tents (Npot) and the c<strong>on</strong>stant of the mineralizati<strong>on</strong><br />
(k) were calculated according to Filep and Tóthné (1980) (Table 5.).<br />
It can be established that the applied treatments has not remarkably effect <strong>on</strong> the values<br />
of Npot, but higher c<strong>on</strong>stant rate of mineralizati<strong>on</strong> was achieved in the limed treatments<br />
than the treatments without lime. The lowest mineralizati<strong>on</strong> rate c<strong>on</strong>stant was<br />
established in the c<strong>on</strong>trol.<br />
Table 5: Calculated Npot and k values<br />
Treatments Npot (mg/kg) k (35ºC)<br />
C<strong>on</strong>trol 277.799 0.056<br />
N1 233.247 0.057<br />
N2 233.451 0.058<br />
N3 234.028 0.058<br />
NPK group 233.551 0.058<br />
Limed group 249.885 0.064<br />
3. Interpretati<strong>on</strong> of results of chemical and biological analysis to yield<br />
Obtained results of biological analysis can be compared to the yield (Figure 2.).<br />
The c<strong>on</strong>stant of the mineralizati<strong>on</strong> was the lowest in the c<strong>on</strong>trol. Increasing dose of <strong>on</strong>esided<br />
N slightly increased the k value and decreased the yield, especially N2 and N3.<br />
This effect can be explained by the increasing acidizati<strong>on</strong> of soil and the increasing<br />
c<strong>on</strong>tent of easily soluble N fracti<strong>on</strong>s in these treatments. The higher k was obtained in<br />
the case of the limed group. It can be explained the above menti<strong>on</strong>ed more favourable<br />
soil c<strong>on</strong>diti<strong>on</strong>s. The result of NPK treatment-group was better than the results of N<br />
treatments and c<strong>on</strong>trol but worse compared to the limed group.<br />
231
Figure 2: Relati<strong>on</strong>ship between the yield and the c<strong>on</strong>stant of the mineralizati<strong>on</strong> (k)<br />
From obtained values of Npot it was evident that liming improves the c<strong>on</strong>diti<strong>on</strong>s of<br />
mineralizati<strong>on</strong> of soil. It can be explained by higher microbiological activity and higher<br />
pH of the soil (Kátai et. al. (1999)). Furthermore the mineralizati<strong>on</strong> potential of soil is<br />
depended not <strong>on</strong>ly the actual mineralized N c<strong>on</strong>tent and pH of soil but the amount of<br />
easily soluble, low molecular weighted and mobilizable organic N fracti<strong>on</strong> as well.<br />
From the results it was evident that the used modified incubati<strong>on</strong> method is a reliable<br />
tool plan and implement of professi<strong>on</strong>al N supplement.<br />
Results of chemical analysis can be compared to the yield as well (Figure 3.).<br />
N (mg/kg)<br />
k value<br />
12<br />
10<br />
8<br />
6<br />
4<br />
2<br />
0<br />
0,066<br />
0,064<br />
0,062<br />
0,06<br />
0,058<br />
0,056<br />
0,054<br />
0,052<br />
C<strong>on</strong>trol N1 N2 N3 NPK Limed<br />
Nmin<br />
Norg<br />
Yield<br />
k<br />
yield<br />
C<strong>on</strong>trol N1 N2 N3 NPK Limed<br />
Figure 3: Relati<strong>on</strong>ship between the yield and the 0.01 M CaCl2 soluble mineral and<br />
organic N<br />
232<br />
7<br />
6<br />
5<br />
4<br />
3<br />
2<br />
1<br />
0<br />
7<br />
6<br />
5<br />
4<br />
3<br />
2<br />
1<br />
0<br />
Yield (t/ha)<br />
Yield (t/ha)
From obtained data, it can be established that the c<strong>on</strong>tent of 0.01 M CaCl2 soluble Nmin<br />
and Norg fracti<strong>on</strong>s were the lowest in the c<strong>on</strong>trol. One-sided N treatments mostly<br />
increased the amount of mineral N forms, but excess doses of N (>50 kg/ha) resulted<br />
significant yield-decrement. Moreover, 150 kg/ha N dose (N3) highly increased the Nmin<br />
in the upper layer. This excess amount of mineral N (mostly nitrate) can be dangerous<br />
for the quality of soil water. Liming treatments produced higher yields without excess<br />
c<strong>on</strong>tent of nitrate.<br />
REFERENCES<br />
Filep Gy. - Tóthné Bíró Á. (1980): Hazai talajok mineralizálható N-készletének és Nszolgáltatásának<br />
mérése és számítása. Agrokémia és Talajtan. 29. 229-244. (in<br />
Hungarian)<br />
Filep Gy. - Ferencz G. (1999): A talaj N-szolgáltató képességének becslésére használt<br />
néhány számítási módszer értékelése. DATE Tudományos Közleményei. Tom.<br />
XXXIV. 73-82. (in Hungarian)<br />
Houba, V. J. G. - Novozamsky, I. - Huijbregts, A. W. M. - Van Der Lee J. J.<br />
(1986): Comparis<strong>on</strong> of soil extrcti<strong>on</strong>s by 0,01 CaCl2 by EUF and by some<br />
c<strong>on</strong>venti<strong>on</strong>al extracti<strong>on</strong> procedures. Plant and Soil 96. 433-437. p<br />
Kádár I. - Németh T. - Szemes I. (1999): Triticale trágyareakciója a nyírlugosi<br />
tartamkísérletben. Növény<strong>term</strong>elés, Tom. 48. No. 6. 647-661. (in Hungarian)<br />
Kátai J. – LazányI J. – Veres E. (1999): Talajmikrobiológiai vizsgálatok a Westsik<br />
vetésforgó tartamkísérletben. DATE Tiszántúli mezőgazdasági tudományos napok.<br />
K<strong>on</strong>ferencia kiadvány (szerk.: Loch J. – Vágó I. – Jávor A.): 175-184. o. (in<br />
Hungarian)<br />
Lazányi J. – Loch J. – Jászberényi I. (2002): Analysis of 0.01 M CaCl2 soluble<br />
organic nitrogen in the treatments of Westsik’s crop rotati<strong>on</strong> experiment.<br />
Agrokémia és Talajtan. 51:79-88. o.<br />
MSZ 20135:1999: De<strong>term</strong>inati<strong>on</strong> of the soluble nutrient element c<strong>on</strong>tent of the soil.<br />
Hungarian Standards Instituti<strong>on</strong>. Budapest (in Hungarian)<br />
Nagy P. T. (2004):<br />
Németh T. (1996): Talajaink szervesanyag-tartalma és nitrogénforgalma. MTA<br />
Talajtani és Agrokémiai kutató Intézete, Budapest (in Hungarian)<br />
Stanford G. - Smith S. J. (1972): Nitrogen mineralizati<strong>on</strong> potentials of soils. Soil Sci.<br />
Soc. Am. Proc. 36:465-472. In. Page A. L. - Miller R. H. - Keeney D. R. (1982):<br />
Methods of soil analysis. Part 2, Agr<strong>on</strong>omy No. 9 (2) p. 711-733. Madis<strong>on</strong>,<br />
Wincosin<br />
Stanford G. (1973): Rati<strong>on</strong>ale for optimum nitrogen fertilizati<strong>on</strong> in corn producti<strong>on</strong>. J.<br />
Envir<strong>on</strong>. Qual. 2:159-166. In. Page A. L. - Miller R. H. - Keeney D. R. (1982):<br />
Methods of soil analysis. Part 2, Agr<strong>on</strong>omy No. 9 (2) p. 711-733. Madis<strong>on</strong>,<br />
Wincosin<br />
Stanford G. - Carter J. N. - Smith S. J. (1974): Estimates of potentially mineralizable<br />
soil nitrogen based <strong>on</strong> short-<strong>term</strong> incubati<strong>on</strong>s. Soil Sci. Soc. Am. Proc. 38:99-102.<br />
In. Page A. L. - Miller R. H. - Keeney D. R. (1982): Methods of soil analysis. Part<br />
2, Agr<strong>on</strong>omy No. 9 (2) p. 711-733. Madis<strong>on</strong>, Wincosin USA<br />
233
EFFECT OF FERTILIZATION SYSTEMS AND LIMING ON MOBILE<br />
PHOSPHORUS CONTENT IN DIFFERENT SOIL TYPES<br />
ABSTRACT<br />
Augusta Olivia Lujerdean<br />
University of Agriculture Science and Veterinary Medicine Cluj-Napoca,<br />
Faculty of Animal Breeding and Biotechnology<br />
3-5 Calea Manastur Street 400732, Romania<br />
Liming and applicati<strong>on</strong> <strong>on</strong> acidic soils of inorganic fertilizers or organic manure had the<br />
c<strong>on</strong>siderable influence <strong>on</strong> the mobile phosphorus c<strong>on</strong>tent and pH of soil. P(AL)<br />
de<strong>term</strong>inati<strong>on</strong>s suggest that differences in the soil physical-chemical properties and the<br />
forming which P is applied influences the effectiveness of fertilizati<strong>on</strong> and P balance.<br />
The phosphorus transformati<strong>on</strong> process was changed under the influence of liming<br />
intensity, soil liming changed phosphate forms and the ratio between mineral fracti<strong>on</strong>s<br />
of phosphates.<br />
The aim of this study was to establish the effect of liming and fertilizati<strong>on</strong> <strong>on</strong> the<br />
amount of available soil phosphorus in different classes of North-Western Romanian<br />
soils: albic luvisoil, clay illuvial brown, clay illuvial chernozem.<br />
Observati<strong>on</strong>s showed a maximum effect of pH increase and P mobile c<strong>on</strong>tent fertilising<br />
inorganic NPK together with manure <strong>on</strong> albic luvisoil.<br />
Keywords: soil liming, mobile phosphorus, inorganic fertilizer, farmyard manure<br />
INTRODUCTION<br />
Different classes of soils in North-West Romania present low productivity inadequate<br />
soil management, irregular supply of nutrients, the acidity being pointed out as the<br />
cause for declined yield.<br />
Phosphorus availability in most soils is optimal at the soil pH of 6.5. Below this value<br />
as in more the soils of this part of Romania phosphorus will react with ir<strong>on</strong> (Fe)/<br />
aluminium (Al) oxides and start forming precipitates which prohibit the plant from<br />
being able to fixate phosphorus. Liming acidic soils will generally increase availability<br />
of phosphorus to plant. Ideally a liming program should be planned so that the pH can<br />
be kept between 5.5 and 6.8 if maximum benefit is to be derived from applied<br />
phosphorus (Havlin et al.2005). Over liming can depress phosphorus solubility due to<br />
the formati<strong>on</strong> of insoluble precipitates with calcium. A decrease in phosphorus<br />
availability because of this formati<strong>on</strong> of Ca-P minerals also happen in an alkaline soil<br />
with a pH value higher than 6.5. Several other characteristics affect the process and<br />
availability of phosphorus fixati<strong>on</strong>. The texture and type of soil str<strong>on</strong>gly influences the<br />
level of mobile phosphorus.<br />
MATERIALS AND METHODS<br />
Soil samples were obtained from l<strong>on</strong>g time experiments established in 1961 at Livada,<br />
Satu Mare County. This experiment includes treatments with different levels of mineral<br />
fertilizers, organic manure and lime inputs.<br />
234
Tops soil (0-30 cm) samples were taken from fields with an average size of 1.0 ha. They<br />
were representative for the following classes of soil: luvisoil, clay illuvial brown, clay<br />
illuvial chernozem. Each type of soil was fertilized as follows: N100, P 70, mineral NPK<br />
(N100 P70 K60) mineral and organic NPK (N100 P70 K60 + 20t ha -1 farmyard) and lime<br />
additi<strong>on</strong> at various rates: 0.5 and 10 t ha -1<br />
Detailed land use and agricultural management data were collected through<br />
Romanian Statistic Annual (2004-2005). Each field was c<strong>on</strong>sidered as a separate unit<br />
because it is not managed in a coherent rotati<strong>on</strong>.<br />
In 2006 five representative points of each type of soil were selected for sampling. The<br />
soils were air dried and ground ≤ 2mm sieve at the time of sampling.<br />
Extracti<strong>on</strong>s with acetate-lactate soluti<strong>on</strong> (AL) were performed using soil to<br />
soluti<strong>on</strong> ratio of 1:20 and the shaking time was 30 minutes. 5ml filtrate was treated with<br />
(NH4)2 MoO4 0,15% and 5 ml fresh prepared 1,5% ascorbic acid standard curve was<br />
prepared using suitable volumes of standard soluti<strong>on</strong> in place of sample. It has been<br />
used a series of 6 standard soluti<strong>on</strong>s within the approximate range of 0.01 – 2.0 mg P/l.<br />
After 10 minutes absorbance was measured at 880 nm using a reagent blank as a<br />
reference soluti<strong>on</strong>.<br />
Soil pH was measured in water using 1:2.5 ratio.<br />
All analyses were carried out in 4 repetiti<strong>on</strong>s.<br />
RESULTS AND DISCUSSIONS<br />
1. Chemical analysis of soils showed that the pH and mobile phosphorus c<strong>on</strong>tent was<br />
exactly related to lime ratio and type of fertilizer. Impact of treatment (liming and<br />
fertilizing) <strong>on</strong> different types of soil is shown in the below tables:<br />
1. ALBIC LUVISOIL (initial pH = 4.90, P=15.4 ppm)<br />
In this experiment the str<strong>on</strong>gest effect <strong>on</strong> improving soil acidity and phosphorus c<strong>on</strong>tent<br />
of albic luvisoil was observed when fertilizing with mineral and organic manure<br />
mixture (N100 P 70 K60 + 20t/ ha -1 FYM) and applying 10t ha -1 limest<strong>on</strong>e.<br />
On the background without organic fertilizer the greatest efficacy was recorded when<br />
albic luvisoil had been applied with N100 P 70 K60 and 10t / ha -1<br />
Table 1: Effect fertilizing and liming 5t/ ha -1 CaCO3 <strong>on</strong> albic luvisoil<br />
Treatment ∆pH P (ppm)<br />
C<strong>on</strong>trol 0.49 21.5<br />
N100 0.21 10.1<br />
P70 0.47 28.0<br />
N100 P70 K60 0.96 40.1<br />
N100 P 70 K60 + FYM 0.89 46.5<br />
FYM 0.35 40.0<br />
∆pH = pH variati<strong>on</strong><br />
FYM = farmyard manure<br />
235
Table 2: Effect of fertilizing and liming 10t/ ha -1 CaCO3 <strong>on</strong> albic luvisoil<br />
Treatment ∆pH P (ppm)<br />
C<strong>on</strong>trol 0.85 20.5<br />
N100 0.24 15.4<br />
P70 0.88 39.0<br />
N100 P70 K60 1.09 85.4<br />
N100 P 70 K60 + FYM 0.90 91.5<br />
FYM 0.90 58.0<br />
2. CLAY ILLUVIAL BROWN SOIL (initial pH = 5.10, P = 18 ppm)<br />
Liming 10t/ ha -1 and fertilizing N100 P 70 K60 + FYM increased soil pH by 1.02 units and<br />
mobile phosphorus raised to 70.1 ppm <strong>on</strong> clay illuvial brown soil.<br />
Table 3: Impact fertilizing and liming 5t/ ha -1 CaCO3 <strong>on</strong> clay illuvial brown soil<br />
Treatment ∆pH P (ppm)<br />
C<strong>on</strong>trol 0.29 24.5<br />
N100 -0.20 19.2<br />
P70 0.29 28.4<br />
N100 P70 K60 0.88 38.4<br />
N100 P 70 K60 + FYM 0.80 45.3<br />
FYM 0.32 33.0<br />
Table 4: Impact fertilizing and liming 10t/ ha -1 CaCO3 <strong>on</strong> clay illuvial brown soil<br />
Treatment ∆pH P (ppm)<br />
C<strong>on</strong>trol 0.65 20.5<br />
N100 -0.10 23.4<br />
P70 0.50 38.2<br />
N100 P70 K60 1.00 60.9<br />
N100 P 70 K60 + FYM 1.02 70.1<br />
FYM 0.70 39.0<br />
3. CLAY-ILLUVIAL CHERNOZEM (initial pH = 5.20, P = 25.4 ppm)<br />
Applying <strong>on</strong>ly mineral fertilizer or together with manure and liming = 10t/ ha -1 pH of<br />
clay illuvial chernozem significantly increased: 1.06, respectively 1 pH units. Lime has<br />
positive effect <strong>on</strong> P c<strong>on</strong>centrati<strong>on</strong>, an increase to 48.9 ppm, respectively 50.4 ppm was<br />
found.<br />
236
Table 5: Impact fertilizing and liming 5t/ ha -1 CaCO3 <strong>on</strong> clay illuvial brown soil<br />
Treatment ∆pH P (ppm)<br />
C<strong>on</strong>trol 0.15 28.9<br />
N100 0.10 21.4<br />
P70 0.35 30.0<br />
N100 P70 K60 0.90 42.3<br />
N100 P 70 K60 + FYM 0.88 48.2<br />
FYM 0.20 34.9<br />
Table 6: Impact fertilizing and liming 5t/ ha -1 CaCO3 <strong>on</strong> clay illuvial brown soil<br />
Treatment ∆pH P (ppm)<br />
C<strong>on</strong>trol 0.55 14.3<br />
N100 0.20 24.5<br />
P70 0.40 30.4<br />
N100 P70 K60 1.06 48.9<br />
N100 P 70 K60 + FYM 1.00 50.4<br />
FYM 0.35 40.9<br />
CONCLUSIONS<br />
1. The phosphorus c<strong>on</strong>tent and pH value were found to be str<strong>on</strong>gly dependent both of<br />
treatment and soil type.<br />
2. Soil supplementati<strong>on</strong> with 10t/ha lime has the greatest effect <strong>on</strong> albic luvisoil pH<br />
that increased with 0.85 units while P c<strong>on</strong>tent of clay illuvial brown soil raised to<br />
31.2 ppm.<br />
3. Neutralizati<strong>on</strong> of soil acidity with limest<strong>on</strong>e not <strong>on</strong>ly increases the soil pH, but also<br />
changes the availability of P. Applying 10t ha -1 CaCO3 decreased to 20.5 ppm<br />
(albic luvisoil), 14.3 ppm (clay illuvial chernozem) and 20.4 ppm (clay illuvial<br />
brown soil).<br />
4. This effect can be offset by fertilizati<strong>on</strong>. The applicati<strong>on</strong> of NPK together with<br />
manure increased the P level to 70.1 ppm (clay illuvial brown), 50.4 ppm (clay<br />
illuvial chernozem) and 91.5 ppm (albic luvisoil).<br />
REFERENCES<br />
Havlin, J.L., James, D. Beat<strong>on</strong>, Samuel L. Tisdale and Werner L. Nels<strong>on</strong>, (2005):<br />
Soil Fertility and Fertilizers: An Introducti<strong>on</strong> to Nutrient Management 7 th Editi<strong>on</strong>.<br />
Ec<strong>on</strong>omic Journal, 68: 302-316 p<br />
Larsen, S. (1967): Soil Phosphorus. Advances in Agr<strong>on</strong>omy 19: 105-207 p<br />
Augusta Olivia Lujerdean (2003): Cercetari privind metodologia agrochimica de<br />
de<strong>term</strong>inare a formelor de fosfati din soluri. Teza de doctorat, Universitatea de<br />
Stiinte Agricole si Medicina Veterinara, Cluj-Napoca<br />
Pierysznski, G.M. (2000): Methods of Phosphorus Analyzis for Soils Sdiments<br />
Residuals and Waters. Southern Coop. Series Bull No 396<br />
237
Rusu, M., Boeriu, I., Kurtinec, P., Botha, P.V. (1981): Aspecte privind chimismul<br />
fosforului in solurile acide din Nord Vestul tarii. Analele I.C.C.P. Fundulea, Vol 47<br />
Irina Vintila, Borlan, Z., Rauta, C., Daniliuc, D., Tiganas, L. (1984): Situatia<br />
agrochimica a solurilor din Romania, Ed. Ceres, Bucuresti.<br />
238
COMPOSTED SLAUGHTERHOUSE BY-PRODUCTS EFFECT<br />
ON CROP YIELD<br />
Péter Ragályi, Imre Kádár<br />
Research Institute for Soil Science and Agricultural Chemistry<br />
H-1022, Budapest, Herman Ottó Str. 15, Hungary. E-mail: ragalyi@rissac.hu<br />
ABSTRACT<br />
Use of treated slaughterhouse waste as fertilizer can be a good way to maintain soil<br />
productivity as well as reduce organic waste. Effect and residual effect of different<br />
composts and meat powder were examined <strong>on</strong> crop development and yield <strong>on</strong> a<br />
calcareous sandy soil. The soil c<strong>on</strong>tained 1-6% CaCO3 and 1-1.5% humus, humus layer<br />
was 60-80 cm and pH(H2O) 7.0-7.4 as an average. The site had a poor N, P and K<br />
supply. Water table was at 6-8 m depth. Trials were set in 2002 and 2003 with 5 doses<br />
(0, 25, 50, 100, 200 t/ha fresh compost and 0; 2,5; 5; 10; 20 t/ha meat powder <strong>on</strong>ce<br />
applied) and 4 replicati<strong>on</strong>s giving a sum of 20 plots with 5x8=40 m 2 area each, arranged<br />
in split-plot design. Sown crops were maize in 2002, mustard in 2003 and triticale<br />
m<strong>on</strong>oculture from 2004.<br />
In the first two years of the trial the yield was negligible due to draught, though<br />
tendencies showed 50t/ha dose mature compost and 25 t/ha immature compost as<br />
optimal treatment.<br />
All of the composts had significant residual effect <strong>on</strong> triticale in 2004 as the weather<br />
was favourable and immature compost increased harvested straw and grain air-dried<br />
yield from 1.6 to 5.3 t/ha. Maximal yields occurred at the maximal compost doses<br />
without depressi<strong>on</strong>. In following years residual effects were moderated, but differences<br />
were significant in the case of str<strong>on</strong>ger composts even in 2006, in the 4 th and 5 th<br />
experimental years.<br />
Keywords: compost, fertilizati<strong>on</strong>, crop yield, slaughterhouse by-product<br />
INTRODUCTION<br />
Fodder use of animal origin wastes is strictly regulated by EU since BSE (bovine<br />
sp<strong>on</strong>giform encephalopathy) disease showed up, so alternative utilities have to be<br />
found. In Hungary 100-120 milli<strong>on</strong> t<strong>on</strong>s of wastes produced of which 5% is so called<br />
“hazardous”. Hazardous wastes from animal bodies reach 300-400 thousand t<strong>on</strong>s of<br />
which 70-90 thousand are composted. After heat treatment these wastes become n<strong>on</strong>hazardous,<br />
and other treatments like composting make land applicati<strong>on</strong> possible. This<br />
practice may improve soil parameters like organic matter, nutrient c<strong>on</strong>tent, water<br />
holding capacity and also reduces the yearly deposited 55 000 t<strong>on</strong>s of slaughterhouse<br />
wastes and sewage sludge (Vermes 1998, Kiss et al. 2001). Burying or incinerati<strong>on</strong> of<br />
these materials is expensive and can result in envir<strong>on</strong>mental polluti<strong>on</strong> (Izsáki 2000).<br />
Debreczeni and Izsáki (1985) set up pot experiments to study the changes of field crops<br />
grown <strong>on</strong> calcareous sandy soil and chernozem meadow clayey soil mixed with various<br />
doses of sewage sludge from a tannery. On calcareous sandy soil 13 g sludge D.M. / kg<br />
soil increased mustard above ground parts with 74% but higher doses caused<br />
depressi<strong>on</strong>. Maize yields (above ground) were doubled in 33 g sludge D.M. / kg soil<br />
239
treatment, while spring barely had 60 and 20% more yield due to 13 and 66 g sludge<br />
D.M. / kg soil resp.<br />
Effect of fertilizati<strong>on</strong> using various rates of tannery sewage sludge was studied in<br />
field experiment <strong>on</strong> crop yields <strong>on</strong> calcareous humic sandy soil. The 60 t/ha (15-20 t/ha<br />
D.M.) sludge dressing applied for four years completed with phosphorus and potassium<br />
fertilizers was found to increase by 23 and 15% the grain yield of spring barely and rye<br />
resp. as compared to the fertilized c<strong>on</strong>trol in the first two years, however in the third and<br />
fourth years yield dropped by 20%. 120 t/ha sludge applied for a four-year period was<br />
found to cause yield depressi<strong>on</strong> in spring barley in first year, in further years rye and<br />
winter wheat grain yields were roughly equal with those obtained in the fertilized<br />
treatment. The 60 + 60 t/ha dose for two-two years did not cause depressi<strong>on</strong> and<br />
resulted in 2-16% higher yields (Izsáki and Debreceni 1987). A slight residual effect<br />
over 2-3 year of this experiment has been observed in 120 t/ha and 60 + 60 t/ha<br />
treatments (Debreczeni and Izsáki 1989).<br />
Sim<strong>on</strong> and Szente (2000) set up field experiments with municipal sewage sludge<br />
compost in maize <strong>on</strong> slightly acidic loamy-sand textured brown forest soil with 0, 10,<br />
40 t/ha loads 2 weeks before sowing (in 1996) and 0, 10, 20, 40 t/ha loads 1 week<br />
before sowing (in 1997). In the first year 9.6 t/ha yield of the c<strong>on</strong>trol plot was increased<br />
with 14% and 4% by 10 and 40 t/ha compost resp. In sec<strong>on</strong>d year 20 t/ha dose gave<br />
68% higher yield while 40 t/ha gave similar yield to 3.7 t/ha of c<strong>on</strong>trol.<br />
Applicati<strong>on</strong> of sewage sludge is often limited by N c<strong>on</strong>tent and not by relatively low<br />
heavy metal c<strong>on</strong>tent. Total N c<strong>on</strong>tent release and utility parallel with decay take more<br />
years. Unfortunately <strong>on</strong>ly a few data is available about N release rate of different<br />
sewage sludge in different soil types, weather c<strong>on</strong>diti<strong>on</strong>s and cultivati<strong>on</strong> practice.<br />
Kádár et al. (2002) set up a field experiment with dried communal sewage sludge<br />
and slaughterhouse compost with 0, 25, 50, 100, 200 t/ha rate <strong>on</strong> clayey brown forest<br />
soil applied 5-6 weeks before sowing of sugar beet. Despite of draught the crop could<br />
develop well <strong>on</strong> treated plots while c<strong>on</strong>trol plants were depressed. The highest 200 t/ha<br />
load of slaughterhouse compost resulted the highest sugar yield of 8.7 t/ha compared to<br />
6.5 t/ha of c<strong>on</strong>trol. Optimal 25 t/ha dose of sewage sludge enhanced yield to 7.2 t/ha,<br />
above this rate N overdose worsened quality parameters. Sludge and compost improved<br />
structure, water management and fertility of soil.<br />
MATERIALS AND METHODS<br />
The l<strong>on</strong>g-<strong>term</strong> field experiment was set up at experimental site of the Research Institute<br />
for Soil Science and Agricultural Chemistry at Őrbottyán <strong>on</strong> a calcareous sandy soil<br />
with 1-6% CaCO3, 1-1.5% humus, 10-15% clay fracti<strong>on</strong>, pH(H2O): 7.0-7.4 in average<br />
in the ploughed layer. The water table was at 6-8 m, the site was pr<strong>on</strong>e to drought and<br />
weakly supplied with N, P and K. Materials were applied at 5 different levels in 4<br />
replicati<strong>on</strong>s making 20 plots for each experiment (compost form). The plots had an area<br />
of 5x8=40m 2 and arranged in split-plot design. In each experiment the applied rates<br />
were 0, 25, 50, 100, 200 t/ha fresh compost. In case of meat powder the doses were 0,<br />
2.5, 5.0, 10.0, 20.0 t/ha. As the soil and the composts were poor in potassium, 200 kg/ha<br />
K2O fertilizati<strong>on</strong> was given in all the experiments in spring 2003. The composts were<br />
processed by ATEVSZOLG Inc.<br />
240
The mature compost became friable, inodorous, homogeneous material after 2-m<strong>on</strong>th<br />
air-exposure and 10-m<strong>on</strong>th maturati<strong>on</strong>. Immature compost was stinking, rough,<br />
heterogeneous, after 6-week air-exposure and without maturati<strong>on</strong>. Meat powder based<br />
semi-mature compost was the material of immature compost maturing 6 m<strong>on</strong>ths more,<br />
but was still stinking and rough. Cooked meat based semi-mature compost received 2m<strong>on</strong>th<br />
air-exposure and 8-m<strong>on</strong>th maturati<strong>on</strong>.<br />
Testing immature and semi-mature composts as well as extreme high applicati<strong>on</strong><br />
doses served experimental purposes. Before applicati<strong>on</strong> 2-2 composite samples (from<br />
20-20 core samples) were taken from the materials. Analyses were made in the<br />
Research Institute for Soil Science and Agricultural Chemistry. The average<br />
compositi<strong>on</strong>s of composts are shown in Table 1.<br />
Table 1: Compositi<strong>on</strong> of composts and meat powder in D.M.<br />
with cc.HNO3+cc.H2O2 digesti<strong>on</strong><br />
Measured Unit Mature Immature Semi-mature Semi-mature Meat<br />
parameters compost compost compost mp* compost cm** powder<br />
Dry Matter % 38.9 45.8 60.0 55.7 95.0<br />
Org. Matter % 26.3 41.7 40.3 43.8 58.6<br />
Organic C % 15.2 24.1 23.3 25.3 33.9<br />
C/N ratio 7.5 7.7 7.1 8.7 5.3<br />
Ca % 9.31 12.65 11.25 11.69 7.02<br />
P % 2.22 5.56 4.26 5.26 4.06<br />
N % 2.04 3.12 3.26 2.89 6.41<br />
K % 0.76 0.76 0.83 0.50 0.41<br />
Mg % 0.70 0.36 0.37 0.54 0.18<br />
Na % 0.52 0.79 0.69 0.63 0.45<br />
S % 0.50 0.70 0.62 0.75 0.60<br />
Zn mg/kg 540 270 164 237 104<br />
Cu mg/kg 109 46 19 42 13<br />
NH4-N mg/kg 169 3006 941 882 167<br />
NO3-N mg/kg 2480 1135 61 122 1<br />
*meat powder based, **cooked meat based<br />
In the first experimental year (2002) maize (Zea mays), in the sec<strong>on</strong>d mustard (Sinapis<br />
alba) and from the third triticale (X Triticosecale) m<strong>on</strong>oculture were grown. The<br />
amount of precipitati<strong>on</strong> <strong>on</strong> sandy soil has str<strong>on</strong>g effect <strong>on</strong> yields. The years 2002 and<br />
2003 were drought. The maize received 237 mm and the mustard 52 mm of<br />
precipitati<strong>on</strong> during their growing seas<strong>on</strong>. Between 2004 and 2006 the triticale had<br />
satisfactory amount of precipitati<strong>on</strong> evenly distributed (Table 2).<br />
241
Table 2: Precipitati<strong>on</strong> of the experimental stati<strong>on</strong>, mm (Calcareous sandy soil,<br />
Őrbottyán, 2002-2006)<br />
2002 2003 2004 2005 2006 45-year<br />
average<br />
January 6 40 46 7 51 35<br />
February 13 27 49 52 39 30<br />
March 14 0 53 8 35 39<br />
1 st quarter 33 67 148 67 125 95<br />
April 30 12 39 96 23 46<br />
May 46 32 42 41 82 59<br />
June 41 8 68 48 83 67<br />
2 nd quarter 117 52 149 185 188 173<br />
July 52 57 35 85 30 61<br />
August 98 13 67 124 118 51<br />
September 59 17 13 74 29 46<br />
3 rd quarter 209 87 115 283 177 168<br />
October 52 79 48 14 14 44<br />
November 32 45 55 33 15 55<br />
December 40 7 36 76 4 45<br />
4 th quarter 124 131 139 123 33 144<br />
Total 483 337 551 658 523 569<br />
RESULTS AND DISCUSSION<br />
The average 20-30 t/ha rate of farmyard manure decays in 3-4 years in soil. About half<br />
of its 2.0-3.0% D.M. nitrogen c<strong>on</strong>tent can be utilized during this time of which 50% in<br />
the first year, 30% in the sec<strong>on</strong>d and 20% in the third (Sarkadi 1975).<br />
In 2002 the 200 t/ha immature compost caused depressi<strong>on</strong> in yield, otherwise no<br />
significant effect could develop. Trends showed that up to 50 t/ha mature compost and<br />
25 t/ha immature compost applicati<strong>on</strong> resulted in slightly better maize stand. Above<br />
ground D.M. yield was 3 t/ha as an average. Drought seas<strong>on</strong> c<strong>on</strong>tinued in 2003 when<br />
mustard yields were ec<strong>on</strong>omically negligible.<br />
Effects of the composts were enhanced in 2004 as the crop received sufficient<br />
precipitati<strong>on</strong> during the 9 m<strong>on</strong>ths growing seas<strong>on</strong>. The plantati<strong>on</strong> could develop better<br />
and reach higher yields even at lower doses of the applied materials, but maximal load<br />
boosted the results. The 2 nd year residual effect of mature compost was moderate, but<br />
significant. Immature compost could increase 3-fold the c<strong>on</strong>trol yield, that is from 1.6<br />
t/ha to 5.3 t/ha grain (table 3).<br />
Highest dose of meat powder based semi-mature compost had also str<strong>on</strong>g after<br />
effects <strong>on</strong> triticale and doubled the yield of c<strong>on</strong>trol, while the cooked meat based <strong>on</strong>e<br />
increased it with nearly 50% (table 4). Meat powder had also similar positive effects,<br />
242
maximal yield could be reached at 5 t/ha load, but c<strong>on</strong>siderably decline did not occur<br />
even at higher levels (table 5). The soil could have 320 kg/ha N through the applied 5<br />
t/ha meat powder, which could cover the N demand of the higher biomass producti<strong>on</strong><br />
(Kádár and Ragályi 2004).<br />
Table 3: Effect of slaughterhouse composts <strong>on</strong> air-dried triticale yield, t/ha<br />
(Calcareous sandy soil, Őrbottyán)<br />
Compost t/ha fresh material LSD5% Mean<br />
0 25 50 100 200<br />
Mature compost (applied 09 May 2002)<br />
Triticale 2004<br />
Grain 2.2 2.2 2.0 2.9 2.6 0.6 2.4<br />
Straw 3.8 3.8 3.3 4.8 4.5 1.0 4.0<br />
Total<br />
Triticale 2005<br />
6.0 6.0 5.3 7.6 7.1 1.4 6.4<br />
Grain 1.7 1.9 1.7 2.2 2.3 0.6 1.9<br />
Straw 2.8 2.9 2.6 3.6 3.5 1.1 3.1<br />
Total 4.5 4.8 4.2 5.8 5.8 1.6 5.0<br />
Immature compost (applied 09 May 2002)<br />
Triticale 2004<br />
Grain 1.6 2.8 3.2 4.5 5.3 1.8 3.5<br />
Straw 2.8 4.5 5.0 6.7 8.0 2.8 5.4<br />
Total<br />
Triticale 2005<br />
4.4 7.3 8.2 11.2 13.4 4.6 8.9<br />
Grain 1.8 1.9 2.2 3.1 3.2 1.2 2.4<br />
Straw 2.6 2.7 3.1 4.9 5.3 1.8 3.7<br />
Total<br />
Triticale 2006<br />
4.4 4.6 5.2 8.0 8.5 3.0 6.1<br />
Grain 0.8 0.8 1.1 1.1 1.5 0.4 1.1<br />
Straw 1.5 1.8 2.1 2.2 3.1 1.2 2.1<br />
Total 2.3 2.6 3.2 3.4 4.6 1.5 3.2<br />
Comparing the composts the most effective was the meat powder based semi mature<br />
<strong>on</strong>e, but the other semi mature and the meat powder had also definitive effects. Mature<br />
and immature composts were applied <strong>on</strong>e year earlier so they had milder effects.<br />
In 2005, in the 3 rd - 4 th years of the treatments mature compost residual effects<br />
weakened and was not able to produce significant differences in yield so further<br />
experiments were <strong>term</strong>inated. Meat powder based semi mature gave the highest yields<br />
even in this year, which was 30% less, than in previous year in average. Maximal doses<br />
were able to rise 5-6 t/ha c<strong>on</strong>trol plot yield to 8-10 t/ha, but the effects are already<br />
significant between 25 and 100 t/ha compost as well as 5 and 20 t/ha meat powder<br />
treatment. Lower doses of the applied materials however caused no significant effect<br />
any more.<br />
243
Average yields in 2006 were <strong>on</strong>ly half of that in 2005 and treatment effects were just<br />
above the significant limit except for meat powder in the case of grain yield.<br />
Table 4: Effect of slaughterhouse composts <strong>on</strong> air-dried triticale yield, t/ha<br />
(Calcareous sandy soil, Őrbottyán)<br />
Compost t/ha fresh material LSD5% Mean<br />
0 25 50 100 200<br />
Meat powder based semi-mature compost (applied 18 Nov 2002)<br />
Triticale 2004<br />
Grain 2.4 3.8 4.3 4.4 5.4 1.7 4.1<br />
Straw 3.9 5.9 6.3 6.5 8.1 2.1 6.2<br />
Total 6.3 9.7 10.6 10.9 13.6 3.8 10.2<br />
Triticale 2005<br />
Grain 2.3 2.2 3.0 3.3 3.2 0.9 2.8<br />
Straw 3.3 3.2 4.3 5.2 5.6 1.3 4.3<br />
Total 5.6 5.4 7.3 8.5 8.8 2.2 7.1<br />
Triticale 2006<br />
Grain 1.2 1.2 1.4 1.5 1.6 0.4 1.4<br />
Straw 2.3 2.4 2.9 3.0 3.4 1.1 2.8<br />
Total 3.5 3.6 4.3 4.5 5.0 1.4 4.2<br />
Cooked meat based semi-mature compost (applied 06 May 2003)<br />
Triticale 2004<br />
Grain 3.2 3.1 3.8 4.7 4.6 0.9 3.8<br />
Straw 4.9 5.0 5.7 6.6 6.7 1.2 5.8<br />
Total 8.1 8.0 9.4 11.3 11.4 2.0 9.6<br />
Triticale 2005<br />
Grain 2.5 2.4 2.3 3.0 3.2 0.5 2.7<br />
Straw 3.9 3.7 3.3 4.5 5.1 0.9 4.1<br />
Total 6.5 6.1 5.6 7.5 8.3 1.3 6.8<br />
Triticale 2006<br />
Grain 1.1 1.0 1.0 1.4 1.4 0.2 1.2<br />
Straw 2.3 2.1 2.0 2.6 2.6 0.5 2.3<br />
Total 3.4 3.2 3.0 4.0 4.0 0.7 3.5<br />
The main results and c<strong>on</strong>clusi<strong>on</strong>s can be summarised as follows:<br />
1. Composts and meat powder are valuable fertilizers having much higher c<strong>on</strong>tent of<br />
N, P, Ca, Zn and Cu compared to the farmyard manure. These materials could be<br />
successfully used in cultures having great fertilizer demand like sugar beet and also<br />
fibre crops or cereals.<br />
2. Composts and meat powder have c<strong>on</strong>siderable effect and even after 3-4 years<br />
residual effect <strong>on</strong> yield.<br />
3. According to results composts can be applied at 25-50 t/ha rate as a fertilizer<br />
similarly to manure. On sandy soils, poorly supplied with K additi<strong>on</strong>al K<br />
fertilizati<strong>on</strong> is recommended. Applicati<strong>on</strong> above 50 t/ha can cause depressi<strong>on</strong> in<br />
244
unfavourable years. 100-200 t/ha doses can also cause envir<strong>on</strong>mental risk because<br />
of the high N and P c<strong>on</strong>tent.<br />
Table 5: Effect of meat powder <strong>on</strong> air-dried triticale yield, t/ha. Applied 18 Nov<br />
2002 (Calcareous sandy soil, Őrbottyán)<br />
Meat powder t/ha LSD5% Mean<br />
0 2.5 5 10 20<br />
Triticale 2004<br />
Grain 2.7 2.8 4.7 4.5 4.2 1.3 3.8<br />
Straw 4.6 5.0 6.9 6.9 7.0 1.7 6.1<br />
Total<br />
Triticale 2005<br />
7.3 7.8 11.6 11.4 11.2 3.0 9.9<br />
Grain 1.9 2.0 1.9 3.0 3.9 1.3 2.6<br />
Straw 3.0 3.2 2.7 4.4 6.7 2.5 4.0<br />
Total<br />
Triticale 2006<br />
4.9 5.2 4.6 7.4 10.6 3.7 6.6<br />
Grain 1.0 0.9 1.0 1.1 1.2 0.4 1.1<br />
Straw 1.6 1.6 1.9 1.8 2.3 0.6 1.8<br />
Total 2.6 2.5 3.0 2.9 3.5 1.0 2.9<br />
REFERENCES<br />
Debreczeni I., Izsáki Z. (1985): Tannery sewage sludge effect <strong>on</strong> crop element<br />
compositi<strong>on</strong>. Agrokémia és Talajtan 34: (3-4) 421-432. (in Hungarian)<br />
Debreczeni I., Izsáki Z. (1989): Tannery sewage sludge fertilizati<strong>on</strong> effect and residual<br />
effect <strong>on</strong> cereals <strong>on</strong> a sandy soil. Növény<strong>term</strong>elés 38:3 231-239. (in Hungarian)<br />
Izsáki Z., (2000): Collecting, disposal and utility of agricultural wastes. Tessedik<br />
Sámuel Főiskola, Szarvas. 94 pp. (in Hungarian)<br />
Izsáki Z. – Debreczeni I. (1987): Tannery sewage sludge fertilizati<strong>on</strong> effect <strong>on</strong> a sandy<br />
soil. Növény<strong>term</strong>elés 36:6 481-489. (in Hungarian)<br />
Thaer, A. (1809): In: Kádár I. (ed.) (1996): Albrecht Thaer: “Basics of rati<strong>on</strong>al<br />
agriculture. Manuring.” MTA TAKI. Budapest. 99 pp. (in Hungarian)<br />
Kádár I., (2003): Report of ATEV compost effect <strong>on</strong> soil fertility. MTA TAKI.<br />
Manuscript. (in Hungarian)<br />
Kádár I., Hámori V., Morvai B., Petróczki F. (2002): Soil load and polluti<strong>on</strong> limit<br />
values; sewage sludge and slaughterhouse waste compost effect <strong>on</strong> sugar beet. In:<br />
Cukorrépa <strong>term</strong>esztési/<strong>term</strong>eltetési tanfolyam és tanácskozás. 37-40. Szerk.:<br />
Várnainé J.A. Cukoripari Egyesülés. Budapest. (in Hungarian)<br />
Kádár I., Ragályi P. (2004): Final report in 2004 about l<strong>on</strong>g-<strong>term</strong> field experiments set<br />
up with slaughterhouse composts. In: MAE Talajtani Társaság Talajvédelmi Klub.<br />
FVM Növény és Talajvédelmi Szolgálat. Budapest. 6 pp. (in Hungarian)<br />
Kiss J., Sim<strong>on</strong> M., Horváth Z., Kádár I., Kriszt B., Szoboszlay S., Morvai B., Csomor<br />
L., Szántó G. (2001): Biological degradati<strong>on</strong> of animal origin fatty wastes. In: XVI.<br />
Országos Környezetvédelmi K<strong>on</strong>ferencia. (Szerk: Elek Gy.– Vécsi B.) 351-360 pp.<br />
Siófok. (in Hungarian)<br />
245
Sarkadi J. (1975): Methods for estimating fertilizer demand. Mezőgazdasági Kiadó,<br />
Budapest. 252 pp. (in Hungarian)<br />
Sim<strong>on</strong> L., Szente K. (2000): Sewage sludge compost effect <strong>on</strong> maize N c<strong>on</strong>tent and<br />
yield. Agrokémia és Talajtan. 49: 231-246. (in Hungarian)<br />
Vermes L. (1998): Waste management, waste utility. Mezőgazda Kiadó. Budapest. 201<br />
pp. (in Hungarian)<br />
246
THE EFFECT OF BENTONITE ON SPECIFIC SOIL PARAMETERS AND<br />
MICROBIAL CHARACTERISTICS OF THE CARBON CYCLE<br />
János Kátai 1 , Magdolna Tállai 1 , János Lazányi 2 ,<br />
Edina Veres Lukácsné 1 , Zsolt Sándor 1<br />
1 University of Debrecen, Centre of Agricultural Sciences,<br />
Department of Agrochemistry and Soil Science<br />
2 University of Debrecen, Centre of Agricultural Sciences,<br />
Department of Rural Development and Land Use<br />
ABSTRACT<br />
The effect of increasing dosages (5, 10, 15, 20 t/ha) of bent<strong>on</strong>ite (Bent<strong>on</strong>ite experiment)<br />
and bent<strong>on</strong>ite composted with manure (Bent<strong>on</strong>ite + manure experiment) was studied in<br />
a small plot experiment <strong>on</strong> acidic sandy soil at the Experimental Farm of the<br />
Nyíregyháza Research Centre, UD CAS. The effect of treatments <strong>on</strong> the physical,<br />
chemical and microbial soil parameters can be summarized as follows:<br />
• Bent<strong>on</strong>ite significantly increased the water-holding capacity of soils. In samples<br />
treated with bent<strong>on</strong>ite moisture c<strong>on</strong>tent was 2-3 % higher than that in treatments<br />
with bent<strong>on</strong>ite and manure.<br />
• The pH value of the soil was shifted towards neutral. In bent<strong>on</strong>ite treatments, pH<br />
was more increased than in treatments of bent<strong>on</strong>ite composted with manure.<br />
Accordingly, hydrolytic acidity was also reduced.<br />
• When studying the nutrient c<strong>on</strong>tent of soil, we found that bent<strong>on</strong>ite increased,<br />
though not always significantly, the readily available potassium c<strong>on</strong>tent of soil. As<br />
a result of the composted treatments, the readily available nitrate, phosphorus and<br />
potassium c<strong>on</strong>tent of the soil significantly increased.<br />
• Regarding the studied microbial parameters, we found that the composted<br />
treatments caused a higher (though not always significantly) increment in the total<br />
number of bacteria, the number of microscopic fungi and cellulose-decomposing<br />
bacteria and the biomass-C c<strong>on</strong>tent of soils. In both series of treatments, the high<br />
dosages caused a reducti<strong>on</strong> in the number of microbes.<br />
High bent<strong>on</strong>ite dosages also reduced carb<strong>on</strong> dioxide formati<strong>on</strong>.<br />
Saccharase activity was significantly increased by small dosages of bent<strong>on</strong>ite, while<br />
composted treatments resulted in a lower, not significant increment in enzyme<br />
activities.<br />
• In the experiment, several relati<strong>on</strong>ships were found between the physical, chemical<br />
and microbial soil parameters.<br />
Keywords: bent<strong>on</strong>ite, moisture c<strong>on</strong>tent, pH, quantitative changes in microbes,<br />
microbial activity<br />
INTRODUCTION<br />
Nowadays, sustainable agriculture is a worldwide known noti<strong>on</strong>, a basis of which is the<br />
protecti<strong>on</strong> of the envir<strong>on</strong>mental elements and reducing the envir<strong>on</strong>mental loading to the<br />
minimum. These aspects should also be c<strong>on</strong>sidered in soil meliorati<strong>on</strong>, therefore,<br />
increasing attenti<strong>on</strong> is paid to the study and applicati<strong>on</strong> of natural, envir<strong>on</strong>mentally<br />
247
enign materials in agriculture. Numerous researchers studied products from mining<br />
and industry for increasing the organic and inorganic colloid c<strong>on</strong>tent of sandy soils and<br />
for improving their water and nutrient management (Blaskó, 2005). Zeolites (Kazó,<br />
1981; Köhler, 2000), ground rhyolite tuff, ground basaltic tuff, and bent<strong>on</strong>ite (Köhler,<br />
2000; Lazányi, 2003) have already been examined.<br />
Carb<strong>on</strong> is an essential element of life, a composer of live and dead organic matter,<br />
the carb<strong>on</strong> cycle is a basic process of the biogeochemical cycle (Jolánkai et al., 2005).<br />
Microorganisms play an extremely important role in transforming organic matter into<br />
inorganic matter and in the cycles of the different elements including carb<strong>on</strong>. For<br />
describing the transforming processes, intensity of cellulose-decompositi<strong>on</strong> (Kádár et<br />
al., 2005), carb<strong>on</strong>-dioxide producti<strong>on</strong> of soils (Zsuposné Oláh, 2005; Varga et al. 2005),<br />
and the amount of carb<strong>on</strong> in the microbial biomass are used (Szili-Kovács et al., 2005;<br />
Szili-Kovács 2006; Lukácsné Veres et al., 2006).<br />
With special regards to the requirements of sustainability, we joined an experiment<br />
of sand meliorati<strong>on</strong>, in which we studied the effect of bent<strong>on</strong>ite, a natural, clay-like<br />
st<strong>on</strong>e rich in colloids, and bent<strong>on</strong>ite composted with manure <strong>on</strong> the moisture c<strong>on</strong>tent,<br />
pH, nutrient management and microbial activity of soil, which is dem<strong>on</strong>strated via<br />
specific elements of the carb<strong>on</strong> cycle.<br />
MATERIAL AND METHODS<br />
The small-plot experiment was set up at the experimental farm of the Nyíregyháza<br />
Research Centre of UD CAS in October 2002. Samples were collected from the top 2-<br />
20cm layer of soil. The in vitro experiments were carried out at the soil microbial<br />
laboratory of the Department of Soil Sciences, UD CAS FA between 2003 and 2005.<br />
The soil type was brown forest soil with alternating layers of clay (”kovárvány”), with<br />
sandy texture and acidic pH (pH(H2O) 5.28; pH(KCl) 4.33 ). The soil was deficient in<br />
nitrogen, the level of phosphorus and potassium supply was medium and satisfactory,<br />
respectively. Treatments were performed with increasing dosages of bent<strong>on</strong>ite and<br />
bent<strong>on</strong>ite composted with manure (Table 1).<br />
From the physical parameters, moisture c<strong>on</strong>tent was de<strong>term</strong>ined after drying to<br />
c<strong>on</strong>stant mass at 105 0 C (Klimes-Szmik, 1962). From chemical soil parameters, pH was<br />
measured in suspensi<strong>on</strong>s with distilled water and KCl (Filep, 1995). The nitrate-N<br />
c<strong>on</strong>tent of soil was de<strong>term</strong>ined with the Na-salicylate method (Felföldy, 1987), while<br />
the easily soluble phosphorus and potassium c<strong>on</strong>tent of soil was measured with 0.1m<br />
amm<strong>on</strong>ium-lactate (AL) extractor (Gerei, 1970).<br />
The amount of microbes in the soil was de<strong>term</strong>ined by the plate diluti<strong>on</strong> method<br />
from soil-water suspensi<strong>on</strong> as the total number of bacteria (<strong>on</strong> Bouill<strong>on</strong> agar) and the<br />
number of microscopic fungi (<strong>on</strong> pept<strong>on</strong>e glucose agar) (Szegi, 1979). The number of<br />
cellulose-decomposing bacteria was de<strong>term</strong>ined by the method of the most probable<br />
number of bacteria according to Poch<strong>on</strong> et al. (1962). Regarding soil microbial activity,<br />
the carb<strong>on</strong>-dioxide producti<strong>on</strong> of soil was measured according to the method of<br />
Witkamp (1966. cit. Szegi, 1979), while the soil biomass-C producti<strong>on</strong> and saccharase<br />
enzyme activity were de<strong>term</strong>ined by the fumigati<strong>on</strong>-incubati<strong>on</strong> method (Jenkins<strong>on</strong> et<br />
al.,1976) and the method of Frankenberger et al., respectively. The statistical analysis<br />
was performed using the SPSS 9.0 program<br />
248
RESULTS<br />
The effect of treatments <strong>on</strong> the studied soil characteristics is presented as a mean of the<br />
three years (2003-2005).<br />
Soil moisture c<strong>on</strong>tent (Figure 1) varied between 12.69 and 14.38 % (mass %) in the<br />
average of three years. In treatments with bent<strong>on</strong>ite, a significant increase was observed<br />
in moisture c<strong>on</strong>tent, 2-3% higher than in treatments with bent<strong>on</strong>ite composted with<br />
manure.<br />
Moisture c<strong>on</strong>tent<br />
(mass %)<br />
15<br />
13<br />
11<br />
9<br />
7<br />
5<br />
1 2 3 4 5 6 7 8 9 10<br />
Treatments<br />
Figure 1: The effect of bent<strong>on</strong>ite <strong>on</strong> soil moisture c<strong>on</strong>tent<br />
The pH value of soils (Figure 2) increased significantly in both treatment series and<br />
soluti<strong>on</strong>s (H2O; KCl). Bent<strong>on</strong>ite treatments increased the pH value more than the<br />
bent<strong>on</strong>ite + manure treatments.<br />
pH<br />
7<br />
6<br />
5<br />
4<br />
3<br />
1 2 3 4 5 6 7 8 9 10<br />
pH (H2O)<br />
Treatments<br />
pH (KCl) y1<br />
18<br />
13<br />
8<br />
3<br />
Hydrolytic<br />
acidity<br />
(y1)<br />
Figure 2: The effect of bent<strong>on</strong>ite <strong>on</strong> the pH and hydrolytic acidity of soil<br />
A c<strong>on</strong>siderable reducti<strong>on</strong> was observed in the hydrolytic acidity (y1) (Figure 2) with<br />
increasing pH. In both treatment series, even small dosages (5 t/ha; 5+9 t/ha) caused a<br />
significant reducti<strong>on</strong>, while the change was smaller in the case of higher dosages.<br />
The NO3-N c<strong>on</strong>tent of soil (Table 1) reduced, though not always significantly, as a<br />
result of bent<strong>on</strong>ite treatments. This cannot be stated about the bent<strong>on</strong>ite + manure<br />
treatments where NO3-N c<strong>on</strong>tent increased in proporti<strong>on</strong> to the increase in dosages. As a<br />
249
esult of higher dosages, no c<strong>on</strong>siderable changes were observed in nitrate c<strong>on</strong>tent.<br />
However, the nitrate c<strong>on</strong>tent of treatments 8, 9 and 10 was significantly higher than that<br />
of the c<strong>on</strong>trol.<br />
We have also measured the readily available phosphorus and potassium c<strong>on</strong>tent.<br />
The AL-soluble phosphorus c<strong>on</strong>tent of soils significantly reduced as a result of<br />
bent<strong>on</strong>ite treatments, regardless of dosage. The phosphorus c<strong>on</strong>tent of composted<br />
treatments, however, increased significantly, and no c<strong>on</strong>siderable reducti<strong>on</strong> was found<br />
even in the high dosage treatments (treatments 9 and 10).<br />
The readily available potassium c<strong>on</strong>tent of soils increased slightly as a result of<br />
bent<strong>on</strong>ite treatments, but the values were not significantly different from those of the<br />
c<strong>on</strong>trol. However, the composted treatments significantly increased the potassium<br />
c<strong>on</strong>tent of soils.<br />
Table 1. The effect of treatments <strong>on</strong> the readily available nutrient c<strong>on</strong>tent of the soil<br />
(average of 2003-2005)<br />
Sample<br />
Dosage<br />
Treatment<br />
Nitrate-N<br />
(mg - kg -1 )<br />
AL-P2O5<br />
(mg kg -1 )<br />
AL-K2O<br />
(mg k g -1 )<br />
Bent<strong>on</strong>ite<br />
1. - 0 (t/ha) 6.50 148.77 250.97<br />
2. 1.× 5 (t/ha) 6.23 119.43 253.77<br />
3. 2.× 10 (t/ha) 5.93 118.67 248.77<br />
4. 3.× 15 (t/ha) 5.90 141.53 277.50<br />
5. 4.× 20 (t/ha) 5.80 124.57 237.60<br />
Bent<strong>on</strong>ite+manure<br />
6. - 0 6.70 145.90 252.20<br />
7. 1.× 5 (t/ha)+9 (t/ha) 7.17 156.87 301.80<br />
8. 2.× 10 (t/ha)+18 (t/ha) 8.20 153.53 312.93<br />
9. 3.× 15 (t/ha)+27 (t/ha) 8.50 163.87 283.53<br />
10. 4.× 20 (t/ha)+36 (t/ha) 8.40 154.87 307.20<br />
Significant Difference (5%) 0.25 3.65 15.6<br />
The impact of treatments <strong>on</strong> soil microbial parameters (Table 2) is dem<strong>on</strong>strated via<br />
specific characteristics of the carb<strong>on</strong> cycle.<br />
Bent<strong>on</strong>ite treatments increased slightly the total number of bacteria and the<br />
amount of microscopic fungi, the increase of the latter was not significant. Higher<br />
dosages resulted in a decrease in the number of microbes. In both groups of microbes,<br />
the medium dosages had an inducing effect. Composted treatments had a higher<br />
increasing effect <strong>on</strong> the amount of microbes, though it was not always significant.<br />
The number of cellulose-decomposing bacteria was increased c<strong>on</strong>siderably by<br />
treatments of bent<strong>on</strong>ite no. 3 and treatment no. 2 of the combined series. High bent<strong>on</strong>ite<br />
250
dosages significantly reduced the amount of cellulose-decomposing microbes. The high<br />
dosage treatments with bent<strong>on</strong>ite composted with manure had a weaker inhibiting<br />
effect.<br />
In treatments with the two high dosages of bent<strong>on</strong>ite, CO2 producti<strong>on</strong> was lower<br />
than that of the c<strong>on</strong>trol. Medium and high dosages of bent<strong>on</strong>ite + manure also<br />
significantly reduced the formati<strong>on</strong> of CO2 in the soil.<br />
The soil biomass-C value reduced under that of the c<strong>on</strong>trol already at applying medium<br />
dosages of bent<strong>on</strong>ite. The small dosages of composted treatments caused a significant<br />
increment, while the two high dosages reduced its values and treatment 10 resulted in a<br />
significant reducti<strong>on</strong>.<br />
Regarding saccharase enzyme activity, a higher value was measured in treatment<br />
2 am<strong>on</strong>g the bent<strong>on</strong>ite treatments. Further treatments reduced the enzyme activity,<br />
though not significantly. Composted treatments resulted in a smaller, n<strong>on</strong>-significant<br />
increase in enzyme activity.<br />
Sample<br />
Table 2. The effect of treatments <strong>on</strong> the amount of microbe groups and their<br />
microbial activity (average of 2003-2005)<br />
Dosage<br />
Total<br />
bacteria<br />
(*10 6 g -1 soil)<br />
Total fungi<br />
(*10 3 g -1 soil)<br />
Cellulose<br />
decomposing<br />
bacteria<br />
(*10 3 g -1 soil)<br />
CO2producti<strong>on</strong><br />
(CO2 mg<br />
100g -1 )<br />
Biomass-C<br />
(µg g -1 )<br />
Saccharase<br />
(glucose<br />
mg 100g -1 )<br />
1. - 3.20 82.01 11.59 8.47 99.14 7,96<br />
2. 1.× 3.80 64.69 9.89 8.73 100.71 9,10<br />
3. 2.× 5.35 82.92 12.79 8.45 85.35 8,75<br />
4. 3.× 3.94 54.91 7.90 5.74 85.27 8,76<br />
5. 4.× 4.14 55.18 4.01 7.98 83.27 7,55<br />
6. - 5.33 82.73 9.03 8.23 117.72 8,01<br />
7. 1.× 6.15 71.21 20.95 8.10 136.18 8,94<br />
8. 2.× 6.07 95.47 9.49 6.89 127.27 8,24<br />
9. 3.× 6.05 88.73 7.03 8.37 118.67 8,34<br />
10. 4.× 6.60 91.37 8.10 7.16 83.05 8,07<br />
(SD5%) 0.25 3.54 4.73 0.30 3.86 0.72<br />
In correlati<strong>on</strong> analysis, we searched for correlati<strong>on</strong>s between the physical, chemical and<br />
microbial soil parameters for both treatment series (Table 3). In our analysis, we<br />
emphasize <strong>on</strong>ly the str<strong>on</strong>g correlati<strong>on</strong>s.<br />
In bent<strong>on</strong>ite treatments, a str<strong>on</strong>g correlati<strong>on</strong> was found between the nitrate-N<br />
c<strong>on</strong>tent and readily available phosphorus c<strong>on</strong>tent of soil (r=0.71). A str<strong>on</strong>g correlati<strong>on</strong><br />
was observed between changes in soil nitrate-N c<strong>on</strong>tent and biomass-C c<strong>on</strong>tent<br />
(r=0.91). Am<strong>on</strong>g the microbial parameters, a str<strong>on</strong>g positive correlati<strong>on</strong> was found<br />
between the total number of bacteria and the number of cellulose-decomposing bacteria<br />
251
(r=0.71), and between the total number of bacteria and CO2 producti<strong>on</strong> of the soil<br />
(r=0.82).<br />
Table 3. Correlati<strong>on</strong> ( r ) between the examined physical, chemical, and<br />
microbiological parameters of soil<br />
* Correlati<strong>on</strong> is significant at 0.05 level (2-tailed)-medium<br />
** Correlati<strong>on</strong> is significant at 0.01 level (2-tailed)-tight<br />
In treatments of bent<strong>on</strong>ite composted with manure, a str<strong>on</strong>g correlati<strong>on</strong> was found<br />
between pH and readily available phosphorus c<strong>on</strong>tent of the soil (r=0.71). Nitrate-N and<br />
the total number of fungi also correlated closely (r=0.80). Am<strong>on</strong>g microbial parameters,<br />
the total number of bacteria showed a str<strong>on</strong>g positive correlati<strong>on</strong> with CO2 producti<strong>on</strong><br />
(r=0.81), similarly to bent<strong>on</strong>ite treatments, and a str<strong>on</strong>g correlati<strong>on</strong> was found between<br />
the changes in the total number of bacteria and the biomass-C c<strong>on</strong>tent of soil (r=0.79).<br />
Composted treatments had a favourable effect <strong>on</strong> the amount of cellulose-decomposing<br />
bacteria, but the number of fungi showed a str<strong>on</strong>g negative correlati<strong>on</strong> with their<br />
increasing numbers (r=-0.71).<br />
252
DISCUSSION<br />
In our experiments, we found, similarly to the earlier observati<strong>on</strong>s of Lazányi (2005),<br />
that bent<strong>on</strong>ite has a favourable effect <strong>on</strong> the soil moisture c<strong>on</strong>tent.<br />
Szegi et al. (2003) studied the effect of bent<strong>on</strong>ite <strong>on</strong> the biomass of pea and sweet<br />
corn. According to them, the fertility of soil improved with improving physical and<br />
chemical soil parameters. Our experiments have c<strong>on</strong>firmed the earlier results (Szegi et<br />
al. 2002, 2003), according to which bent<strong>on</strong>ite reduced the acidic pH of the soil ( pH(H2O)<br />
5.28 ) and simultaneously the values of hydrolytic acidity were also reduced. Bent<strong>on</strong>ite<br />
composted with manure increased the readily available phosphorus, potassium and<br />
nitrate c<strong>on</strong>tent of the soil.<br />
No data were found in the literature about the changes in the amount and activity<br />
of soil microbes. According to our experience, the amount of bacteria and fungi were<br />
increased primarily by treatments of medium dosage. There were <strong>on</strong>ly slight changes in<br />
CO2 producti<strong>on</strong> and saccharase enzyme activity, while the amount of microbial<br />
biomass-C was c<strong>on</strong>siderably higher in treatments with bent<strong>on</strong>ite composted with<br />
manure.<br />
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DE ATC Debrecen, április 1-2. 4-8. p.<br />
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de Cultura Pentru Grau Si Porumb Sympozi<strong>on</strong> <str<strong>on</strong>g>Internati<strong>on</strong>al</str<strong>on</strong>g>. 7-8 iulie 2005<br />
Oradea-Romania. 293-300. p.<br />
Lukácsné Veres E. - Zsuposné Oláh Á. (2006): Examinati<strong>on</strong> of biological activity in<br />
chernozems. Cereal Research Communicati<strong>on</strong>s, Vol. 34. No. 1. 363-366. p.<br />
Poch<strong>on</strong>, J. - Tardieux, P. (1962): Tecniques D ’ Analyse en Micobiologie du Sol.<br />
Collecti<strong>on</strong> „Technivues de Base”. 102. p.<br />
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Budapest. 250-256. p.<br />
Szegi T. - Makádi M. (2003): Bent<strong>on</strong>it hatása a talajok tulajd<strong>on</strong>ságára, különös<br />
tekintettel az aggregátum stabilitásra és ezáltal a <strong>term</strong>éseredményekre, zöldborsó és<br />
csemegekukoricaesetében.www.phd.hu/tavasz2003/tsz_2003/tudomanyosszekciok/<br />
tavasziszel 2003_szegitamas.htm - 17k.<br />
Szegi T. - Michéli E. –Tombácz E. – Lazányi J. (2002): Improving rheological and<br />
adsorpti<strong>on</strong> characteristics of sandy soil by mineral colloids applicati<strong>on</strong>.<br />
Proceedings of the Alps-Adria Scientific Workshop. Opatija, 4-8 March 2002. 219-<br />
223. p.<br />
Szili-Kovács T. - Török K. (2005): Szénforráskezelés hatása a talaj mikrobiális<br />
akivitására és biomasszájára felfagyott homoki szántók<strong>on</strong>. Agrokémia és Talajtan,<br />
Vol. 54. No. 1-2. 149-162. p.<br />
Szili-Kovács T. (2006): A talaj mikrobiális biomassza meghatározása kloroform<br />
fumigációs módszerrel. Agrokémia és Talajtan. Vol. 55. No. 2. 515-530. p.<br />
Varga, Cs. - Helmeczi, B. - Buban, T. (2005): Effect of black polyethylene mulching<br />
<strong>on</strong> cellulose decompositi<strong>on</strong> of sand soil. Zeszyty Naukowe Uniwersytetu<br />
Rzeszowskiego, Seria Rolnicza, Rrodukcja Roslinna 2. Zeszyt 27/2005. 21-25. p.<br />
Witkamp, M. (1966): Decompositi<strong>on</strong> of leaf litter in relati<strong>on</strong> to envir<strong>on</strong>ment<br />
microflore and microbial respirati<strong>on</strong>. Ecology, 47. 194-201. p.<br />
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cellulose <strong>on</strong> meadow chernozem soil. Cereal Research Communicati<strong>on</strong>s, Vol. 33.<br />
No. 1. 341-343. p.<br />
254
INVESTIGATION OF MINERAL CONTENT IN WINTER WHEAT GRAIN<br />
SAMPLES FROM LONG-TERM FIELD EXPERIMENTS<br />
Zoltán Győri – Árpád Tóth – Diana Ungai<br />
University of Debrecen Centre of Agricultural Sciences Faculty of Agr<strong>on</strong>omy<br />
Institute of Food Science, Quality Assurance and Microbiology<br />
ABSTRACT<br />
Besides the technological quality of the wheat flour c<strong>on</strong>sumers are primarily interested<br />
in the nutriti<strong>on</strong>-biological value (for example minerals, vitamins, etc.) of the final<br />
products.<br />
The assay dem<strong>on</strong>strates the results of the researches in different soils in Hungary<br />
within the framework of the Nati<strong>on</strong>al L<strong>on</strong>g-<strong>term</strong> Field Trials. The winter wheat grain<br />
samples are from three research sites: Karcag, Nagyhörcsök and Iregszemcse. The<br />
analyzed winter wheat grain samples are from the corn – corn - winter wheat - winter<br />
wheat crop rotati<strong>on</strong>. The applied fertilizer doses were the following: 0 – 150 – 200<br />
kg/ha N, 0 – 100 – 120 kg/ha P, 0 – 100 – 200 kg/ha K, the other two active substances<br />
remained the same. The c<strong>on</strong>centrati<strong>on</strong>s of calcium, magnesium and manganese of wheat<br />
samples are shown in this article.<br />
The results of the analysis c<strong>on</strong>ducted in our accredited laboratory dem<strong>on</strong>strate that<br />
the effect of the K and P fertilizers <strong>on</strong> the Mg and Mn c<strong>on</strong>centrati<strong>on</strong> depends rather <strong>on</strong><br />
the producti<strong>on</strong> site, while the effect of the N fertilizer depends <strong>on</strong> the crop year. The<br />
applicati<strong>on</strong> of N fertilizer increased the Ca c<strong>on</strong>tent of the wheat grain samples in all<br />
three research sites in every statistically proved case.<br />
Keywords: L<strong>on</strong>g-<strong>term</strong> field experiment, winter wheat grain, NPK fertilizati<strong>on</strong>, Ca-,<br />
Mg-, Mn-c<strong>on</strong>tent<br />
INTRODUCTION<br />
Besides the technological quality of the wheat flour c<strong>on</strong>sumers are primarily interested<br />
in the quality of the final product, i.e. food and the attractiveness of its exterior,<br />
nutriti<strong>on</strong>-biological value and their healthiness and usefulness for the human organism.<br />
Most of the attenti<strong>on</strong> is usually directed at micro-nutrients, such as vitamins and<br />
minerals. In human nutriti<strong>on</strong> in Europe, winter wheat c<strong>on</strong>stitutes <strong>on</strong>e of the most<br />
important sources of micro-, and macro-elements (Mahan et al., 1996., Lagua et al.,<br />
1996).<br />
Pais (1981) claims that wheat grains c<strong>on</strong>tain 6.8 mg kg -1 of copper, 62 mg kg -1 of<br />
ir<strong>on</strong>, 34 mg kg -1 of manganese and 31 mg kg -1 of zinc. Sugiura et al. (1998) obtained the<br />
following average results for wheat flours: protein c<strong>on</strong>tent 15.5%, ash c<strong>on</strong>tent 1%,<br />
potassium c<strong>on</strong>tent 4800 mg kg -1 , calcium c<strong>on</strong>tent 300 mg kg -1 , phosphorous c<strong>on</strong>tent<br />
3200 mg kg -1 , magnesium c<strong>on</strong>tent 1400 mg kg -1 , copper c<strong>on</strong>tent 2.5 mg kg -1 , ir<strong>on</strong><br />
c<strong>on</strong>tent 27 mg kg -1 , manganese c<strong>on</strong>tent 21.6 mg kg -1 and zinc c<strong>on</strong>tent 13 mg kg -1 .<br />
The variability of mineral c<strong>on</strong>tent of wheat flours can be attributed to<br />
envir<strong>on</strong>mental and genetic factors and their interacti<strong>on</strong>. Several mineral elements show<br />
high positive correlati<strong>on</strong> with protein c<strong>on</strong>tent in both flours and bran (Posner and Hibbs,<br />
1997).<br />
255
Western Canadian hard red spring wheat crops were characterized for cadmium, copper,<br />
ir<strong>on</strong>, manganese, selenium and zinc by Gawalko et al. (2002). Samples were received<br />
from crop districts in Manitoba, Saskatchewan and Alberta for the 1996, 1997 and 1998<br />
crops. Year-to-year variati<strong>on</strong>s in grain chemistry are small for Cd, Mn, Se and Zn, but<br />
Cu and Fe c<strong>on</strong>tents show 12% and 9% decreases respectively over the three years. The<br />
overall variability for the plant-essential trace elements, Cu, Fe, Mn and Zn, is low in<br />
comparis<strong>on</strong> with Cd and Se. It is dem<strong>on</strong>strated that the spatial variati<strong>on</strong> in crop<br />
chemistry across the Canadian Prairie wheat producing regi<strong>on</strong> is greater than the year to<br />
year variati<strong>on</strong>s.<br />
To compare the effects of organic manuring and inorganic fertilizati<strong>on</strong>, the<br />
c<strong>on</strong>tents of trace elements in wheat harvested from a field experiment were analysed.<br />
Inorganic fertilizati<strong>on</strong> compared to organic manuring increased the c<strong>on</strong>tent of<br />
manganese in wheat. Kirkham (1983) studied the effect of the sewage sludge <strong>on</strong> the<br />
n<strong>on</strong>-essential elements (Cd, Cr, Ni, Pb), as well as nine essential elements (N, P, K, Ca,<br />
Mg, Cu, Fe, Mn, Zn) c<strong>on</strong>tent of wheat samples in Manhattan, Kansas. Soil and winter<br />
wheat were sampled in 1980 after a total of 128 t ha −1 of sludge (dry-weight basis) had<br />
been added to the soil (32 t ha −1 year −1 ) since 1976. Soil in sludge-fertilized fields had<br />
more N, P, K, Mg, Fe, Cu, Mn, Zn, Cd, Cr and Ni than did soil from the c<strong>on</strong>trol fields.<br />
In general, c<strong>on</strong>centrati<strong>on</strong>s of essential elements in plants grown <strong>on</strong> sludge were similar<br />
to c<strong>on</strong>centrati<strong>on</strong>s of essential elements in plants grown with inorganic fertilizer.<br />
Based <strong>on</strong> the results of l<strong>on</strong>g-<strong>term</strong> field trials, Kádár (1991) found that after<br />
fertilizati<strong>on</strong> the following proved changes occurred in the c<strong>on</strong>centrati<strong>on</strong>s of the<br />
individual elements: N fertilizati<strong>on</strong> increased the Al and Mn c<strong>on</strong>tent; P fertilizati<strong>on</strong><br />
increased the following: Al, B, Ca, P, Sr and decreased Zn c<strong>on</strong>tent. K fertilizati<strong>on</strong><br />
increased the Ba and K c<strong>on</strong>tent and decreased Mg c<strong>on</strong>tent.<br />
The nitrogen fertilizers decrease the soil pH (Debreczeni and Debreczeniné, 1994)<br />
and it is affect the uptake of mineral elements with special regard to the Mn-uptake.<br />
MATERIALS AND METHODS<br />
The assay dem<strong>on</strong>strates the results of the researches in different soils in Hungary within<br />
the framework of the Nati<strong>on</strong>al L<strong>on</strong>g-<strong>term</strong> Field Trials. Field trials started in 5 fields in<br />
1966 and in 4 fields in 1967. The fundamental c<strong>on</strong>cepti<strong>on</strong> was to use 4-year crop<br />
rotati<strong>on</strong>: winter wheat – corn – corn – peas, and winter wheat – corn – corn – winter<br />
wheat, out of which peas were excluded in 2000. The researches are d<strong>on</strong>e in split-splitspot<br />
arrangement, in four repetiti<strong>on</strong>s.<br />
The analyzed winter wheat grain samples are from the corn – corn - winter wheat -<br />
winter wheat crop rotati<strong>on</strong>. Out of the different fertilizati<strong>on</strong> doses we used the results of<br />
the nutrients in the c<strong>on</strong>trol and high-dose plots in our calculati<strong>on</strong>s. The applied fertilizer<br />
doses were the following: 0 – 150 – 200 kg/ha N, 0 – 100 – 120 kg/ha P, 0 – 100 – 200<br />
kg/ha K, the other two active substances remained the same. When applying the<br />
fertilizer, we have to c<strong>on</strong>sider that besides active substances additi<strong>on</strong>al elements are<br />
given as well. The following includes the quantity of the additi<strong>on</strong>al elements in<br />
Amm<strong>on</strong>ium-nitrate, Superphosphate and Potassium-chloride fertilizers that are<br />
important in our analysis (mg/kg; %):<br />
256
Element Amm<strong>on</strong>ium-nitrate Superphosphate Potassium-chloride<br />
Ca 6,0 % 17,4 % 3470<br />
Mg 23,8 570 302<br />
Mn 0,299 152 19<br />
Source: Győri (1998).<br />
The winter wheat grain samples in our analysis are from three research sites: Karcag,<br />
Nagyhörcsök and Iregszemcse. The soil type is meadow chernozem in Karcag and<br />
calcareous chernozem at the other two research sites. Table 1 shows the characteristics<br />
of the cultivated layer of the soil at the different research sites. The dates of c<strong>on</strong>trol<br />
plots are calculated from the averages of results of the 10 th , the 16 th and the 20 th years.<br />
Table 1: Nutrient supply in different research sites<br />
Parameters Karcag Nagyhörcsök Iregszemcse<br />
Humus c<strong>on</strong>tent (%) 2,7 (medium) 2,7 (good) 2,4 (medium)<br />
AL-P2O5 (mg/kg) 34 (very weak) 81 (weak) 103 (medium)<br />
AL-K2O (mg/kg) 270 (good) 147 (medium) 150 (medium)<br />
pHKCl 4,7 7,2 7,4<br />
KA 47 38 37<br />
Source: Debreczeni and Debreczeniné (1994)<br />
The analyses were carried out by the accredited laboratory of the Institute of Food<br />
Science, Quality Assurance and Microbiology of the Faculty of Agriculture of the<br />
Centre for Agricultural Sciences of the University of Debrecen. The mineral element<br />
c<strong>on</strong>tents of the samples were de<strong>term</strong>ined with OPTIMA 3300 DV ICP-OES (Perkin-<br />
Elmer Ltd. Wellesley, USA) using No. BCR CRM 189 certified reference material<br />
(Kovács et al., 1996, 1998). The c<strong>on</strong>centrati<strong>on</strong>s of calcium, magnesium and manganese<br />
of wheat samples are shown in this article.<br />
The data were evaluated by using <strong>on</strong>e-way ANOVA, and there were carried out average<br />
and standard deviati<strong>on</strong> calculati<strong>on</strong>s run <strong>on</strong> SPSS 11.5 for Windows and Excel 6.0 for<br />
Windows.<br />
RESULTS<br />
The effect of N fertilizati<strong>on</strong> <strong>on</strong> the mineral c<strong>on</strong>tent of winter wheat samples<br />
The applied N fertilizati<strong>on</strong> significantly increased the Ca c<strong>on</strong>tent of wheat samples in<br />
every studied crop year in Karcag (Figure 1). At the Iregszemcse research site, the<br />
effect of N fertilizati<strong>on</strong> <strong>on</strong> the Ca c<strong>on</strong>tent was proved by variance analysis <strong>on</strong>ly in some<br />
cases, however, this time the 150 and 200 kg/ha N doses– similarly to the results in<br />
Karcag– increased the Ca c<strong>on</strong>tent in each case. In the Nagyhörcsök research site,<br />
similarly to the effects in Karcag, the effect of the N fertilizati<strong>on</strong> was clear: the various<br />
doses of N fertilizers had a positive effect <strong>on</strong> the Ca c<strong>on</strong>tent of wheat samples.<br />
According to Győri’s (1998) measurements the Amm<strong>on</strong>ium-nitrate has 6% Ca as<br />
additi<strong>on</strong>al element in the fertilizer. The amount of Ca was 0 kg/ha in the c<strong>on</strong>trol<br />
257
treatments and it was 12 kg/ha in the case of 200 kg/ha applied N-fertilizer. Take this<br />
fact into account is important to evaluate accurately the results.<br />
Ca c<strong>on</strong>ent (mg/kg)<br />
800<br />
700<br />
600<br />
500<br />
400<br />
300<br />
200<br />
100<br />
0<br />
2001 2004 2005 2001 2004 2005 2001 2004 2005<br />
Karcag Iregszemcse Nagyhörcsök<br />
C<strong>on</strong>trol 200 kg/ha<br />
Figure 1: Ca c<strong>on</strong>tent of winter wheat samples as an effect of N fertilizati<strong>on</strong><br />
The effect of N fertilizati<strong>on</strong> <strong>on</strong> the Mg c<strong>on</strong>tent was inc<strong>on</strong>sistent in all the three research<br />
sites, depending <strong>on</strong> the crop year; it both decreased and increased the Mg c<strong>on</strong>centrati<strong>on</strong>.<br />
For example in 2002, applying 200 kg/ha N in Nagyhörcsök after winter wheat forecrop<br />
significantly increased while in 2005 after corn forecrop decreased the Mg c<strong>on</strong>tent of<br />
the wheat grain samples.<br />
In Karcag in 2001, after corn forecrop it significantly decreased while in 2005 after<br />
winter wheat forecrop increased the Mg c<strong>on</strong>tent of wheat grain samples. In the<br />
Iregszemcse research site we found the opposite effect: N fertilizati<strong>on</strong> after corn<br />
forecrop increased, while after winter wheat decreased the Ca c<strong>on</strong>tent of wheat. In the<br />
Nagyhörcsök research site in 2001 and 2002, the applicati<strong>on</strong> of 200 kg/ha N doses<br />
significantly increased the Mn c<strong>on</strong>tent of winter wheat grain samples, while significant<br />
effect was not fund in 2003-2005.<br />
The effect of phosphorus fertilizati<strong>on</strong> <strong>on</strong> the mineral c<strong>on</strong>tent of winter wheat<br />
samples<br />
The effect of phosphorus fertilizati<strong>on</strong> <strong>on</strong> the Mg and Mn c<strong>on</strong>tent of winter wheat grain<br />
samples were not significant either at 120 kg/ha dose, or in dry or humid crop years in<br />
Karcag. The effect of phosphorus <strong>on</strong> the Ca c<strong>on</strong>tent was statistically proved in <strong>on</strong>e case:<br />
in 2003, after corn forecrop the applicati<strong>on</strong> of 120 kg/ha P2O5 active substance<br />
decreased the Ca c<strong>on</strong>tent of the wheat grain samples from 602,3±23,5 mg/kg to<br />
544,0±24,0 mg/kg compared to the c<strong>on</strong>trol plots. As regards samples from Iregszemcse<br />
and Nagyhörcsök, the effect of phosphorus was significant in <strong>on</strong>e crop year,<br />
respectively. Applying 100 kg/ha P2O5 fertilizer significantly increased the Ca c<strong>on</strong>tent<br />
of wheat grain samples after winter wheat forecrop in Iregszemcse in 2002, and after<br />
corn forecrop in Nagyhörcsökön in 2001.<br />
258
According to our results, the effect of phosphorus fertilizati<strong>on</strong> <strong>on</strong> the Mg and Mn<br />
c<strong>on</strong>tent of the samples was not significant in Karcag.<br />
In the Iregszemcse research site, the effect of phosphorus fertilizer <strong>on</strong> the Mg c<strong>on</strong>tent of<br />
the samples was <strong>on</strong>ly significant in 2002, while <strong>on</strong> the Mn c<strong>on</strong>tent of samples was<br />
significant in 2001, 2002 and 2004. In these cases the phosphorus doses increased the<br />
c<strong>on</strong>centrati<strong>on</strong> of these elements.<br />
We found that in Nagyhörcsök the applicati<strong>on</strong> of 100 or 120 kg/ha P2O5 active<br />
substance– irrespectively of the forecrop – increased the Mg and Mn c<strong>on</strong>tent of the<br />
wheat grain samples in almost every crop year. (Table 2).<br />
Table 2: The effect of phosphorus fertilizati<strong>on</strong> <strong>on</strong> the Magnesium and Manganese<br />
c<strong>on</strong>tent of winter wheat samples from Nagyhörcsök<br />
Mg c<strong>on</strong>tent (mg/kg) Mn c<strong>on</strong>tent (mg/kg)<br />
2001 2004 2005 2001 2004 2005<br />
c<strong>on</strong>trol 775,8±25,8 1013,3±29,8 959,0±29,8 24,8±1,4 29,2±2,0 10,7±0,3<br />
100 kg/ha<br />
933,3±24,5 1219,5±19,5 1086,3±48,7 38,3±1,5 45,1±1,0 12,5±0,1<br />
P2O 5<br />
p value 0,000 0,000 0,011 0,000 0,000 0,000<br />
The effect of K fertilizati<strong>on</strong> <strong>on</strong> the mineral c<strong>on</strong>tent of winter wheat samples<br />
In 2004 in Karcag, the applicati<strong>on</strong> of 200 kg/ha K fertilizer significantly increased the<br />
Ca, Mg and Mn c<strong>on</strong>tent of wheat grain samples after corn forecrop, while in the same<br />
year this effect was not experienced after winter wheat forecrop.<br />
In the Iregszemcse research site, the effect of K <strong>on</strong> the nutrient c<strong>on</strong>tent of the samples<br />
was statistically proved in <strong>on</strong>e crop rotati<strong>on</strong> in <strong>on</strong>e year (2004): the average manganese<br />
c<strong>on</strong>tent of the c<strong>on</strong>trol wheat samples was 12,2±0,1 mg/kg, and the applicati<strong>on</strong> of 100<br />
kg/ha K2O increased this value to 12,9±0,1 mg/kg. The effect was proved at P=1%<br />
significance level.<br />
C<strong>on</strong>trary to the results from the Karcag and Iregszemcse research sites, (Figure 2)<br />
the applicati<strong>on</strong> of K fertilizer decreased the Ca and Mg c<strong>on</strong>tent of the wheat grain<br />
samples in each case in Nagyhörcsök, the results are statistically proved. The<br />
applicati<strong>on</strong> of K fertilizer decreased the Ca c<strong>on</strong>tent of wheat grain samples from<br />
525,5±32,6 mg/kg to 442,3±39,8 mg/kg in 2001, and from 682,3±47,4 mg/kg to<br />
321,3±12,8 mg/kg in 2005. In Nagyhörcsök, the changes in the Mg c<strong>on</strong>tent proved to be<br />
significant <strong>on</strong>ly in 2001, when the Mg c<strong>on</strong>tent of the c<strong>on</strong>trol wheat samples decreased<br />
from 1022,8±20,2 mg/kg to 890,0±32,7 mg/kg.<br />
259
Ca and Mg c<strong>on</strong>tent (mg/kg)<br />
1400<br />
1200<br />
1000<br />
800<br />
600<br />
400<br />
200<br />
0<br />
2001 2004 2005 2001 2004 2005<br />
Ca c<strong>on</strong>tent Mg c<strong>on</strong>tent<br />
C<strong>on</strong>trol 200 kg/ha<br />
Figure 2: The effect of K fertilizati<strong>on</strong> <strong>on</strong> the Ca and Mg c<strong>on</strong>tent of winter wheat<br />
samples<br />
DISCUSSIONS<br />
In the calcerous chernozem soils in Iregszemcse and Nagyhörcsök the applied N<br />
increased the Mg and Mn c<strong>on</strong>tent of the wheat grain samples in dry years and decreased<br />
in humid years. We cannot draw the same c<strong>on</strong>clusi<strong>on</strong> from the results of the meadow<br />
chernozem soil with salt in the deeper layers in Karcag. The increase of the Mnc<strong>on</strong>centrati<strong>on</strong><br />
is probably related to the decrease of soil pH in c<strong>on</strong>sequence of nitrogenapplying.<br />
The applicati<strong>on</strong> of N fertilizer increased the Ca c<strong>on</strong>tent of the wheat grain<br />
samples in all three research sites in every statistically proved case. The unique Caincreasing<br />
effect might be the result of the c<strong>on</strong>siderable amount of Ca as additi<strong>on</strong>al<br />
element in the fertilizer.<br />
As regards phosphorus fertilizati<strong>on</strong>, we found that its effect <strong>on</strong> the nutrient c<strong>on</strong>tent<br />
depends <strong>on</strong> the producti<strong>on</strong> site and the soil type. In Nagyhörcsök and Iregszemcse, the<br />
applicati<strong>on</strong> of phosphorus increased the Mg and Mn c<strong>on</strong>tent of the wheat grain samples<br />
in every statistically proved cases, while in Karcag it had no effect <strong>on</strong> the c<strong>on</strong>centrati<strong>on</strong><br />
of these elements.<br />
The K c<strong>on</strong>tent of the soil was sufficient in Karcag, while in Nagyhörcsök and<br />
Iregszemcse it was medium, therefore we expected that the effect of K fertilizati<strong>on</strong><br />
would have higher effect at these cropping sites. C<strong>on</strong>trary, the nutrient-increasing effect<br />
of K fertilizati<strong>on</strong> was proved <strong>on</strong>ly in Nagyhörcsök. At the Iregszemcse research site,<br />
although the K supply was <strong>on</strong>ly medium, the K fertilizati<strong>on</strong> had no effect <strong>on</strong> the nutrient<br />
c<strong>on</strong>tent probably due to other soil-chemical factors.<br />
The results of the analysis c<strong>on</strong>ducted in our accredited laboratory dem<strong>on</strong>strate that<br />
the effect of the K and P fertilizers <strong>on</strong> the Mg and Mn c<strong>on</strong>centrati<strong>on</strong> depends rather <strong>on</strong><br />
the producti<strong>on</strong> site, while the effect of the N fertilizer depends <strong>on</strong> the crop year.<br />
260
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Debreczeni, B., Debreczeni, B.-né. (1994): Trágyázási kutatások 1960-1990.<br />
Akadémiai Kiadó, Budapest. 40-60 p.<br />
Gawalko, E.J., Garrett, R.G., Nowicki, T.W. (2002): Cadmium, Copper, Ir<strong>on</strong>,<br />
Manganese, Selenium, and Zinc in Canadian Spring Wheat. Communicati<strong>on</strong>s in<br />
Soil Science and Plant Analysis, 33: 3121-3133 p.<br />
Győri, Z. (1998): A <strong>term</strong>esztési tényezők hatása egyes gab<strong>on</strong>afélék és maghüvelyesek<br />
minőségére. MTA doctoral thesis.<br />
Kádár, I. (1991): A talajok és növények nehézfémtartalmának vizsgálata.<br />
Környezetvédelmi és Területfejlesztési Minisztérium és MTA Talajtani és<br />
Agrokémiai Kutató Intézete, Budapest.<br />
Kirkham, M. B. (1983): Elemental c<strong>on</strong>tent of soil, sorghum and wheat <strong>on</strong> sludgeinjected<br />
agricultural land. Agriculture, Ecosystems and Envir<strong>on</strong>ment, 9: 281-292 p.<br />
Kovács, B., Dániel, P., Győri, Z., Loch, J., Prokisch, J. (1998): Studies <strong>on</strong> Parameters<br />
of Inductively Coupled Plasma Spectrometer. Communicati<strong>on</strong>s in Soil Science and<br />
Plant Analysis, 29: 2035-2054.<br />
Kovács, B., Győri, Z., Prokisch, J., Loch, J., Dániel, P. (1996): A study of plant<br />
sample preparati<strong>on</strong> and inductively coupled plasma emissi<strong>on</strong> spectrometry<br />
parameters. Communicati<strong>on</strong>s in soil science and plant analysis, 27: 1177-1198.<br />
Lagua, R.T. Claudio, V.S. (1996): Nutriti<strong>on</strong> and diet therapy reference dicti<strong>on</strong>ary.<br />
Chapman & Hall. New York etc.<br />
Mahan, L.K., Escott, S.S. (1996): Krause’s food, nutriti<strong>on</strong>, diet therapy. W. B.<br />
Saunders Company. Phiadelphia etc.<br />
Pais, I. (1981): The role of the trace elements in foods. Acta Alimentaria, 10: 244–245<br />
p.<br />
Posner, E.S., Hibbs, A.N. (1997): Wheat flour milling. American Associati<strong>on</strong> of<br />
Cereal Chemists, Inc., St. Paul, Minnesota, U. S. A.<br />
Srikumar, T.S., Öckerman, P.A. (1991): The effects of organic and inorganic<br />
fertilizati<strong>on</strong> <strong>on</strong> the c<strong>on</strong>tent of trace elements in cereal grains. Food Chemistry, 42:<br />
225-230 p.<br />
Sugiura, S.H., D<strong>on</strong>g, F.M., Rathb<strong>on</strong>e, C.K., Hardy, R.W. (1998): Apparent protein<br />
digestibility and mineral availabilities in various feed in gredients for salm<strong>on</strong>id<br />
feeds. Aquaculture, 159: 177-202 p.<br />
261
COMPARISON OF SOIL-TEST EXTRACTANTS FOR ZINK AND COPPER<br />
Rita Kremper - Sándor Berényi - Jakab Loch – Andrea Balla Kovács<br />
Debrecen University, Centre for Agricultural Sciences<br />
Department of Agrochemistry and Soil Science<br />
ABSTRACT<br />
KCl-EDTA, Lakanen Erviö and CaCl2- DTPA soluti<strong>on</strong>s were compared as soil Cu and<br />
Zn extractant. The former two methods are the Hungarian official methods, while<br />
CaCl2- DTPA is a frequently studied method word-wide. 27 agricultural soil samples<br />
were collected for the study from the Soil Informati<strong>on</strong> and M<strong>on</strong>itoring System (SIMS)<br />
database. LE and KCl-EDTA extracted similar amounts of Zn and Cu. CaCl2-DTPA<br />
extracted much smaller amounts both for Zn and Cu (approximately half-third as much)<br />
than the other two extractant. It can be attributed to the pH of the extractants. We<br />
suppose that KCl-EDTA and LE solved Zn and Cu oxides-hydrates, Zn and Cu<br />
carb<strong>on</strong>ates at pH 4.36 and pH 4.6 respectively which can not be solved by CaCl2-<br />
DTPA at pH 7.2. We found good correlati<strong>on</strong> between the three methods despite of the<br />
small sample number.<br />
Keywords: extractrant, DTPA, KCl-EDTA, Lakanen-Erviö, microelements<br />
INTRODUCTION<br />
The three sequential steps generally followed to develop micr<strong>on</strong>utrient cati<strong>on</strong> soil tests<br />
are extractant selecti<strong>on</strong>, greenhouse evaluati<strong>on</strong> and field calibrati<strong>on</strong> (Martens and<br />
Lindsay, 1990). The first step is to select an extractant that will solubilize a<br />
proporti<strong>on</strong>ate part of labile forms of micr<strong>on</strong>utrient cati<strong>on</strong>s from different soils. The<br />
sec<strong>on</strong>d step is to evaluate if amounts of cati<strong>on</strong>s extracted are related to quantities<br />
absorbed by plants from different soils. This step comm<strong>on</strong>ly is completed int he<br />
greenhouse rather than in the field. The third step field calibrati<strong>on</strong> of a soil extractant is<br />
c<strong>on</strong>ducted if a suitable relati<strong>on</strong>ship is established during the greenhouse evaluati<strong>on</strong>.<br />
During field test scientist separate soils into resp<strong>on</strong>sive and n<strong>on</strong>-resp<strong>on</strong>sive categories,<br />
or de<strong>term</strong>ine the insufficient, sufficient and transiti<strong>on</strong>al z<strong>on</strong>e of the available<br />
micr<strong>on</strong>utrient cati<strong>on</strong>s in the soil.<br />
Most of the Zn and Cu in soils is unavailable to plants and may be divided by chemical<br />
extracti<strong>on</strong> into five chemical pools (McLaren and Crawford, 1973)<br />
1. as free and complexed i<strong>on</strong>s in soil soluti<strong>on</strong><br />
2. as n<strong>on</strong>-specifically and specifically adsorbed cati<strong>on</strong>s<br />
3. as i<strong>on</strong>s occluded mainly in soil carb<strong>on</strong>ates and hydrous oxides<br />
4. in biological residues and living organisms<br />
5. in the lattice structure of primarily and sec<strong>on</strong>dary minerals.<br />
In soils Zn and Cu cati<strong>on</strong>s would be specifically adsorbed by carb<strong>on</strong>ates, hydrous<br />
oxides of Al, Fe and Mn, soil organic matter and phyllosilicates (Udo et al., 1970)<br />
Soil extractants for plant available Cu and Zn forms can be divided into water and<br />
neutral salts, weak and str<strong>on</strong>g acids and chelating agents. Neutral salts by itself (as<br />
CaCl2) proved to extract too small amounts for Zn and Cu which are not measurable<br />
accurately. Dilute acids as (0.025-0.1M HCl) has been used <strong>on</strong> acidic soils as<br />
262
micr<strong>on</strong>utrient soil tests for many years. The disadvantage of this method is to solve<br />
such Zn and Cu forms which are not available for plants, and unsuitable for calcareous<br />
soils.<br />
One of the major advances in micr<strong>on</strong>utrient soil testing has been the development<br />
of extracting soluti<strong>on</strong>s that c<strong>on</strong>tain DTPA and EDTA. Chelates reduce the activity of<br />
free metal i<strong>on</strong>sin soluti<strong>on</strong> through the formati<strong>on</strong> of soluble metal-chelate complexes.<br />
Free i<strong>on</strong> c<strong>on</strong>centrati<strong>on</strong> in the soil soluti<strong>on</strong> is replenished from solid phases in resp<strong>on</strong>se<br />
to this micr<strong>on</strong>utrient chelati<strong>on</strong>. The quantity of micr<strong>on</strong>utrient extracted by a chelate<br />
reflects both the initial c<strong>on</strong>centrati<strong>on</strong> in the soil soluti<strong>on</strong> (intensity factor) and the ability<br />
of the soil to maintain this c<strong>on</strong>centrati<strong>on</strong> (capacity factor), (Viets and Lindsay, 1973).<br />
Therefore, chelating agents may simulate nutrient removal by plant roots and the<br />
replenishment of the micr<strong>on</strong>utrient from labile phases in the soil.<br />
In this study we focus <strong>on</strong> the first step i.e. laboratory test of the extractant<br />
selecti<strong>on</strong>. We chose KCl-EDTA, Lakanen Erviö extractants as official Hungarian<br />
methods and DTPA-CaCl2 as a frequently studied extractant word-wide. Molnáros et al.<br />
have already made similar investigati<strong>on</strong>s <strong>on</strong> west Hungarian soils, but they have not<br />
involved DTPA- CaCl2 in their studies.<br />
Our aim was to examine the relati<strong>on</strong>ship am<strong>on</strong>g Cu and Zn extracted by the three<br />
extractants <strong>on</strong> 27 soil samples with different soil properties.<br />
MATERIALS AND METHODS<br />
Charactaristics of the applied extractants are represented in Table 1. 27 soil samples<br />
from Soil Informati<strong>on</strong> and M<strong>on</strong>itoring System (SIMS) of Hungary were used. The soil<br />
samples were collected from Hajdú-Bihar and Szabolcs-Szatmár-Bereg counties. The<br />
soils varied greatly in their physical and chemical properties and included sand, loam<br />
and clay as well. Table 2 summarizes the most important soil properties.<br />
Extactant<br />
KCl-EDTA<br />
DTPA-CaCL2<br />
Lakanen<br />
Erviö (LE)<br />
Table 1. Some physical and chemical characteristics of soils<br />
Extractant<br />
compositi<strong>on</strong><br />
0.05 M EDTA<br />
(as Komplex<strong>on</strong> III)<br />
0.1 M KCl<br />
0.005 M DTPA<br />
0.1 M CaCl2<br />
0.1 M TEA<br />
0.02 M EDTA<br />
(as Komplex<strong>on</strong> II)<br />
0.5 M NH4OAc<br />
0.5 M CH3COOH<br />
Soil<br />
extractant<br />
ratio<br />
Shaking<br />
time<br />
(min)<br />
pH Reference<br />
1:2 120 4.36 MSZ 20135<br />
1:2 120 7.2<br />
Lindsay et al.<br />
1978<br />
1:10 0 4.65 MSZ 20135<br />
263
Table 2. Some physical and chemical characteristics of soils<br />
Soil<br />
number<br />
pHKCl KA CaCO3% Hu% Texture<br />
1 6.37 31
LE and KCl-EDTA extracted similar amounts of Zn and Cu and CaCl2- DTPA extracted<br />
much smaller amounts both for Zn and Cu (approximately half-third as much) than the<br />
other two extractant. The thirst two extractant were acidic (pH 4.6 and pH 4.35<br />
respectively), thus KCl-EDTA and LE solved such micr<strong>on</strong>utrient forms e. g. Zn and Cu<br />
oxides-hydrates, Zn and Cu carb<strong>on</strong>ates which can not be solved by CaCl2- DTPA at pH<br />
7.2.<br />
As we expected for each extractants the extracted Cu c<strong>on</strong>centrati<strong>on</strong>s were the<br />
largest in case of clay, followed by loam and the smallest for sand. Zn c<strong>on</strong>centrati<strong>on</strong><br />
followed the same tendency for each extractant, but here we got greater values of<br />
averages for sand than for loam.<br />
By he investigated methods the different soil soluti<strong>on</strong> ratio and pH influence the<br />
accurate measurement of low c<strong>on</strong>centrati<strong>on</strong>s. Extracts of KCl-EDTA with 1:2 soil<br />
soluti<strong>on</strong> ratio are better measurable than LE extracts with 1:10 soil soluti<strong>on</strong> ratio.<br />
DTPA-CaCl2 soluti<strong>on</strong> was buffered to pH=7.2, thus the extracted amounts were smaller<br />
than that of KCl-EDTA soluti<strong>on</strong>.<br />
Results of the correlati<strong>on</strong> analysis show (Table 4, Table 5) that in spite of the small<br />
number of samples the extractant c<strong>on</strong>centrati<strong>on</strong> of the three methods vary<br />
proporti<strong>on</strong>ately. Former similar results were experienced by Balla Kovács et al. Though<br />
the extracted amounts of Cu are similar for KCl-EDTA and LE, the correlati<strong>on</strong> between<br />
these two extractants is not greater than the correlati<strong>on</strong> between these and DTPA-CaCl2<br />
according to the summarised data. When we sorted the soil samples by texture in some<br />
cases the correlati<strong>on</strong> data were better than the summarized data, but in case of Zn we<br />
got worse correlati<strong>on</strong> for loam, and in case of Cu the correlati<strong>on</strong> coefficients were<br />
smaller for sand and loam.<br />
Classifying the soils by lime c<strong>on</strong>tent, for calcareous soils in case of Cu correlati<strong>on</strong><br />
coefficient between LE and KCl-EDTA was greater than between DTPA-CaCl2 and the<br />
other two extractant. C<strong>on</strong>trasted with the other two extractants, buffered DTPA-CaCl2<br />
do not solve Cu forms attached to carb<strong>on</strong>ates, which causes the differences in the<br />
correlati<strong>on</strong> coefficients.<br />
265
Soil<br />
Number<br />
Table 3. Copper and Zn extracted by three extractants for various soils<br />
KCl-<br />
EDTA<br />
Zn (mg/kg) Cu (mg/kg)<br />
LE<br />
CaCl2-<br />
DTPA<br />
KCl-<br />
EDTA<br />
LE<br />
CaCl2-<br />
DTPA<br />
Sand<br />
1 2.02 1.52 0.51 0.62 7.45 2.11<br />
2 0.50 0.32 0.13 2.20 0.37 0.15<br />
3 0.58 0.51 0.20 0.84 1.12 0.37<br />
4 1.04 1.71 0.46 2.46 3.35 1.04<br />
5 1.86 2.66 1.17 1.64 1.86 1.07<br />
6 2.46 2.86 1.09 4.52 5.59 1.89<br />
7 2.42 2.92 1.37 3.82 4.72 2.31<br />
8 6.80 13.71 2.16 4.40 9.07 2.24<br />
Average 2.21 3.28 0.89 2.35 3.50 1.28<br />
Scattering 2.01 4.33 0.69 1.45 2.57 0.85<br />
CV% 90.95 132.25 78.06 61.99 73.52 66.46<br />
Loam<br />
9 2.02 2.92 0.46 6.90 8.45 2.06<br />
10 1.42 1.65 0.33 4.46 5.59 1.07<br />
11 0.84 1.84 0.34 1.34 3.23 0.77<br />
12 0.64 2.03 0.13 4.28 6.21 2.73<br />
13 1.58 2.73 0.90 2.40 4.84 1.54<br />
14 1.42 2.35 0.44 2.88 5.71 1.09<br />
15 1.18 3.93 1.37 6.16 7.70 2.01<br />
16 2.52 2.47 0.98 6.02 6.71 2.46<br />
17 0.70 0.70 0.47 3.92 4.47 1.89<br />
18 2.52 2.54 0.84 7.78 9.44 2.46<br />
19 2.82 1.90 0.63 5.14 5.22 1.91<br />
Average 1.61 2.28 0.63 4.59 6.49 1.85<br />
Scattering 0.77 0.82 0.36 1.97 1.99 0.65<br />
CV% 79.86 52.59 90.65 315.95 217.56 252.62<br />
Clay<br />
20 2.72 3.43 1.74 4.54 5.96 2.98<br />
21 3.12 2.22 0.47 10.56 11.18 4.62<br />
22 1.34 1.65 0.39 5.74 9.19 2.34<br />
23 0.92 2.16 0.27 3.46 6.83 2.21<br />
24 5.44 6.09 2.70 19.36 11.44 6.58<br />
25 4.44 3.81 2.25 12.9 14.04 5.47<br />
26 8.28 7.23 3.42 12.9 13.17 4.72<br />
27 3.44 3.55 1.71 9.68 9.44 3.48<br />
Average 3.71 3.77 1.62 9.33 9.63 3.85<br />
Scattering 2.37 1.97 1.16 5.55 3.35 1.73<br />
CV% 47.88 36.06 57.58 42.92 30.64 35.03<br />
Summarised<br />
Average 2.41 3.02 1.00 5.60 6.75 2.35<br />
Scattering 1.90 2.60 0.85 4.30 3.43 1.51<br />
CV% 79.11 86.34 85.36 76.70 50.81 64.32<br />
266
Table 4. Correlati<strong>on</strong> coefficient between extracting soluti<strong>on</strong>s sorted by texture<br />
Aspects of sort Extractant Lakanen-Erviö CaCl2-DTPA Summarized<br />
n=27<br />
KCl-EDTA<br />
CaCl2-DTPA<br />
0.82<br />
0.73<br />
0.89<br />
Sand KCl-EDTA 0.98<br />
0.91<br />
Zn<br />
n=8<br />
Loam<br />
CaCl2-DTPA<br />
KCl-EDTA<br />
0.87<br />
0.26<br />
0.38<br />
n = 11 CaCl2-DTPA 0.69<br />
Clay KCl-EDTA 0.94<br />
0.92<br />
n=8 CaCl2-DTPA 0.95<br />
Summarized KCl-EDTA 0.81<br />
0.91<br />
n=27 CaCl2-DTPA 0.87<br />
Sand KCl-EDTA 0.17<br />
0.36<br />
Cu<br />
n=8<br />
Loam<br />
CaCl2-DTPA<br />
KCl-EDTA<br />
0.91<br />
0.83<br />
0.69<br />
n = 11 CaCl2-DTPA 0.66<br />
Clay KCl-EDTA 0.79<br />
0.96<br />
n=8 CaCl2-DTPA 0.82<br />
Table 5. Correlati<strong>on</strong> coefficient between extracting soluti<strong>on</strong>s sorted by lime c<strong>on</strong>tent<br />
Aspects of sort Extractant Lakanen-Erviö CaCl2-DTPA Summarized<br />
n=27<br />
KCl-EDTA<br />
CaCl2-DTPA<br />
0.82<br />
0.73<br />
0.89<br />
Zn<br />
N<strong>on</strong>-calcareous<br />
soils n = 18<br />
KCl-EDTA<br />
CaCl2-DTPA 0.88<br />
0.93<br />
0.87<br />
Calcareous soils<br />
n=9<br />
KCl-EDTA<br />
CaCl2-DTPA 0.98<br />
0.95<br />
0.96<br />
Cu<br />
Summarized<br />
n=27<br />
N<strong>on</strong>-calcareous<br />
soils n = 18<br />
KCl-EDTA<br />
CaCl2-DTPA<br />
KCl-EDTA<br />
CaCl2-DTPA 0.81<br />
0.87<br />
0.83<br />
0.89<br />
0.91<br />
0.92<br />
Calcareous soils<br />
n=9<br />
KCl-EDTA 0.86 0.69<br />
267
SUMMARY<br />
27 soil samples with various soil properties were extracted by three different methods,<br />
LE and KCL-EDTA extracted nearly the same Zn and Cu amounts, while the buffered<br />
DTPA-CaCl2 extracted <strong>on</strong>ly half - third of that. We suppose that LE and KCl-EDTA<br />
solve such micr<strong>on</strong>utrients forms that are not available for plants. The extracted Zn and<br />
Cu amounts showed good correlati<strong>on</strong> in most of the cases. To de<strong>term</strong>ine the exact<br />
relati<strong>on</strong>ship between the extractants further studies are required.<br />
REFERENCES<br />
Andrea Balla Kovács és Sándor Berényi- Imre Vágó 2006 Comparis<strong>on</strong> of 0.01M<br />
CaCl2-0.005M DTPA and 0,1M KCl-0,01M EDTA extractants for de<strong>term</strong>inati<strong>on</strong><br />
of Cu and Zn in soils in greenhouse experiment Acta Agr<strong>on</strong>omica Óváriensis<br />
Vol.47. No.1<br />
Brennan R. F. and Best, E. 1993: Copper in Soil Analysis an Interpretati<strong>on</strong> Manual<br />
commissi<strong>on</strong>ed by Australian Soil and Plant Analysis Council Inc 1993 CSIRO<br />
publishing<br />
Lidsay, W.L., and W.AQ. Norvell, 1978. Development of DTPA soil test for zinc,<br />
ir<strong>on</strong>, manganese and copper. Soil Sci. Soc. Am. J. 42:421-428<br />
Maftoun, M., V. Mohaselli, N. Karimian, and A. M. R<strong>on</strong>aghi 2003: Laboratory and<br />
Greenhouse Evaluati<strong>on</strong> of Five Chemical Extractants for Estimating Available<br />
Copper in Selected Calcareous Soils of Iran Communicati<strong>on</strong> in Soil Science and<br />
Plant Analysis Vol. 34. Nos. 9-10, 1451-1463, 2003<br />
Martens, D.C., Lindsay, W.L., 1990 Testing soils for copper, ir<strong>on</strong>, manganese and<br />
zinc in Soil testing and plant analysis , 3 rd ed. SSSA Book Series, no.3 edited by<br />
Wes<strong>term</strong>an, R. L.<br />
McLaren, R.G., and D.V. Crawford, 1973. Studies <strong>on</strong> soil copper. I. The fracti<strong>on</strong>ati<strong>on</strong><br />
of copper in soils. J. Soil Sci. 24:172-181<br />
Molnáros, I., Gráczol, Cs., 2000 A talajok réz-, cink- és mangántartalmának<br />
összehas<strong>on</strong>lítása KCl-EDTA, Lakanen-Erviö és töménysavas feltárással a<br />
Talajvédelmi Információs és M<strong>on</strong>itoring Rendszer vizsgálatai alapján. Agrokémiai<br />
és Talajtan Tom. 49. No.1-2.<br />
Udo, E.J., H.L. Bohn, and T. C. Tuckerm, 1970 Zinc adsorpti<strong>on</strong> by calcareous soils.<br />
Soil Sci. Soc. Am. J. 34:405-407<br />
Viets, F.G. and Lindsay, W. L., 1973 Testing soils for Zn, Cu, Mn, and Fe In Soil<br />
Testing and Plant Analysis (eds L.M. Walsh and J. D. Beat<strong>on</strong>.) pp. 162-8 (Soil<br />
Science Society of America Inc.: Wiscosin, USA)<br />
268
YIELD OF RYE AS AFFECTED BY THE CROPYEAR IN THE WESTSIK`S<br />
CROP ROTATION LONG-TERM FIELD EXPERIMENT<br />
István Henzsel – Gyulané Györgyi<br />
University of Debrecen, Centre of Agricultural Sciences,<br />
Research Centre Nyíregyháza<br />
Rye is grown <strong>on</strong> different types of soils, including sandy soils low in humus, brown<br />
forest soils and <strong>on</strong> soils with shallow fertile layer, often eroded. Early growth and<br />
tillering is favored by a mild, rainy and l<strong>on</strong>g autumn, and a cool, l<strong>on</strong>g spring. A critical<br />
period for rye is the time when late frosts occur in May during flowering, accompanied<br />
by cool, rainy days (Kruppa-Szabó, 2005). Cool and rainy weather at flowering may<br />
result in reduced seed set. In case June is too hot, grains will not develop well, which<br />
also results in reduced yield. The best yields can be expected when rainfall is abundant<br />
in May and the weather is cool in June (Szabó, 1992).<br />
The objective of the present study was to de<strong>term</strong>ine the effects of the yearly<br />
rainfall and temperature <strong>on</strong> the yield comp<strong>on</strong>ent of rye under different regimes of<br />
organic manure supply.<br />
MATERIALS AND METHODS<br />
The study was c<strong>on</strong>ducted in the Westsik’s crop rotati<strong>on</strong> l<strong>on</strong>g-<strong>term</strong> field experiment<br />
located at the Nyíregyháza Research Center of the University of Debrecen. The crop<br />
rotati<strong>on</strong> experiment was established by Vilmos Westsik in 1929 with the overall<br />
objective to improve the fertility of sandy soils. The crop rotati<strong>on</strong> experiment offers<br />
possibility to study different regimes of organic matter supply. The experiment c<strong>on</strong>sists<br />
of 14 rotati<strong>on</strong>-treatments of three-year-cycle and <strong>on</strong>e treatment of four-year-cycle.<br />
The soil of the experiment is a loose sandy soil, slightly acid, low in O.M. This<br />
study was c<strong>on</strong>ducted in four rotati<strong>on</strong> regimes including <strong>on</strong>e with no nutrient supply, <strong>on</strong>e<br />
with straw manure treatment, <strong>on</strong>e with farmyard manure treatment, and <strong>on</strong>e with a<br />
sec<strong>on</strong>d crop green manure treatment.<br />
The design of the selected treatments is as follows:<br />
Sand-improvement with the periodical use of fallow (I):<br />
Crop 1: fallow (weeds are ploughed under before flowering)<br />
Crop 2: rye without fertilizati<strong>on</strong><br />
Crop 3: potato without fertilizati<strong>on</strong><br />
Sand-improvement with fermented straw manure (V):<br />
Crop 1: rye with N, P and K fertilizers after the applicati<strong>on</strong> of straw manure<br />
fermented with calcium-amm<strong>on</strong>ium-nitrate<br />
Crop 2: potato with N, P and K fertilizati<strong>on</strong><br />
Crop 3: rye without fertilizati<strong>on</strong><br />
Sand-improvement with farmyard manure (XI):<br />
Crop 1: fodder crop with farmyard manure, P and K fertilizati<strong>on</strong><br />
Crop 2: rye with P and K fertilizati<strong>on</strong><br />
Crop 3: potato with N fertilizati<strong>on</strong><br />
269
Sand-improvement with lupin green manure as sec<strong>on</strong>d crop ploughed under in autumn<br />
(XVI):<br />
Crop 1: rye with N, P and K fertilizati<strong>on</strong>, followed by lupin green manure<br />
Crop 2: potato with P and K fertilizati<strong>on</strong><br />
Crop 3: rye with N, P and K fertilizati<strong>on</strong><br />
Rye yields from the above regimes were analyzed in the years 2002, 2003, 2004, 2005<br />
and 2006.<br />
Sampling areas were designated in each plot before harvesting the crop.<br />
Measurements were performed <strong>on</strong> the height of rye plants, length of the spikes, number<br />
of spikes, thousand grain weight, and rye yield. We were seeking for possible<br />
correlati<strong>on</strong>s between yield and yield comp<strong>on</strong>ents of rye, and data of rainfall and<br />
temperature at Nyíregyháza.<br />
RESULTS<br />
The yearly and m<strong>on</strong>thly rainfall totals at Nyíregyháza are presented in Table 1. On the<br />
basis of the yearly rainfall, the year 2003 can be characterized as deficient in rainfall<br />
(432 mm), while in the year 2004 there was an abundance of rainfall (704 mm). Rainfall<br />
in the rest of the years was around the average.<br />
The yearly and m<strong>on</strong>thly mean temperature values measured at Nyíregyháza are<br />
presented in Table 2. On the basis of the yearly mean temperature, the coolest year was<br />
2005 (9.6 °C). Warmer than 2005 and close to each other were 2003 (9.9 °C), 2004 and<br />
2006 (10.1 and 10.2 °C). The hottest year was 2002 (11.1 °C).<br />
Table 1: Rainfall at Nyíregyháza (mm)<br />
Year 2001 2002 2003 2004 2005 2006<br />
January 8.5 32.1 43.2 21.7 12.5<br />
February 15.0 26.0 42.0 38.5 56.4<br />
March 35.5 4.3 79.4 8.0 59.5<br />
April 24.8 18.0 22.9 76.8 60.4<br />
May 62.0 35.7 51.7 60.3 91.6<br />
June 55.5 41.2 115.9 29.9 118.1<br />
July 76.0 54.7 99.0 84.2 10.0<br />
August 79.0 19.8 80.5 120.7 110.9<br />
September 64.5 66.0 53.1 43.8 44.6 5.0<br />
October 8.0 60.5 107.1 34.8 16.5 21.0<br />
November 32.5 29.0 22.5 55.8 29.6 18.7<br />
December 11.0 40.0 17.5 35.5 37.0 12.0<br />
Total (mm) 552 432 704 568 576<br />
270
Table 2: Mean temperature at Nyíregyháza (°C)<br />
Year 2001 2002 2003 2004 2005 2006<br />
January -1,3 -2,8 -3,0 -0,9 -4,5<br />
February 4,0 -5,3 0,1 -3,3 -1,9<br />
March 7,0 3,3 5,7 2,9 3,2<br />
April 11,2 10,3 11,6 11,3 12,0<br />
May 20,4 19,4 14,6 16,1 15,6<br />
June 20,1 21,1 18,9 18,6 19,3<br />
July 23,1 21,8 21,0 21,2 22,5<br />
August 20,6 22,4 20,3 19,8 19,4<br />
September 14,0 14,8 15,0 15,0 16,2 16,9<br />
October 12,4 9,1 7,9 11,2 10,2 11,1<br />
November 1,9 5,9 6,1 4,8 2,7 5,9<br />
December -5,2 -2,4 -0,3 0,8 0,0 2,5<br />
Mean (°C) 11,1 9,9 10,1 9,6 10,2<br />
Rye yield data taken in the selected crop rotati<strong>on</strong> regimes are presented in Figure 1.<br />
Crop rotati<strong>on</strong> regime with no nutrient supply:<br />
The lowest rye yield was obtained in 2002 and 2006 (1.26 t/ha and 1.27 t/ha<br />
respectively), years with an average amount of rainfall. Higher yield was obtained in<br />
2003, a drier year (1.56 t/ha) and in 2005, a year with an average amount of rainfall<br />
(1.73 t/ha). The highest yield was obtained in 2004, a year with high amount of rainfall<br />
(2.03 t/ha).<br />
Crop rotati<strong>on</strong> regime with straw manure treatment:<br />
Also in this regime, the lowest yield was obtained in 2002 (1.74 t/ha) and in 2006 (1.74<br />
t/ha), followed by the years 2003 (2.56 t/ha) 2005 (2.68 t/ha). The highest yield was<br />
obtained again in 2004 (2.87 t/ha).<br />
Crop rotati<strong>on</strong> regime with farmyard manure:<br />
The lowest rye yield was obtained in 2002 (1.93 t/ha), followed by the year 2003 (2.52<br />
t/ha). Close to each other were 2005 and 2006 with yields of 3.13 and 3.02 t/ha,<br />
respectively. The highest yield was obtained in 2004, with a value of 3.82 t/ha, which is<br />
1.9 t/ha more than that obtained in 2002.<br />
Crop rotati<strong>on</strong> regime with lupin green manure as sec<strong>on</strong>d crop ploughed under in<br />
autumn:<br />
The lowest yield was obtained again in 2002 (1.44 t/ha), followed by 2006 (1.96 t/ha),<br />
2003 (2.26 t/ha) and 2005 (2.60 t/ha) similarly to regimes with no nutrient supply and<br />
with straw manure. The highest yield was obtained in 2004 (3.93 t/ha). Also in this<br />
regime, there was a substantial difference between the lowest and the highest yield<br />
obtained. The yield difference between 2004 and 2004 was greater than 100 %.<br />
271
Yield (t/ha) .<br />
4,5<br />
4,0<br />
3,5<br />
3,0<br />
2,5<br />
2,0<br />
1,5<br />
1,0<br />
0,5<br />
0,0<br />
I V XI XIV<br />
Crop rotati<strong>on</strong> regime<br />
Figure 1: Yield of rye in the Westsik’s crop rotati<strong>on</strong> l<strong>on</strong>g-<strong>term</strong> experiment<br />
2002 was the least favorable year for rye. The weather was unfavourable for tillering,<br />
which resulted in a low number of spikes/m 2 . There was sufficient rain in the critical<br />
period for flowering, but during grain development the water requirement of rye was<br />
not met, and in additi<strong>on</strong>, both periods were hotter than usual. The spikes were shorter<br />
and the thousand grain weight was less than the average. As a result, rye yield was the<br />
lowest of the years studied. It is also unfavourable for rye, if rainfall is in excess during<br />
flowering and grain development, as it was the case in 2006. Even if the number of<br />
spikes, spike length, and the thousand grain weight is around the average, yield tends to<br />
be lower under such weather c<strong>on</strong>diti<strong>on</strong>s.<br />
If the weather is dry, but c<strong>on</strong>diti<strong>on</strong>s are favourable for tillering, resulting in an<br />
above-average number of spikes per unit area, then an average level of yield can be<br />
expected even with shorter spikes and lower thousand grain weight, as it was the case in<br />
2003. The highest rye yield can be expected in cases, when the weather is favourable for<br />
tillering. When rain is abundant in autumn, there will be sufficient amount of water in<br />
the soil after sowing, and if the weather is mild, the soil will not get frozen too early. In<br />
this case, an above average number of spikes per unit area can be expected. It is<br />
favourable if rain is abundant in spring and early summer, and also, if weather is cool<br />
during flowering and grain development. This will result in l<strong>on</strong>ger spikes with higher<br />
number of grains and higher thousand grain weight, which all will produce an<br />
outstanding yield.<br />
SUMMARY<br />
The objective of the study was to de<strong>term</strong>ine the effects of the yearly rainfall and<br />
temperature <strong>on</strong> the yield and yield comp<strong>on</strong>ents of rye under different regimes of organic<br />
manure supply.<br />
272<br />
2002<br />
2003<br />
2004<br />
2005<br />
2006
It was found that either way of applying nutrients affected the height of rye. When May<br />
is cool, the amount of rainfall has less influence <strong>on</strong> the height of rye with farmyard<br />
manure than with other ways of nutrient supply.<br />
The number of rye spikes varies as a functi<strong>on</strong> of the ways of nutrient supply. There<br />
was a smaller variati<strong>on</strong> in the number of spikes in treatments where farmyard manure<br />
was applied as compared with the applicati<strong>on</strong> of green manure or manuring with straw.<br />
However, if weather c<strong>on</strong>diti<strong>on</strong>s are unfavorable during the tillering of rye, the number<br />
of spikes will be lower, without respect to the way of nutrient supply.<br />
If the weather is warm and dry in spring and in early-summer there is a great<br />
variati<strong>on</strong> in the length of spikes between rotati<strong>on</strong> regimes with and without nutrient<br />
supply. In case the spring is rainy, there is no difference in the length of spikes between<br />
rotati<strong>on</strong> regimes.<br />
If the weather is warm and dry in spring and in early-summer there is a negative<br />
correlati<strong>on</strong> between spike length and spike number per unit area irrespective to nutrient<br />
supply. In rainy and cool weather this negative correlati<strong>on</strong> may not be evident.<br />
The thousand seed weight was not significantly affected by the method of nutrient<br />
supply, but the effect of the cropyear was evident. Warm weather during flowering and<br />
graining may result in a lower value of thousand seed weight as compared with values<br />
obtained in cool weather. When the weather is cooler in May and June, with sufficient<br />
rainfall at flowering and graining, the thousand seed weight will be high, irrespective to<br />
the way of nutrient supply. Even when the weather is cooler in late spring and early<br />
summer with less-than-average amount of rainfall at graining, a higher value of<br />
thousand seed weight can be expected than in spring with average rainfall and warm<br />
weather. When May and June are cool and rainfall is in excess of the amount that rye<br />
requires, thousand grain weight may be depressed.<br />
Comparing the rotati<strong>on</strong> regimes it turns out that each way of nutrient supply<br />
increased the yield of rye over the rotati<strong>on</strong> regime with no nutrient. In most of the cases,<br />
the use of farmyard manure produced higher yield than manuring with straw or using<br />
green manure. In treatments where straw manure was used, the effect of the cropyear <strong>on</strong><br />
the variati<strong>on</strong> of rye yield was smaller than in treatments where farmyard manure or<br />
green manure was used. In case weather c<strong>on</strong>diti<strong>on</strong>s are favorable for rye, an outstanding<br />
level of yield can be attained, similarly to the use of farmyard manure.<br />
REFERENCES<br />
Kruppa J. – Szabó M.: Rozs és évelő rozs. In: Növény<strong>term</strong>esztéstan 1. Gab<strong>on</strong>afélék.<br />
2005. Szerk. Jolánkai M., Mezőgazda Kiadó, Budapest, 228-237. p.<br />
Szabó M.: Rozs. In: Szántóföldi növény<strong>term</strong>esztés. 1992. Szerk. Bocz E. Mezőgazda<br />
Kiadó, Budapest. 283-292. p.<br />
273
1<br />
RESULT OF ALFALFA CROPPING BY THE BASIC OF TIME- AND<br />
VILLAGE TYPE DATA FOR COUNTY OF SZABOLCS SZATMÁR BEREG<br />
László Nagy<br />
Nyiregyháza Research Centre of DUASC<br />
4-6. str. Westsik V. Nyíregyháza<br />
ABSTRACT<br />
The volume of alfalfa (Medicago sativa L.) cropping in Hungary depends <strong>on</strong> the district<br />
of producti<strong>on</strong> but mostly <strong>on</strong> the possibilities of utilizati<strong>on</strong>s, because this species can<br />
growth almost all over the country , (Késmárki 2005).<br />
In Hungary <strong>on</strong> the first place by the producti<strong>on</strong> area volume stands the next<br />
counties – Jász-Nagykun-Szolnok, Békés and Hajdú independently by the type of<br />
utilizati<strong>on</strong>. The importance of the counties has altered in the different periods observed,<br />
1st. table. By the average- and the safety of grain yield excellent districs are in Tolna,<br />
Pest, Komárom-Esztergom counties for many years. To see <strong>on</strong> the hay producti<strong>on</strong>, the<br />
excellent counties are in Györ-Sopr<strong>on</strong>, Vas and Baranya if we observ the intensiv<br />
period of alfalfa producti<strong>on</strong> in Hungary (Nagy 1989, 1996).<br />
The purposes of my presentati<strong>on</strong> firstly are, to show the formati<strong>on</strong> of the districts<br />
and villages of lucerne producti<strong>on</strong>, in the sec<strong>on</strong>d instance - the c<strong>on</strong>necti<strong>on</strong> of the first<br />
<strong>on</strong>e with the soil background.<br />
The task was completed with data of three different courses: the basic <strong>on</strong>e is 1936-<br />
1962; the sec<strong>on</strong>d <strong>on</strong>e is 1961-1985; the third <strong>on</strong>e is 2004-2006 period.<br />
The main results are the next: the producti<strong>on</strong> increased with forthy percent in the<br />
period of 1961-1960 and 1971-1980. In the course of 1981-1985 there was no change in<br />
the producti<strong>on</strong> area relative to the basic period, but decreased relative to both previous<br />
<strong>on</strong>es (Nagy 1989, 1996). Present (last) period can also characterized by the decreasing.<br />
The value of this is <strong>on</strong>ly sixthy percent relative to the basic <strong>on</strong>e.<br />
By the data of basic period at some villages there are c<strong>on</strong>siderable percent alfalfa<br />
producti<strong>on</strong> area relative to the whole (counties) <strong>on</strong>e. These are: Nyíregyháza,<br />
Tiszavasvári, Újfehértó independently the year observed. In the instances of some<br />
villages the percent of lucerne relative to whole villages producti<strong>on</strong> area is very high,<br />
more than 7,0%. These are: Milota, Tiszadada, Szamossályi, Hermánszeg<br />
In the instances of the so called statistical districts show hudge values by two<br />
cases. One of them is Tiszavasvári-, the another <strong>on</strong>e is Fehérgyarmat dictrict. In both<br />
cases the value of accumulated cropping area are more, than ten percent of the countie’s<br />
<strong>on</strong>e. Observing the area c<strong>on</strong>centrati<strong>on</strong> relative to the soil characteristics of the district<br />
given can see, there is 0,8 correlati<strong>on</strong> coefficient betwen acid soil and the value of<br />
producti<strong>on</strong> area.<br />
Key words. Alfalfa, producti<strong>on</strong>, l<strong>on</strong>g <strong>term</strong> data, soil properties<br />
INTRODUCTION<br />
The volume of alfalfa (Medicago sativa L.) cropping in Hungary depends <strong>on</strong> the district<br />
of producti<strong>on</strong> and the possibilities of utilizati<strong>on</strong>s purposes.<br />
274
2<br />
The main modificati<strong>on</strong> properties are until now the animal husbandary and the seed<br />
producti<strong>on</strong> for abroad, marketing line.<br />
The most important producti<strong>on</strong> areas by the volume for hay and seed producti<strong>on</strong> are the<br />
districts of the next counties – Jász-Nagykun-Szolnok, Békés, Hajdú, and recently Pest,<br />
1st. table.<br />
By the level of average- and the safety of grain (seed) yield excellent districts are<br />
in Tolna, Pest, Komárom-Esztergom counties for many years. Also excellent districts<br />
for good level of the hay producti<strong>on</strong>, are Györ-Sopr<strong>on</strong>, Vas and Baranya counties if we<br />
observ the intensiv (1961-1985) period of alfalfa producti<strong>on</strong> in, Hungary,(Nagy 1989,<br />
1996). It was very good effect <strong>on</strong> to the level of seed- and hay producti<strong>on</strong> to develop<br />
hungarian varieties which are much better in some indxes, than foreign <strong>on</strong>es nowdays<br />
too.<br />
The purposes of my presentati<strong>on</strong> are: to show the formati<strong>on</strong> of the districts and<br />
villages of lucerne producti<strong>on</strong> and the importance of the soil background.<br />
MATERIAL AND METHODS<br />
The basic of the paper are statistical data have been collected from the Central<br />
Statistical Bureau proceedings, from three different periods. The first (basic) is origins<br />
from 1936-1962 years with the data of 1936,1937, 1938, 1948 1962 years; the sec<strong>on</strong>d<br />
<strong>on</strong>e comes from 1961-1985 (25 years l<strong>on</strong>g period), the third <strong>on</strong>e origins from 2004-<br />
2006 (3 years l<strong>on</strong>g period). The (soil) data of the statistical district bel<strong>on</strong>g to the<br />
Szabolcs-Szatmár Bereg counties origins from the book of Geczy published in 1968.<br />
The villages bel<strong>on</strong>g to the different districts origins from the inter net. The correlati<strong>on</strong><br />
coefficient was accounted by the programme of excell.<br />
RESULT AND DISCUSSION<br />
Until now the producti<strong>on</strong> area of alfalfa in Hungary has changed relative to the period<br />
of 1936-1962. In the meantime, in the period of 1961-1985, area covered by alfalfa<br />
increased with 64%. From this time the area, until nowdays has decreased with 41,5%<br />
relative to the basic data. Relative to 1961-1985’s data the decreasing is 64,3%.<br />
The positi<strong>on</strong> of alfalfa a bit altered by the excellent plantig activity of some county,<br />
eg. Szabocs Szatmár Bereg, betwen 2004-2006. The percent proporti<strong>on</strong> of some county<br />
in the view point of alfalfa producti<strong>on</strong> also has changed in this period, because, nine<br />
county has increased the producti<strong>on</strong> area. In ten county has been decreasedt it, 1.<br />
táblázat<br />
Am<strong>on</strong>g the statistical district of Szabolcs - Szatmár – Bereg county there are two<br />
with very high value in the basic period, <strong>on</strong>e of them is the Tiszavasvári, the another<br />
<strong>on</strong>e is the Fehérgyarmati.<br />
The alfalfa producti<strong>on</strong> value both of them is more than 10 percent. To the first <strong>on</strong>e<br />
bel<strong>on</strong>g nine villages, to the sec<strong>on</strong>d <strong>on</strong>e bel<strong>on</strong>g forthy nine.<br />
Studying the soil properties of the statistical districts, can establish, in Szabolcs<br />
Szatmár Bereg, the positi<strong>on</strong> of the acid soils relates to the proporti<strong>on</strong> of lucerne<br />
utilizati<strong>on</strong>. The coefficient of correlati<strong>on</strong> betwen them is 0,8, 2nd table.<br />
There are high variability in the village range data. In the basic period some village has<br />
c<strong>on</strong>siderable proporti<strong>on</strong> relative to the total utilzati<strong>on</strong> area. More than fithy percent of<br />
275
3<br />
the total alfalafa area bel<strong>on</strong>g to the forthy four villages of different statitistical districts.<br />
The list and their proporti<strong>on</strong> from the whole area of the important villages can see in the<br />
3rd and 4th tables.<br />
Table 1: Proporti<strong>on</strong> of alfalfa producti<strong>on</strong> area of the counties in Hungary in different<br />
period (by the data of CSB)<br />
1936-1962 1961-1985 2004-2006<br />
County %<br />
Jász-Nagykun.-Sz. 12,3 11,9 11,2<br />
Békés 11,3 10,7 8,1<br />
Hajdú 8,0 8,1 9,7<br />
Pest 6,5 7,1 9,8<br />
Baranya 5,9 4,8 2,8<br />
Borsod 5,8 5,8 9,1<br />
Cs<strong>on</strong>grád 5,8 6,4 6,9<br />
Fejér 5,1 5,3 3,4<br />
Heves 5,1 4,0 2,3<br />
Somogy 4,9 3,8 2,4<br />
Győr 4,8 4,0 5,6<br />
Szabolcs-Sz.-B. 4,5 3,8 3,2<br />
Bács-Kiskun 4,1 7,8 8,5<br />
Tolna 3,9 5,0 3,4<br />
Veszprém 3,6 3,0 3,9<br />
Nográd 2,2 2,1 1,5<br />
Vas 2,2 2,0 2,6<br />
Komárom 2,0 2,9 3,9<br />
Zala 1,8 1,6 1,6<br />
Total % 100,0 100,0 100,0<br />
Total ha 249 942 409566 146534<br />
North Great Plain % 24,86 23,76 24,12<br />
r 1,0 0,92 (a/b) 0,73 (a/c)<br />
Note a b c<br />
The most important <strong>on</strong>es bel<strong>on</strong>g to this group of are the next: Nyíregyháza,<br />
Tiszavasvári, Nagyhalász, Tyukod, Ökörítófülpös, Fehérgyarmat, Dombrád,<br />
Vásárosnamény, Nagykálló, Nyírbogát, Nyírtass.<br />
An another group with villages Milota, Tiszadada, Szamossályi, Hermánszeg is<br />
important because here is very high, 7-8% the proporti<strong>on</strong> of the lucerne in their total<br />
sowing area. It can be stated, 29,1 percent of the villages have 3-5% lucerne proporti<strong>on</strong>,<br />
52,6 percent have 1-3% proporti<strong>on</strong> and 9,1 percent have less than 1% lucerne area<br />
proporti<strong>on</strong> in 1936-1962 period. Recent statistical data isn’t c<strong>on</strong>venient for this purpose<br />
because of the missing of wide scale data collecti<strong>on</strong>. Because of this fact this data base<br />
is very importan for the people dealing with this theme in any time.<br />
276
4<br />
Table 2: Percent of alfalfa and- the soils types in the district of Szabolcs Szatmár<br />
Bereg county (by the data of Géczy 1968)<br />
Soil pH properties<br />
alfalfa Str<strong>on</strong>gly alkaline saturated acid Str<strong>on</strong>gly<br />
District<br />
alkaline<br />
acid<br />
%<br />
Tiszavasvári 15,2 0,3 21,9 1,5 75,9 0,1<br />
Fehérgyarmati 12,6 0,1 0,8 0,1 64,9 34,3<br />
Mátészalkai 9,9 2,2 13,5 0,9 70,1 13,3<br />
Vásárosnaményi 8,6 0,1 0,1 14,4 61,4 23,8<br />
Nyíregyházi 8,6 2,1 14,5 28,8 54,7 0,1<br />
Kisvárdai 8,1 3,0 12,3 14,3 57,6 12,8<br />
Nagykállói 5,9 3,4 6,8 33,0 52,0 1,8<br />
Csengeri 5,6 0,1 0,1 1,4 58,9 39,7<br />
Nyírbátori 5,0 0,1 20,6 16,1 57,8 5,4<br />
Baktalórántházi 4,8 7,5 11,6 11,1<br />
-<br />
58,2 11,6<br />
r alfalfa –a,b,c,d,e 1,0 -0,43(a/b) 0,16(a/c) 0,47(a/d) 0,81(a/e) -0,03(a/f)<br />
Note a b c d e f<br />
Sz.Sz.B. county 100,0 1,8 10,8 13,7 60,9 12,5<br />
Table 3: The most important villages for alfalfa producti<strong>on</strong> in Szabolcs Szatmár<br />
Bereg county (by the basic of CSB 1976)<br />
District<br />
Villages name with highest producti<strong>on</strong> area <strong>on</strong><br />
1st 2nd 3rd 4th<br />
place<br />
Total<br />
number<br />
of<br />
villages<br />
Tiszavasvári Tiszavasvári Tiszadada Tiszalök Tiszaeszlár 9<br />
Nyíregyházi Nyíregyháza Nyírpaz<strong>on</strong>y Kótaj Nyírtelek 9<br />
Ibrány –<br />
Nagyhalászi Nagyhalász Gávavencsellő Ibrány Kemecse 17<br />
Csengeri Tyukod Porcsalma Csenger Ura 11<br />
Mátészalkai Ököritófülpös Szamosszeg Vállaj Mérk 26<br />
Fehérgyarmati Fehérgyarmat Tunyogmatolcs Szatmárcseke Milota 49<br />
Kisvárdai Dombrád Fényeslitke Kisvárda Zsurk 30<br />
Vásárosnaményi Vásárosnamény Tarpa Aranyosapáti Gulács 27<br />
Nagykállói Nagykálló Balkány Szakoly Kállósemjén 8<br />
Nyírbátori Nyírbogát Nyírgyulaj Nyírbátor Nyírbéltek 20<br />
Baktalórántházi Nyírtass Apagy Nyírmada Nyíribr<strong>on</strong>y 19<br />
Total observed 11 11 11 11 225<br />
277
5<br />
Table 4: Of the most important villages in Szabolcs Szatmár Bereg county<br />
by the alfalfa producti<strong>on</strong> areas (by the basic of CSB 1976)<br />
Producti<strong>on</strong> area (ha)<br />
Sum of<br />
District 1st 2nd 3rd 4th the area<br />
placed villages<br />
ha<br />
Tiszavasvári 597,6 278,8 258,4 224,4 1359,2<br />
Nyíregyházi<br />
Ibrány -<br />
623,0 66,6 66,2 65,2 821,0<br />
Nagyhalászi 168,2 164,4 113,0 91,6 537,2<br />
Csengeri 231,2 129,6 74,2 72,5 507,6<br />
Mátészalkai 228,2 104,4 71,8 62,3 466,7<br />
Fehérgyarmati 131,7 99,8 96,6 94,6 422,6<br />
Kisvárdai 143,3 114,6 74,0 68,8 400,7<br />
Vásárosnaményi 177,4 59,0 55,0 50,9 342,3<br />
Nagykállói 117,2 84,6 63,0 60,6 325,4<br />
Nyírbátori 72,0 61,2 57,7 53,8 244,8<br />
Baktalórántházi 72,4 67,4 40,8 33,2 213,8<br />
Total 2562,1 1230,4 970,8 878,0 5641,3<br />
Total of the county 11239<br />
REFERENCES:<br />
Géczy G. 1968: Magyarország mezőgazdasági területe. Akadémiai Kiadó, Budapest.<br />
284. p<br />
Késmárki I. 2005. Lucerna. p:357-384. In Antal J. (szerk). Növény<strong>term</strong>esztéstan 2.<br />
Mezőgazda kiadó. Budapest.<br />
KSH. 1976: Növény<strong>term</strong>elés. 2. kötet. Községsoros adatok 1936-1962. Történeti<br />
Statisztikai Kötetek. Budapest. KSH Könyvtár és dokumentációs Szolgálat.<br />
Nagy L. 1989: A nagyüzemi szarvaskerep mag<strong>term</strong>esztés eredményei, 25 éves<br />
tapasztalatok a fejlesztés lehetőségei. Vetőmag folyóírat, Budapest. November, 11-<br />
13.<br />
Nagy L. 1996: Szarvakerep vetőmag és takarmány <strong>term</strong>esztés fejlesztése. Kandidátusi<br />
értekezés.129 p.<br />
278
THE IMPORTANCE OF LONG-TERM FIELD EXPERIMENTS IN THE<br />
QUALITY ASSURANCE OF CROP PRODUCTION<br />
ABSTRACT<br />
Éva Széles, Zsuzsanna Szathmáry, Péter Sipos, Dóra Hovánszki,<br />
Csilla Uri, Zoltán Győri<br />
University of Debrecen, Centre for Agricultural Sciences<br />
Institute of Food Science, Quality Assurance and Microbiology<br />
The quality assurance and traceability of food products has become a major issue of<br />
food safety during the previous years. L<strong>on</strong>g-<strong>term</strong> filed experiments can give well<br />
applicable data for food safety and food processing.<br />
The objectives of l<strong>on</strong>g-<strong>term</strong> field trials are to measure the effectiveness of different<br />
fertilizers and types of farmyard manure in different cropping envir<strong>on</strong>ments, and to<br />
de<strong>term</strong>ine the accumulati<strong>on</strong> of various elements in the soil or the plant. This is the <strong>on</strong>ly<br />
way to obtain statistically reliable data about the interacti<strong>on</strong> between the effect of<br />
fertilizers and other management techniques (soil cultivati<strong>on</strong>, crop density, etc.), as well<br />
as to set the limit values for the chemical soil examinati<strong>on</strong>.<br />
In this work, applicability and necessity of l<strong>on</strong>g-<strong>term</strong> field experiments to the food<br />
safety through a Nati<strong>on</strong>al L<strong>on</strong>g-Term Field Trials was studied.<br />
Keywords: Nati<strong>on</strong>al L<strong>on</strong>g-Term Field Trials, element correlati<strong>on</strong>s<br />
INTRODUCTION<br />
As food producti<strong>on</strong> is getting more and more global, the multiple sources, suppliers and<br />
process steps al<strong>on</strong>g producti<strong>on</strong> pose a threat to the safety of the product. Therefore, the<br />
quality assurance and traceability of food products has become a major issue of food<br />
safety during the previous years. Participants in the food chain are expected to know the<br />
origin of the raw materials they use in order to comply with new regulati<strong>on</strong>s and with<br />
c<strong>on</strong>sumer expectati<strong>on</strong>s. Regulati<strong>on</strong> EC/178/2002, known comm<strong>on</strong>ly as the food law of<br />
the EU has taken up a new integrated approach, stating that the regulati<strong>on</strong>s for food and<br />
feed cannot be handled separately. The “farm to fork” or “stable to table” principle<br />
covers all sectors of the food chain, including feed producti<strong>on</strong>, primary producti<strong>on</strong>, food<br />
processing, storage, transportati<strong>on</strong> and retail sale. Hungary, as an export-oriented and<br />
EU member country also has to meet the new EU requirements in order to be able to<br />
produce goods that meet the requirements of other countries.<br />
Since the requirements in the crop producti<strong>on</strong> sector are less strict than in animal<br />
producti<strong>on</strong> and the issue of the quality assurance of feed grain crops and forages is even<br />
more neglected, efforts are c<strong>on</strong>tinuous in the EU to involve primary producti<strong>on</strong> in the<br />
scope of food regulati<strong>on</strong>. Primary producti<strong>on</strong> (the agricultural sector) has a critical role<br />
in ensuring safe raw materials, since deteriorati<strong>on</strong>s in the raw materials cannot be<br />
eliminated later during the processing stage. Therefore, the quality assurance of food<br />
products should start with the certificati<strong>on</strong> of safe seed and cultivati<strong>on</strong> techniques.<br />
This requirement can be met by using management techniques during which the<br />
level of the possible c<strong>on</strong>taminating factors in the soil and their intake by the crop is<br />
measured and c<strong>on</strong>trolled. L<strong>on</strong>g-<strong>term</strong> field trials are a system of researches c<strong>on</strong>ducted at<br />
279
the same plots for several years or decades. The most famous field trials are in<br />
Rothamsted (UK), Halle (Germany) since 1878, Askov (Denmark) since 1894, Poltava<br />
(Ukraine) since 1885 (Sárdi, 2003). The objectives of l<strong>on</strong>g-<strong>term</strong> field trials are to<br />
measure the effectiveness of different fertilizers and types of farmyard manure in<br />
different cropping envir<strong>on</strong>ments, and to de<strong>term</strong>ine the accumulati<strong>on</strong> of various elements<br />
in the soil or the plant. This is the <strong>on</strong>ly way to obtain statistically reliable data about the<br />
interacti<strong>on</strong> between the effect of fertilizers and other management techniques (soil<br />
cultivati<strong>on</strong>, crop density, etc.), as well as to set the limit values for the chemical soil<br />
examinati<strong>on</strong> (Loch-Nosticzius, 2004).<br />
MATERIAL AND METHODS<br />
Examined samples are from the NLTFT (Nati<strong>on</strong>al L<strong>on</strong>g-Term Field Trials) established<br />
in 1966 and the main objective was to create a uniform trial network that gives points of<br />
reference for the nati<strong>on</strong>al planning of nutrient demand by involving the different micro-<br />
and macro ecological features of different cropping sites. On the other hand, it provides<br />
a good opportunity to examine the l<strong>on</strong>g <strong>term</strong> effect of mineral fertilizer levels <strong>on</strong> the<br />
quality characteristics of yield, such as its nutrient status.<br />
The samples used are from Bicsérd, Karcag, Nagyhörcsök, Iregszemcse,<br />
Keszthely, Hajdúböszörmény, Kompolt, Putnok and Mos<strong>on</strong>magyaróvár research sites.<br />
The characteristic of these sites according to their soil, agrochemical and climatic<br />
properties was presented by Debreczeni and Debreczeniné (1994) in details. Analysed<br />
winter wheat, maize and pea samples were produced from 1999 to 2003. Different NPK<br />
fertilizer doses and ratios were applied in the experiment. 3 levels of nitrogen (150, 200,<br />
250 kg/ha), 4 levels of phosphorus (200, 100 150 and 200 kg/ha) and 2 levels of<br />
potassium (100 and 200 kg/ha) fertilizer doses were used for winter wheat and maize in<br />
different ratios. For peas, 2 levels of nitrogen (40 and 120 kg/ha), 2 levels of<br />
phosphorus (100 and 150 kg/ha) and 1 level of potassium (100 kg/ha) were applied in<br />
different ratios.<br />
The analysis of the grain samples was performed in the Institute of Food Science,<br />
Quality Assurance and Microbiology of the University of Debrecen, Centre of<br />
Agricultural Sciences. De<strong>term</strong>inati<strong>on</strong> of element c<strong>on</strong>tents was performed by Perkin-<br />
Elmer Optima 3300 DV inductively coupled plasma optical emissi<strong>on</strong> spectrometer<br />
(ICP-OES) after HNO3 and H2O2 digesti<strong>on</strong> (Kovács et al., 1996; Kovács et al., 1998).<br />
In plant samples underg<strong>on</strong>e wet destructi<strong>on</strong> by HNO3 and H2O2 we de<strong>term</strong>ined B,<br />
Ca; Cr; Cu; Fe; K; Mg; Mn; Ni; P; S; Zn c<strong>on</strong>tent in the dry matter by Perkin Elmer,<br />
Optima 3300 DV, ICP-OES (USA).<br />
Samples were de<strong>term</strong>ined by Perkin-Elmer, Optima 3300 DV inductively coupled<br />
plasma emissi<strong>on</strong> spectrometer.<br />
RESULTS AND DISCUSSIONS<br />
In our experiment, more than 3000 pea, wheat and corn samples from the NLTFT<br />
(Nati<strong>on</strong>al L<strong>on</strong>g-Term Field Trials) from 1999 to 2003 have been analysed. The database<br />
with over 35000 data allowed for drawing several important c<strong>on</strong>clusi<strong>on</strong>s.<br />
The values of the elements represent the value of the plant as raw material for food<br />
or feed when c<strong>on</strong>sidering the supplementati<strong>on</strong> of the plant with a given element, and<br />
280
essential to the de<strong>term</strong>inati<strong>on</strong> of the supplementati<strong>on</strong> of the plant with specific nutrients.<br />
Furthermore, they have an important role in the extensi<strong>on</strong> services and quality<br />
assurance. Table 1. shows the values with the corresp<strong>on</strong>ding deviati<strong>on</strong>s.<br />
Table 1: Basic data for the nutrient c<strong>on</strong>tent of peas, corn and wheat<br />
Pea Corn Wheat<br />
Mean Dev. Mean Dev. Mean Dev.<br />
B mg kg -1 6.48± 0.7 2.79± 0.6 0.748± 0.3<br />
Ca mg kg -1 1015± 193 119± 47 510± 69<br />
Cu mg kg -1 7.56± 1.9 1.57± 0.3 5.05± 0.8<br />
Fe mg kg -1 63.4± 20 27.8± 7 59.4± 36<br />
K mg kg -1 9820± 852 3158± 276 3695± 354<br />
Mg mg kg -1 1195± 107 1112± 118 1146± 118<br />
Mn mg kg -1 12.1± 2.8 7.18± 1.4 37.1± 4.1<br />
Ni mg kg -1 3.50± 2.8 0.88± 0.3 0.59± 0.3<br />
P mg kg -1 3897± 774 3057± 454 3319± 529<br />
S mg kg -1 2120± 135 1407± 179 1725± 193<br />
Sr mg kg -1 5.09± 1.7 0.659± 0.2 2.73± 1.0<br />
Zn mg kg -1 36.4± 14.0 15.2± 4.2 23.7± 8.2<br />
Protein % % 27.0± 2.2 8.56± 1.1 14.6± 2.1<br />
Figure 1: Deviati<strong>on</strong> in % of the specific elements at pea, corn and wheat samples<br />
The average and standard deviati<strong>on</strong> values are resulted from the different fertilizati<strong>on</strong>s<br />
and from the average of the 50-100 samples from several cropping sites. The extent of<br />
281
the standard deviati<strong>on</strong>, the % deviati<strong>on</strong> represents the extreme nutrient c<strong>on</strong>centrati<strong>on</strong>s of<br />
a given plant, as a result of the different soil types and different nutrient supply. The<br />
high % deviati<strong>on</strong> indicates that the intake of the specific nutrient is primarily influenced<br />
by the envir<strong>on</strong>mental c<strong>on</strong>diti<strong>on</strong>s, at the same time, the small standard deviati<strong>on</strong><br />
indicates that the intake of the element is independent of the envir<strong>on</strong>mental factors and<br />
the plant is supplied with the sufficient nutrients, but the amount built-in is significantly<br />
c<strong>on</strong>trolled by genetics. Figure 1 dem<strong>on</strong>strates the % deviati<strong>on</strong>.<br />
Apparently, the standard deviati<strong>on</strong> of some elements, which have an important role<br />
in the build-up of the grain is small and are hardly de<strong>term</strong>ined by envir<strong>on</strong>mental factors.<br />
As de<strong>term</strong>ined by genetics, the c<strong>on</strong>centrati<strong>on</strong> of these elements in the grain have to be<br />
small, they might have an impact <strong>on</strong> the yield. One of them may be the parameter that<br />
can limit the yield, according to Liebig’s minimum theory. These elements include<br />
sulphur, potassium, magnesium, manganese, nitrogen. Calcium, bor<strong>on</strong> (especially at<br />
wheat), ir<strong>on</strong>, nickel, str<strong>on</strong>tium and zinc are not expected to influence the yield of the<br />
examined plants in the soils in Hungary. However, these elements can improve the<br />
quality of the yield, as supplied in higher amounts, their c<strong>on</strong>centrati<strong>on</strong> in the grain will<br />
increase. C<strong>on</strong>sequently, zinc has no influence <strong>on</strong> the quantity of the yield, but supplied<br />
in higher amount, the plant can build more zinc into the grain, thus the raw material<br />
produced will meet food and feed requirements. If these elements are not supplied, the<br />
c<strong>on</strong>sumers of the products made from this raw material will develop malnutriti<strong>on</strong><br />
symptoms of the specific microelement.<br />
It is important to compare the microelement c<strong>on</strong>tent of the specific plants in the<br />
dietetics point of view. To compare the different data, we have normalized the nutrient<br />
c<strong>on</strong>tent of corn and peas to that of the wheat, as shown in Figure 3.<br />
Figure 3: Comparing the nutrient c<strong>on</strong>tent of the crops normalized to the element<br />
c<strong>on</strong>tent of the wheat<br />
282
The comparis<strong>on</strong> of the nutrient c<strong>on</strong>tent of the crops shows that the sulphur, phosphorus<br />
and magnesium c<strong>on</strong>tent is similar, however, several nutrients show significant<br />
deviati<strong>on</strong>s. The bor<strong>on</strong> and nickel c<strong>on</strong>tent of pea, being 6-8 times higher than that of<br />
wheat is noticeable. At the same time, the manganese c<strong>on</strong>tent of wheat is 5 times higher<br />
than that of pea or corn.<br />
To de<strong>term</strong>ine the protein c<strong>on</strong>tent of the grain, we used the nitrogen c<strong>on</strong>tent. The<br />
sulphur c<strong>on</strong>tent of the grain is de<strong>term</strong>ined by amount of the amino acids c<strong>on</strong>taining<br />
sulphur (cystein, cystine). The amount and quality of protein in the grain significantly<br />
influences the quality of the yield and the nutriti<strong>on</strong>al and feeding value.<br />
It is noticeable that <strong>on</strong>ly at wheat was the correlati<strong>on</strong> between the nitrogen and sulphur<br />
c<strong>on</strong>tent close, as shown in Figure 4.<br />
Figure 4: Correlati<strong>on</strong> of the nitrogen and sulphur c<strong>on</strong>tent of the crops<br />
The amino acids c<strong>on</strong>taining sulphur have an important role in the build-up of the protein<br />
structure, therefore, the analysis of the nitrogen-phosphorus ratio as regards their buildin<br />
the grain is of great importance in our further research. The samples from the NLTFT<br />
(Nati<strong>on</strong>al L<strong>on</strong>g-Term Field Trials) are an important basis for this work.<br />
CONCLUSIONS<br />
Summarizing, we can c<strong>on</strong>clude that the results of the analysis of the samples from the<br />
NLTFT (Nati<strong>on</strong>al L<strong>on</strong>g-Term Field Trials) helped us to identify several new relati<strong>on</strong>s<br />
and supports the scientific work in the fields of plant nutriti<strong>on</strong>, sustainable development,<br />
maintenance of soil fertility and the producti<strong>on</strong> of healthy raw materials. C<strong>on</strong>sequently,<br />
l<strong>on</strong>g <strong>term</strong> field trials are essential in quality assurance, since it takes l<strong>on</strong>g years to<br />
identify the optimal plants and nutrient doses in a given crop rotati<strong>on</strong> and their<br />
c<strong>on</strong>centrati<strong>on</strong> in the grain, in order to obtain the maximum quality for a given<br />
producti<strong>on</strong> purpose.<br />
283
MORPHOLOGICAL AND AFLP VARIATION IN SOME GENOTYPES OF<br />
POA ANGUSTIFOLIA L. AND POA HUMILIS EHRH. EX HOFFM.<br />
Zsuzsa Lisztes-Szabó 1 , Ákos A. Zubor 2 , Béla Tóthmérész 3 ,<br />
Mária Papp 4 , József Prokisch 2<br />
1 Institute of Plant Sciences, University of Debrecen, H-4015, Debrecen, P. O. Box 36<br />
2 Department of Food Science and Quality Assurance, University of Debrecen,<br />
3 Department of Ecology, University of Debrecen, H-4015, Debrecen, P. O. Box 36<br />
4 Department of Botany, University of Debrecen, H-4015, Debrecen, P. O. Box 36<br />
ABSTRACT<br />
Quantitative morphological characteristics of P. humilis and P. angustifolia individuals<br />
collected from different habitats in Eastern Hungary were compared after growing at the<br />
same site for three years. The individuals were also investigated using AFLP technique.<br />
The study showed that the morphological plasticity of these species is c<strong>on</strong>siderable and<br />
the quantitative morphological characteristics measured were of <strong>on</strong>ly limited usefulness<br />
for tax<strong>on</strong>omic separati<strong>on</strong>. There was no difference in utility between the vegetative and<br />
reproductive features. AFLP analysis of genomic DNA showed a high degree of<br />
polymorphism within the species. Eastern Hungarian populati<strong>on</strong>s of P. humilis and P.<br />
angustifolia could not be distinguished <strong>on</strong> the basis of molecular characters using 3<br />
AFLP primer combinati<strong>on</strong>s. The Poa pratensis L. species group dem<strong>on</strong>strates that in<br />
cases of polymorphic taxa the genetic variability allows effective adaptati<strong>on</strong> to slight<br />
envir<strong>on</strong>mental changes.<br />
Keywords: morphological characters, AFLP, transplantati<strong>on</strong>, multivariate analysis<br />
INTRODUCTION<br />
Poa angustifolia L. and Poa humilis Ehrh. ex Hoffm. are am<strong>on</strong>g the most widespread<br />
native grass species in Hungary and are widely used in cool seas<strong>on</strong> turfs and forage<br />
grasses, with many known cultivars. P. angustifolia (or P. pratensis subsp. angustifolia)<br />
and P. humilis bel<strong>on</strong>g to Poa pratensis L. aggregati<strong>on</strong>. The species of this aggregati<strong>on</strong><br />
differ in their leaf width and a few other – in some cases not obvious – morphological<br />
features. Many individuals with in<strong>term</strong>ediate morphological characters can be observed.<br />
The morphological variability of P. pratensis genotypes is dem<strong>on</strong>strated by several<br />
authors (B<strong>on</strong>os et al., 2000; Frenot et al., 1999; Rua, 1996; Reader and B<strong>on</strong>ser, 1993).<br />
The identificati<strong>on</strong> of the species within the P. pratensis aggregati<strong>on</strong> is difficult and<br />
accurate and reliable methods are necessary for distinguishing and characterising them.<br />
Since the influence of envir<strong>on</strong>mental factors can be eliminated using molecular<br />
approaches, the use of DNA based molecular markers is an effective method for the<br />
separati<strong>on</strong> of species. Recently, a PCR-based assay for plant DNA fingerprinting,<br />
AFLP, has been developed which reveals significant levels of DNA polymorphism (Vos<br />
et al., 1995).<br />
The aim of this study is to c<strong>on</strong>trast the morphological and genetic variability of P.<br />
angustifolia and P. humilis populati<strong>on</strong>s collected from East Hungary to provide<br />
additi<strong>on</strong>al informati<strong>on</strong> <strong>on</strong> the extent of genetic divergence. Individuals of P. humilis and<br />
P. angustifolia collected from different habitats were cultivated in the same<br />
284
envir<strong>on</strong>ment for three years, then the morphological features of the c<strong>on</strong>trol individuals<br />
(which remained in the original habitat) were compared with the transplanted <strong>on</strong>es. The<br />
study looks for answers to the following questi<strong>on</strong>s: (1) whether there are differences in<br />
morphological features between the two species, excluding those used for identificati<strong>on</strong><br />
and collecti<strong>on</strong>, (2) whether the transplanted individuals differ from the c<strong>on</strong>trols in their<br />
morphological features (3) whether reproductive features more readily reveal tax<strong>on</strong>omic<br />
differences than do vegetative features (4) whether there is any obvious separati<strong>on</strong><br />
between the two species - and am<strong>on</strong>g other Poa species - based <strong>on</strong> AFLP markers.<br />
MATERIALS AND METHODS<br />
Plant material: The shoot clusters for transplantati<strong>on</strong> were collected in May 2003 from<br />
sandy, sandy loam and alkaline soil pastures in Hajdú-Bihar County, Eastern Hungary<br />
(Table 1). Two individuals of P. pratensis and an individual of P. compressa were also<br />
included in the molecular part of this study because the genetic distance am<strong>on</strong>g P.<br />
pratensis, P. angustifolia and P. humilis may give more informati<strong>on</strong> about their<br />
tax<strong>on</strong>omic status.<br />
Table 1: Origins of studied P. humilis, P. angustifolia, P. pratensis and<br />
P. compressa populati<strong>on</strong>s.<br />
Populati<strong>on</strong> Origin Lat. (N) L<strong>on</strong>g. (E) Associati<strong>on</strong><br />
P. humilis (1) Józsa 47°34’ 21°36' Agrosti-Alopecuretum<br />
P. humilis (2) Pallag 47°36’ 21°40' Cynod<strong>on</strong>ti-Poetum<br />
P. humilis (3) Martinka 47°35’ 21°46' Astragalo-Festucetum<br />
P. humilis (4) Mikepércs 47°28’ 21°39' Trifolio-Lolietum<br />
P. humilis (5) Hajdúbagos 47°25’ 21°41' Agropyro-C<strong>on</strong>volvuletum<br />
P. angustifolia (1) Pallag 47°36’ 21°40' Cynod<strong>on</strong>ti-Poetum<br />
P. angustifolia (2) Martinka 47°35’ 21°48' Astragalo-Festucetum<br />
P. angustifolia (3) Martinka 47°36’ 21°50' Cynod<strong>on</strong>ti-Festucetum<br />
P. angustifolia (4) Zsáka 47°09’ 21°26' Agrosti-Alopecuretum<br />
P. pratensis (1) Bagamér 47°28’ 22°00' Caricetum acutiformis-ripariae<br />
P. pratensis (2) Martinka 47°35’ 21°51' Caricetum vulpinae<br />
P. compressa Mikepércs 47°29’ 21°39' Trifolio-Lolietum<br />
Additi<strong>on</strong>ally, to estimate the genetic relati<strong>on</strong> within the aggregati<strong>on</strong> it is informative to<br />
compare the three species with P. compressa as a distinct species outside the<br />
aggregati<strong>on</strong>.<br />
The shoots of the 12 populati<strong>on</strong>s were separated to small tiller units and planted in<br />
the Botanical Garden of University of Debrecen in the same plot. There were 10 rows in<br />
the plot, <strong>on</strong>e row for each populati<strong>on</strong>. A row c<strong>on</strong>tained 15 tillers. The distance between<br />
tillers was about 100 mm, the width of a row is 500 mm. Plastic dividing walls of 200<br />
285
mm depth were sinked into the soil between the rows to prevented spreading of stol<strong>on</strong>s<br />
to the neighbouring rows.<br />
Morphometrical analysis: In May 2006, shoots were collected from the labelled shoot<br />
clusters of the original habitats (c<strong>on</strong>trols) and others were collected from the<br />
experimental plot. The following characteristics of the vegetative organs were<br />
measured: (1) height at flowering, (2) lengths of the l<strong>on</strong>gest leaf-blade of lateral tillers,<br />
(3) maximum widths of these leaves, (4) length of the flag leaf-blade, (5) and the flag<br />
leaf sheath, (6) width of the flag leaf-blade, (7) length of the ligule of the flag leaves.<br />
The following characters of the reproductive organs were measured: (8) length of the<br />
panicle, (9) length of the l<strong>on</strong>gest panicle branch, (10) length of the spikelets from the<br />
end of the lowest panicle branches, (11) length of the lower and (12) upper glume, (13)<br />
length of the palea, (14) length of the lemma. We measured 15 individuals from each<br />
populati<strong>on</strong>. The average of 10 data per individual was used for the following features: 2,<br />
11, 12, 13 and 14. A slide-gauge was used for measuring.<br />
AFLP method: For AFLP, each sample c<strong>on</strong>sisted of about <strong>on</strong>e leaf per plant for ten<br />
plants per populati<strong>on</strong>. ZenoGene kit (Zen<strong>on</strong>Bio Ltd, Szeged, Hungary) was used for<br />
DNA extracti<strong>on</strong> following manufacturer’s instructi<strong>on</strong>s. Extracted DNA samples were<br />
stored at –20 °C until used. After the AFLP procedure (K. Szabó et al., 2006) the final<br />
amplified and selected PCR fragments were appeared by capillary electrophoresis with<br />
ABI Prism 3100 Genetic Analyzer in fragment analysis mode at the Agricultural<br />
Biotechnology Centre (Gödöllő, Hungary) using 360 mm length capillaries and POP-4<br />
gel. GeneScan HD-400 Rox Size was applied as standard marker. AFLP fragments were<br />
identified by GeneScan 3.7 computer program.<br />
Data analysis: Morphological data were analysed by principal comp<strong>on</strong>ent analysis<br />
using the R statistical program (R Development Core Team, 2005). For cluster analysis<br />
of AFLP data each detected band above 80 threshold limit was scored as present (1) or<br />
absent (0) in all samples. The unweighted pair group method using arithmetic means<br />
(UPGMA) was used and a dendrogram was c<strong>on</strong>structed based <strong>on</strong> Jaccard’s similarity<br />
coefficient (Jaccard, 1908) by the statistical software package SPSS 11.0 for Windows<br />
(SPSS Inc., USA). Genetic diversity and significant differences in the populati<strong>on</strong>s were<br />
estimated by the proporti<strong>on</strong> of polymorphic loci. The allele frequencies and<br />
heterozygosity were estimated based <strong>on</strong> square root of recessive genotypes. These<br />
estimates and Nei-genetical distances (1978) were analysed by TFPGA (Miller and<br />
Mark, 1997).<br />
RESULTS<br />
Morphometrical analysis: We did not observe a sharp separati<strong>on</strong> <strong>on</strong> the PCA biplot<br />
diagram based <strong>on</strong> all features of the two species (Fig. 1). Clusters of the individuals of<br />
the two species show complete overlap. However, the variables show unambiguously<br />
two different tendencies, depending <strong>on</strong> whether they are vegetative (v1-v8, v14), or<br />
reproductive (v9-v13) characteristics.<br />
286
PC 2<br />
5<br />
4<br />
3<br />
2<br />
1<br />
0<br />
-1<br />
-2<br />
-3<br />
-4<br />
V2<br />
V5<br />
V4<br />
V10<br />
V11<br />
V14<br />
V9<br />
V1<br />
V6<br />
V12 V13<br />
V7<br />
-6 -4 -2 0 2 4 6<br />
PC 1<br />
Figure 1: PCA biplot of the two species based <strong>on</strong> all variables.<br />
(1-8, 14 vegetative, 9-13 reproductive variables.) □: transplanted P. angustifolia, ○:<br />
transplanted P. humilis, �: c<strong>on</strong>trol P. angustifolia, �: c<strong>on</strong>trol P. humilis. v1: height,<br />
v2: panicle length, v3: panicle width, v4: flag leaf sheath length, v5: flag leaf blade<br />
length, v6: ligule length of the flag leaves, v7: the widest width of leaves, v8: flag leaf<br />
blade width, v9: spikelet length, v10: upper glume length, v11: lower glume length,<br />
v12: palea length, v13: lemma length, v14: l<strong>on</strong>gest leaf blade lengths of lateral tillers.<br />
There was no separati<strong>on</strong> based <strong>on</strong> the features of reproductive organs (Fig. 1). However,<br />
the individuals were c<strong>on</strong>centrated into two, partly overlapping groups based <strong>on</strong> the<br />
characteristics of their vegetative organs. The first is the group of transplanted<br />
individuals which have larger vegetative organs (Fig. 1). The sec<strong>on</strong>d is the group of<br />
individuals with smaller organs which were collected from the original turf cl<strong>on</strong>es. An<br />
explanati<strong>on</strong> for this is that in the experimental envir<strong>on</strong>ment the interspecific competiti<strong>on</strong><br />
was suppressed as seedlings of other species were eliminated and the transplanted plants<br />
were able to spread and occupy all of the available area.<br />
The PCA results separated the transplanted and c<strong>on</strong>trol populati<strong>on</strong>s of P.<br />
angustifolia into two groups but the transplanted and c<strong>on</strong>trol populati<strong>on</strong>s of P. humilis<br />
did not show such separati<strong>on</strong> (Fig. 1). The quantitative morphological characteristics of<br />
P. angustifolia resp<strong>on</strong>ded more sensitively to the transplantati<strong>on</strong>.<br />
Neither species nor transplanted populati<strong>on</strong>s were separated based <strong>on</strong> the<br />
reproductive characters. The two species did not show differences c<strong>on</strong>sidering the<br />
vegetative characteristics, but the transplanted and c<strong>on</strong>trol individuals of P. angustifolia<br />
separated slightly.<br />
In the AFLP analysis three primer combinati<strong>on</strong>s were used and generated clear banding<br />
patterns. A total of 338 DNA fragments were produced across 12 genotypes of four Poa<br />
287
species (plus P. pratensis and P. compressa) with 333 fragments being polymorphic<br />
(98.52% polymorphism) (Table 2). A representative example of the amplificati<strong>on</strong><br />
products obtained at P. angustifolia genotypes using the primer combinati<strong>on</strong> EcoRI-<br />
ACC+Tru1I-CAG is shown in Fig. 5. The number of bands from each primer<br />
combinati<strong>on</strong>s ranged from 84 (EcoRI-ACC+Tru1I-CAA) to 159 (EcoRI-ACC+Tru1I-<br />
CAG). In the present study, a wide variati<strong>on</strong> in the number of polymorphic bands (83-<br />
157) and the percentages of polymorphic bands within a primer combinati<strong>on</strong> (98.80-<br />
98.74 %) were observed. The mean polymorphic rate was 98.47 %.<br />
Table 2: The number of bands and degree of polymorphism revealed<br />
by AFLP primer combinati<strong>on</strong>s.<br />
Primer combinati<strong>on</strong>s Total bands Polymorphic bands Polymorphism rate (%)<br />
EcoRI-ACC+Tru1I-CAG 159 157 98.74<br />
EcoRI-ACC+Tru1I-CAC 95 93 97.89<br />
EcoRI-ACC+Tru1I-CAA 84 83 98.80<br />
Total 338 333 98.52<br />
A presence (1) or absence (0) binary data matrix c<strong>on</strong>taining 333 polymorphic AFLP<br />
fragments was used to generate the genetic similarity estimates. The Nei-genetical<br />
similarity, between 0.87 and 0.95, shows remarkable genetic variati<strong>on</strong> am<strong>on</strong>g the<br />
studied Poa populati<strong>on</strong>s. At the same time, the species of the P. pratensis group are<br />
close to each other, <strong>on</strong>ly P. compressa shows some separati<strong>on</strong>. Inside the group, the<br />
similarity is the biggest between P. pratensis and P. humilis, and the smallest between<br />
P. pratenis and P. angustifolia.<br />
UPGMA clustering analysis was carried out in order to dem<strong>on</strong>strate graphically the<br />
genetic similarities am<strong>on</strong>g the Poa genotypes (Fig. 2), to present a dendrogram of 338<br />
AFLP fragments obtained from three primer combinati<strong>on</strong>s at 12 Poa populati<strong>on</strong>s. P.<br />
humilis (2) is separated at a high level from other populati<strong>on</strong>s. P. humilis (1) and P.<br />
humilis (3) are close to each other. P. angustifolia (2) and P. angustifolia (3) is similar,<br />
as are P. angustifolia (1) and P. angustifolia (4). But the populati<strong>on</strong>s of different species<br />
are not separated from each other as would be expected. The two populati<strong>on</strong>s of P.<br />
pratensis are in different clusters.<br />
288
Figure 2: Genetic similarity am<strong>on</strong>g Poa genotypes revealed by UPGMA cluster<br />
analysis based <strong>on</strong> AFLP data.<br />
The heterozygosity is bigger (25.6 %) in P. humilis than in P. angustifolia (23.4 %).<br />
The heterozygosity and polymorphism in P. humilis populati<strong>on</strong>s are bigger than P.<br />
angustifolia populati<strong>on</strong>s (Table 3).<br />
Table 3: Mean heterozygosity and polymorphism rate in populati<strong>on</strong>s of P.<br />
angustifolia and P. humilis.<br />
Heterozygosity (%) Polymorphism (%)<br />
P. humilis 25.60 94.40<br />
P. angustifolia 23.40 83.80<br />
The mean theta value of 0.2581 (0.2307 lower limit and 0.2827 upper limit) does not<br />
mean significant differences am<strong>on</strong>g the populati<strong>on</strong>s of studied species with 95%<br />
c<strong>on</strong>fidence interval and 1000 bootstrap repeat.<br />
Populati<strong>on</strong>s were found to be genetically similar and there was a wide range of genetic<br />
variability am<strong>on</strong>g germplasm sources. The different leaf blade width of the tillers of the<br />
two species is apparent, but sharp separati<strong>on</strong> cannot be seen between the two species in<br />
the clusters <strong>on</strong> the basis of AFLP markers. This questi<strong>on</strong>s the value of the leaf blade<br />
width as a tax<strong>on</strong>omic character. The species level difference is ambiguous between the<br />
four species.<br />
289
DISCUSSION<br />
The species level segregati<strong>on</strong> of the two observed grass species is not supported by the<br />
morphological features measured. Rua (1996) reported similar results, as the<br />
quantitative morphological features did not reveal sharp statistical segregati<strong>on</strong> between<br />
some Poa species. In our study the basis of the identificati<strong>on</strong> was the width of the leaf<br />
blades whilst other features, such as the length values, also used in identificati<strong>on</strong> keys,<br />
did not prove to be useful.<br />
Our previous observati<strong>on</strong>s suggested that the habitat adaptati<strong>on</strong>, due to the habitat<br />
variati<strong>on</strong>s, destroyed the sharp border in morphological features between the two<br />
species. The results of the present study also supported that there was no phenotypic gap<br />
between these species, based <strong>on</strong> the measured characteristics. After three years growing<br />
in the same plot, tillers showed differences in morphological characteristics compared<br />
to individuals of original habitats. In some cases it seems there are not corresp<strong>on</strong>dences<br />
between morphological characters and genotypes (McElroy, et al. 2002; Gillespie and<br />
Boles, 2001). These observati<strong>on</strong>s are c<strong>on</strong>sistent with the results of our study.<br />
The present study c<strong>on</strong>firms the previous results that behind the habitat-sensitive<br />
features of grass shoots there is high within-species genetic variability. This could be<br />
due to a relatively high proporti<strong>on</strong> of potentially sexually produced off-types (Lars<strong>on</strong> et<br />
al., 2001; Curley and Jung, 2004). AFLP variati<strong>on</strong> of populati<strong>on</strong> in Poa pratensis L.<br />
aggregati<strong>on</strong> does not show species level separati<strong>on</strong> between the studied species based<br />
<strong>on</strong> the generated AFLP markers. The Poa pratensis L. group serves as an example of<br />
the fact that in cases of polymorphic taxa the genetic variability results in adaptati<strong>on</strong> to<br />
small envir<strong>on</strong>mental changes and that separable ecotypes exist. Thus, this study<br />
provides further evidence for the utility of AFLP for turfgrass genetics as well as<br />
important genetic informati<strong>on</strong> for turf breeders and managers.<br />
Our study showed that the morphological plasticity of our species was<br />
c<strong>on</strong>siderable, which was related to the small scale variability of the habitats. Thus, the<br />
measured quantitative morphological characters were not useful for tax<strong>on</strong>omic<br />
separati<strong>on</strong>. There was no difference in the usefulness of the vegetative and generative<br />
features c<strong>on</strong>sidering for the separati<strong>on</strong> of the species.<br />
The study of AFLP variati<strong>on</strong> of populati<strong>on</strong>s of the two species suggests complex<br />
populati<strong>on</strong> structures with high levels of introgressi<strong>on</strong>. Such results are c<strong>on</strong>sistent with<br />
the morphological variati<strong>on</strong> seen in genera Poa generally. The high degree of<br />
polymorphism in Eastern Hungarian populati<strong>on</strong>s resulted in c<strong>on</strong>tinuous forms from<br />
apparently pure P. humilis to apparently pure P. angustifolia. The transplantati<strong>on</strong><br />
experiment showed that, despite c<strong>on</strong>siderable phenotypic plasticity, the morphology is<br />
de<strong>term</strong>ined by genotype (Howland et al., 1995). Poa taxa have high genetic potential<br />
for microevoluti<strong>on</strong>ary development.<br />
REFERENCES<br />
B<strong>on</strong>os, S. A., Meyer, W. A., Murphy, J. A. (2000): Classificati<strong>on</strong> of Kentucky<br />
bluegrass genotypes grown as spaces-plants. Hortscience 35 (5): 910-913.<br />
Curley, J., Jung, G. (2004): RAPD-based genetic relati<strong>on</strong>ships in Kentucky bluegrass.<br />
Crop Sciences 44: 1299-1306.<br />
290
Frenot, Y., Aubry, M., Misset, M. T., Gloaguen, J. C., Gourret, J. P., Lebouvier,<br />
M. (1999): Phenotypic plasticity and genetic diversity in Poa annua L. (Poaceae)<br />
at Crozet and Kerguelen Islands (subantarctic). Polar Biology 22 (5): 302-310.<br />
Gillespie, L. J., Boles, R. (2001): Phylogenetic relati<strong>on</strong>ships and infraspecific variati<strong>on</strong><br />
in Canadian Arctic Poa based <strong>on</strong> chloroplast DNA restricti<strong>on</strong> site data. Canadian<br />
Journal of Botany 79: 679-701.<br />
Howland, D. E:, Oliver R. P., Davy, A. J. (1995): Morphological and molecular<br />
variati<strong>on</strong> in natural populati<strong>on</strong>s of Betula. New Phytol. 130: 117-124.<br />
K. Szabó, Zs.; B. Tóthmérész, Á. A. Zubor, M. Papp, Z. Győri, J. Prokisch (2006):<br />
AFLP variati<strong>on</strong> in Poa pratensis L. agg. in Eastern Hungary. In: Proceedings of<br />
41st Croatian and 1st <str<strong>on</strong>g>Internati<strong>on</strong>al</str<strong>on</strong>g> Symposium <strong>on</strong> Agriculture, ISBN 953-6331-<br />
39-X, p. 225-226.<br />
Lars<strong>on</strong>, S. R., Waldr<strong>on</strong>, B. L., M<strong>on</strong>sen, S. B., St. John, L., Palazzo, A. J.,<br />
Mccracken, C. L., Harris<strong>on</strong>, R. D. (2001): AFLP variati<strong>on</strong> in agamospermous<br />
and dioecious bluegrasses of Westers North America. Crop Sciences 41: 1300-<br />
1305.<br />
McElroy, J. S., Walker, R. H., van Santen, E. (2002): Patterns of variati<strong>on</strong> in Poa<br />
annua populati<strong>on</strong>s as revealed by can<strong>on</strong>ical discriminant analysis of life history<br />
traits. Crop Science 42 (2): 513-517.<br />
Miller, Mark P. (1997): Tools for populati<strong>on</strong> genetic analysis (TFPGA) 1.3: A<br />
Windows program for the analysis of allozyme and molecular populati<strong>on</strong> genetic<br />
data. Computer software distributed by author.<br />
R Development Core Team (2005): R: A language and envir<strong>on</strong>ment for statistical<br />
computing. R Foundati<strong>on</strong> for Statistical Computing, Vienna, Austria. ISBN 3-<br />
900051-07-0, URL http://www.R-project.org.http://her.bio.nau.edu/miller/tfpga.htm.<br />
Reader, R. J., B<strong>on</strong>ser, S. P. (1993): C<strong>on</strong>trol of plant frequency <strong>on</strong> an envir<strong>on</strong>mental<br />
gradient – effects of abiotic variables, neighbors, and predators <strong>on</strong> Poa pratensis<br />
and Poa compressa (Gramineae). Canadian Journal of Botany - Revue Canadienne<br />
de Botanique 71 (4): 592-597.<br />
Rua, G. H. (1996): The genus Poa in Patag<strong>on</strong>ia: a phenetic analysis of species of Poa<br />
subgenus Poa. Botanical Journal of the Linnean Society 121 (3) 229-241.<br />
Vos, P., Hogers, R., Bleeker, M., Reijans, M., van de Lee, T., Hornes, M., Frijters,<br />
A., Pot, L., Peleman, J., Kuiper, M., Zabeau, M. (1995): AFLP: A new<br />
technique for DNA fingerprinting. Nucleic Acids Res. 23: 4407-4414.<br />
291
THE EFFECT OF SOME FOLIAR COMPOSITIONS ON THE PRODUCTION, NUMBER OF<br />
INFLORESCENCES AND FRUIT IN THE TECHNOLOGY OF CULTIVATING<br />
TOMATO IN THE FIELD<br />
Popa Alina-Grigoriţa<br />
University of Oradea, Faculty of Enviromental Protecti<strong>on</strong><br />
ABSTRACT<br />
The experiments proved the influence certain foliar compositi<strong>on</strong>s have <strong>on</strong> the producti<strong>on</strong> of<br />
tomato fruit, the number of inflorescences and fruit <strong>on</strong> the plant in the technology of<br />
cultivating tomato in the field, with an organic-mineral fertilizati<strong>on</strong> applied <strong>on</strong> the soil<br />
Key words: Foliar compositi<strong>on</strong>s, nutritive elements, producti<strong>on</strong>, number of inflorescences,<br />
number of tomato fruit.<br />
INTRODUCTION<br />
Compared to other horticultural soils (especially vine and tree growing), the soils cultivated<br />
with vegetables in the field are usually more fertile. This initial fertility must be maintained<br />
and developed in order to sustain the intensive character of the technology and nutriti<strong>on</strong> of<br />
tomato culture.<br />
The real needs of the field tomatoes towards the main nutritive elements (Cs= specific<br />
c<strong>on</strong>sumpti<strong>on</strong> N, P, K kg/ t product) are very important in the fertilizati<strong>on</strong> system of field<br />
tomatoes, the relati<strong>on</strong>s between the de<strong>term</strong>ining nutrients and the total quantity of nutritive<br />
elements “exported” from the soil together with the crops, at the area unit (Cg= global<br />
c<strong>on</strong>sumpti<strong>on</strong> N, P, K kg/ ha).<br />
As regards the specific and global c<strong>on</strong>sumpti<strong>on</strong>s, the tomatoes take part in the group of<br />
species with an average c<strong>on</strong>sumpti<strong>on</strong>. The fertilizati<strong>on</strong> systems must ensure a management<br />
of the nutrients, which, by sorts, doses and methods, must de<strong>term</strong>ine the accomplishment of<br />
great quality producti<strong>on</strong>s (Rusu M. et. al, 2005).<br />
Taking into account the intensive character of the technology of tomato culture,<br />
according to the fertilizati<strong>on</strong> <strong>on</strong> soil, it was studied the effect of some foliar compositi<strong>on</strong>s in<br />
order to finish some systems of fertilizing tomato, based <strong>on</strong> the positive interacti<strong>on</strong> of the<br />
fertilizati<strong>on</strong> <strong>on</strong> soil to the foliar <strong>on</strong>e; the latter having an integrated character.<br />
MATERIAL AND RESEARCH METHOD<br />
The field culture was initiated <strong>on</strong> argic faeoziom soil.<br />
The experiments were carried out in the year 2003, in locality Oradea, using Unirea soil.<br />
The experiment was d<strong>on</strong>e with the blocks method, in 3 repetiti<strong>on</strong>s, the area of a variant<br />
being 6, 4 mp.<br />
The effect of 4 foliar fertilizers was studied, and the witness was showered with water.<br />
292
Three foliar fertilizati<strong>on</strong>s were d<strong>on</strong>e: the first treatment at the first inflorescence and the<br />
following <strong>on</strong>es at an interval of 14 days.<br />
The effect of the foliar fertilizers <strong>on</strong> the producti<strong>on</strong> of tomato fruit and <strong>on</strong> the number of<br />
inflorescence and fruit <strong>on</strong> the plant was carried out.<br />
The foliar compositi<strong>on</strong>s sorts were applied <strong>on</strong> an agrochemical background resulted from<br />
the interacti<strong>on</strong> of the organic fertilizati<strong>on</strong> (50 t/ ha semi fermented manure) to the complex<br />
applicati<strong>on</strong> of the mineral manure (N120P120K120).<br />
RESULTS AND DISCUSSIONS<br />
It can be noticed in table 1 that significant results were obtained at all foliar fertilized<br />
variants, particularly at the variant fertilized with Bi<strong>on</strong>utrifag F (table no.1, figure no.1).<br />
Table 1 The effect of the foliar fertilizati<strong>on</strong> up<strong>on</strong> the producti<strong>on</strong> in the crop of tomatoes<br />
(UNIREA) from the field (Oradea, 2003)<br />
No.<br />
var.<br />
Foliar assortment<br />
Producti<strong>on</strong><br />
C<strong>on</strong>centrati<strong>on</strong><br />
of the<br />
soluti<strong>on</strong> t/ha Difference<br />
Significance of<br />
difference<br />
1 Martor stropit cu apă - 35,82 - -<br />
2 Nutrifag 1% 41,50 5,68 ***<br />
3 Bi<strong>on</strong>utrifag F 1% 44,60 8,78 ***<br />
4 Ferticare 24-8-16 1% 42,10 6,28 ***<br />
5 Polyfeed 19-19-19 1% 42,16 6,34 ***<br />
DL (5%) 1,74 DL (1%)<br />
2,53 DL (0,1%) 3,79<br />
293
Producti<strong>on</strong> t/ha<br />
50<br />
45<br />
40<br />
35<br />
30<br />
25<br />
20<br />
15<br />
10<br />
5<br />
0<br />
C<strong>on</strong>trol sprinkled<br />
with water<br />
Nutrifag Bi<strong>on</strong>utrifag F Ferticare 24-8-16 Polyfeed 19-19-19<br />
Foliar assortment<br />
Producţia t/ha<br />
Figure 1 The effect of the foliar fertilizati<strong>on</strong> up<strong>on</strong> the producti<strong>on</strong> in the crop of tomatoes<br />
(UNIREA) from the field (Oradea, 2003)<br />
The significant results of the applicati<strong>on</strong> of foliar manure <strong>on</strong> the tomato field culture are due<br />
to an organic mineral under layer and to its recovery by an extra-radicular applicati<strong>on</strong> of<br />
manure as phase/additi<strong>on</strong>al fertilizati<strong>on</strong>s (three treatments at an interval of 14 days).<br />
In additi<strong>on</strong>, the foliar fertilizati<strong>on</strong>s have de<strong>term</strong>ined a greater number of inflorescences and<br />
fruit <strong>on</strong> the plant, fact that prol<strong>on</strong>ged the period of fructificati<strong>on</strong> and cropping (table no. 2).<br />
Tabel 2 The influence of foliar fertilizati<strong>on</strong> up<strong>on</strong> the number of inflorescences and plant<br />
fruits<br />
No.<br />
var.<br />
Foliar assortment<br />
Number of<br />
inflorescences<br />
Număr fructe<br />
number of fruit<br />
1 Martor stropit cu apă 12,1 36,3<br />
2 Nutrifag 13,0 39<br />
3 Bi<strong>on</strong>utrifag F 16,0 48<br />
4 Ferticare 24-8-16 14,7 44,1<br />
5 Polyfeed 19-19-19 15,2 45,6<br />
From the efficient foliar sorts, in the first place Bi<strong>on</strong>utrifag F and Polyfeed 19-19-19 are<br />
focused at the field tomato culture, because they c<strong>on</strong>tain a balanced representati<strong>on</strong> of<br />
nutriti<strong>on</strong> elements and a superior recovery of the fertilizati<strong>on</strong> <strong>on</strong> soil.<br />
294
CONCLUSIONS<br />
The rigorous experimentati<strong>on</strong> of the foliar manure, with an intensive technology for<br />
tomatoes, shows that these compositi<strong>on</strong>s with extra radicular applicati<strong>on</strong> can be framed in<br />
the c<strong>on</strong>text of fertilizing <strong>on</strong> soil, as a measure of an “integrated” character. In the intensivec<strong>on</strong>venti<strong>on</strong>al<br />
fertilizing technologies, the foliar fertilizati<strong>on</strong> c<strong>on</strong>tributes more intensely at the<br />
productive recovery, of the quantity and quality of fruit, due to the nutrients’ c<strong>on</strong>tributi<strong>on</strong> <strong>on</strong><br />
the soil.<br />
The field cultivated tomatoes, <strong>on</strong> an agrochemical organic-mineral background (50t/ha<br />
manure + N120P120K120), present significant producti<strong>on</strong> increases at the applied foliar<br />
sorts and in the first place at the complex product Bi<strong>on</strong>utrifag F (with macro- micro<br />
elements and biological active substances). These effects are due especially to the<br />
accomplishment of some productivity elements (number of inflorescences, number of fruit),<br />
and also to prol<strong>on</strong>ging the ripeness and cropping.<br />
REFERENCES<br />
1. Alina Grigoriţa Popa, 2007, Optimizarea agrochimică a sistemului sol-plantă în<br />
tehnologia de cultivare în spaţii protejate a tomatelor, Teză de doctorat, USAMV Cluj-<br />
Napoca, 168-170;<br />
2. Mihai Rusu, Marilena Mărghitaş, Ioan Oroian, Tania Mihăiescu, Adelina Dumitraş,<br />
2005, Tratat de Agrochimie, Ed. Ceres, Bucureşti, 80-82, 569-572.<br />
295
METHODS AND MEASURES FOR AN AGROCHEMICAL OPTIMIZATION OF THE GREENHOUSE<br />
SOIL AT THE TOMATOES CULTURE<br />
Popa Alina Grigoriţa<br />
University of Oradea, Faculty of Enviromental Protecti<strong>on</strong><br />
ABSTRACT<br />
The suggested subject represents a synthesis of the methods for agrochemical optimizati<strong>on</strong><br />
of greenhouse soil cultivated with tomatoes, included in the doctorate thesis called: Agro-<br />
Chemical Optimizati<strong>on</strong> of the Soil-Plant System in the Protected Space Cultivati<strong>on</strong><br />
Technologies of the Tomatoes<br />
The optimizati<strong>on</strong> of the c<strong>on</strong>tent of greenhouse soil cultivated with tomatoes in organic<br />
matter c<strong>on</strong>stitutes an essential objective, because an optimized agrochemical support<br />
prevents and reduces some disturbing factors as: salinity-salinizati<strong>on</strong>, acidity-acidificati<strong>on</strong>,<br />
toxicity of other i<strong>on</strong>s, polluting effects.<br />
Key words: Agrochemical optimizati<strong>on</strong>, organic matter, soil reacti<strong>on</strong>, soluble salts,<br />
nutritive elements.<br />
1./ The optimizati<strong>on</strong> of the organic matter c<strong>on</strong>tent from the greenhouse soil at<br />
tomatoes culture<br />
The agrochemical optimizati<strong>on</strong> of soils implies a c<strong>on</strong>comitant solving of problems related to<br />
correcting the extreme reacti<strong>on</strong> states and the requests of fertilizati<strong>on</strong><br />
The system of vegetables nutriti<strong>on</strong> in protected areas differs fundamentally from the<br />
vegetables nutriti<strong>on</strong> and fertilizati<strong>on</strong> <strong>on</strong> the field because of certain particularities such as:<br />
� The intensive growth of the vegetal mass, influenced by the envir<strong>on</strong>ment factors<br />
(temperature, humidity);<br />
� The relati<strong>on</strong> between the aerial part and roots is in great favor of the aerial part;<br />
� Using the soil mixtures with an increased c<strong>on</strong>tent of organic matter and nutritive<br />
elements, with very diverse physical-chemical characteristics;<br />
� The immobilizati<strong>on</strong> of some i<strong>on</strong>s by the organic matter (bore, cuprum, manganese);<br />
� The intense microbiological activity in the soil that favors the development of some<br />
pathogen agents, imposes the taking of some preventi<strong>on</strong> and c<strong>on</strong>trol measures;<br />
� The possibility to c<strong>on</strong>trol the envir<strong>on</strong>ment factors and soil;<br />
� Difficulties in ensuring a nutritive balance in the soil, according to the plant’s age and<br />
metabolism.<br />
Because of the intensive character of the system of greenhouse culture, the applicati<strong>on</strong> of<br />
organic manure <strong>on</strong> the basic fertilizati<strong>on</strong> is necessary, for obtaining an optimum c<strong>on</strong>tent of<br />
organic matter in soil, which in fact is the main nutritive support of the culture (Tisdale S.,<br />
1993).<br />
For the greenhouse culture of tomatoes, decomposed manure is used and applied <strong>on</strong> the<br />
basic fertilizati<strong>on</strong>, with about 2 weeks before the seed planting. Because of the presence of<br />
296
some pathogen agents, a chemical and thermic disinfecti<strong>on</strong> is necessary, with hot vapors<br />
(150 o C), at 90 o -95 o C, for 6-7 hours, at 30cm deep.<br />
The manure doses are 80-100t/ha <strong>on</strong>ce a year in greenhouses with older cultures and<br />
twice a year (cycle I and cycle II) in new greenhouses (Davidescu D. and Velicica<br />
Davidescu, 1992; Ciofu et al, 2003).<br />
Composted birds manure can be also applied with 3 m<strong>on</strong>ths in advance in doses of 15-<br />
25t/ha and forestry composts from resins bark or beech (Rusu, 1993).<br />
In greenhouse c<strong>on</strong>diti<strong>on</strong>s, the mobility of nutritive elements from the organic manure is<br />
more intense than at the same culture in the field, thus c<strong>on</strong>sidering that for each 100t manure<br />
in a vegetati<strong>on</strong> cycle in the soil it is liberated in average: 250-300 kg N, 120-170 kg P2O5<br />
and 350-450 kg K2O (after Davidescu D., 1992, cited by Avarvarei, 1997).<br />
The technological steps for optimizing the c<strong>on</strong>tent of organic matter (MO) is a<br />
compulsory request for this technology, so that every interpretati<strong>on</strong> and practical measure<br />
takes into account this value of the organic comp<strong>on</strong>ent. An optimized agrochemical support<br />
solves the nutriti<strong>on</strong> effects and prevents or diminishes some disturbing factors (salinitysalinizati<strong>on</strong>,<br />
acidity-acidificati<strong>on</strong>, toxicity of other i<strong>on</strong>s, polluting effects) (Rusu, 1968,<br />
1988).<br />
2./ The agrochemical optimizati<strong>on</strong> of the reacti<strong>on</strong> state and of the c<strong>on</strong>tent of soluble<br />
salts in the soil from the greenhouse at tomato culture<br />
As regards the optimizati<strong>on</strong> of the c<strong>on</strong>tent of soil in nutritive elements, the c<strong>on</strong>centrati<strong>on</strong>s of<br />
the nutritive i<strong>on</strong>s have to be guided towards the levels of optimum sufficiency in soil, after<br />
fertilizati<strong>on</strong>s.<br />
The greenhouse soils are frequently pr<strong>on</strong>e to salinizati<strong>on</strong> and alkalizati<strong>on</strong>, following<br />
the accumulati<strong>on</strong> of soluble salts obtained from the fertilizati<strong>on</strong> with organic and chemical<br />
manure, irrigati<strong>on</strong> water, water from the phreatic layer and the process of chemical<br />
alterati<strong>on</strong> of the soil mineral fracti<strong>on</strong>.<br />
The soil disinfecti<strong>on</strong> using vapors in greenhouses has the effect of increasing<br />
salinizati<strong>on</strong> due to the accelerati<strong>on</strong> of the transforming process of the amm<strong>on</strong>iac azoth<br />
(NH +<br />
4 ), which inhibits the activity of nitrificati<strong>on</strong> bacteria. These two processes lead to a<br />
temporary accumulati<strong>on</strong> of NO2 (Davidescu D. and Velicica Davidescu, 1992).<br />
Heavy irrigati<strong>on</strong> water, as well as the superficial phreatic layer, c<strong>on</strong>stitutes a source of<br />
enriching the surface soil layer with mineral salts (bicarb<strong>on</strong>ate, carb<strong>on</strong>ate, chlor, sodium<br />
sulphate, magnesium).<br />
Sweet irrigati<strong>on</strong> waters (rain water) diminish the salinizati<strong>on</strong> effect with the diluti<strong>on</strong> of<br />
the soluble salts.<br />
For this reas<strong>on</strong>, shortly after being in use, the reacti<strong>on</strong> of the greenhouse soil tends to<br />
become neuter-alkaline or even alkaline, fact that has negative influences <strong>on</strong> the cultures<br />
development.<br />
The applicati<strong>on</strong> of the amendments is imposed when the value of pH exceeds 8.5.<br />
297
The total c<strong>on</strong>tent of soluble salts has to be adjusted and a correct system of soil fertilizati<strong>on</strong><br />
has to be established, for the greenhouse soil to corresp<strong>on</strong>d from the agrochemical point of<br />
view (Neaţă Gabriela, 2002).<br />
As regards the salinity tolerance it is well known that tomatoes do not support<br />
c<strong>on</strong>centrati<strong>on</strong>s over 0.17% in light soils, over 0.20% in medium soils and over 0.25% in<br />
compact soils (Maria Apahideanu, 1998).<br />
The salinity tolerance of the main greenhouse cultures is appreciated <strong>on</strong> the limits of the<br />
c<strong>on</strong>centrati<strong>on</strong>s (table 1):<br />
Culture<br />
Table 1 Salinity tolerance of the main greenhouse cultures<br />
(mineral residue, g/100g sol) (after Bernstein, 1987 cited by Rusu, 1993)<br />
The limit where the culture starts to<br />
be affected<br />
Fine<br />
texture<br />
Thick<br />
texture<br />
Middle<br />
texture<br />
The limit where the producti<strong>on</strong><br />
decreases with 50 %<br />
Thick<br />
texture<br />
Middle<br />
texture<br />
Fine<br />
texture<br />
1. Cucumbers 0,15 0,20 0,23 0,25 0,30 0,35<br />
2. Green<br />
peppers<br />
0,15 0,18 0,20 0,23 0,30 0,43<br />
3. Lettuce 0,15 0,18 0,20 0,23 0,30 0,43<br />
4. Cabbage 0,25 0,20 0,23 0,26 0,40 0,55<br />
5. Tomatoes 0,27 0,20 0,25 0,30 0,45 0,60<br />
6. Spinach 0,20 0,25 0,30 0,30 0,45 0,60<br />
The preventi<strong>on</strong> of the exaggerated increase of the salts c<strong>on</strong>tent is solved by taking some<br />
preventive and curative measures:<br />
� Applying fertilizers in best ec<strong>on</strong>omic doses, especially those with azoth;<br />
� Using some fertilizers with well balanced relati<strong>on</strong>s between the nutritive elements<br />
depending <strong>on</strong> the requests of the species with the needed reports;<br />
� Applicati<strong>on</strong> of extra-radicular fertilizati<strong>on</strong> during the vegetati<strong>on</strong> period as a measure of<br />
diminishing the dose applied <strong>on</strong> the soil;<br />
298
� The pH decrease in the greenhouse soil can be obtained by using oligotrophic peat<br />
(highly acid, with pH 3-4,5 and Ah of 90m.e./100g peat) or mesotrophic peat (with pH<br />
5-5,5 and Ah of 50-60m.e./100g peat)(Rusu, 1982);<br />
� Washing the soluble salts by irrigati<strong>on</strong>s, when the soil is permeable and the phreatic<br />
layer is under the critical level. In the greenhouse soils with difficult draining where<br />
prophylactic washings cannot be d<strong>on</strong>e, the low mineralized oligotrophic peat, which<br />
c<strong>on</strong>tains 90-95% organic matter, is applied;<br />
� Correcting the pH with gyps, when it exceeds the value of 8,2-8,4 and the PSA value is<br />
greater than 10% (Rusu, 1993);<br />
� On the soils with a low c<strong>on</strong>tent of mineral azoth it is necessary to apply 0,25-0,50 kg<br />
azoth for every t<strong>on</strong> of applied peat, in order to prevent a temporary immobilizati<strong>on</strong> of<br />
the soluble azoth.<br />
Depending <strong>on</strong> the applied doses and knowing the c<strong>on</strong>tributi<strong>on</strong> of the peat’s nutritive<br />
elements, it can be calculated the quantities of mineral fertilizers that have to be applied to<br />
ensure the plants’ needs regarding the adaptati<strong>on</strong> to different c<strong>on</strong>centrati<strong>on</strong>s of salts.<br />
REFERENCE<br />
1. AVARVAREI I., VELICICA DAVIDESCU, R. MOCANU, M.GOIAN, C. CARAMETE, M. RUSU,<br />
1997, Agrochimie, Ed. Sitech, Craiova;<br />
2. DAVIDESCU D., VELICICA DAVIDESCU, 1992, Agrochimie horticolă, Ed.Academiei<br />
Române, Bucureşti, 91-497;<br />
3. MARIA APAHIDEAN, 1998, C<strong>on</strong>tribuţii la stabilirea unei metode noi de cultură, “fără<br />
sol”, a tomatelor în seră, Teză de doctorat, Cluj-Napoca;<br />
4. R. CIOFU, NISTOR STAN, VICTOR POPESCU, PELAGHIA CHILOM, SILVIU APAHIDEAN,<br />
ARSENIE HORGOŞ, VIOREL BERAR, KARL FRITZ LAUER, NICOLAE ATANASIU, 2004,<br />
Tratat de legumicultură, Ed. CERES, Bucureşti, 129-136, 617-643;<br />
5. RUSU M, V. MUNTEANU, N. METEŞ, J. JANCU, 1988, Probleme ale optimizării<br />
agrochimice a solurilor sub influenţa măsurilor agrochimice, Analele ICCPT, vol.LVI,<br />
261-267;<br />
6. RUSU M., I. BOERIU, I. VLĂDUŢU, 1968, Potenţialele activităţii ale unor i<strong>on</strong>i în solurile<br />
acide calcarizate, Stiinţa Solului, nr.4, vol.VI, 26-32;<br />
7. RUSU M., V. MUNTEANU, I. IANCU, DANIELA DAVID, ALEXANDRA IONESCU, MARIA<br />
FLOREA, ANA STANCIU, 1982, Cercetări privind regimul microelementelor din sol în<br />
interacţiune cu îngrăşămintele şi amendamentele, Analele ICCPT, vol.XLIX, 121, 130;<br />
8. RUSU M., V. MUNTEANU, I. IANCU, MARILENA MĂRGHITAŞ, S. ALB, 1993, Efectul<br />
magneziului pe solurile modificate agrochimic, Bul. USAMV, A-H, Cluj-Napoca, 47/2,<br />
117-124;<br />
9. TISDALE S.L., 1993, Soil fertility and fertilizers, Macmillan Publ.Co. New-York, USA.<br />
299
THE INFLUENCE OF THE SEED’ BIOLOGICAL LINKS ON THE<br />
WHEAT PRODUCTION QUALITY<br />
Mariana Popovici<br />
University of Oradea, Faculty for Envir<strong>on</strong>mental Protecti<strong>on</strong><br />
ABSTRACT<br />
The quality of wheat producti<strong>on</strong> is a complex characteristic, depending <strong>on</strong> both quantitative<br />
elements (c<strong>on</strong>tent of useful substances: proteins, fats, carbohydrates etc.) and <strong>on</strong> their<br />
chemical compositi<strong>on</strong> (essential aminoacids, saturated and n<strong>on</strong>-saturated fat acids, etc.) but<br />
also <strong>on</strong> quantitative elements (physical, biochemical, mechanical and technological<br />
characteristics of the final products).<br />
INTRODUCTION<br />
The quality characteristic of the wheat is much more complex than the producti<strong>on</strong> capacity<br />
and it is influenced by several factors having a specific c<strong>on</strong>tributi<strong>on</strong> in its completi<strong>on</strong>; if <strong>on</strong>e<br />
adds the influence of the envir<strong>on</strong>mental factors and the modificati<strong>on</strong>s that occur in the<br />
technological process <strong>on</strong>e may complete this overall picture. Thus, the wheat quality cannot<br />
be expressed in <strong>term</strong>s depending <strong>on</strong> a single trait but it depends <strong>on</strong> numerous characteristics,<br />
each having its precise importance in assessing the value of the final product.<br />
The main way through which <strong>on</strong>e can take acti<strong>on</strong> <strong>on</strong> the quality and substance of the<br />
wheat proteins c<strong>on</strong>sists in their ameliorati<strong>on</strong>, all these characteristics being genetically<br />
c<strong>on</strong>trolled. The agricultural researches and practice showed that even the technological<br />
factors are making their c<strong>on</strong>tributi<strong>on</strong> to largely impact <strong>on</strong> the c<strong>on</strong>tent and the amount of<br />
protein producti<strong>on</strong> per hectare. Am<strong>on</strong>g these factors, <strong>on</strong>e can place first the chemical<br />
fertilisers, especially those based <strong>on</strong> azote (nitrogen) which c<strong>on</strong>diti<strong>on</strong> the aminoacids and<br />
proteins biosynthesis and, implicitly, the harvest’ quality. An important role is detained by<br />
the crops’ rotati<strong>on</strong>, irrigati<strong>on</strong>s, soil workings, which all compete to increase the<br />
efficaciousness of the fertilizers appliance.<br />
Within the performed experiments, the indices for quality of the autumns wheat are<br />
been showed <strong>on</strong> the basis of the analysis regarding the protein c<strong>on</strong>tent in gluten.<br />
MATERIAL AND METHOD<br />
Am<strong>on</strong>g the wheat quality indices, there have been analysed the following: the protein<br />
c<strong>on</strong>tent of the B,SC1,SC2 biological links and other “replicati<strong>on</strong>s” during the 2004-2005 and<br />
2005-2006 periods and the amount of gluten present in the biological links of “Dropia”<br />
autumn wheat, employed in the 2004-2006 two-year research period within the premises of<br />
the Leş producti<strong>on</strong> farm, Bihor County.<br />
300
RESULTS AND DISSCUSSIONS<br />
1. The protein c<strong>on</strong>tent influence of the biological links.<br />
The protein c<strong>on</strong>tent of the wheat grains is a hereditary trait, str<strong>on</strong>gly influenced both by the<br />
envir<strong>on</strong>mental c<strong>on</strong>diti<strong>on</strong>s and by the crop <strong>on</strong>es. The research carried-out clearly shows the<br />
str<strong>on</strong>g reacti<strong>on</strong> of the protein c<strong>on</strong>tent functi<strong>on</strong> of the ecological circumstances, the variati<strong>on</strong><br />
amplitude from <strong>on</strong>e locati<strong>on</strong> to another and, especially, from <strong>on</strong>e year to another is reaching<br />
5 up to 7%.<br />
Within the framework of the carried-out experiences, there have were carried out<br />
analysis <strong>on</strong> the protein c<strong>on</strong>tent of the B,SC1,SC2 biological links and other “replicati<strong>on</strong>s”<br />
and the results achieved are presented in the table 1 bellow.<br />
Table1: The impact of the biological links of wheat seed <strong>on</strong> the protein c<strong>on</strong>tent of grains<br />
Biological 2004-2006 2005-2006 2004-2006 Average<br />
category/class Protein % Protein % Protein %<br />
Base (B) mt 12.3 100.0 11.8 100.0 12.1 100.0<br />
SC1 12.0 97.6 11.8 100.0 11.9 98.3<br />
SC2<br />
Other<br />
11.2 91.1 11.3 95.7 11.3 93.3<br />
replicati<strong>on</strong>s I 8.4 70.7 000<br />
7.3 61.8 000<br />
Other<br />
7.9 65.3<br />
replicati<strong>on</strong>s II 7.1 57.7 000<br />
7.3 61.8 000<br />
7.2 59.5<br />
DL 5% 5.3 10.4<br />
DL 1% 14.4 21.5<br />
DL0.1% 25.4 30.4<br />
According with the obtained data, two main aspects are mainly emphasised, namely:<br />
� The biological link of the seed puts its mark <strong>on</strong> the protein c<strong>on</strong>tent. The differences<br />
between the biological links obtained and maintained through a c<strong>on</strong>servative<br />
ameliorati<strong>on</strong> process (B,SC1,SC2) are not significant <strong>on</strong>es, as they are maintained<br />
within some high-levelled parameters during the two-year research period. An evident<br />
depreciati<strong>on</strong> can be observed in the case of the “other replicati<strong>on</strong>s” in which case, the<br />
protein percentage diminishes to the statistically ensured quotas, but which are negative<br />
in comparis<strong>on</strong> with the Base (Mt) biological link. The striking differences are due to<br />
the lack of selecti<strong>on</strong> pressure exercised <strong>on</strong> this biological material, the biological n<strong>on</strong>purified<br />
amount thus negatively marks the wheat grains’ quality.<br />
� The differences, although less relevant, show the influence of the climatic year <strong>on</strong> the<br />
protein percentage, soil factors making their impact <strong>on</strong> the wheat harvest’ quality.<br />
301
2. The influence of the biological links of the autumn wheat’ seeds <strong>on</strong> the gluten<br />
c<strong>on</strong>tent.<br />
Am<strong>on</strong>g the comp<strong>on</strong>ent traits of the bread-making quality, a key-role is detained by the<br />
protein quantity present in the grain, especially of those proteins comp<strong>on</strong>ents of the gluten.<br />
The efficaciousness (force) of dough, the water absorpti<strong>on</strong>’ capacity and the bread tolerance<br />
to obsolescence, all largely rely <strong>on</strong> these latest proteins comp<strong>on</strong>ent.<br />
The bread volume depends in an essential manner <strong>on</strong> the gluten quality which, unlike<br />
the quality, is <strong>on</strong> the first hand genetically established, being very little influenced by the<br />
envir<strong>on</strong>mental c<strong>on</strong>diti<strong>on</strong>s. At last, the quality is influenced by the bio-chemical properties of<br />
the glutenins, main comp<strong>on</strong>ents of the gluten.<br />
The gluten has some properties which c<strong>on</strong>fer to it a unique positi<strong>on</strong> am<strong>on</strong>g the vegetal<br />
proteins, namely: cohesi<strong>on</strong>, elasticity and extensibility in hydrated state. As a c<strong>on</strong>sequence,<br />
the (wheat) flour characteristics are, <strong>on</strong> the first hand, a functi<strong>on</strong> of gluten and of its<br />
behaviour in aqueous envir<strong>on</strong>ment. The gluten analysis supposes an assessment of its<br />
c<strong>on</strong>tent and its quality.<br />
The data <strong>on</strong> gluten amount c<strong>on</strong>tained in the biological links of the<br />
(“Dropia”) autumn type of wheat for the 2004-2006 two-year period are<br />
being presented in the table 2 bellow.<br />
Table 2: The impact of the biological links of wheat seed <strong>on</strong> the gluten c<strong>on</strong>tent of the<br />
grains<br />
Biological 2004-2006 2005-2006 2004-2006 Average<br />
category/class Gluten % Gluten % Gluten %<br />
Base (B) mt 28.0 100.0 26.8 100.0 27.4 100.0<br />
SC1 28.0 100.0 26.0 97.0 27.0 98.5<br />
SC2 27.5 98.2 26.0 97.0 26.8 97.8<br />
Other<br />
replicati<strong>on</strong>s I 21.3 76.1 000<br />
19.9<br />
74.2 000<br />
20.6 75.2<br />
Other<br />
replicati<strong>on</strong>s II 20.4 72.8 000<br />
19.2<br />
71.6 000<br />
19.8 72.3<br />
DL 5% 12.3 11.4<br />
DL 1% 20.1 21.4<br />
DL0,1% 25.8 28.5<br />
The c<strong>on</strong>tent of humid gluten has a similar range with the protein c<strong>on</strong>tent. Thus, at the level<br />
of both years of research, the issue of the positive influence of the c<strong>on</strong>servative ameliorati<strong>on</strong><br />
process positively occurs. The gluten c<strong>on</strong>tent, given the c<strong>on</strong>diti<strong>on</strong>s of a c<strong>on</strong>trolled<br />
replicati<strong>on</strong> of seed’ producti<strong>on</strong>, varies almost imperceptibly; this small variati<strong>on</strong> de<strong>term</strong>ines<br />
us to take into c<strong>on</strong>siderati<strong>on</strong> that, in order to reach at a good quality bread producti<strong>on</strong>, <strong>on</strong>e<br />
302
may employ (wheat) flour produced from the SC2 biological link, having the same value of<br />
the humid gluten as in the case of the B and SC1 (biological) links.<br />
At the level of “other replicati<strong>on</strong>s” (biological) links the discrepancies are striking. The<br />
lack of selecti<strong>on</strong> pressure at the level of these replicati<strong>on</strong>s, leads inevitably to the qualitative<br />
decrease of the flour, which, as above signalised, is associated with a producti<strong>on</strong>. From all<br />
above menti<strong>on</strong>ed, it is occurs the fact that the proper seed for obtaining some top-quality<br />
bread-making products must be placed at the level of the SC2 seed type.<br />
CONCLUSIONS<br />
1. The gluten c<strong>on</strong>tent of the wheat is str<strong>on</strong>gly influenced by the biological category (class)<br />
of the seed used, high differences c<strong>on</strong>cerning the c<strong>on</strong>tent occur between the biological<br />
links obtained through ameliorati<strong>on</strong> (B,SC1,SC2) and “other replicati<strong>on</strong>s”.<br />
2. The gluten c<strong>on</strong>tent, in a similar manner with the protein c<strong>on</strong>tent, has increased values in<br />
the case of B,SC1,SC2 biological categories (classes) while the “other replicati<strong>on</strong>s”<br />
category has quite reduced values, from which it results the fact that the seed used to<br />
obtain bakery products must be placed at the level of the SC2 seed.<br />
3. The protein c<strong>on</strong>tent and the gluten <strong>on</strong>e show less significant differences in the two-year<br />
research period; these are due to the soil and climatic factors.<br />
REFERENCES<br />
1. Haş, Ivan (2006)-Producerea seminţelor la plantele agricole, Editura Academic Pres,<br />
Cluj-Napoca<br />
2. Munteanu, S.L., I.Borceanu, M.Axinte, G.V.Roman(2001) - Fitotehnie, Editura I<strong>on</strong><br />
I<strong>on</strong>escu de la Brad, Iaşi<br />
3. Savatti,M., G.Nedelea, M.Ardelean,(2004)-Tratat de ameliorare a plantelor,Editura<br />
Marineasa,Timişoara<br />
303
THE INFLUENCE OF SOIL OIL POLLUTION ON AGRICULTURAL CROPS<br />
Maria Şandor, Nicu Cornel Sabău, Cornel Domuţa, Cristian Domuţa, Radu Brejea<br />
Agricultural Research Stati<strong>on</strong> Oradea<br />
The Faculty for Envir<strong>on</strong>mental Protecti<strong>on</strong>, University of Oradea<br />
Calea Aradului St. No. 5. email: scdaoradea@yahoo.com.<br />
Gen Magheru Blvd., No. 26.email: nsabau@uoradea .ro.<br />
ABSTRACT<br />
The paper presents the results of researches reguarding oil polluted soils, that took place<br />
at the Agricultural Research stati<strong>on</strong> in Oradea, from 1993 to 2002.<br />
The experimental device was made out of 1 m² microparcels, spread out in a random<br />
order in a Latin square; these parcels were polluted under c<strong>on</strong>trol with petroleum from<br />
Suplacu de Barcău, Bihor County, with the following c<strong>on</strong>centrati<strong>on</strong>s : 0, 1, 3, 5 and 10<br />
% petroleum <strong>on</strong> the ploughed layer, with 4 repetiti<strong>on</strong>s.<br />
The experience was set out <strong>on</strong> a luvosoil , and the soil was cultivated with millet, a<br />
plant which is c<strong>on</strong>sidered to be tolerant to soil polluti<strong>on</strong>, in the first 3 years, and with<br />
spring wheat in the last 7 yeras of research.<br />
The results of the research have shown that the yield losses are proporti<strong>on</strong>al with<br />
the petroleum c<strong>on</strong>centrati<strong>on</strong>, and had a descending evoluti<strong>on</strong>. For instance, in the case<br />
of the 1% polluti<strong>on</strong>, losses are insignificant after 7 years of crops.<br />
Keywords: polluti<strong>on</strong>, petroleum, biodegradati<strong>on</strong>.<br />
INTRODUCTION<br />
Soil polluti<strong>on</strong> with oil residue is a very complex phenomen<strong>on</strong> which involves knowing<br />
the chemical nature and c<strong>on</strong>centrati<strong>on</strong> of the pollutive agent and the soil c<strong>on</strong>diti<strong>on</strong>s.<br />
Polluti<strong>on</strong> with oil residue is manifested especially in the upper layer of the soil, but in<br />
more seriuos cases of polluti<strong>on</strong>, effects were encountered at 80 cm depth <strong>on</strong> the soil<br />
profile, the depth at which the pollutive agent got into the soil was influenced by<br />
quantity, time of acti<strong>on</strong> <strong>on</strong> the soil, local microenvir<strong>on</strong>ment, phisycal and chemical soil<br />
properties.<br />
Oil extracti<strong>on</strong>, processing and transport in Bihor took place at the sites in Suplacu<br />
de Barcău, Marghita and Oradea, which have become nowadays stati<strong>on</strong>s for OMV and<br />
Petrolsub SA Suplacu de Barcău Refinery, today in c<strong>on</strong>servati<strong>on</strong>. Following these<br />
activities, the soil is affected by histortical polluti<strong>on</strong> <strong>on</strong> a surface of 200 ha, and is in<br />
need of measurements of ecological rehabilitati<strong>on</strong>. [3.]<br />
The researches carried out in Romania by Toti Mh (2003) c<strong>on</strong>cerning the polluti<strong>on</strong><br />
effects <strong>on</strong> agricultural land from the Southern part of Romania , have prooved that the<br />
plant`s average life expectati<strong>on</strong> diminished after a polluti<strong>on</strong> of 1kg waste / m² (0,3%) in<br />
the ploughed layer. [4.] The authors c<strong>on</strong>sider that a polluti<strong>on</strong> of 1,5 – 3,0 kg waste / m²<br />
is a moderate <strong>on</strong>e, between 3 kg – 15 kg waste / m² the polluti<strong>on</strong> becomes str<strong>on</strong>g, and<br />
between 15 – 30 kg waste / m² it is extremely str<strong>on</strong>g, and thus the plants seeds no l<strong>on</strong>ger<br />
germinate, and over 30 kg waste / m² it is excessive.<br />
304
For the c<strong>on</strong>diti<strong>on</strong>s in from Western Romania, Colibaş I publishes in 1995 the first<br />
partial results of researches regarding yield losses in millet, in the first year of<br />
c<strong>on</strong>trolled polluti<strong>on</strong> with different doses of petroleum. [2.]<br />
MATERIALS AND METHOD<br />
The researches carried out in Oradea wanted to establish the effects c<strong>on</strong>trolled polluti<strong>on</strong><br />
with petroleum from Suplacu de Barcău had <strong>on</strong> agricultural yield and <strong>on</strong> the<br />
biodegradati<strong>on</strong> period, without any ameliorative measures.<br />
The oil reserve in Suplacu de Barcău is located at relatively small depth, in a layer<br />
of P<strong>on</strong>tian sands, and has a high c<strong>on</strong>tent of asphaltines.<br />
Almost half the soils in Romania (49,397%) which are affected by polluti<strong>on</strong> with<br />
oil are luvosoils, and that the soil from Suplacu de Barcău is also a luvosoil, the<br />
experience carried out was placed also <strong>on</strong> a luvosoil. (Table 1.)<br />
Table 1: Some physical and chemical properties of the luvosoil from Oradea<br />
Horiz<strong>on</strong>t Depth Textural DA<br />
(cm) Class (g/cm 3 pH V Humus N C/N<br />
) (H2O) (%) (%) (%)<br />
Ap 0-25 LP 1,35 5,51 64,3 2,40 0,116 13,0<br />
El 25-44 LP 1,48 5,78 67,3 2,23 0,111 13,5<br />
Bt1w 44-60 TT 1,56 6,24 77,9 1,91 0,096 13,5<br />
Bt2 60-93 TT 1,58 6,46 83,4 1,73 0,087 13,4<br />
BC 93-110 TT 1,62 6,51 86,6 0,70 0,036 13,3<br />
C 110-152 TT 1,61 6,60 92,4 0,50 0,026 13,2<br />
The soil is dusty and with a c<strong>on</strong>tent of clay, in the worked layer and in the eluvial <strong>on</strong>e,<br />
and with medium clay in the layers underneath these <strong>on</strong>es. The values of the bulk<br />
density are medium in the eluvial layer and becomes large in the layer where clay<br />
gathers, and at the base of the profile, which indicates that the soil is poorly ploughed at<br />
the surface and poorly settled from a 25 – 44 cm depth , and moderately settled <strong>on</strong> the<br />
rest of the profile.[1.] The soil reacti<strong>on</strong> is moderately acid in the first two horiz<strong>on</strong>s and<br />
weak acid deeper. The total humus and N c<strong>on</strong>tent is small in the ploughed layer, and the<br />
values of the C/N index are high, which comes to prove a poor N supply in the soil.<br />
The experimental field set out in 1993 is made out of parcels of 1m² set out in a<br />
latin square, randomised, in four repetiti<strong>on</strong>s, which were willingly polluted with<br />
petroleum from Suplacu de Barcău with 0, 3, 9, 15, and 30 l/m², thus resulting<br />
c<strong>on</strong>centrati<strong>on</strong>s of 0 in the ploughed layer (unpolluted witness) and 1, 3, 5, 10 %.<br />
The field was cultivated in the first three years with millet (1993 – 1995) a plant<br />
with a high tolerance to polluti<strong>on</strong>, and afterwards for the next seven years with spring<br />
wheat, Speranţa breed.<br />
RESULTS AND DISCUSSIONS<br />
1. The climate c<strong>on</strong>diti<strong>on</strong>s, characterised by rainfall and by annual temperatures,<br />
show that the period of the ten years observed had more rainfall than the average multi<br />
annual with 26,2 m and was warmer with 0,5 ºC.<br />
305
The annual rainfall was between 367 mm in 2000 and 886 mm in 1996, the deviati<strong>on</strong>s<br />
from the normal standard values ranged from – 268mm to +251 mm. Temperatures<br />
varied between 9,6 – 12 ºC, these values were registered in 1996 and 2000, and in<br />
comparis<strong>on</strong> with the multi annual average they had a -0,9 – +1,5 ºC variati<strong>on</strong>.<br />
2. The annual millet producti<strong>on</strong> of the unpolluted parcels, vary <strong>on</strong>e from another,<br />
because of the different climate c<strong>on</strong>diti<strong>on</strong>s in the three years of research, starting at a<br />
very small yield (1993 – 17,6 q/ha hay) and grew to 44,2 q/ha. The producti<strong>on</strong>s from<br />
1995 are smaller than the <strong>on</strong>es in the previous year, but the difference of 3,5 q/ha is<br />
insignificant.<br />
The various petroleum c<strong>on</strong>centrati<strong>on</strong> in the ploughed layer de<strong>term</strong>ined millet hay<br />
producti<strong>on</strong>s, correlated linearly. The inverse linearly correlati<strong>on</strong>s are very significant<br />
statistically, for every <strong>on</strong>e of the years researched, the correlati<strong>on</strong> coefficients are : R =<br />
0,9045 in the first year of c<strong>on</strong>trolled polluti<strong>on</strong>, R = 0,8456 for the 2 nd year of c<strong>on</strong>trolled<br />
polluti<strong>on</strong> and R = 0,8615 for the 3 rd year. (Figure 1.)<br />
Millet yields (q/ha)<br />
60<br />
50<br />
40<br />
30<br />
20<br />
10<br />
0<br />
1993 y = -1.026x + 15.994<br />
1994 y = -2.6898x + 37.401<br />
1995 y = -1.8137x + 38.192<br />
0 2 4 6 8 10<br />
Petroleum c<strong>on</strong>centrati<strong>on</strong> in ploughed layer (%)<br />
1993 1994 1995<br />
Figure 2: The influence of petroleum polluti<strong>on</strong> <strong>on</strong> millet hay producti<strong>on</strong> (1993-1995)<br />
The slope of the regressi<strong>on</strong> lines for the years points out that the average yield losses,<br />
registered with the increase in the petroleum c<strong>on</strong>centrati<strong>on</strong> with 1 % , are of 1,026 q/ha<br />
millet hay, in the first year of c<strong>on</strong>trolled polluti<strong>on</strong>, 2,6898 q/ha millet hay in 1994 and<br />
1,8137 q/ha in the 3 rd year of research. In order to compare the yields from the polluted<br />
and the unpolluted witness parcel, seeing that the yields in the witness were very<br />
different in the 3 years of research, the differences between the polluted and unpolluted<br />
parcels were used, expressed in percentages.<br />
The relative yield losses of the polluted parcels, from the three years of study have<br />
had an evoluti<strong>on</strong> described best by a polinominal curve of the II degree. The<br />
correlati<strong>on</strong>s that were established like this show correlati<strong>on</strong> coefficients with no<br />
statistical insurance for the 1% c<strong>on</strong>centrati<strong>on</strong> R= 0,06790, statistically significant for the<br />
c<strong>on</strong>centrati<strong>on</strong> 3 %, R=8890 and R=0,9520, distinctively significant, for the<br />
c<strong>on</strong>centrati<strong>on</strong>s of 5 % and 10 %. (Figure 2.).<br />
306
Yield losses millet hay (%)<br />
80<br />
70<br />
60<br />
50<br />
40<br />
30<br />
20<br />
10<br />
0<br />
3% y = -14.385x 2 + 57364x - 6E+07<br />
R = 0.6790 *<br />
10 % y = -10.806x 2 + 43086x - 4E+07<br />
R = 0.9520**<br />
5 % y = -22.876x 2 + 91227x - 9E+07<br />
R = 0.8890**<br />
1993 1994 1995<br />
3% 5% 10%<br />
Figure 2. The evoluti<strong>on</strong> of relative yield losses (%) 1993 – 1995<br />
If we analyse the shape of evoluti<strong>on</strong> curves for the yield losses we notice that these<br />
curves have grown in the sec<strong>on</strong>d year of experiments, for all the variants of petroleum<br />
c<strong>on</strong>centrati<strong>on</strong>, and that year also marked significant yield losses : 23,08 % for the 1 % ,<br />
42,31 % for the 3 %, 61,54 % for 5 % and 65,61 % for the maximum c<strong>on</strong>centrati<strong>on</strong> of<br />
10 %. In the 3 rd year of c<strong>on</strong>trolled polluti<strong>on</strong>, the yield losses diminish indicating, the<br />
possibility of the soil naturally recovering from the polluti<strong>on</strong>.<br />
The average yield for an experience cultivated with millet (1993 – 1995) was of<br />
34,2 q/ha in the unpolluted witness variant, and ranged between 28,3 – 14,5 q/ha in the<br />
variants with c<strong>on</strong>trolled polluti<strong>on</strong>, bigger where the c<strong>on</strong>centrati<strong>on</strong> was 1% and smaller<br />
at a c<strong>on</strong>centrati<strong>on</strong> of 10 %. (Table 2).<br />
The statistical analysis of the losses from the medium yields (5,9 – 19,7 q/ha) show<br />
that they are all significant, in all the c<strong>on</strong>centrati<strong>on</strong>s studied. The annual yield<br />
differences for 1 % polluti<strong>on</strong> in the ploughed layer are significant in the first year (-34<br />
q/ha), significantly different in the sec<strong>on</strong>d year (-10,2 q/ha) and do not have any<br />
statistical importance in the 3 rd year (-5,9 q/ha). The variant polluted with 3 % shows<br />
significant differences <strong>on</strong>ly in the first two years (-5,5 and – 18,7 q/ha), just to become<br />
<strong>on</strong>ly significant from a statistical point of view in the 3 rd year (-10 q/ha).<br />
The differences from the 3 rd year of observati<strong>on</strong>, insignificant for the 1%<br />
c<strong>on</strong>centrati<strong>on</strong>, suggest that most of the oil residue have been biodegraded and the soil is<br />
recovering ecologically. The fact that the 3 % shows also <strong>on</strong>ly some small differences<br />
can suggest that the biological recovery has started.<br />
These speculati<strong>on</strong>s are also sustained by the statistical analysis of the interacti<strong>on</strong>.<br />
Variants (c<strong>on</strong>centrati<strong>on</strong>s) x years indicates that the average yield losses are insignificant<br />
when talking about the polluti<strong>on</strong> of the ploughed land with 1 % and distinctively<br />
significant when it comes to 3 and 5 % c<strong>on</strong>centrati<strong>on</strong> of petroleum.<br />
307
Table 2: Yield losses millet hay due to petroleum poluti<strong>on</strong>. (q/ha) 1993-1995<br />
Variant<br />
Years (Y) Average Semnificati<strong>on</strong><br />
(V) 1993 1994 1995 1993-1995<br />
(%) (q/ha) (q/ha) (q/ha) q/ha Diference V VxY<br />
0 - - - 34,2 - - -<br />
1 -3,4 -10,2 -4,0 28,3 -5,9 ooo -<br />
3 -5,5 -18,7 -10,0 22,2 -11,4 ooo oo<br />
5 -7,4 -27,2 -14,3 17,9 -16,3 ooo oo<br />
10 -11,2 -29,2 -18,7 14,5 -19,7 ooo ooo<br />
1993 1994 1995 Variant (V) Variant x Years (VxY)<br />
DL 5 % 2,46 5,25 5,56 2,49 7,83<br />
DL 1 % 3,46 7,37 7,80 3,33 11,39<br />
DL 0,1 % 4,88 10,41 11,02 4,39 17,09<br />
3. Average annual yield in spring wheat from the unpolluted witness parcels, were<br />
between 21,8 q/ha , in the 4 th year of observati<strong>on</strong> (1996) and 4 q/ha in 2000 (which was<br />
c<strong>on</strong>sidered to be <strong>on</strong>e of the years with most drought ever) (Figure 3.)<br />
Yields (q/ha)<br />
25<br />
20<br />
15<br />
10<br />
5<br />
0<br />
2002<br />
2001<br />
2000<br />
1999<br />
1998<br />
1997<br />
0 1 3 5 10 1996<br />
Petroleum c<strong>on</strong>centrati<strong>on</strong> (%)<br />
Figure 3. Annual spring wheat yield (q/ha)<br />
The yields from the variant`s repetiti<strong>on</strong> had a linearly evoluti<strong>on</strong> in each and every <strong>on</strong>e<br />
of the seven years with spring wheat crop (1996-2002), inverse proporti<strong>on</strong>al with the<br />
petroleum c<strong>on</strong>centrati<strong>on</strong> from the ploughed layer. The linearly correlati<strong>on</strong>s thus<br />
established are significant statistically, in the first five years of the interval (R = 0,8096<br />
308
– 0,6093) and extremely significant in the last two years (R = 0,4539 – 0,4058) (Table<br />
3.)<br />
Table 3. Regressi<strong>on</strong> equati<strong>on</strong>s between spring wheat yield and petroleum<br />
c<strong>on</strong>centrati<strong>on</strong> (1996-2002)<br />
Nr. Anul Ecuaţia Coeficient de Semnificaţie<br />
crt.<br />
corelaţie<br />
1 1996 Y = - 1,4818 X + 18,271 R = 0,8723 oo<br />
2 1997 Y = - 1,2357 X + 19,696 R = 0,8906 oo<br />
3 1998 Y = - 0,4377 X + 7,9734 R = 0,7833 oo<br />
4 1999 Y = - 0,4169 X + 7,7041 R = 0,7531 oo<br />
5 2000 Y = - 0,1430 X + 3,9634 R = 0,6093 oo<br />
6 2001 Y = - 0,0885 X + 5,6811 R = 0,4539 o<br />
7 2002 Y = - 0,3566 X + 10,0100 R = 0,4058 o<br />
The relative producti<strong>on</strong> losses for spring wheat range between 24,3 % for 1 %<br />
c<strong>on</strong>centrati<strong>on</strong> to 74,7 % for 10 % c<strong>on</strong>centrati<strong>on</strong>. These losses are bigger than the <strong>on</strong>es<br />
registered during the previous year in millet. This is because wheat is less tolerant to<br />
polluti<strong>on</strong> than millet.<br />
The evoluti<strong>on</strong> of relative yield losses in spring wheat from the last seven years of<br />
observati<strong>on</strong> has a rising tendency, described in polynomial equati<strong>on</strong>s of the 2 nd degree,<br />
extremely significant for high c<strong>on</strong>centrati<strong>on</strong>s (5 and 10 %), significant for 3 % and with<br />
no importance for a 1 % c<strong>on</strong>centrati<strong>on</strong> of petroleum. (Figure 4.)<br />
The average values of relative yield losses decrease annually and reach about 10 –<br />
15 % for the small 1-3 % c<strong>on</strong>centrati<strong>on</strong>s, 25 % for the 5 % c<strong>on</strong>centrati<strong>on</strong> and 37 % for<br />
the maximum 10 % c<strong>on</strong>centrati<strong>on</strong>. The average yield differences in spring wheat from<br />
the last seven years of research in spring wheat polluti<strong>on</strong> are of 1,1 q/ha for a 1 %<br />
polluti<strong>on</strong> – significant statistically, 3,3 q/ha for 3 %, 5,0 q/ha at 5 % and 6,3 % for 10 %,<br />
the last two are very significant. (Table 4.)<br />
The interacti<strong>on</strong> (petroleum c<strong>on</strong>centrati<strong>on</strong>) x years does not have any statistical<br />
value for the variant polluted under c<strong>on</strong>trol with 1 %, and is significant for the<br />
c<strong>on</strong>centrati<strong>on</strong> of 3 % and extremely significant for the 5 % and 10 % c<strong>on</strong>centrati<strong>on</strong> in<br />
the first 20 cm of the soil profile.<br />
If we analyse the annual yield losses evoluti<strong>on</strong> from the period in which spring<br />
wheat was cultivated we notice that in the variant polluted with 1 % petroleum, the yield<br />
losses are extremely significant statistically in 1996, significant in 1997 – 1999, <strong>on</strong>ly be<br />
become insignificant in the last three years of the research.<br />
In the variant with a 10 % polluti<strong>on</strong>, the yield in the last year is not significant<br />
statistically, is significant in the ninth year, distinctively significant in the eighth year,<br />
and very significant in the first seven years.<br />
309
Yields losses, spring wheat (%)<br />
90<br />
80<br />
70<br />
60<br />
50<br />
40<br />
30<br />
20<br />
10<br />
0<br />
-10<br />
-20<br />
-30<br />
-40<br />
-50<br />
-60<br />
3% y = 0.4101x 2 - 1645.2x + 2E+06<br />
R = 0.4791 *<br />
10 % y = 1.0655x 2 - 4267.5x + 4E+06<br />
R = 0.7760 **<br />
1996 1997 1998 1999 2000 2001 2002<br />
5% y = 0.8161x 2 - 3269.5x + 3E+06<br />
R = 0.7144 **<br />
3% 5% 10%<br />
Figure 4. The evoluti<strong>on</strong> of relative yield losses (1996 – 2002)<br />
Tabel 4 Yield losses due polluti<strong>on</strong> with petroleum in spring wheat (q/ha)<br />
1996 - 2002<br />
Years<br />
No. Variant<br />
Yield<br />
Semnificati<strong>on</strong><br />
Crt. (V)<br />
1996-2002<br />
(%) q/ha Difference V V x Y<br />
1 0 11,5 - - -<br />
2 1 9,6 -1,1 o -<br />
3 3 8,2 -3,3 ooo o<br />
4 5 6,5 -5,0 ooo ooo<br />
5 10 5,2 -6,3 ooo ooo<br />
D.L. Variant Variant x Years<br />
(V)<br />
(VxY)<br />
5 % 1,0 2,7<br />
1 % 1,5 3,6<br />
0,1 % 2,1 4,8<br />
If we sum up all this data we see that the annual yield losses loose their statistical value<br />
after the 7 th year of c<strong>on</strong>trolled polluti<strong>on</strong>, for a 1 % c<strong>on</strong>centrati<strong>on</strong>, after the 8 th year in 3<br />
% and 5 % petroleum c<strong>on</strong>centrati<strong>on</strong>, and in the 9 th year for a 10 % c<strong>on</strong>centrati<strong>on</strong>.<br />
310
CONCLUSIONS<br />
The experimental field with microparcels, set out in a randomised latin square, from the<br />
luvosoil at the Agricultural Research Stati<strong>on</strong> Oradea, was polluted in a c<strong>on</strong>trolled<br />
manner with petroleum brought from Suplacu de Barcău, in different c<strong>on</strong>centrati<strong>on</strong>s of<br />
0, 1 ,3, 5 and 10 % in four repetiti<strong>on</strong>s. Between 1993 – 1995 the field was cultivated<br />
with millet, a plant which is c<strong>on</strong>sidered to be tolerant to polluti<strong>on</strong> (first 3 years) and<br />
with spring wheat Speranţa for the next 7 years.<br />
The annual yields in the variants with different c<strong>on</strong>centrati<strong>on</strong>s of polluti<strong>on</strong> are<br />
linearly correlated both for millet and wheat, the correlati<strong>on</strong>s are distinctively<br />
significant for the first three years of millet crop and for the next 5 years of wheat crop.<br />
The evoluti<strong>on</strong> of relative yield losses from the research period are described by a<br />
polynomial regressi<strong>on</strong> of the sec<strong>on</strong>d degree, both for wheat and millet. They are<br />
significant for the variant polluted with 3 %, distinctively significant for the variants<br />
polluted with 5 and 10 % and with no statistical significance for the 1 % c<strong>on</strong>centrati<strong>on</strong>.<br />
The biggest yield losses in millet crop were registered in the 2 nd year of research, <strong>on</strong>ly<br />
to decrease in the 3 rd .<br />
The average yield losses in three years, in millet were very significant, and ranged<br />
between 5,9 q/ha hay for the variant of 1% petroleum and 19,7 q/ha hay for the 10 %<br />
petroleum. The influence of the interacti<strong>on</strong> variant x years of growing show that these<br />
differences are insignificant for the 1 %, distinctively significant for the 3 and 5 % and<br />
very significant for the 10 % c<strong>on</strong>centrati<strong>on</strong>.<br />
In wheat, the average yield losses for seven years are significant for the small 1 %<br />
c<strong>on</strong>centrati<strong>on</strong> and very significant for the other variants. The statistical significance of<br />
the yield losses from the 4 th year of experience come to prove that wheat is less tolerant<br />
to petroleum polluti<strong>on</strong> than millet.<br />
The evoluti<strong>on</strong> of annual yield losses show that the oil residue in the soil are<br />
biodegraded, the periods of time needed to do that are direct proporti<strong>on</strong>al with the<br />
initial c<strong>on</strong>centrati<strong>on</strong>s. Annual yield losses loose their statistical significance after 7<br />
years in crop at a 1 % c<strong>on</strong>centrati<strong>on</strong>, 8 years in average c<strong>on</strong>centrati<strong>on</strong> of 3-5 % and 9<br />
years in big c<strong>on</strong>centrati<strong>on</strong>s of 10 %. The yields from the 10 th year of experiments are<br />
still smaller than in the unpolluted variant with 12 – 13 % at the variant polluted with 1<br />
– 3 %, 25,6 % at the 5 % and 36,9 % for the 10 %.<br />
REFERENCES<br />
1. Colibaş I, Colibaş Maria, Tirpe Gh (2000): Solurile brune luvice, caracterizare şi<br />
ameliorare – Ed. Mirt<strong>on</strong> Timişoara.<br />
2. Colibaş I., Colobaş Maria, Şandor Maria (1995): Măsuri de ameliorare a solurilor<br />
poluate cu rezidii petroliere. – Cum să cultivăm pământul în z<strong>on</strong>a centrală din<br />
vestul ţării – Staţiunea de Cercetări Agrozootehnice Oradea.<br />
3. Sabău N.C., Domuţa C., Berchez O. (2002): Geneza degradarea şi poluarea solului,<br />
Partea a II-a Degradarea şi poluarea solului – Editura Uniersităţii din Oradea;<br />
4. Toti Mh., Dumitru Mh., Rovena Voiculescu Anca, Mihalache Mh., Mihalache<br />
Gabi, C<strong>on</strong>stantinescu Carolina (2003): Metodologia de biodegradare a solurilor<br />
poluate cu ţiţei, cu ajutorul microorganismelor specifice selecţi<strong>on</strong>ate din microflora<br />
autoht<strong>on</strong>ă – Edituta GNP Minischool;<br />
311
EFFECT OF BIOFERTILIZATION ON PARSLEY YIELD AND N CONTENT<br />
ON TWO SOILS<br />
Ida Kincses 1 - Tibor Filep 2 - Andrea B. Kovács 1 - Péter T. Nagy 1 – Imre Vágó 1<br />
1 University of Debrecen, Department of Agricultural Chemistry, Debrecen<br />
2 Research Institute for Soil Science and Agricultural Chemistry of the Hungarian<br />
Academy of Sciences, Budapest<br />
ABSTRACT<br />
A pot experiment was carried out to investigate effect of N fertilizer and biofertilizer <strong>on</strong><br />
parsley (Petroselinum tuberosum) yield and N c<strong>on</strong>tent of plant, as well as the 0.01 M<br />
CaCl2 soluble N c<strong>on</strong>centrati<strong>on</strong>s in soils. Two soils were used, <strong>on</strong>e of them is a<br />
chernozem with slightly acidic pH and with high c<strong>on</strong>tent of organic matter, and the<br />
other <strong>on</strong>e is an acidic sandy soil with low organic matter c<strong>on</strong>tent.<br />
On chernozem, the N fertilizati<strong>on</strong> significantly increased the yield of plant, while<br />
there was no significant biofertilizer effect. On sandy soil, the dry matter of plants<br />
increased with increasing N doses comparing to c<strong>on</strong>trol, but no correlati<strong>on</strong> has been<br />
found between biofertilizer and dry matter weight.<br />
Investigating N c<strong>on</strong>tent of plants <strong>on</strong> chernozem soil, it was found that N<br />
fertilizati<strong>on</strong> enhanced the N c<strong>on</strong>centrati<strong>on</strong> in plant, but there was no relati<strong>on</strong>ship<br />
between N c<strong>on</strong>tent and biofertilizer rates at all. For sandy soil, at high rates of N the<br />
biofertilizati<strong>on</strong> caused an increase in N c<strong>on</strong>centrati<strong>on</strong> of plant, while at lower doses just<br />
the opposite was noticed. So between two soils there was a remarkable difference: in<br />
case of acidic sandy soil, the adding of biofertilizer caused an increase in both plant<br />
biomass and in N c<strong>on</strong>tent of leaf. These effects are not appeared for chernozem. Due to<br />
the very low N supply ability of sandy soil, the increment of microbes due to<br />
biofertilizati<strong>on</strong> caused N immobilizati<strong>on</strong>, because the enhanced microbial biomass take<br />
up soluble N.<br />
Based <strong>on</strong> data from experiment, it can be c<strong>on</strong>cluded that positive effect of<br />
biofertilizati<strong>on</strong> <strong>on</strong> yield, N c<strong>on</strong>tent of plant and soil displayed <strong>on</strong>ly <strong>on</strong> chernozem soil<br />
during that short period of investigati<strong>on</strong>. On the acidic soil with low organic matter<br />
c<strong>on</strong>tent, these effects are no appeared because of N immobilizati<strong>on</strong> by microbes.<br />
Keywords: N fertilizati<strong>on</strong>, biofertilizati<strong>on</strong>, parsley, pot experiment, 0,01M CaCl2<br />
soluble N-forms<br />
INTRODUCTION<br />
In horticulture cropping, the main goal is to reach appropriate quality and quantity of<br />
crops, which mainly de<strong>term</strong>ined by genetics parameters of plants, ecological<br />
circumstances and the nutrient status of soil. The nutrient supply is the <strong>on</strong>e of the most<br />
important factor to get maximum yields. In our country, the nutrient supply is based <strong>on</strong><br />
adding to soils chemical fertilizers.<br />
Biofertilizers are an alternative way to substitute mineral fertilizers for increasing<br />
soil fertility and crop producti<strong>on</strong>. Biofertilizers are products c<strong>on</strong>taining living cells of<br />
different types of microorganisms, which have an ability to c<strong>on</strong>vert important elements<br />
from unavailable to available form through biological processes (HEGDE et al., 1999).<br />
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Evaluati<strong>on</strong> of effect of fertilizati<strong>on</strong> <strong>on</strong> plants needs an examinati<strong>on</strong> <strong>on</strong> plant-soil system.<br />
Amount, biodiversity and activity of living beings in soils depend <strong>on</strong> physical and<br />
chemical parameters, while soil microorganism impact soil processes (SZABÓ, 1986).<br />
Applicati<strong>on</strong> of chemical fertilizers may cause acidificati<strong>on</strong> in soils due to its<br />
chemical attributes (CHANDER – ABROL, 1972; FELIZARDO et al., 1972; POWER,<br />
1972). To decrease the harmful effects of fertilizati<strong>on</strong>, farmers are trying to decrease the<br />
amount of chemicals, and to substitute that to manure or biofertilizers, which are<br />
permitted in biofarming. In biofarming, the sustainability and improving of soil fertility<br />
plays the greatest role (AUBERT, 1981; NAGY et al.2001).<br />
Our experimental data can be used, in the main objective of crop producti<strong>on</strong>, in<br />
improving of envir<strong>on</strong>mental protecti<strong>on</strong>, and the site-specific fertilizati<strong>on</strong>.<br />
MATERIALS AND METHODS<br />
An experiment was carried out with parsley in spring of 2006 with 12 treatments and 4<br />
replicati<strong>on</strong>s at the Experimental Site of Debrecen University. Soils used were from<br />
Latókép (calcouorus chernozem) and Újfehértó (acid arenosol). Important parameters of<br />
soils are the followings:<br />
Látókép: pH (H2O): 6.7; pH(KCl): 5.9; pH(CaCl2): 6.23; Humus (%): 2.5; KA: 42;<br />
Újfehértó: pH (H2O): 4.1; pH(KCl): 3.7; pH(CaCl2): 6.23; Humus (%): 1.3; KA: 27;<br />
10 kg of Látókép and 12 kg of Újfehértó soil was dispensed to pots. The N, P and K<br />
treatments were added as NH4NO3, KH2PO4, KCl soluti<strong>on</strong>. The amount of applied<br />
biofertilizer was in accordance with the prescribed volume and two times of that.. The<br />
Phylaz<strong>on</strong>it MC biofertilizer c<strong>on</strong>tained carboxi-methyl-cellulose (CMC), microelements,<br />
Azotobacter croococcum and Bacillus megaterium soil bacteria, heteroauxin, gibberelin,<br />
vitamin B. Treatmnets applied were in Table 1.<br />
Table 1: Combinati<strong>on</strong> of treatments<br />
Biofertilizer ml/pot<br />
Treatment N g/pot Látókép Újfehértó P g/pot K g/pot<br />
1 0.2 70 80 0.1 0.3 N1*B1<br />
2 0.2 35 40 0.1 0.3 N1*B2<br />
3 0.2 - - 0.1 0.3 N1*B3<br />
4 0.1 70 80 0.1 0.3 N2*B1<br />
5 0.1 35 40 0.1 0.3 N2*B2<br />
6 0.1 - - 0.1 0.3 N2*B3<br />
7 0.05 70 80 0.1 0.3 N3*B1<br />
8 0.05 35 40 0.1 0.3 N3*B2<br />
9 0.05 - - 0.1 0.3 N3*B3<br />
10 - 70 80 0.1 0.3 N4*B1<br />
11 - 35 40 0.1 0.3 N4*B2<br />
12 - - - 0.1 0.3 N4*B3<br />
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Seeds were sown <strong>on</strong> 5th may 2006. Pots were irrigated daily with dei<strong>on</strong>ized water<br />
during the experiment, according to 60 % of maximum water capacity of soils. Plants<br />
were harvested 20 th of July and soil samples were taken.<br />
Beside plant biomass, the C, N and S c<strong>on</strong>tent of plant also were measured by a dry<br />
combusti<strong>on</strong> method. The 0.01 M CaCl2 soluble N and P c<strong>on</strong>tent of soil (HOUBA et al.,<br />
1994) was de<strong>term</strong>ined by a c<strong>on</strong>tinues flow equipment.<br />
RESULTS AND DISCUSSIONS<br />
We would like to get results with this experiment that how the N and biofertilizati<strong>on</strong><br />
affect dry matter of parsley, and N c<strong>on</strong>tent of plant. Treatment effects <strong>on</strong> 0.01 M CaCl2soluble<br />
total N and NO3-N c<strong>on</strong>centrati<strong>on</strong> at the end of growing period were also<br />
examined. The experiment was set up <strong>on</strong> a chernozem soil with good N supply, high<br />
organic carb<strong>on</strong> c<strong>on</strong>tent, around neutral pH, which gives good circumstances for<br />
microbes; and <strong>on</strong> an another <strong>on</strong>e, which have opposite properties, with poor N supply,<br />
low c<strong>on</strong>tent of organic C, and with very acidic character. The experimental design<br />
opens the door to compare plant parameters <strong>on</strong> two soils.<br />
As you can see from data of Table 2, in which dry matter weight of parsley was<br />
presented, the N fertilizati<strong>on</strong> significantly increased the yield of plant. Although there<br />
was no significant biofertilizer effect, but it can be c<strong>on</strong>cluded that bio-treatments<br />
enhanced the dry matter.<br />
Table 2: Dry matter c<strong>on</strong>tent of parsley <strong>on</strong> chernozem<br />
Leaf dry matter (g pot -1 )<br />
Phylaz<strong>on</strong>it doses (cm 3 pot -1 N (g pot<br />
)<br />
-1 )<br />
70 35 0<br />
Mean LSD5% (N)<br />
0.2 17.7 17.5 17.5 17.6<br />
0,1 15.6 15.1 15.1 15.3<br />
0,05 14.6 14.4 14.3 14.4 1.4<br />
0 15.6 15.6 14.7 15.3<br />
Mean 15.9 15.7 15.4 15.7<br />
LSD5% (phylaz<strong>on</strong>it): 1.2<br />
According to data presented in Table 3, the dry matter of plants increased with<br />
increasing N doses comparing to c<strong>on</strong>trol <strong>on</strong> sandy soil. However, there were no<br />
significant differences am<strong>on</strong>g N treatments.<br />
No correlati<strong>on</strong> has been found between biofertilizer and dry matter weight <strong>on</strong> sandy<br />
soil. It is also true that biofertilizati<strong>on</strong> increased dry matter of parsley at high N doses,<br />
while at lower <strong>on</strong>es or at c<strong>on</strong>trol the applicati<strong>on</strong> of biofertilizati<strong>on</strong> decreased.<br />
314
Table 3: Dry matter c<strong>on</strong>tent of parsley <strong>on</strong> sandy soil<br />
Leaf dry matter (g pot -1 )<br />
Phylaz<strong>on</strong>it doses (cm 3 pot -1 N (g pot<br />
)<br />
-1 )<br />
80 40 0<br />
Mean LSD5% (N)<br />
0.2 5.4 5.6 4.4 5.1<br />
0.1 5.8 4.5 4.9 5.1<br />
0.05 4.7 5.0 5.0 4.9 0.8<br />
0 4.0 4.1 4.3 4.1<br />
Mean 5.0 4.8 4.6 4.8<br />
LSD5% (phylaz<strong>on</strong>it): 0.7<br />
Investigating N c<strong>on</strong>tent of plants <strong>on</strong> chernozem soil, it was found that N fertilizati<strong>on</strong><br />
enhanced the N c<strong>on</strong>centrati<strong>on</strong> in plant, but there was no relati<strong>on</strong>ship between N c<strong>on</strong>tent<br />
and biofertilizer rates at all (Table 4).<br />
Table 4: Nitrogen c<strong>on</strong>tent of parsley <strong>on</strong> chernozem soil<br />
Leaf N c<strong>on</strong>tent (%)<br />
Phylaz<strong>on</strong>it doses (cm 3 pot -1 N (g pot<br />
)<br />
-1 )<br />
70 35 0<br />
Mean LSD5% (N)<br />
0.2 3.4 2.6 2.7 2.9<br />
0.1 2.9 2.5 2.9 2.8<br />
0.05 2.9 3.0 2.3 2.8 0.3<br />
0 2.2 2.0 2.5 2.2<br />
Mean 2.9 2.5 2.6 2.7<br />
LSD5% (phylaz<strong>on</strong>it): 0.3<br />
Similarly to chernozem, with increasing of N fertilizer rates the plant N c<strong>on</strong>centrati<strong>on</strong><br />
increased, and this treatment effect was c<strong>on</strong>firmed statistically (Table 5).<br />
Relati<strong>on</strong>ship between biofertilizer rates and N c<strong>on</strong>tent has been shown a very similar<br />
trend like in case of dry matter c<strong>on</strong>tent of parsley. At high rates of N, the biofertilizati<strong>on</strong><br />
gave an increase in N c<strong>on</strong>centrati<strong>on</strong> of plant, while at lower doses just the opposite was<br />
noticed.<br />
315
Table 5: N c<strong>on</strong>tent in plant <strong>on</strong> sandy soil<br />
Leaf N c<strong>on</strong>tent (%)<br />
Phylaz<strong>on</strong>it doses (cm 3 pot -1 N (g pot<br />
)<br />
-1 )<br />
80 40 0<br />
Mean LSD5% (N)<br />
0.2 3.2 2.8 2.8 2.9<br />
0.1 2.5 2.5 3.2 2.7<br />
0.05 2.1 2.4 2.0 2.2 0.4<br />
0 1.9 1.9 2.6 2.1<br />
Mean 2.4 2.4 2.6 2.5<br />
LSD5% (phylaz<strong>on</strong>it):0.3<br />
Between two soils there was a remarkable difference: in case of acidic sandy soil, the<br />
adding of biofertilizer caused an increase in both plant biomass and in N c<strong>on</strong>tent of leaf.<br />
These effects are not appeared for chernozem. Due to the very low N supply ability of<br />
sandy soil, the increment of microbes amount due to biofertilizati<strong>on</strong> may cause N<br />
immobilizati<strong>on</strong>, because the enhanced microbial biomass take up soluble N.<br />
To take further investigati<strong>on</strong> <strong>on</strong> this problem, 0.01 M CaCl2 soluble total N and nitrate<br />
was measured from soil samples taken at the end of growing period. Both N fertilizati<strong>on</strong><br />
and biofertilizati<strong>on</strong> significantly increased the soluble total nitrogen c<strong>on</strong>centrati<strong>on</strong> <strong>on</strong><br />
the chernozem soil (Figure 1).<br />
25<br />
20<br />
15<br />
10<br />
5<br />
0<br />
B1 B2 B3<br />
N1 N2 N3 N4<br />
Figure 1: Effect of N and biofertilizati<strong>on</strong> <strong>on</strong> 0.01 M CaCl2 soluble<br />
total N c<strong>on</strong>tent of chernozem (LSD5% (N): 2.90 LSD5% (B): 2.51)<br />
316
On sandy soil, the total N c<strong>on</strong>tent increased with increasing N doses, while adding of<br />
biofertilizer to soil caused no significant increase in soil CaCl2-soluble total nitrogen<br />
c<strong>on</strong>centrati<strong>on</strong> (Figure 2).<br />
15<br />
10<br />
5<br />
0<br />
Figure 2 Effect of N doses and biofertilizati<strong>on</strong> <strong>on</strong> 0.01 M CaCl2<br />
soluble total N c<strong>on</strong>tent <strong>on</strong> sandy soil (LSD5% (N): 3.9 LSD5% (B):<br />
3.37)<br />
Moreover, biofertilizer have a negative influence during the investigated period <strong>on</strong><br />
0.01M CaCl2-soluble total N c<strong>on</strong>tent of sandy soil with acidic character and low c<strong>on</strong>tent<br />
of organic matter.<br />
The nitrogen fertilizati<strong>on</strong>, beside the total N, enhanced the nitrate c<strong>on</strong>centrati<strong>on</strong>s in<br />
0.01 M CaCl2 extracti<strong>on</strong>s (Figure 3). Biofertilizer also have an increasing effect <strong>on</strong><br />
nitrate, but <strong>on</strong>ly at high doses of N.<br />
12<br />
10<br />
8<br />
6<br />
4<br />
2<br />
0<br />
B1 B2 B3<br />
N1 N2 N3 N4<br />
B1 B2 B3<br />
N1 N2 N3 N4<br />
Figure 3: Effect of nitrogen rates and biofertilizati<strong>on</strong> <strong>on</strong> nitrate<br />
c<strong>on</strong>centrati<strong>on</strong> <strong>on</strong> chernozem (LSD5% (N): 3.78 LSD5% (B): 3.28)<br />
317
There was a significant increase in nitrate-N comparing to c<strong>on</strong>trol after N fertilizer<br />
adding, while biofertilizer doses decreased (not significantly) the nitrate c<strong>on</strong>centrati<strong>on</strong>s<br />
<strong>on</strong> the sandy soil (Figure 4).<br />
1<br />
0,8<br />
0,6<br />
0,4<br />
0,2<br />
0<br />
B1 B2 B3<br />
N1 N2 N3 N4<br />
Figure 4: Effect of nitrogen and bio-fertilizati<strong>on</strong> <strong>on</strong> nitrate-N <strong>on</strong><br />
sandy soil (LSD5% (N): 0.36 LSD5% (B): 0.31)<br />
Based <strong>on</strong> data from experiment, it can be c<strong>on</strong>cluded that positive effect of<br />
biofertilizati<strong>on</strong> <strong>on</strong> yield, N c<strong>on</strong>tent of plant and soil displayed <strong>on</strong>ly <strong>on</strong> chernozem soil<br />
during that short period of investigati<strong>on</strong>. On the acidic soil with low organic matter<br />
c<strong>on</strong>tent, these effects are no appeared because of N immobilizati<strong>on</strong> by microbes.<br />
REFERENCES<br />
Aubert, C. (1981): Organischer Landbau. Verlag Eugen Ulmer, Stuttgart<br />
Chander,-H. - Abrol,-I. P. (1972): Effect of three nitrogenous fertilizers <strong>on</strong> the<br />
soluti<strong>on</strong> compositi<strong>on</strong> of a saline sodic soil. Commun. Soil Sci. Pl. Anal., 3. 1. 51-56. p.<br />
Felizardo,-B. C. - Bens<strong>on</strong>,-N. R. - Cheng,-H. H. (1972): Nitrogen, salinity, and acidity<br />
distributi<strong>on</strong> in an irrigated orchard soil as affected by placement of nitrogen<br />
fertilizers. Soil Sci. Soc. Amer. Proc., 36. 803-808. p.<br />
Hegde, D. M. – Dwived, B. S. – Sudhakara, S. N.: 1999. Biofertilizers for cereals<br />
producti<strong>on</strong> in India – a review – Indian J. Agric. Sci. vol. 69 73-83 pp.<br />
Houba, V. J. G. - Novozamsky, I. - Temminghoff, E. (1994): Soil analysis procedures<br />
Extracti<strong>on</strong> with 0,01 M CaCl2. (syllabus) Dept. Of Soil Sci. And Plant Nutriti<strong>on</strong><br />
Wageningen Agricultural University – The Nederlands<br />
Nagy J. -Német T. -Szabó J. -Pásztor L. -Dobos A. (2001): Tájgazdasági körzetek<br />
kialakítása a Kreybig-féle „Átnézetes Talajismereti Térképsorozat” alapján.<br />
Agrártudományi Közlemények 1. Acta Agraria Debreceniensis, 20-25.<br />
Nagy P.T. (2000): Égetéses elven működő elemanalizátor alkalmazhatósága talaj- és<br />
növényizsgálatokban. Agrokémia és Talajtan. Tom. 49. No. 3-4. 521-534.<br />
Power,-J. F. et al. (1972): Effect of nitrogen source <strong>on</strong> corn and bromegrass<br />
producti<strong>on</strong>, soil pH, and inorganic soil nitrogen. Agr<strong>on</strong>. J., 64. 341-344. p.<br />
Szabó I. M. (1986): Az általános talajtan biológiai alapjai. Mezőgazdasági Kiadó,<br />
Budapest.<br />
318
THE INFLUENCE OF CHEMICAL AND BIOFERTILIZERS ON THE YIELD<br />
AND NITROGEN CONTENT OF LETTUCE (LACTUCA SATIVA L.)<br />
Andrea Balla Kovács, Ida Kincses, Imre Vágó, Rita Kremper<br />
University of Debrecen Faculty of Agricultural Science<br />
Department of Agricultural Chemistry and Soil Science<br />
ABSTRACT<br />
The greenhouse pot experiment <strong>on</strong> calcareous chernozem soil from Debrecen-Látókép<br />
and <strong>on</strong> sandy soil from Újfehértó was performed to study the effects of biofertilizer in<br />
the presence of reduced doses of chemical fertilizers <strong>on</strong> the yield and nitrogen c<strong>on</strong>tent<br />
of lettuce and the water soluble nitrogen forms (NO3 - and total-N) measured in 0.01 M<br />
CaCl2 extracts. The experimental plant selected was cabbage head lettuce (Lactuca<br />
sativa L.). The bi-factorial trials were arranged in a randomized complete block design<br />
with four replicati<strong>on</strong>s, applying four levels (0, 0.05, 0.1, 0.2 g pot -1 ) of N as NH4NO3<br />
with or without applicati<strong>on</strong> of biofertilizer as Phylaz<strong>on</strong>it MC. P and K doses applied<br />
were identical in all pots.<br />
The results obtained suggested: Nitrogen doses markedly increased growth (fresh<br />
and dry weight) of lettuce, had significant positive influence <strong>on</strong> N c<strong>on</strong>tent of plant and<br />
<strong>on</strong> values of NO3 - , total N measured in 0.01 M CaCl2 soil extract . Its increasing effect<br />
was the same <strong>on</strong> two types of soils, <strong>on</strong>ly significance of variance varied.<br />
The applicati<strong>on</strong> of biofertilizer was not as effective as N doses and its effect was<br />
dependent <strong>on</strong> soil caracteristics. The Phylaz<strong>on</strong>it MC had an increasing influence, but<br />
not significant effect <strong>on</strong> the fresh and dry weight of lettuce leaves and slightly changed<br />
N c<strong>on</strong>tent of plant. On chernozem soil, we have found an increasing effect in values of<br />
N c<strong>on</strong>tent of plant, NO3 - and total N measured in 0.01 M CaCl2 extracts. On the<br />
c<strong>on</strong>trary, <strong>on</strong> sandy soil a slight decreasing effect of biofertilizer was found in N c<strong>on</strong>tent<br />
of plant and total N measured in 0.01 M CaCl2. It seems that in our experiment, (over<br />
the entire growing seas<strong>on</strong>) applicati<strong>on</strong> of phylaz<strong>on</strong>it <strong>on</strong> sandy soil might cause N<br />
immobilizati<strong>on</strong>, in c<strong>on</strong>sequence decreased the mineral N fracti<strong>on</strong> available for plant and<br />
so decreased the N c<strong>on</strong>tent of plant. To gain more, reliable applicability of Philaz<strong>on</strong>it<br />
MC, further studies are needed.<br />
Keywords: lettuce, mineral fertilizers, nitrogen, biofertilizers<br />
INTRODUCTION<br />
Nitrogen as a major c<strong>on</strong>stituent of all plants is <strong>on</strong>e of the most important nutrients. It<br />
has got a unique positi<strong>on</strong> because relatively high amounts are required by most<br />
agricultural crops for optimal yields (Black 1968, Stevens<strong>on</strong> and Cole 1999). Most<br />
plants take up N as nitrate (NO3 - ) and amm<strong>on</strong>ium (NH4 + ) from the soil soluti<strong>on</strong><br />
(Blackmer 2000). The use of nitrogen fertilizers is standard practice in vegetable<br />
producti<strong>on</strong> systems. Increasing c<strong>on</strong>cern over nitrate c<strong>on</strong>taminati<strong>on</strong> of soils, nitrate<br />
leaching in groundwater and nitrate c<strong>on</strong>tent in vegetable may require a reducti<strong>on</strong> in N<br />
applicati<strong>on</strong> in crop producti<strong>on</strong>, while maintaining optimal productivity (Gutezeit and<br />
Fink 1999). Lettuce is an important horticultural crop. High nitrate c<strong>on</strong>centrati<strong>on</strong> in<br />
leafy vegetables c<strong>on</strong>stitutes a health hazard and is therefore undesirable. Applicati<strong>on</strong>s of<br />
319
iofertilizers may help to avoid envir<strong>on</strong>mental hazards for plants, animals and human<br />
beings.<br />
The present work was planned to study the effect of biofertilizer, Philaz<strong>on</strong>it MC in<br />
the presence of reduced doses of chemical fertilizers <strong>on</strong> the yield and nitrogen c<strong>on</strong>tent<br />
of lettuce and the water soluble nitrogen forms (NO3 - and total N) measured in 0.01 M<br />
CaCl2 extracts.<br />
MATERIALS AND METHODS<br />
The greenhouse pot experiment was performed using soil samples from two regi<strong>on</strong>s,<br />
calcareous chernozem soil from Debrecen-Látókép (1) and sandy soil from Újfehértó<br />
(2). The important properties of soils used are included in Table 1.<br />
Table 1. Properties of experimental soils<br />
Soil no. pHKCl pHH2O pHCaCl2 Hu% KA<br />
1 5.9 6.7 6.2 2.5 42<br />
2 3.7 4.1 4.8 1.3 27<br />
KA: Plasticity index according to Arany<br />
10 kg and 12 kg soil (chernozem soil from Debrecen-Látókép and sandy soil from<br />
Újfehértó, respectively) were weighed into Mitscherlich type pots. The experimental<br />
plant selected was cabbage head lettuce (Lactuca sativa L.). Four seed of lettuce were<br />
sown in the soil per pot. I<strong>on</strong> exchanged water was added to all pots to keep the soil at<br />
c<strong>on</strong>stant moisture (60% of the water-holdig capacity) using daily weighing.<br />
The bi-factorial trials were arranged in a randomized complete block design with four<br />
replicati<strong>on</strong>s, applying four levels of N with or without applicati<strong>on</strong> of biofertilizer<br />
(Table 2.). P and K doses applied were identical in all pots (0.2 g P2O5 pot -1 and 0.2 g<br />
K2O pot -1 ).<br />
Treatment N doses<br />
g pot -1<br />
Table 2. Scheme of treatments applied<br />
Phylaz<strong>on</strong>it doses in<br />
sandy soil<br />
cm 3 pot -1<br />
Phylaz<strong>on</strong>it doses<br />
in chernosem soil<br />
cm 3 pot -1<br />
code<br />
1. 0 0 0 c<strong>on</strong>trol<br />
2. 0 40 35 N0+phyl<br />
3. 0.05 0 0 N1<br />
4. 0.05 40 35 N1+phyl<br />
5. 0.1 0 0 N2<br />
6. 0.1 40 35 N2+phyl<br />
7. 0.2 0 0 N3<br />
8. 0.2 40 35 N3+phyl<br />
The nitrogen, phosphorus and potassium were added in soluti<strong>on</strong> of NH4NO3, KH2PO4<br />
and KCl, respectively. The applied biofertilizer was Phylaz<strong>on</strong>it MC (phyl), which<br />
c<strong>on</strong>tains carboxi-methyl-cellulose (CMC), microelements, Azotobacter croococcum,<br />
320
Bacillus megatherium soil bacteria, heteroauxin, gibberelin and vitamin B. The soil in<br />
each pot was stirred well to ensure uniform mixing of soil and nutrients.<br />
At the end of vegetati<strong>on</strong> period soil samples were taken, air dried and sieved<br />
(
The fresh and dry yields of lettuce grown <strong>on</strong> calcareous chernozem soil were much<br />
higher in all pots than in the case of plants cultivated <strong>on</strong> sandy soil. The fresh and dry<br />
weight <strong>on</strong> chernozem soil ranged 210.6 - 248.4 g pot -1 and 11.3 - 13.6 g pot -1 , <strong>on</strong> sandy<br />
soil ranged 55.5 - 77.8 g pot -1 and 4.3 – 6.2 respectively.<br />
Table 5. Mean fresh weight of lettuce cultivated <strong>on</strong> calcareous chernozem soil<br />
(Látókép) as influenced by N and phylaz<strong>on</strong>it doses<br />
fresh weight (g pot -1 )<br />
phylaz<strong>on</strong>it doses (cm 3 pot -1 N<br />
)<br />
g pot -1 0 40<br />
mean<br />
0 210.6 217.4 213.9<br />
0.05 239.8 242.3 241.1<br />
0.1 244.1 245.9 245.0<br />
0.2 245.3 248.4 246.8<br />
mean 234.9 238.5<br />
LSD5% (phylaz<strong>on</strong>it): n.s.<br />
236.7<br />
LSD5% (N)<br />
15.02<br />
Table 6. Mean dry weight of lettuce cultivated <strong>on</strong> calcareous chernozem soil<br />
(Látókép) as influenced by N and phylaz<strong>on</strong>it doses<br />
dry weight (g pot -1 )<br />
phylaz<strong>on</strong>it doses (cm 3 pot -1 N<br />
)<br />
g pot -1 0 40<br />
mean<br />
0 11.3 11.4 11.4<br />
0.05 13.3 13.6 13.5<br />
0.1 13.2 11.3 12.3<br />
0.2 13.5 13.6 13.6<br />
mean 12.8 12.5<br />
LSD5% (phylaz<strong>on</strong>it): n.s.<br />
12.7<br />
Source of<br />
variati<strong>on</strong><br />
LSD5% (N)<br />
0.73<br />
Table 7. Summary of ANOVA (F-test) for different source of variance<br />
fresh weight dry weight fresh weight fresh weight<br />
<strong>on</strong> sandy soil <strong>on</strong> chernozem soil<br />
N fertilizati<strong>on</strong> *** * *** ***<br />
Phylaz<strong>on</strong>it doses n. s. n. s. n. s. n. s.<br />
n.s.:n<strong>on</strong>-significant; *:significant at P
N c<strong>on</strong>tent of plant: Studying the nitrogen c<strong>on</strong>tent of lettuce leaves we found that there<br />
was significant relati<strong>on</strong>ship between N treatments and plant N c<strong>on</strong>tent. Increasing<br />
applicati<strong>on</strong> of N from 0.05 g pot -1 up to 0.2 g pot -1 had significant positive influence <strong>on</strong><br />
N c<strong>on</strong>tent of plant <strong>on</strong> both types of soils (Figure 1, Table 8). Biofertilizer did not cause<br />
marked effect <strong>on</strong> N c<strong>on</strong>tent of plant <strong>on</strong> chernozem soil. Inoculati<strong>on</strong> of this type of soil<br />
with Philaz<strong>on</strong>it slightly increased N c<strong>on</strong>tent with the excepti<strong>on</strong> of treatments with no<br />
mineral fertilizer.<br />
On sandy soil applicati<strong>on</strong> of Phylaz<strong>on</strong>it significantly (P
CaCl2- NO3 - -N (mg kg -1 )<br />
9<br />
8<br />
7<br />
6<br />
5<br />
4<br />
3<br />
2<br />
1<br />
0<br />
5.7<br />
6.1<br />
5.8<br />
7.0<br />
5.5<br />
0.4 0.4 0.6 0.6 0.5 0.8<br />
C<strong>on</strong>trol<br />
N0+phyl<br />
Újftó<br />
Látókép<br />
N1<br />
N1+phyl<br />
N2<br />
N2+phyl<br />
8.1<br />
N3<br />
2.0<br />
7.2<br />
2.2<br />
N3+phyl<br />
Figure 2. Mean NO3 - - N (mg kg -1 ) measured in CaCl2 soil extracts from Látókép<br />
and Újfehértó as influenced by N and phylaz<strong>on</strong>it applicati<strong>on</strong> rates. For interpretati<strong>on</strong><br />
of treatments see Table 2.<br />
totalN(mg kg -1 )<br />
16<br />
14<br />
12<br />
10<br />
8<br />
6<br />
4<br />
2<br />
0<br />
4.1<br />
C<strong>on</strong>trol<br />
11.3<br />
3.7<br />
N0+phyl<br />
12.7<br />
N1<br />
Újftó<br />
Látókép<br />
4.0<br />
11.7<br />
3.8<br />
N1+phyl<br />
12.7<br />
N2<br />
4.5<br />
14.3 14.7 14.5 14.8<br />
4.2<br />
N2+phyl<br />
N3<br />
8.5<br />
6.0<br />
N3+phyl<br />
Figure 3. Mean total N (mg kg -1 ) measured in CaCl2 soil extracts from Látókép and<br />
Újfehértó as influenced by N and phylaz<strong>on</strong>it applicati<strong>on</strong> rates. For<br />
interpretati<strong>on</strong> of treatments see Table 2.<br />
By the end of the growing seas<strong>on</strong> nitrate c<strong>on</strong>tent of sandy soil becomes very low (0.4-<br />
0.8 mg kg -1 ). Only with the applicati<strong>on</strong> of highest N doses (0.2 g pot -1 ) caused increased<br />
CaCl2-NO3 - values (2.0-2.2 mg kg -1 ). This increasing effect of N doses <strong>on</strong> NO3 - c<strong>on</strong>tent<br />
was significant. We measured higher c<strong>on</strong>centrati<strong>on</strong>s of NO3 - by turns <strong>on</strong> chernosem soil.<br />
This might be due to higher c<strong>on</strong>tent of organic matter which could mineralize during the<br />
growing seas<strong>on</strong>. Both N doses and biofertilizati<strong>on</strong> significantly increased the CaCl2-<br />
NO3 - <strong>on</strong> chernozem soil. The effect of chemical fertilizer was higher. Measured data<br />
show that similarly to CaCl2-NO3 - , the values of CaCl2-total N were higher in<br />
324<br />
8.2
chernozem soil in all pots either. The increased N fertilizer doses caused significant<br />
improvement in CaCl2-total N c<strong>on</strong>centrati<strong>on</strong> measured <strong>on</strong> both types of soils. Effect of<br />
Phylaz<strong>on</strong>it slightly increased values of CaCl2-total N <strong>on</strong> chernozem soil, but<br />
significantly decreased these values <strong>on</strong> sandy soil.<br />
Table 9. Summary of ANOVA (F-test) for different source of variance for NO3 - - N<br />
measured in CaCl2 soil extracts<br />
Source of<br />
variati<strong>on</strong><br />
NO3 - - N<br />
<strong>on</strong> sandy soil <strong>on</strong> chernozem soil<br />
significance LSD5% significance LSD5%<br />
N fertilizati<strong>on</strong> *** 0.41 * 1.04<br />
Phylaz<strong>on</strong>it doses n. s. - ** 0.73<br />
n.s.:n<strong>on</strong>-significant; **:significant at P
Jászberényi, I. – Loch, J. – Sarkadi, J. (1994): Experiences with 0.01 M CaCl2 as an<br />
extracti<strong>on</strong> reagent for use as a soil testing procedure in Hungary – Comm. Soil Sci.<br />
vol. 25 1771-1777 pp.<br />
Nagy P.T. (2000): Égetéses elven működő elemanalizátor alkalmazhatósága talaj- és<br />
növényvizsgálatokban. Agrokémia és Talajtan. Tom. 49. No. 3-4. 521-534.<br />
326
THE INFLUENCE OF USING BIOLOGICAL ADDITIVES ON THE<br />
NUTRITIONAL QUALITIES OF CORN SILAGE AND ON THE FATTENING<br />
PERFORMANCES OF BULL CALVES<br />
Daniel Mierlita; I. Chereji; Cristina Maerescu<br />
University of Oradea, Romania, 26, Magheru Street, Oradea;<br />
ABSTRACT<br />
Ensiling corn by using a biological additive based <strong>on</strong> homofermentative lactic acid<br />
bacteria (Lactobacillus plantarum + Pediococcus pentosaceus) ensures, by comparis<strong>on</strong><br />
with natural fermentati<strong>on</strong>, a forage with higher nutriti<strong>on</strong>al qualities, with a lower pH<br />
(acid), a lower c<strong>on</strong>tent of amm<strong>on</strong>iacal N and raw cellulose and more undegraded<br />
protein. The use of corn silage treated with inoculant in the food of young bull calves<br />
intended for fattening, has ensured a significant growth in weight gain, DM<br />
c<strong>on</strong>sumpti<strong>on</strong> and degree of the food’s capitalizati<strong>on</strong>, as opposed to the lot which<br />
received naturally fermented forage in the rati<strong>on</strong>s. Thus by using inoculant to ensile<br />
corn, the period of fattening was decreased by 26 days and 82 kg of c<strong>on</strong>centrates and<br />
120 kg of corn silage were saved. The biological additive used for the ensiling has not<br />
influenced the output of slaughtering, but it has led to the increase of the muscle area,<br />
which is positively correlated with the proporti<strong>on</strong> of superior quality meat in the<br />
carcass.<br />
Keywords: corn silage, inoculants, fattening up bull calves.<br />
INTRODUCTION<br />
For fattening up young bulls, using fermented forage (especially corn) represents the<br />
most efficient form of feeding, taken into account the technical and ec<strong>on</strong>omical<br />
advantages that it provides (Halga, 2005). The process of ensiling the forage depends <strong>on</strong><br />
the dominance of the lactic acid bacteria over the rest of the microflora, which, in<br />
anaerobic c<strong>on</strong>diti<strong>on</strong>s, increase the acidity of the envir<strong>on</strong>ment, thus reducing the<br />
competiti<strong>on</strong> between microorganisms. The efficiency of the ensiling process is given by<br />
the way in which sugars are fermented in acid and by how the altering processes<br />
de<strong>term</strong>ined by the clostridium bacteria (butyric acid bacteria) are avoided (Woolford-<br />
2004). It is important that the decrease of pH in the ensiled forage be d<strong>on</strong>e as quickly as<br />
possible (during the first 2-3 days) in order to avoid the degradati<strong>on</strong> and loss of the<br />
protein (Webb, 1998). During the unc<strong>on</strong>trolled processes of fermenting the silage, 45%<br />
of the protein is decomposed up to NH3N (J<strong>on</strong>es, 1995; 1996).<br />
The c<strong>on</strong>trol of the silage’s fermenting was accomplished by using inorganic acids<br />
for a l<strong>on</strong>g time (sulfuric, phosphoric and hydrochloric acids) or external organic acids<br />
(formic and propi<strong>on</strong>ic), which had a series of negative effects in the short run (ruminal<br />
and metabolic acidoses, ruminal parakeratosis) but especially in the l<strong>on</strong>g run,<br />
materialized usually in the reducti<strong>on</strong> of fertility as an effect of the acids collecting the<br />
microelements (Suttle, 1998). At the present time, because of modern biotechnologies,<br />
the guiding of silage fermentati<strong>on</strong> is performed by using biological additives that<br />
c<strong>on</strong>tain selected strains of lactic acid bacteria, especially lactobacillus, or lactic acid<br />
bacteria and enzymes (cellulase, hemicellulase, amylase). In the USA <strong>on</strong>ly the<br />
327
iological additives (the inoculants) have been approved by the FDA and can be used in<br />
ensiling forages (An<strong>on</strong>, 1997).<br />
The research performed in this domain (Wilkins, 1996; Steen, 1997 ; Keady, 1998,<br />
2000 ; Mierlita 2002) has shown that using inoculants for corn ensiling has led to a<br />
significant reducti<strong>on</strong> of the amm<strong>on</strong>iac level (a criteri<strong>on</strong> for evaluating proteolysis), to a<br />
greater c<strong>on</strong>sumpti<strong>on</strong> (3.8 – 5.2%) and a better digestibility of DM (1.7-4.0%), to a<br />
greater weight gain in bulls intended for fattening (3.8-12.2%), to a better quality and<br />
quantity of cow milk (1.7 -2.2 l/day and an increase of the protein level by 0,05%) as<br />
well as to the improvement in the degree of food capitalizati<strong>on</strong> (by 6%).<br />
In our research we have analyzed the effect of using an inoculant with mixed types<br />
of homofermentative lactic acid bacteria with high tolerance to acidity, which would be<br />
able to c<strong>on</strong>trol the fermentati<strong>on</strong>s throughout the ensilaging period (Lactobacillus<br />
plantarum + Pediococcus pentosaceus – 10 6 units forming col<strong>on</strong>ies/g), <strong>on</strong> the<br />
nutriti<strong>on</strong>al qualities of corn silage as well as <strong>on</strong> the fattening performances of bull<br />
calves.<br />
MATERIALS AND METHODS<br />
The corn used in the ensiling process was harvested in an advanced state of vegetati<strong>on</strong><br />
(kernel: wax-glass-like) with a c<strong>on</strong>tent of 33-34% DM. Two silages were created : <strong>on</strong>e<br />
by natural fermentati<strong>on</strong> and the other by using an inoculant with mixed types of<br />
homofermentative lactic acid bacteria with high tolerance to acidity, composed of<br />
Lactobacillus plantarum + Pediococcus pentosaceus (10 6 units forming col<strong>on</strong>ies/g).The<br />
research c<strong>on</strong>cerning the productive effect was performed <strong>on</strong> two lots of weaned bull<br />
calves of the Romanian Baltata breed (12 heads / lot), aged about 3 m<strong>on</strong>ths give or take<br />
14 days. The fattening period lasted 426 days (14 m<strong>on</strong>ths), respectively from the age of<br />
3 m<strong>on</strong>ths (after the weaning of the calves) and up to the age of 17 m<strong>on</strong>ths. Of the two<br />
lots, <strong>on</strong>e was c<strong>on</strong>sidered a c<strong>on</strong>trol group (GC) and received naturally fermented corn<br />
silage in its food, and the other was c<strong>on</strong>sidered an experimental group (GE) and<br />
received corn silage obtained by adding an inoculant (treated). The corn silage ensured<br />
50% of the rati<strong>on</strong>’s DM in both groups; the rati<strong>on</strong> was completed by hashed alfalfa hay<br />
(10% of DM) and a mix of c<strong>on</strong>centrates (40% of DM) (table1). The food was<br />
distributed in abundance in the form of unique forage mixtures.<br />
Table 1: Experimental scheme<br />
Associated activities<br />
Objective C<strong>on</strong>trol lot (GC) Experimental lot<br />
(GE)<br />
1. The influence of<br />
using inoculants with<br />
mixed types of lactic<br />
acid bacteria <strong>on</strong> the<br />
nutriti<strong>on</strong>al qualities of<br />
corn silage.<br />
Corn ensiled by<br />
means of natural<br />
fermentati<strong>on</strong>.<br />
328<br />
Corn ensiled by<br />
adding inoculant :<br />
Lactobacillus<br />
plantarum +<br />
Pediococcus<br />
pentosaceus (10 6<br />
units forming<br />
col<strong>on</strong>ies/g).<br />
Surveyed aspects<br />
- The chemical<br />
compositi<strong>on</strong> of the<br />
fermented forage<br />
(DM, raw protein,<br />
raw cellulose) ;<br />
- pH ;<br />
- amm<strong>on</strong>iacal N;<br />
- lactic acid (g/kg<br />
DM);
2. The influence of<br />
using inoculants in corn<br />
ensiling <strong>on</strong> the fattening<br />
performances of bull<br />
calves.<br />
Structure of the rati<strong>on</strong> (% of DM)<br />
Corn silage - 50<br />
Alfalfa hay – 10<br />
C<strong>on</strong>centrates* - 40<br />
Corn silage - 50<br />
Alfalfa hay – 10<br />
C<strong>on</strong>centrates* - 40<br />
- evoluti<strong>on</strong> of<br />
weight gain;<br />
- c<strong>on</strong>sumpti<strong>on</strong> of<br />
DM;<br />
- degree of food<br />
capitalizati<strong>on</strong>;<br />
- metabolic<br />
indices in the blood;<br />
- ruminal<br />
parametres (pH , no.<br />
of bacteria/ml,<br />
structure of AGV);<br />
-the quality of the<br />
carcass and meat.<br />
* Mixture of c<strong>on</strong>centrates composed of : corn, barley, soy bean groats, sunflower groats and<br />
cereal-vitamin premix. It ensured 19.8% Pb in the 3-6 m<strong>on</strong>ths period, 17.5% Pb. in the 6-12<br />
m<strong>on</strong>ths period and 15.4% in the 12-17 m<strong>on</strong>ths period.<br />
At the end of the experiment, c<strong>on</strong>trol slaughterings were performed (n=3), establishing<br />
the main butchery indices. The obtained data was processed and interpreted statistically<br />
by using the ‘Student’ test.<br />
RESULTS AND DISCUSSIONS<br />
The use of the homofermentative lactic acid bacteria based inoculant for ensiling corn,<br />
resulted in a forage having nutriti<strong>on</strong>al qualities superior to the <strong>on</strong>e obtained through<br />
natural fermentati<strong>on</strong> (table 2), because the inoculant de<strong>term</strong>ined a rapid reducti<strong>on</strong> of the<br />
pH in the forage, which led to the quick reducti<strong>on</strong> of the level of Coliform bacteria, thus<br />
reducing the proteolysis process. This aspect is emphasized by the lower level of<br />
amm<strong>on</strong>iacal N and the higher level of whole protein in the inoculant treated forage.<br />
Moreover, the inoculant caused a significant decrease in the level of raw cellulose and a<br />
rise in the quantity of lactic acid (g/kg DM).<br />
Table 2: The effect of the inoculant (Lactobacillus plantarum + Pediococcus<br />
pentosaceus) <strong>on</strong> the quality of the corn silage<br />
Parameters<br />
C<strong>on</strong>trol lot<br />
(GC)<br />
Experimental<br />
lot (GE)<br />
Changes due to<br />
the inoculant<br />
treatment (%)<br />
DM (%) 33.75 34.19 + 1.30<br />
pH 4.4 3.8* + 13.64<br />
Raw protein (g/kg DM) 79.2 87.5* + 10.47<br />
Amm<strong>on</strong>iacal N (%of total N) 6.2 4.1** - 33.88<br />
Raw cellulose (% of DM) 26.9 23.8* - 11.53<br />
Lactic acid (g/kg DM) 128.4 135.2* + 5.29<br />
* p< 0.05; ** p
The positive effects of using the inoculant in the producti<strong>on</strong> of fermented forage <strong>on</strong> the<br />
fattening performances of bull calves were materialized in the weight gain increase of<br />
about 6.38%, the DM c<strong>on</strong>sumpti<strong>on</strong> improved 5.23% and the degree of food<br />
capitalizati<strong>on</strong> increased 4.94% (fig. 1 and table 3). Therefore, the period of fattening<br />
was reduced by 26 days (6.1%) when inoculated silage was administered, by<br />
comparis<strong>on</strong> with the naturally fermented silage, thus saving 82 kg of c<strong>on</strong>centrates and<br />
120 kg of corn silage.<br />
12<br />
11<br />
10<br />
9<br />
8<br />
7<br />
6<br />
5<br />
4<br />
3<br />
2<br />
488.1 515.0<br />
940 1000<br />
Final weight (kg) Weight gain<br />
(g/day)<br />
Group C Group Exp.<br />
9.36<br />
9.85<br />
C<strong>on</strong>sumpti<strong>on</strong><br />
DM (kg/day)<br />
7.91 7.52<br />
Specific<br />
c<strong>on</strong>sumpti<strong>on</strong><br />
(UNC/kg gain<br />
Figure 1: The influence of the biological additive used for ensiling corn <strong>on</strong> the<br />
fattening performances of bull calves<br />
330
Table 3: The evoluti<strong>on</strong> of weight gain, DM c<strong>on</strong>sumpti<strong>on</strong> and degree of food<br />
capitalizati<strong>on</strong><br />
C<strong>on</strong>trol lot Experimental<br />
C<strong>on</strong>trol Experim<br />
Specificati<strong>on</strong> (GC) lot (GE) Specificati<strong>on</strong> lot ental lot<br />
(GC) (GE)<br />
Evoluti<strong>on</strong> of<br />
Evoluti<strong>on</strong> of<br />
body mass<br />
average daily<br />
(kg/head):<br />
gain (g/day):<br />
- 3 m<strong>on</strong>ths 84.5±1.31 85.3±1.13 - 3-6 m<strong>on</strong>ths 773 833*<br />
- 6 m<strong>on</strong>ths 154.8±3.15 161.1±2.97 - 6-9 m<strong>on</strong>ths 916 970<br />
- 9 m<strong>on</strong>ths 239.1±4.73 250.3±5.45* - 9-12 m<strong>on</strong>ths 976 1013<br />
- 12 m<strong>on</strong>ths 326.9±6.17 341.5±7.02* - 12-15 m<strong>on</strong>ths 1016 1100*<br />
- 15 m<strong>on</strong>ths 420.4±5.39 442.7±6.17* - 15-17 m<strong>on</strong>ths 1110 1186*<br />
- 17 m<strong>on</strong>ths 488.1±7.34 515.0±5.98** Total<br />
period:<br />
average<br />
Total gain 403.6±5.75 429.7±4.41* 3-17 m<strong>on</strong>ths 940 1000*<br />
Average daily<br />
Specific<br />
c<strong>on</strong>sumpti<strong>on</strong> of<br />
c<strong>on</strong>sumpti<strong>on</strong><br />
DM (kg/head):<br />
(UNC/kg gain):<br />
- 3-6 m<strong>on</strong>ths 3.65 3.79 - 3-6 m<strong>on</strong>ths 4.47 4.30<br />
- 6-9 m<strong>on</strong>ths 6.46 6.59 - 6-9 m<strong>on</strong>ths 6.66 6.42<br />
- 9-12 m<strong>on</strong>ths 8.94 8.76 - 9-12 m<strong>on</strong>ths 8.65 8.17<br />
- 12-15 m<strong>on</strong>ths 10.81 11.47 - 12-15 m<strong>on</strong>ths 10.04 9.85<br />
- 15-17 m<strong>on</strong>ths 13.18 12.99 - 15-17 m<strong>on</strong>ths 11.21 10.34<br />
Total average<br />
Total average<br />
period:<br />
period:<br />
3-17 m<strong>on</strong>ths 9.36 9.85 3-17 m<strong>on</strong>ths<br />
UNC = 6.2 MJ EN (1481 kcal EN) * p
The positive effect of the silage’s biological additive can be observed in the rumen,<br />
where an improvement of pH and of the relati<strong>on</strong> between the acetic and propi<strong>on</strong>ic acids<br />
in favor of the latter, which is beneficial to the producti<strong>on</strong> of meat, can be noticed (table<br />
4). This aspect is due to the fact that lactic acid bacteria in the inoculant at the level of<br />
the rumen present probiotic effects (Weinberg, 2003).<br />
There were no noticed differences between the two lots regarding the output at<br />
slaughter, but the surface of the muscle c<strong>on</strong>tent, which is positively correlated with the<br />
quantity of superior quality meat in the carcass (McD<strong>on</strong>ald, 2002) and the chemical<br />
compositi<strong>on</strong> of the meat (leg) were better in the lot that received inoculated silage in the<br />
food (table 5).<br />
Tabel 5: The effect of the biological additive used in the ensiling of corn <strong>on</strong> the<br />
quality of the carcass and meat<br />
Specificati<strong>on</strong> C<strong>on</strong>trol lot Experimental lot<br />
(GC)<br />
(GE)<br />
Output at slaughter, % 56.2 56.4<br />
Weight of renal suet, g<br />
Dimensi<strong>on</strong> of muscle area, cm<br />
3210 3740<br />
2<br />
- rib secti<strong>on</strong> 8-9<br />
- rib secti<strong>on</strong> 11-12<br />
Chemical compositi<strong>on</strong> of the meat:<br />
- DM, %<br />
- Raw protein, %<br />
- Raw fat, %<br />
61.29<br />
80.62<br />
26.66<br />
21.81<br />
1.88<br />
72.16<br />
81.26<br />
27.81<br />
23.23<br />
1.68<br />
CONCLUSIONS<br />
1. The ensiling of corn by using a biological additive based <strong>on</strong> homofermentative<br />
lactic acid bacteria (Lactobacillus plantarum + Pediococcus pentosaceus) ensures,<br />
by comparis<strong>on</strong> with natural fermentati<strong>on</strong>, a forage of higher nutriti<strong>on</strong>al quality,<br />
having a lower pH (acid), a lower c<strong>on</strong>tent of amm<strong>on</strong>iacal N and raw cellulose and<br />
more undegraded protein.<br />
2. The usage of inoculated corn silage in the food of bull calves intended for fattening<br />
ensured a significant improvement in the weight gain, DM c<strong>on</strong>sumpti<strong>on</strong> and degree<br />
of food capitalizati<strong>on</strong>, as opposed to the lot which received naturally fermented<br />
silage in the food. Thus by using inoculant for the corn silage, the period of<br />
fattening decreased by 26 days and 82 kg of c<strong>on</strong>centrates as well as 120 kg of corn<br />
silage were saved.<br />
3. The biological additive used for ensiling did not influence the output at<br />
slaughtering, but it has led to an increase in the dimensi<strong>on</strong> of the muscle area,<br />
which is positively correlated with the proporti<strong>on</strong> of superior quality meat in the<br />
carcass.<br />
332
REFERENCES<br />
4. An<strong>on</strong> D.L. (1997): – Silage additives: making the best choice. L<strong>on</strong>d<strong>on</strong>: Milk<br />
Development Council, 5-17 pp.<br />
5. Halga P.; I.M. Pop; Te<strong>on</strong>a Avarvarei; Viorica Popa (2005): – Nutritie si<br />
alimentatie animala. Ed. ALFA, Iasi.<br />
6. J<strong>on</strong>es R. (1995): – Forage additive supplement. Farmers Weekly, 26 th November,<br />
24-43 pp.<br />
7. J<strong>on</strong>es R.; Winters A.; Cockburn J. (1996): – Changes in amino acid c<strong>on</strong>tent of<br />
inoculated grass silage and its effect <strong>on</strong> animal producti<strong>on</strong>. In: Biotechnology in the<br />
Feed Industry, Proceedings of the 12 th Annual Symposium. Nottingham University<br />
Press, UK, 249-264 pp.<br />
8. Keady T.W.J. (1998): – The producti<strong>on</strong> of high feed value grass silage and the<br />
choice of compound feed type to maximize animal performance. In: Biotechnology<br />
in the Feed Industry, Proceedings of the 14 th Annual Symposium. Nottingham<br />
University Press, UK, 439-452 pp.<br />
9. Keady T.W.J. (2000): – Bey<strong>on</strong>d the Science: what the farmer looks for in the<br />
producti<strong>on</strong> of silage. In: Biotechnology in the Feed Industry, Proceedings of the<br />
16 th Annual Symposium. Nottingham University Press, UK, 157-174 pp.<br />
10. McD<strong>on</strong>ald P.; Edwards R.A.; Greenhalgh JED; Morgan CA. (2002): – Animal<br />
nutriti<strong>on</strong>. Pears<strong>on</strong>, Prentice Hall.<br />
11. Mierlita D.; B Nagy; Gh. Salajan, Ant<strong>on</strong>ia Odagiu; Mariana Dinea (2000): –<br />
Bioefficiency of bacteria cultures used for corn ensiling. Buletinul USAMV, Cluj-<br />
Napoca, seria Zootehnie, Biotehnologii si MV, vol. 54; 91-102 pp.<br />
12. Steen R.W. (1997): – Factors affecting the utilisati<strong>on</strong> of grass silage for beef<br />
producti<strong>on</strong>. In: Efficient beef producti<strong>on</strong> from grass. Symposium of the British<br />
Grassland Society, No. 24, 129-137 pp.<br />
13. Suttle N.F. (1998): – Sulphur in forages. Proceedings of Symposium An Foras<br />
Taluntais, Dablin, 197-206 pp.<br />
14. Webb K.E.; Matthews J.K. (1998): – Peptide absorpti<strong>on</strong> and its significance in<br />
ruminant protein metabolism. In: Peptides in Mammalian Protein Metabolism.<br />
Portland, L<strong>on</strong>d<strong>on</strong>; 104-142 pp.<br />
15. Weinberg ZG.; Muck ME.; Weimer RJ. (2003): – The survival of silage<br />
inoculant lactic acid bacteria in rumen fluid. Journal of Applied Microbiology, 94:<br />
1066-1078 pp.<br />
16. Wilkins RJ (1996): – Silage producti<strong>on</strong> and utilizati<strong>on</strong>-Perspectives from the<br />
Silage Research <str<strong>on</strong>g>C<strong>on</strong>ference</str<strong>on</strong>g>s. Proceedings of the 11 th <str<strong>on</strong>g>Internati<strong>on</strong>al</str<strong>on</strong>g> Silage<br />
<str<strong>on</strong>g>C<strong>on</strong>ference</str<strong>on</strong>g>, Aberystwyth 37-48 pp.<br />
17. Woolford M.K. (2004): – Inoculantii pentru silozuri: aspecte moderne. Alltech<br />
Technical Publicati<strong>on</strong>s, 73-108 pp.<br />
333
PHYTOPHTORA INFESTANS - ECOLOGICAL CONTROL WITHIN<br />
ROMANIAN (TRANSYLVANIAN PLANE) POTATO CROP<br />
Ioan Oroian, Liviu Hol<strong>on</strong>ec, Viorel Florian, Laura Paulette, Ant<strong>on</strong>ia Odagiu<br />
University of Agricultural Sciences and Veterinary Medicine, Faculty of Agriculture,<br />
3-5 Mănăştur St., 400372 Cluj-Napoca, Romania, e-mail: neluoroian@gmail.com<br />
ABSTRACT<br />
The research aimed the following objectives: recording blight appearance, evoluti<strong>on</strong> and<br />
acti<strong>on</strong> within a technology of organic culture; testing new products and/or products used<br />
in other cultures for potato pathogens; the identificati<strong>on</strong> of other natural products used<br />
in disease c<strong>on</strong>trol; recording the behavior of different potato varieties against mycotic<br />
pathogens in organic agriculture and establishing the sorts of varieties in areas of<br />
interest. Treatments with nine organic products, used for blight c<strong>on</strong>trol, were performed<br />
in experimental field: nutritive clay 1%, 1/1whey, copper sulfate in associati<strong>on</strong> with<br />
lime 1%, bent<strong>on</strong>ite 0.5%, copper oxychloride 4kg/ha, degreased milk 1/1, copper<br />
hydroxide 4kg/ha, cosmetic clay 1% and potassium permanganate 0.3%. The<br />
experimental results and statistical interpretati<strong>on</strong> reveal the validity of the well known<br />
classical products in potato blight c<strong>on</strong>trol. Those based <strong>on</strong> copper (4 kg/ha copper<br />
oxychloride, 4 kg/ha copper hydroxide, 1% copper sulfide in associati<strong>on</strong> with lime), and<br />
other products tested within this trial and susceptible for favorable effects due to their<br />
pH, pellicular protecti<strong>on</strong>, or other properties recoded no significant efficacy, except<br />
whey1/1.<br />
Key words: Phytophora infestans, potato, treatment<br />
INTRODUCTION<br />
The organic agriculture appeared as alternative to intensive, c<strong>on</strong>venti<strong>on</strong>al<br />
(industrialized) agricultural practice, based <strong>on</strong> producti<strong>on</strong> maximizati<strong>on</strong> using inputs,<br />
and high quantities of the energo-intensive stimulators of producti<strong>on</strong>, with the aim of<br />
c<strong>on</strong>tinuous increase of agricultural producti<strong>on</strong> for a prep<strong>on</strong>derantly urban populati<strong>on</strong> in<br />
full process of development (Morar et al., 2003). Within organic agriculture, an<br />
assembly of balanced culture techniques (technological chains) in c<strong>on</strong>cordance with<br />
natural envir<strong>on</strong>mental rules describes the potato culture, with the aim of obtaining crops<br />
of high nutriti<strong>on</strong>al and health quality. Because in our country, <strong>on</strong>ly the first stages of the<br />
organic agriculture are developing and elaborated studies <strong>on</strong> potato culture within<br />
organic system were not yet performed, the study of the suitability of potato culture<br />
within this agricultural system is imposed. The c<strong>on</strong>trol of the main mycotic pathogenic<br />
agent, which produces blight in this culture, will be focused (D<strong>on</strong>escu et al., 1996;<br />
Orian, 2003; Oroian et al., 2003; Plămădeală, 1999).<br />
MATERIAL AND METHOD<br />
The experiments were performed in pedo-climatic c<strong>on</strong>diti<strong>on</strong>s of the Transylvanian<br />
Plane, with a transiti<strong>on</strong> temperate clime and specific traits de<strong>term</strong>ined by the general<br />
orientati<strong>on</strong> of the summits and slopes of Jucu village. This area is part of the wet and<br />
colder specific spreading area of the Transylvanian Plane, Someş Plane. The vertic clay<br />
334
chernozem represents the dominant soil, which exhibits the successi<strong>on</strong> of the Am-Bty-<br />
Cca horiz<strong>on</strong>s characteristic for the forest steppe areas, with marls rich in CaCO3. They<br />
c<strong>on</strong>fer the soil two essential traits: heavy texture and high degree of basic saturati<strong>on</strong>.<br />
The de<strong>term</strong>ined agro-chemical indices are characterized by the neutral soil reacti<strong>on</strong>,<br />
with values between 6.9 - 7.1 (in water), 3.56 - 3.92% humus c<strong>on</strong>tent in arable layer,<br />
0.183 - 0.196% total nitrogen supplies, very high phosphorus c<strong>on</strong>tent (15 ppm) and<br />
mobile potassium (240 ppm).<br />
The potato varieties used within this trial were planted at 70 cm distance between<br />
rows using 4SaBP equipment. Two rows by variety were planted. The following<br />
varieties were used: Tentant, Frumoasa, Robusta, Rozal, Redsec, Roclas, Dacia, Rodas,<br />
Amelia, Agatha, Sante and Cristian.<br />
In experimental filed, treatments with nine blight c<strong>on</strong>trol organic products were<br />
performed: nutritive clay 1%, whey 1/1, copper sulfate in associati<strong>on</strong> with lime 1%,<br />
bent<strong>on</strong>ite 0,5%, copper oxychloride 4kg/ha, degreased milk 1/1, copper hydroxide<br />
4kg/ha, cosmetic clay 1% and potassium permanganate 0,3%.<br />
The experimental data were statistically processed and interpreted using the dispersi<strong>on</strong><br />
analyzes (ANOVA) and Duncan test.<br />
RESULTS<br />
The results c<strong>on</strong>cerning the blight attack degree (AD%) <strong>on</strong> leafs in studied varieties are<br />
presented in figures 1 - 11. The observati<strong>on</strong> was performed after two weeks from<br />
treatment applicati<strong>on</strong>.<br />
Figure 1 represents the c<strong>on</strong>trol diagram, and it is the reference point for the results<br />
obtained after treatment with unc<strong>on</strong>venti<strong>on</strong>al products used in fight against potato<br />
blight. Within c<strong>on</strong>trol variant, Robusta, Rozal, Dacia, Sante and Cristian varieties<br />
recorded the lowest blight attack degree, while Roclas variety recorded the highest.<br />
AD%<br />
5<br />
4,5<br />
4<br />
3,5<br />
3<br />
2,5<br />
2<br />
1,5<br />
1<br />
0,5<br />
0<br />
Tentant<br />
Frumoasa<br />
Robusta<br />
Rozal<br />
Redsec<br />
Roclas<br />
Figure 1: The behavior against mushroom Phytophthora infestans attack in c<strong>on</strong>trol<br />
Positive results after treatment with nutritive clay were recorded in Tentant, Roclas and<br />
Robusta varieties (figure 2). The attack degree recorded in Robusta variety was 0 (zero),<br />
while Agatha and Rivera varieties had negative reacti<strong>on</strong>.<br />
335<br />
Dacia<br />
Amelia<br />
Agatha<br />
Rivera<br />
Sante<br />
Cristian
AD%<br />
4<br />
3,5<br />
3<br />
2,5<br />
2<br />
1,5<br />
1<br />
0,5<br />
0<br />
Tentant<br />
Frumoasa<br />
Robusta<br />
Rozal<br />
Redsec<br />
Roclas<br />
Dacia<br />
Amelia<br />
Agatha<br />
Rivera<br />
Sante<br />
Cristian<br />
Figure 2: The influence of the nutritive clay treatment <strong>on</strong> the attack degree<br />
One positive reacti<strong>on</strong> was recorded when whey treatment was applied in Roclas variety<br />
(figure 3). 10% decrease was obtained compared to c<strong>on</strong>trol. Neutral reacti<strong>on</strong> was<br />
obtained in Tentant variety.<br />
AD%<br />
5<br />
4,5<br />
4<br />
3,5<br />
3<br />
2,5<br />
2<br />
1,5<br />
1<br />
0,5<br />
0<br />
Tentant<br />
Frumoasa<br />
Robusta<br />
Rozal<br />
Redsec<br />
Roclas<br />
Dacia<br />
Amelia<br />
Agatha<br />
Rivera<br />
Sante<br />
Cristian<br />
Figure 3: The evoluti<strong>on</strong> of the blight attack when whey was used<br />
After treatment with copper sulfate in associati<strong>on</strong> with lime, the Roclas variety recorded<br />
a positive reacti<strong>on</strong>, with 20% decrease of blight attack degree compared to c<strong>on</strong>trol. In<br />
Tentant, Agatha and Rivera varieties a neutral reacti<strong>on</strong> was recorded (figure 4).<br />
The bent<strong>on</strong>ite treatment de<strong>term</strong>ined positive reacti<strong>on</strong> in two varieties, meaning<br />
Roclas, where the attack degree decreased by 20% compared to c<strong>on</strong>trol and Agatha<br />
where the attack degree decreased by 10% compared to c<strong>on</strong>trol. In Tentant, Dacia and<br />
Rivera varieties a neutral reacti<strong>on</strong> was recorded (figure 5).<br />
336
AD%<br />
AD%<br />
4<br />
3,5<br />
3<br />
2,5<br />
2<br />
1,5<br />
1<br />
0,5<br />
0<br />
Tentant<br />
Frumoasa<br />
Robusta<br />
Rozal<br />
Redsec<br />
Roclas<br />
Figure 4: The effect of the copper sulfate in associati<strong>on</strong> with lime <strong>on</strong> the attack degree<br />
4<br />
3,5<br />
3<br />
2,5<br />
2<br />
1,5<br />
1<br />
0,5<br />
0<br />
Tentant<br />
Frumoasa<br />
Robusta<br />
Rozal<br />
Redsec<br />
Roclas<br />
Figure 5: The effect of the treatment with copper bent<strong>on</strong>ite <strong>on</strong> the attack degree<br />
Only in two varieties a significant positive reacti<strong>on</strong> was recorded when the treatment<br />
with copper oxychloride was applied: in Roclas variety the attack degree decreased by<br />
20% compared to c<strong>on</strong>trol and in Tentant the attack degree was 10% lower compared to<br />
c<strong>on</strong>trol. In the other variants, a neutral (Agatha and Rivera) or negative reacti<strong>on</strong> was<br />
recorded (figure 6).<br />
337<br />
Dacia<br />
Dacia<br />
Amelia<br />
Amelia<br />
Agatha<br />
Agatha<br />
Rivera<br />
Rivera<br />
Sante<br />
Sante<br />
Cristian<br />
Cristian
AD%<br />
4<br />
3,5<br />
3<br />
2,5<br />
2<br />
1,5<br />
1<br />
0,5<br />
0<br />
Tentant<br />
Frumoasa<br />
Robusta<br />
Rozal<br />
Redsec<br />
Roclas<br />
Figure 6: The effect of the treatment with copper oxychloride <strong>on</strong> the attack degree<br />
The degreased milk treatment de<strong>term</strong>ined a positive reacti<strong>on</strong> in Roclas variety where the<br />
blight attack degree decreased by 20% compared to c<strong>on</strong>trol. In Tentant, Agatha and<br />
Rivera varieties, a neutral reacti<strong>on</strong> was de<strong>term</strong>ined. In Robusta, Rozal, Redsec and<br />
Cristian varieties, de attack degree increased by 20% compared to c<strong>on</strong>trol (figure 7).<br />
AD%<br />
4<br />
3,5<br />
3<br />
2,5<br />
2<br />
1,5<br />
1<br />
0,5<br />
0<br />
Tentant<br />
Frumoasa<br />
Robusta<br />
Rozal<br />
Redsec<br />
Roclas<br />
Figure 7: The influence of the treatment wioth degreased milk <strong>on</strong> the attack degree<br />
When copper hydroxide treatment was applied, a positive reacti<strong>on</strong> was recorded in<br />
Tentant and Roclas varieties, with a 10% decrease of the attack compared to c<strong>on</strong>trol in<br />
each variety. In Rivera variety a neutral reacti<strong>on</strong> was recorded (figure 8).<br />
A positive reacti<strong>on</strong> was recorded in Roclas and Tentant varieties when cosmetic<br />
clay treatment was applied. The blight attack degree decreased by 40% compared to<br />
c<strong>on</strong>trol in Roclas variety and by 10% in Tentant variety (figure 9).<br />
338<br />
Dacia<br />
Dacia<br />
Amelia<br />
Amelia<br />
Agatha<br />
Rivera<br />
Agatha<br />
Rivera<br />
Sante<br />
Cristian<br />
Sante<br />
Cristian
Negative reacti<strong>on</strong> was recorded in the following varieties: Cristian, where the blight<br />
attack degree increased by 30% compared to c<strong>on</strong>trol, and Frumoasa, Robusta, Rozal,<br />
Redsec, Dacia, Amelia and Sante varieties, where the blight attack degree increased by<br />
10% compared to c<strong>on</strong>trol.<br />
AD%<br />
4<br />
3,5<br />
3<br />
2,5<br />
2<br />
1,5<br />
1<br />
0,5<br />
0<br />
Tentant<br />
Frumoasa<br />
Robusta<br />
Rozal<br />
Redsec<br />
Roclas<br />
Figure 8: The effect of the treatment with copper hydroxide <strong>on</strong> the attack degree<br />
AD%<br />
4<br />
3,5<br />
3<br />
2,5<br />
2<br />
1,5<br />
1<br />
0,5<br />
0<br />
Tentant<br />
Frumoasa<br />
Robusta<br />
Rozal<br />
Redsec<br />
Roclas<br />
Figure 9: The effect of the treatment with cosmetic clay <strong>on</strong> the attack degree<br />
The potassium permanganate treatment de<strong>term</strong>ined a positive reacti<strong>on</strong> in Tentant and<br />
Roclas varieties, where the blight attack degree decreased by 10% compared to c<strong>on</strong>trol.<br />
The negative reacti<strong>on</strong> was recorded in: Cristian variety, where the attack degree<br />
increased by 30% compared to c<strong>on</strong>trol, and Rozal, Dacia and Sante varieties, where the<br />
blight attack degree increased by 20% compared to c<strong>on</strong>trol (figure 10).<br />
339<br />
Dacia<br />
Dacia<br />
Amelia<br />
Amelia<br />
Agatha<br />
Agatha<br />
Rivera<br />
Rivera<br />
Sante<br />
Cristian<br />
Sante<br />
Cristian
AD%<br />
4<br />
3,5<br />
3<br />
2,5<br />
2<br />
1,5<br />
1<br />
0,5<br />
0<br />
Tentant<br />
Frumoasa<br />
Robusta<br />
Rozal<br />
Redsec<br />
Roclas<br />
Figure 10: The effect of the treatment with potassium permanganate <strong>on</strong> the attack degree<br />
The treatments applied with the aim of blight c<strong>on</strong>trol de<strong>term</strong>ined a different reacti<strong>on</strong> in<br />
studied varieties, except Roclas variety, which was the <strong>on</strong>ly variety that positively<br />
reacted to all applied treatments (figure 11).<br />
C<strong>on</strong>trol<br />
Nutritive clay<br />
Whey<br />
Copper sulphate in associati<strong>on</strong> with lime<br />
Bent<strong>on</strong>ite<br />
Copper oxychloride<br />
Degreased milk<br />
Copper hydroxide<br />
Cosmetic clay<br />
Potassium permanganate<br />
Figure 11: The evoluti<strong>on</strong> of the blight atack within the interacti<strong>on</strong> between<br />
variety and treatment<br />
340<br />
Dacia<br />
Tentant<br />
Amelia<br />
Frumoasa<br />
Robusta<br />
Agatha<br />
Rozal<br />
Redsec<br />
Rivera<br />
Roclas<br />
Sante<br />
Dacia<br />
Amelia<br />
Cristian<br />
Agatha<br />
Rivera<br />
Sante<br />
Cristian<br />
5<br />
4<br />
3<br />
2<br />
1<br />
0
The Tentant variety positively reacted to five treatments, and had a neutral reacti<strong>on</strong> to<br />
the other applied treatments. The Robusta variety positively reacted to the treatment<br />
with nutritive clay, where recorded the attack degree was 0 (zero), to other two<br />
products, and a negative reacti<strong>on</strong> to the other applied treatments. The Agatha variety<br />
positively reacted <strong>on</strong>ly to <strong>on</strong>e product, negatively to <strong>on</strong>e product, too, and neutral to the<br />
others. The Rivera variety positively reacted to <strong>on</strong>e product, negatively to <strong>on</strong>e product,<br />
and neutral for the rest of them. In the other varieties a negative reacti<strong>on</strong> was recorded.<br />
DISCUSSIONS<br />
The well-known copper based classical products (copper oxychloride 4kg/ha, copper<br />
hydroxide 4kg/ha, copper sulfate in associati<strong>on</strong> with lime 1%) are efficient in potato<br />
blight c<strong>on</strong>trol. The other tested products susceptible of favorable effects due to their pH,<br />
pellicle protecti<strong>on</strong> or other properties were not efficient, except the variant with whey<br />
1/1.<br />
Within experimental c<strong>on</strong>diti<strong>on</strong>s, the lowest potato blight attack was emphasized in<br />
Amelia and Robusta variety, and the lowest alternariosis attack in Sante, Robusta and<br />
Rozal varieties, which recommend them as suitable for organic agriculture.<br />
Am<strong>on</strong>g the tested varieties, five are suitable to be used in organic agriculture: Robusta,<br />
Rozal, Redsec, Amelia and Sante. High producti<strong>on</strong> and degree of blight and alteriosis<br />
resistance characterize these varieties during testing c<strong>on</strong>diti<strong>on</strong>s. Further research is<br />
needed in order to identify other varieties suitable to be used within organic agriculture<br />
system.<br />
The use of unc<strong>on</strong>venti<strong>on</strong>al products susceptible to have properties against potato<br />
pathogenic agents or capacity of their attack limitati<strong>on</strong> represents an alternative which<br />
can be developed within the c<strong>on</strong>text of implementati<strong>on</strong> and extensi<strong>on</strong> of the sustainable<br />
and organic agriculture system.<br />
REFERENCES<br />
D<strong>on</strong>escu D., Enoiu, M., Frâncu G., Plămădeala, B., Popescu, A. (1996): Ghid practic<br />
de protecţie a cartofului. Ed. Ceres, Bucureşti<br />
Morar, G., Fiţiu, A., Cernea, S., Vâtcă, S. D., Oltean, M. I., Sârbu Camelia (2003):<br />
Tehnologii în agricultura ecologică, Ed. Risoprint, Cluj-Napoca<br />
Oroian, I. (2003): Protecţia şi nutriţia plantelor, Ed. AcademicPress, Cluj-Napoca<br />
Oroian, I., Fiţiu, A., Florian, V., Carmen Puia, Dumitraş Adelina, Roiban, G.,<br />
Paulette Laura (2003): C<strong>on</strong>trolul patogenilor în agricultura ecologică,<br />
Ed. AcademicPress, Cluj-Napoca<br />
Plămădeală, B. (1999): Mana cartofului. Biologie şi c<strong>on</strong>trol, Ed. Ceres, Bucureşti<br />
341
THE DYNAMICS OF THE POPULATIONS OF CEREAL PESTS AND<br />
PATHOGENS IN THE ROMANIAN WESTERN PLAIN<br />
Elena Bucurean, Nicolae Csép<br />
University of Oradea, Faculty for Envir<strong>on</strong>mental Protecti<strong>on</strong> Oradea<br />
SUMMARY<br />
In a given agroecosystem, the exact de<strong>term</strong>inati<strong>on</strong> of pest species, meaning primary,<br />
occasi<strong>on</strong>al, potential and migrating pests, the identificati<strong>on</strong> of harmful plant pathogens,<br />
requires particular attenti<strong>on</strong>: these species are different for each crop or z<strong>on</strong>e, an aspect<br />
that imparts certain characteristics to the fighting schemes.<br />
Over the years, it has been noticed that the damages made by pests and diseases in<br />
the western part of the country are not always of equal magnitude: these damages<br />
greatly depend <strong>on</strong> the envir<strong>on</strong>mental and biotic factors’ influence <strong>on</strong> the species present<br />
in the area (Bucurean, 2004; Csep, 2006). C<strong>on</strong>sequently, there are some species that are<br />
present and cause damages every year, species of both cyclical and seas<strong>on</strong>al ec<strong>on</strong>omic<br />
importance and there are other species that are simply part of the area’s fauna.<br />
The spectrum of plant-specific pathogens also undergoes changes, especially under<br />
the influence of the regime of temperature and humidity during the crop’s vegetati<strong>on</strong><br />
period. Performing the meteorological observati<strong>on</strong>s and establishing the inoculum<br />
reserve holds special importance in the applicati<strong>on</strong> of envir<strong>on</strong>ment-friendly<br />
technologies.<br />
Key words: wheat, pests, diseases, damages.<br />
INTRODUCTION<br />
Taking into account the fact that winter wheat is cultivated in over 45 countries, feeding<br />
35–40% of the world’s populati<strong>on</strong>, as well as the fact that it ensures about 40% of the<br />
total need of calories in human diet, it is absolutely imperative to be aware of the<br />
entomofauna and pathogens present in cereal crops every year, in order to foresee and<br />
then prevent the appearance of pest invasi<strong>on</strong>s (Ghizdavu et al.,1998; Perju et al.,1993)<br />
and epiphytotic infecti<strong>on</strong>s due the harmful pathogens. This aspect also stands with<br />
regard to other local crops, with the additi<strong>on</strong> that it is necessary to be aware of the<br />
populati<strong>on</strong>s’ tendencies and the reserve of inoculum, in relati<strong>on</strong> to the evoluti<strong>on</strong> of the<br />
abiotic (temperature, humidity, soil, air currents etc.) and traffic factors (food and the<br />
relati<strong>on</strong>s between predators, parasites and host pests).<br />
RESULTS<br />
a.) Pest populati<strong>on</strong>s: In cereal crops, the tendency of the existing pest populati<strong>on</strong>s in<br />
the area is of slight increase in cereal bugs (Eurygaster spp, Aelia spp), wheat thrips<br />
(Haplotrips tritici), wheat chafer (Anisoplia spp), cereal leaf beetle (Oulema<br />
melanopus), cereal green aphid (Schyzaphis graminum) and various species of<br />
phytophagous diptera (Oscinella sp, Agr<strong>on</strong>yza sp, Phytomiza sp), wire worms (Agriotes<br />
spp) as well as C<strong>on</strong>tinental field vole (Microtus arvalis), whose attack however has a<br />
cyclical and seas<strong>on</strong>al character.<br />
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Table 1. The density of harmful pest species in wheat crops from Oradea in the 2005<br />
– 2006 agricultural year<br />
Species Date of c<strong>on</strong>trol Density<br />
Zabrus tenebrioides October 8 2.7 larvae/ m²<br />
Eurygaster maura May 11 6.2 adults / m²<br />
Eurygaster integriceps May 11 3.1 adults / m²<br />
Aelia rostrata May 11 4.1 adults / m²<br />
Oulema melanopus April 9 12 adults / m²<br />
Oulema melanopus May 13 220 larvae / m²<br />
Schyzaphis graminum June 24 56 aphids / plant<br />
Haplotrips tritici June 16 27 adults and larvae / ear<br />
Anisoplia spp June 29 9 adults / m²<br />
Oscinella frit October 22 11.3% attacked plants<br />
Cephus pygmaeus June 29 3.4% attacked plants<br />
Macrosteles laevis October 28 11 adults / m²<br />
Agriotes spp April 5 6 larvae / m²<br />
Naturally, some of these species hold a minor ec<strong>on</strong>omic importance in the area, partly<br />
as a result of the fact that plants rebuild their vegetative mass, thus no producti<strong>on</strong> losses<br />
due exclusively to pests are registered.<br />
Of the total number of pests collected in cereal ecosystems, the dominant species were<br />
the <strong>on</strong>es whose frequency is presented in table 2. The numbers represent the average<br />
values of mowings at different dates.<br />
Table 2. The frequency of pest fauna (%) in cereal crops<br />
Crop Wheat Barley Oats Triticale<br />
Species % % % %<br />
Phyllotreta spp 29.7 22.7 26.7 42.9<br />
Eurygaster maura 4.5 2.8 0.7 0.2<br />
Aelia acuminata 1.9 1.4 0.7 0.5<br />
Oulema melanopus 58.9 11.8 77.8 10.7<br />
Cephus pygmaeus 12.4 6.7 2.9 16.5<br />
Haplotrips tritici 71.0 41.6 - 0.2<br />
Schyzaphis graminum 46.2 44.4 45.7 2.9<br />
Anisoplia spp 4.9 2.6 - 1.5<br />
Agromyza nigra 18.0 2.3 - -<br />
As to maize crops, taking into account the fact that the majority of soils in the area have<br />
an acid or weak acid reacti<strong>on</strong>, particular difficulties are caused by the larvae of Agriotes<br />
spp, which populate these soils. Their density fluctuates from <strong>on</strong>e year to another,<br />
depending <strong>on</strong> the quantity of rainfall during autumn – spring and <strong>on</strong> the humidity of the<br />
343
soil during the sprouting period, this density varying between 12 – 16 larvae of different<br />
ages / m². Moreover, important damages are caused by the larvae of the turnip moth<br />
(Agrotis segetum), which, due to their accentuated polyphagism, bring about damages<br />
not <strong>on</strong>ly in maize crops, but also in beet, potato and leguminous plants. The density of<br />
the butterflies caught by the pherom<strong>on</strong>al traps varied from <strong>on</strong>e m<strong>on</strong>th to another,<br />
reaching a peak during the flight period.<br />
b.) Plant pathogens: Regarding the culture of winter wheat, the structure of the<br />
complex of pathogens underwent modificati<strong>on</strong>s as to the cultivated species, as well as to<br />
the climatic changes. The last 5 years, having had a rather droughty character, have<br />
been less favorable to pathogen infecti<strong>on</strong>s, favoring the proliferati<strong>on</strong> of insects instead.<br />
Table 3. The main winter wheat diseases in the Romanian Western Plain<br />
Symptomps Disease<br />
Cause agent Ec<strong>on</strong>omical<br />
expressed <strong>on</strong>: comm<strong>on</strong> name<br />
importance<br />
Leaves Barley Yellow<br />
Dwarf<br />
BYDV medium<br />
Leaves Powdery Erysiphe graminis f.sp. low<br />
mildew tritici<br />
Leaves leaf spot H. tritici-repentis medium<br />
Leaves leaf rust Puccinia rec<strong>on</strong>dita low<br />
Leaves stripe rust P. striiformis variable<br />
Seed / head head blight Fusarium spp. medium<br />
Head comm<strong>on</strong> bunt T. foetida, T. Caries variable<br />
Lower stem eyespot Pseudocercosporella<br />
herpotrichoides<br />
variable<br />
Lower stem sharp eyespot Rhizoct<strong>on</strong>ia sp. low<br />
The symptoms of barley yellow dwarf virus were frequently identified between the<br />
years 2000-2002, when the installati<strong>on</strong> of vectors (Aphidae) in autumn was noticed in<br />
str<strong>on</strong>gly developed crops. Because no c<strong>on</strong>trol is available for many viral diseases, such<br />
as barley yellow dwarf, we must supervise the installati<strong>on</strong> (and, when needed, the<br />
fighting) of vectors.<br />
Of the foliar diseases, the importance of the powdery mildew (Erysiphe graminis)<br />
decreased, while that of the yellow leaf spot (Helminthosporium tritici-repentis Died.)<br />
increased, the symptoms of the latter <strong>on</strong> the upper leaves being easily mistaken for the<br />
effects of the drought. The pathogen is causing, in the current c<strong>on</strong>diti<strong>on</strong>s, the greatest<br />
yield losses (up to 250-350 kg/ha), associated with the reducti<strong>on</strong> of seminal quality.<br />
Fungicidal treatment with systemic fungicides, according to regi<strong>on</strong>al advice at heading,<br />
balanced fertilisati<strong>on</strong> can c<strong>on</strong>tribute effectively to the limitati<strong>on</strong> of losses.<br />
The brown leaf rust (Puccinia rec<strong>on</strong>dita) has been present every year, without<br />
affecting more than 5 % of the leaves’ area of assimilati<strong>on</strong>. The stripe rust (Puccinia<br />
striiformis) appeared <strong>on</strong> some very susceptible wheat species, at the end of the<br />
344
vegetati<strong>on</strong> period, without affecting more than 3% of the area of upper leaves. However,<br />
the species represents a potential danger, due to possible damages.<br />
The evoluti<strong>on</strong> of droughty climate during the past few years has significantly<br />
reduced head blight due to infecti<strong>on</strong>s with Fusarium spp. The infecti<strong>on</strong>s are tightly<br />
c<strong>on</strong>nected to the evoluti<strong>on</strong> of the climatic factor in the post – heading period.<br />
In the years following the re-establishment of former properties, as a result of not<br />
performing or inadequately performing seed dressing, the losses in producti<strong>on</strong> caused<br />
by the comm<strong>on</strong> bunt have resurfaced. Of the species of Tilletia sp., the most frequent in<br />
this ecological area are T. caries and T. foetida. During those years, frequencies of<br />
attack <strong>on</strong> the ears of up to 15-20% were recorded. The correct applicati<strong>on</strong> of fungicides<br />
has reduced this indicator below the frequency of 0.2%. Although it is present in many<br />
Transylvanian counties, T. nanifica has not been recorded in this area.<br />
Lower stem diseases (eyespot - Pseudocercosporella herpotrichoides; sharp<br />
eyespot -Rhizoct<strong>on</strong>ia sp.; fusarium rot – Fusarium sp.) are present <strong>on</strong>ly occasi<strong>on</strong>ally,<br />
drawing attenti<strong>on</strong> <strong>on</strong> some technological causes that have favored their appearance:<br />
m<strong>on</strong>ocultures, unbalanced fertilizati<strong>on</strong>, and very susceptible wheat species. Crop<br />
rotati<strong>on</strong> (avoid planting wheat and barley after take-all infested wheats, balanced<br />
fertilisati<strong>on</strong>) are the most efficient preventive measures.<br />
Table 4. The main winter barley diseases in the Romanian Western Plain<br />
Symptoms Disease comm<strong>on</strong> Cause agent Ec<strong>on</strong>omical<br />
expressed <strong>on</strong>: name<br />
importance<br />
Leaves Barley yellow<br />
dwarf virus<br />
BYDV medium<br />
Leaves Powdery mildew Erysiphe graminis<br />
f.sp.hordei<br />
low<br />
Leaves / head Leaf stripe Pyrenophora<br />
graminea<br />
medium<br />
Leaves Rhynchosporium Rhynchosporium medium<br />
leaf-spot<br />
secalis<br />
Head True loose smut Ustilago nuda variable<br />
Head Covered smut Ustilago hordei low<br />
As to the cultures of winter barley that have profited from adequate seed dressing, the<br />
presence of seed-born pathogens, true loose smut (Ustilago nuda), covered smut (U.<br />
hordei), barley leaf stripe (Pyrenophora graminea), was below the limit of provoking<br />
some important yield losses.<br />
CONCLUSIONS<br />
1. The pests of cereal crops represent limitative factors for the producti<strong>on</strong> and<br />
supervising them in the area is a necessity. Therefore, it is imperative to m<strong>on</strong>itor<br />
their numerical evoluti<strong>on</strong> in time, <strong>on</strong> groups of cultures. The pest fauna is widely<br />
345
epresented in this area, and annual damages are recorded as a result of the attacks<br />
of the cereal leaf beetle, cereal bugs and wire worms.<br />
2. The largest number of harmful pest species was registered in wheat crops, in<br />
comparis<strong>on</strong> with the other cereal crops (barley, oats, triticale). The cereal leaf<br />
beetle’s presence corresp<strong>on</strong>ded to percentages between 10.7 and 77.8%, which<br />
proves that this species represents a real problem for the culture of cereals.<br />
3. The specific evoluti<strong>on</strong> of climatic factors has caused a temporary reducti<strong>on</strong> of some<br />
pathogens c<strong>on</strong>sidered dangerous (head blight, eyespot, fusarium rot), but which,<br />
due to their high infectious potential, must be under c<strong>on</strong>stant surveillance in order<br />
to avoid possible damages.<br />
4. Seed dressing , avoiding early seedling, balanced fertilisati<strong>on</strong>, crop rotati<strong>on</strong>,<br />
fungicidal treatments especially after heading, ensure the most effective and safe<br />
protecti<strong>on</strong> of cereals against pathogens.<br />
REFERENCES<br />
1. Bucurean Elena, 2004: Prognoza si avertizare in combaterea daunatorilor la<br />
culturile agricole, Ed. Universităţii din Oradea.<br />
2. Bucurean Elena et al., 2006: Protecţia eficientă şi sigură a plantelor faţă de<br />
dăunători. Ed. Universităţii din Oradea.<br />
3. Csep N.,2006: Protecţia eficientă şi sigură a plantelor faţă de boli.Ed.Universităţii<br />
din Oradea.<br />
4. Ghizdavu I. et al., 1998: Entomologie agricola, E.D.P. Bucuresti<br />
5. Perju T. et al., 1993: Protectia integrata a culturilor de leguminoase, Ed. Ceres<br />
Bucuresti<br />
346
THE IMPORTANCE OF CONTROLLING FORMALDEHYDE FUMIGATION<br />
Lucian Bara, Camelia Bara<br />
University of Oradea, Faculty of Envir<strong>on</strong>mental Protecti<strong>on</strong><br />
26, General Magheru Street, Oradea<br />
ABSTRACT<br />
A comparative study of formaldehyde (HCHO) fumigati<strong>on</strong> was carried out by c<strong>on</strong>trolled<br />
vaporizati<strong>on</strong>, using an electric vapor generator, and by the Formalinpermanganate.method.<br />
De<strong>term</strong>inati<strong>on</strong> of vapor levels as well as bactericidal acti<strong>on</strong> showed the generator to be more<br />
effective. Maximum achievable fumigant levels were temperature dependent and related to<br />
the equilibrium vapor c<strong>on</strong>centrati<strong>on</strong> of HCHO. At a room temperature of 21°C, vaporizati<strong>on</strong><br />
of more than 2,000 ytg of HCHO per liter resulted in c<strong>on</strong>versi<strong>on</strong> of HCHO to<br />
paraformaldehyde, which c<strong>on</strong>densed <strong>on</strong> surfaces and c<strong>on</strong>tributed to prol<strong>on</strong>ged residual<br />
vapor levels.<br />
An electr<strong>on</strong>ic m<strong>on</strong>itor is described which is capable of detecting HCHO levels as low<br />
as 10 Ag/liter and can be used to m<strong>on</strong>itor the complete fumigati<strong>on</strong> process.<br />
INTRODUCTION<br />
Formaldehyde fumigati<strong>on</strong> has l<strong>on</strong>g been an accepted method of sterilizati<strong>on</strong> for areas where<br />
microbiological cleanliness is required. Fumigati<strong>on</strong> is usually carried out by mixing<br />
potassium permanganate and an excess of Formalin (40% formaldehyde) soluti<strong>on</strong> in a<br />
suitable c<strong>on</strong>tainer; sufficient heat is generated by oxidati<strong>on</strong> of part of the formaldehyde<br />
(HCHO) with permanganate to vaporize the remaining formaldehyde and water.<br />
This method of fumigati<strong>on</strong> is violent, messy, and potentially explosive. In this paper,<br />
an alternative procedure is described for fumigati<strong>on</strong> by c<strong>on</strong>trolled vaporizati<strong>on</strong>, in an<br />
electrical vapor generator, of either Formalin or paraformaldehyde, a polymer of HCHO<br />
which is c<strong>on</strong>verted to HCHO vapor up<strong>on</strong> heating. Formaldehyde levels were compared in<br />
fumigati<strong>on</strong>s of several work areas by the c<strong>on</strong>trolled vaporizati<strong>on</strong> and Formalinpermanganate<br />
methods, and the effectiveness of each procedure was de<strong>term</strong>ined from the<br />
survival of resistant Bacillus subtilis spores.<br />
MATERIALS AND METHODS<br />
Reagents. A commercial Formalin soluti<strong>on</strong> c<strong>on</strong>sisting of 40% HCHO c<strong>on</strong>taining 10%<br />
methanol as a depolymerizing agent was used throughout. Paraformaldehyde powder was a<br />
white polymer c<strong>on</strong>sisting of 95% paraformaldehyde.<br />
Areas fumigated. A low-security area and a highsecurity area were m<strong>on</strong>itored throughout<br />
the study. The low-security area was a large processing laboratory; during fumigati<strong>on</strong> the air<br />
c<strong>on</strong>diti<strong>on</strong>ing was turned off, but no attempt was made to seal air supply or exhaust ducts.<br />
The high-security area was a large sealed laboratory whose air supply was carefully<br />
c<strong>on</strong>trolled; exhaust air was passed through a gas-fired furnace to ensure microbiological<br />
347
security. The air c<strong>on</strong>diti<strong>on</strong>ing for this area was turned off during fumigati<strong>on</strong>, and the air<br />
supply and exhaust outlet valves were closed.<br />
Assay of HCHO. Initially, formaldehyde vapor c<strong>on</strong>centrati<strong>on</strong>s were measured by taking air<br />
samples through rubber tubes protruding into the room at various sample points. Four<br />
samples were taken per sampling time, using an evacuated liter flask. Forty milliliters of 0.5<br />
M (NH4)2SO4 was added to each flask, which was then cooled to 4'C to dissolve the<br />
formaldehyde. One milliliter of a suitable diluti<strong>on</strong> of the resulting soluti<strong>on</strong> was then mixed<br />
with 1 ml of a freshly prepared chromotropic acid reagent (1% chromotropic acid in 18 M<br />
H2SO4), and 8 ml of 18 M H2SO4 was added. After standing for 10 min, the mixture was<br />
diluted to a final volume of 25 ml, and the optical density was de<strong>term</strong>ined at 540 nm.<br />
HCHO was estimated from a standard curve in the range 5 to 40 yg/ ml, and the mean<br />
c<strong>on</strong>centrati<strong>on</strong> from the four samples was de<strong>term</strong>ined.<br />
Biological test systems. The spore suspensi<strong>on</strong> used throughout was prepared from B.<br />
subtilis NCTC 8233 prepared by culture for 7 days <strong>on</strong> sporulati<strong>on</strong> agar and suspensi<strong>on</strong> of<br />
the resultant growth in water. The suspensi<strong>on</strong> was subjected to two cycles of heating to<br />
65°C for 30 min followed by centrifugati<strong>on</strong> and resuspensi<strong>on</strong> in distilled water. A sample<br />
was then stained and examined microscopically for spores. Glass rods 5 mm in diameter and<br />
150 mm l<strong>on</strong>g, with a loop at <strong>on</strong>e end and a graduati<strong>on</strong> mark 50 mm from the other, were<br />
dipped into the spore suspensi<strong>on</strong> to the mark. They were then suspended in a laminar flow<br />
cabinet to dry and stored in a sterile jar. Before fumigati<strong>on</strong>, rods were placed in different<br />
parts of the areas to be tested and transferred to a sterile screwcapped test tube at the end of<br />
each procedure, immediately after evacuati<strong>on</strong> of the fumigant. Twenty milliliters of saline<br />
was added to each tube to remove the spores from the rods, and an estimate of viable spores<br />
was obtained by plating diluti<strong>on</strong>s of the suspensi<strong>on</strong> <strong>on</strong>to nutrient agar. This was carried out<br />
within 2 h of removal of rods from the area. Plates were incubated at 37°C, and then<br />
col<strong>on</strong>ies were counted after 3 days and checked after 7 days.<br />
Electric vapor generator. Initially, HCHO was vaporized <strong>on</strong> a 500-W heating plate in a<br />
stainless-steel beaker. The apparatus used subsequently, c<strong>on</strong>sists of a stainless-steel 5-liter<br />
vessel with a 1,200-W heating element. Sufficient capacity was present to allow the additi<strong>on</strong><br />
of water for humidificati<strong>on</strong> and to c<strong>on</strong>tain the resulting foam. A copper plate was interposed<br />
between the element and the bottom of the vessel to ensure a uniform distributi<strong>on</strong> of heat at<br />
a thermostatically c<strong>on</strong>trolled temperature of approximately 170°C.<br />
Electr<strong>on</strong>ic formaldehyde m<strong>on</strong>itor. The apparatus used was based <strong>on</strong> a Figaro 812 N-type<br />
semic<strong>on</strong>ductor device, the c<strong>on</strong>ductivity of which changes in the presence of adsorbed<br />
molecules. No significant interference was observed from substances other than HCHO in<br />
the areas fumigated or by temperature changes normally encountered. The m<strong>on</strong>itor was<br />
calibrated in an enclosed system against HCHO released by paraformaldehyde vaporizati<strong>on</strong>.<br />
RESULTS<br />
HCHO levels by the Formalin-permanganate and hot plate vaporizati<strong>on</strong> methods. For<br />
Formalin-permanganate fumigati<strong>on</strong>s, quantities of Formalin recommended have ranged<br />
from 12 to 59 ml/m 3 of air space , and ratios for Formalin-permanganate have ranged from<br />
348
3:1 to 5:3 . In the following experiments, fumigati<strong>on</strong>s were carried out by using a mixture of<br />
17.8 ml of Formalin and 8.9 g of potassium permanganate per m 3 of air space in a lowsecurity<br />
area at room temperatures of 16, 24, and 300C. In preliminary experiments, room<br />
air samples were taken from each of four positi<strong>on</strong>s, including the ceiling and the floor, and<br />
HCHO levels were de<strong>term</strong>ined by chemical assay. Because no differences in c<strong>on</strong>centrati<strong>on</strong><br />
were observed between any positi<strong>on</strong>s, all results from the sampling points were averaged.<br />
HCHO levels were also measured after vaporizati<strong>on</strong> of 12.5 ml of Formalin (70% of that<br />
used in the Formalin-permanganate fumigati<strong>on</strong>s to allow for losses due to permanganate<br />
oxidati<strong>on</strong>) <strong>on</strong> a hot plate at a room temperature of 27°C.<br />
Survival of B. subtilis after fumigati<strong>on</strong> by the Formalin-permanganate method. B.<br />
subtilis spores survived <strong>on</strong> some rods located at various places in a low-security area during<br />
a 24- h Formnalin-permanganate fumigati<strong>on</strong> at an initial temperature of 1800. Viable counts<br />
of 2 x 106 were obtained from c<strong>on</strong>trol rods, but <strong>on</strong>ly two of nine rods exposed in the treated<br />
area were sterile; viable counts of 20 to 600 were obtained from the others. It is clear that,<br />
although this procedure was fairly effective, sterilizati<strong>on</strong> was not achieved.<br />
Two further experiments were carried out in a sealed jar at 20°C in c<strong>on</strong>trolled HCHO vapor<br />
levels of 150 and 300 Ag/liter of air, with relative humidity maintained at approximately<br />
100%. Exposure to 150, ug of HCHO per liter of air did not result in sterilizati<strong>on</strong> even after<br />
exposure for 6 h. Exposure to 300, Ag/liter was much more effective, and no viable spores<br />
were detected at 3 h.<br />
Use of electric vapor generator. The results from several fumigati<strong>on</strong>s are presented, using<br />
the electric vapor generator and different amounts of Formalin in a low-security area at<br />
different initial temperatures. In procedure 1, persistent HCHO levels similar to those<br />
obtained by the hot plate vaporizati<strong>on</strong> method were achieved. No viable B. subtilis spores<br />
were detected after 24 h of exposure <strong>on</strong> the 18 spore rods located in the main laboratory<br />
area, but a 0.5% survival was detected <strong>on</strong> a rod located in a closed cupboard. The fumigant<br />
level in the cupboard was 160, ug/liter after 6 h. Procedure 2 was carried out with the same<br />
volume of Formalin and an initial room temperature of 15°C. No surviving spores could be<br />
detected after 7 h of exposure <strong>on</strong> 12 spore rods located around the room, but two located in<br />
cupboards showed a survival of 1%. Procedures 3 through 5, which were carried out for<br />
different times and temperatures and with varying amounts of Formalin, are also described.<br />
In procedure 6, a mixture of paraformaldehyde and water equivalent to 8.9 ml of Formalin<br />
per m 3 of air was used. Again, no surviving spores could be detected from rods distributed<br />
throughout the main laboratory area in any of the procedures. All subsequent fumigati<strong>on</strong>s<br />
were carried out with 3.7 g of paraformaldehyde per m3 of room volume, which is<br />
c<strong>on</strong>verted to HCHO equivalent to that present in 8.9 ml of Formalin.<br />
Humidity in each area was increased to 80%, a level which has been shown to increase<br />
the effectiveness of fumigati<strong>on</strong> by causing HCHO to dissolve in a film of moisture around<br />
microorganisms, where it is more bactericidal than in vapor form .Use of Formalin at a<br />
room temperature of 200C would have c<strong>on</strong>tributed approximately 30% to the relative<br />
humidity. More reliable humidity c<strong>on</strong>trol could be achieved with paraformaldehyde, which<br />
was also more c<strong>on</strong>venient to use. Use of electr<strong>on</strong>ic HCHO m<strong>on</strong>itor. The electr<strong>on</strong>ic m<strong>on</strong>itor<br />
was developed because of the time involved in chemical assays of air samples. When the<br />
349
output of the m<strong>on</strong>itor was plotted against HCHO vapor levels obtained by chemical<br />
analysis, a power-law relati<strong>on</strong>ship was obtained. Such a relati<strong>on</strong>ship is particularly useful,<br />
since good sensitivity was achieved at the lower vapor levels and the total fumigati<strong>on</strong> range<br />
was covered <strong>on</strong> the same scale.<br />
Relati<strong>on</strong>ship between temperature and equilibrium vapor c<strong>on</strong>centrati<strong>on</strong>. The<br />
relati<strong>on</strong>ship between temperature and the equilibrium vapor c<strong>on</strong>centrati<strong>on</strong> is shown in. For<br />
normal air-c<strong>on</strong>diti<strong>on</strong>ed laboratories at 20 o C, the equilibrium vapor c<strong>on</strong>centrati<strong>on</strong> is<br />
approximately 2000 ,ug of HCHO per liter. HCHO vapor in excess of the equilibrium vapor<br />
c<strong>on</strong>centrati<strong>on</strong> will rec<strong>on</strong>dense <strong>on</strong> the coldest surfaces, producing a paraformaldehyde film<br />
over surfaces within the room. This film slowly evaporates, producing an unpleasant odor.<br />
HCHO levels after fumigati<strong>on</strong>. The persistence of HCHO vapor after completi<strong>on</strong> of the<br />
fumigati<strong>on</strong> and restorati<strong>on</strong> of ventilati<strong>on</strong> is a significant problem, especially in high-security<br />
laboratories. The extent of the problem can be seen from chart recordings from the<br />
electr<strong>on</strong>ic m<strong>on</strong>itor during fumigati<strong>on</strong> of a high- and a low-security area.<br />
DISCUSSION<br />
The data presented above dem<strong>on</strong>strate several advantages of the HCHO generator over the<br />
Formalin-permanganate method in HCHO fumigati<strong>on</strong>. Rapidly falling vapor levels in the<br />
permanganate method are due, in part, to the formati<strong>on</strong> of water c<strong>on</strong>densate <strong>on</strong> cold surfaces<br />
which, because of the high partiti<strong>on</strong> of coefficient of HCHO in water , rapidly takes up the<br />
HCHO vapor. High-localized HCHO levels generated by rapid vaporizati<strong>on</strong> in this method<br />
also lead to excessive c<strong>on</strong>densati<strong>on</strong> of paraformaldehyde.<br />
From the standpoints of safety, c<strong>on</strong>venience, and effectiveness, the HCHO generator<br />
method was more successful, particularly where large areas were to be fumigated. It is not<br />
necessary to rapidly leave the area after additi<strong>on</strong> of Formalin. Instead, it is <strong>on</strong>ly necessary to<br />
adjust a timer well in advance of the fumigati<strong>on</strong>, and any necessary sealing of doors and<br />
other outlets can then proceed at leisure and without the discomfort of leaking vapor. The<br />
importance of adequate sealing is illustrated by the rapid loss of fumigant from a lowsecurity<br />
area. Slow diffusi<strong>on</strong> of vapor into closed cupboards was dem<strong>on</strong>strated in procedure<br />
1. Cupboards, drawers, and other such fittings in the room must be opened to allow<br />
adequate penetrati<strong>on</strong> of fumigant.<br />
The value of maintenance of a uniform temperature was particularly apparent at a<br />
fumigant c<strong>on</strong>centrati<strong>on</strong> of 1.500, ug of HCHO per liter. At this level, the restorati<strong>on</strong> of<br />
ventilati<strong>on</strong> has rapidly removed all traces of the fumigant down to the level of human<br />
detecti<strong>on</strong>, comm<strong>on</strong>ly accepted as 1 ug/liter. This level complies with most of the<br />
internati<strong>on</strong>al standards currently in use.<br />
350
REFERENCES<br />
1. Bartzokas, C. A., K. McCarthy, W. B. Shacklet<strong>on</strong>, and B. F. Baker. 1978, Observati<strong>on</strong>s<br />
of the effects of formaldehyde <strong>on</strong> cockroaches and their flora. 1. Survival<br />
of vaccinia virus infected cockroaches during fumigati<strong>on</strong> with formaldehyde, J. Hyg.<br />
80:125-129.<br />
2. BraswelI, J. R., D. R. Spiner, and R. K. Hoffman. 1970, Adsorpti<strong>on</strong> of formaldehyde by<br />
various surfaces during gaseous dec<strong>on</strong>taminati<strong>on</strong>. Appl. Microbiol. 20: 765-769.<br />
3. Tucker, J. F., E. G. Harry, and H. E. Wainman. 1975, The effect of fumigati<strong>on</strong> with methyl<br />
bromide or formaldehyde <strong>on</strong> the infectivity of poultry house litter naturally<br />
c<strong>on</strong>taminated with Salm<strong>on</strong>ella virchow. Br. Vet. J. 131:474-485.<br />
4. Wade, A. 1977. Disinfectants and antiseptics, p. 520. In W. Martindale (ed.), The extra<br />
pharmacopoeia, 27th ed. The Phannaceutical Press, L<strong>on</strong>d<strong>on</strong>.<br />
351
THE CA AND MG SUPPLY OF THE SOIL EXTRACTED WITH DIFFERENT<br />
SOLVENT SOLUTIONS<br />
István Kocsis 1 Lajos Szabó 1 Simándi Péter 1 László Pásztor 2<br />
1 Tessedik Sámuel College, Faculty of Agricultural Water- and Envir<strong>on</strong>ment Management<br />
H-5540 Szarvas, Szabadság str. 1-3., Hungary<br />
2 Plant and Soil Protecti<strong>on</strong> Service of the Jász-Nagykun-Szolnok County, Szolnok<br />
INTRODUCTION<br />
Informati<strong>on</strong> up<strong>on</strong> Ca c<strong>on</strong>tent of the soil used to be crucial<br />
• if in the arable layer, calcium carb<strong>on</strong>ate is lacking, and<br />
• if calcium c<strong>on</strong>tent of the soil is excessive in relati<strong>on</strong> to the colloidal capacity of the<br />
respective layer (Filep, 1988, Németh, 1971, Filep, 2002.).<br />
There are alternative methods for characterising the Ca c<strong>on</strong>tent of soils. The soils are well<br />
supplied with calcium:<br />
• if the c<strong>on</strong>tent of CaCO3 is between 1 and 5 % measured with the method of Scheibler<br />
• if the measured AL-Ca values are about 50-100 mg/100 mg in sandy soils, 100-200<br />
mg/100 g in loam soils, and 500-700 mg/100 g in clayey soils.<br />
• if the Ca c<strong>on</strong>tent is exchangeable at a 70-80 % rate of the S value<br />
• if the Ca values obtained with the EUF method are within the interval 35.0-60.0 mg/100<br />
(Filep 2002)<br />
Results of the method according to Scheibler are not informative for soils c<strong>on</strong>taining little<br />
Ca, whereas extracti<strong>on</strong> with AL gives err<strong>on</strong>eous results for calcareous soils. De<strong>term</strong>inati<strong>on</strong><br />
of exchangeable cati<strong>on</strong>s is performed with different solvents than with other nutritive<br />
elements. The EUF method measures Ca together with other elements, however, it requires<br />
a special device.<br />
Excessive or insufficient Ca disturbs the uptake of other elements in the soil. The rule<br />
of calcium-potassium influencing the absorpti<strong>on</strong> of phosphorus by Ca as well as the<br />
antag<strong>on</strong>isms of Ca:Mg and Ca:K are widely recognised. The role of Ca and Mg influencing<br />
the uptake of meso- and micro-elements is also known (e.g. abundance of Ca may cause<br />
chlorosis due to ir<strong>on</strong> deficiency). The Ca-EUF Mn ratio indicates that two agricultural<br />
elements, the liming and the loosening of the soil could be applied.<br />
The ratio between the c<strong>on</strong>tents of lime and colloids is c<strong>on</strong>sidered as an important parameter<br />
of the soil. The Ca status should reas<strong>on</strong>ably reflect the availability of both macro- and<br />
microelements. Informati<strong>on</strong> related to the Mg status is necessary mainly in sandy and in<br />
alkaline soils. Relevant methods are largely similar to those applied in the case of Ca<br />
de<strong>term</strong>inati<strong>on</strong>. The solvent soluti<strong>on</strong> however should not c<strong>on</strong>tain the element in questi<strong>on</strong>:<br />
(e.g. Ca salt is suitable for assessing Mg e.g. 0.01 M CaCl2). For Ca and Mg extracti<strong>on</strong> KCl<br />
soluti<strong>on</strong> is recommended. In assessing the Ca c<strong>on</strong>tent available for the plants, mild solvents<br />
give more reliable informati<strong>on</strong> than the more c<strong>on</strong>centrated <strong>on</strong>es. For the de<strong>term</strong>inati<strong>on</strong> of Ca<br />
c<strong>on</strong>tent, we used mild soluti<strong>on</strong>s, which facilitated the measurement of other elements too.<br />
352
For micro-elements, EDTA+KCl 15 times diluted was applied. Instead of EDTA,<br />
EDTE+NH4Cl at the same c<strong>on</strong>centrati<strong>on</strong> proved to be useful to measure Na and KCl<br />
c<strong>on</strong>tent. For the de<strong>term</strong>inati<strong>on</strong> of Ca c<strong>on</strong>tent, mild solvents completed with a complexforming<br />
substance have been employed.<br />
MATERIAL AND METHOD<br />
The macerate with HNO3+H2O2 was prepared according to the standard MSZ 21470-<br />
50:1998 (For all elements). Many elements of the soil are indifferent from the point of view<br />
of plant nutriti<strong>on</strong>; however, their registrati<strong>on</strong> fixes the “upper point” or the maximum value<br />
for comparing it with the data obtained with other extracti<strong>on</strong> methods.<br />
Ethylene-diamine-tetra-acetate + KCl, or EDTA according to MSZ 20135:1999, serves for<br />
the measurement of micro-elements but it is less used in the internati<strong>on</strong>al practice. The KCl<br />
is an active solvent that carries a complex forming effect and keeps the micro-elements in a<br />
dissolved status.<br />
Amm<strong>on</strong>ium-lactate, MSZ 20135:1999 or AL. The acid buffering system (pH = 3.75)<br />
overestimates substantially the available P and Mg c<strong>on</strong>tent of alkaline (carb<strong>on</strong>atec<strong>on</strong>taining)<br />
soils, whereas underestimates them in acid soils.<br />
Amm<strong>on</strong>ium-chloride with EDTE (EDTE+NH4Cl) soluti<strong>on</strong>.<br />
It means 0.0033 M ethylene-diamine-tetra-acetic-acid, 0.0067 M amm<strong>on</strong>ia-soluti<strong>on</strong> and<br />
0.0067 M amm<strong>on</strong>ium chloride per 1 dm 3 .<br />
Preparati<strong>on</strong>: a 1000 cm 3 test-tube is filled with 0.96 g ethylene-diamine-tetra-acetic-acid<br />
(selecti<strong>on</strong>-B) and 40 cm 3 distilled water added. Then 0.9 cm 3 25 m/m % amm<strong>on</strong>ia-soluti<strong>on</strong><br />
is mixed to the selecti<strong>on</strong>-B until being completely dissolved (3-4 minutes).<br />
Further 0.33 g amm<strong>on</strong>ium-chloride is dissolved and filled to about 950 cm 3 with distilled<br />
water. The pH is adjusted to 7.00 ± 0.05 with 0.01 M hydrochloric acid or amm<strong>on</strong>ia<br />
soluti<strong>on</strong>, if necessary, and completed until the caesura with water.<br />
RESULTS AND DISCUSSION<br />
For the characterisati<strong>on</strong> of soil profiles, the important moment is the assessment of the Ca<br />
and Mg profile. From the point of view of fertility, a deeply penetrating humus layer and a<br />
thin, upper, lime-less layer are advantageous.<br />
The Ca and Mg supply of the arable layer signalises a favourable nutriti<strong>on</strong>al status and<br />
granular structure. Ca c<strong>on</strong>tent of the soil is not <strong>on</strong>ly c<strong>on</strong>sidered in the upper layer but also in<br />
the deeper strata. Especially calciphilous plants with a large root system are affected, e.g.<br />
alfalfa. Subsequently, 5 soil profiles are presented from the point of view of Ca c<strong>on</strong>tent<br />
using different solvent soluti<strong>on</strong>s (Table 1).<br />
353
Table 1. Ca supply of 5 soil profiles measured by using different soluti<strong>on</strong>s<br />
for extracti<strong>on</strong><br />
Samples pH(KCl) CaCO3(%) AL-Ca EDTE-Ca KCl-Ca Exch. Ca H2O-Ca<br />
Cs1 7.27 3.2 16284 698 2992 515.0 25.30<br />
Cs2 7.31 15.0 58494 768 2636 497.2 25.70<br />
Cs3 7.55 26.0 98023 712 2030 392.2 20.90<br />
Cs4 7.54 22.0 90226 648 1661 350.2 20.80<br />
Cs5 7.84 12.0 47998 575 1076 334.0 20.10<br />
KR1 7.05 1.1 29539 852 3676 594.6 32.40<br />
KR2 7.20 6.2 51742 689 3136 570.6 16.10<br />
KR3 7.32 13.0 65102 587 2820 459.8 19.10<br />
KR4 7.30 12.0 7588 557 2400 379.8 23.00<br />
KMR1 4.92 0.0 4439 374 3408 436.8 8.57<br />
KMR2 5.03 0.0 4528 417 4037 496.8 5.20<br />
KMR3 5.59 0.0 4841 559 4175 524.0 2.74<br />
KMR4 6.38 0.1 4791 571 3495 361.4 1.68<br />
KMR5 6.91 4.9 19683 610 3590 389.0 3.35<br />
KH1 7.17 3.8 18902 726 2499 411.8 24.80<br />
KH2 8.10 18.0 72984 454 688 437.8 13.30<br />
KH3 8.39 12.0 43129 355 253 114.2 8.70<br />
KH4 8.37 10.1 31476 347 168 100.4 9.31<br />
KMH1 6.94 0.0 1436 389 483 60.4 7.03<br />
KMH2 7.11 0.0 588 329 302 50.8 3.30<br />
KMH3 7.07 0.0 591 324 185 53.4 3.23<br />
KMH4 7.49 0.0 464 280 197 35.2 2.58<br />
The measurements prove that the soluti<strong>on</strong>s used for extracti<strong>on</strong> produced highly variable data<br />
of Ca c<strong>on</strong>tent. Most quantities appeared in c<strong>on</strong>centrated acid extracts, in<strong>term</strong>ediate in<br />
extracts with neutral soluti<strong>on</strong>s, the less was extracted with watery soluti<strong>on</strong>s forming<br />
complexes. The results obtained could be also c<strong>on</strong>verted from <strong>on</strong>e method to the other.<br />
354
Ca CO3(%) = AL − Cax2,5<br />
10000<br />
EDTE − CaxKA<br />
Exch.-Ca (mg/100 g) =<br />
75<br />
The data obtained with mild complex forming solvents approach the quantities of<br />
exchangeable Ca, moreover, they are informative in a mathematical sense. That means,<br />
results obtained with EDTE-Ca signify the values of exchangeable Ca c<strong>on</strong>tent.<br />
With Mg, the same test has been performed with different solvents. Results are shown in<br />
Table 2.<br />
Table 2. Mg supply of 5 soil profiles measured by using different soluti<strong>on</strong>s for extracti<strong>on</strong><br />
Samples pH(KCl) AL-Mg EDTE-Mg KCl-Mg Exch.-Mg H2O-Mg<br />
Cs1 7.27 485 34.20 174 28.56 2.43<br />
Cs2 7.31 747 26.00 105 23.84 1.54<br />
Cs3 7.55 1194 37.20 138 25.53 1.84<br />
Cs4 7.54 2123 108.00 335 60.50 5.51<br />
Cs5 7.84 2867 226.00 572 86.88 12.40<br />
KR1 7.05 645 70.30 349 45.62 4.25<br />
KR2 7.20 931 57.10 332 55.66 2.25<br />
KR3 7.32 1335 59.10 410 66.67 3.08<br />
KR4 7.30 1920 80.00 526 80.83 4.16<br />
KMR1 4.92 809 93.30 747 78.65 3.18<br />
KMR2 5.03 934 94.50 938 99.83 3.20<br />
KMR3 5.59 1068 104.00 1019 115.07 2.14<br />
KMR4 6.38 837 92.60 758 80.22 1.84<br />
KMR5 6.91 847 71.00 581 77.44 1.21<br />
KH1 7.17 588 38.60 186 24.68 2.91<br />
KH2 8.10 1793 111.00 252 54.21 5.42<br />
KH3 8.39 1080 54.30 88 18.15 3.17<br />
KH4 8.37 1020 63.60 79 19.36 3.70<br />
KMH1 6.94 109 25.40 54 8.59 2.13<br />
KMH2 7.11 61 18.20 39 8.59 1.63<br />
KMH3 7.07 85 24.70 36 10.77 2.07<br />
KMH4 7.49 53 14.10 33 7.62 2.38<br />
355
Measurements prove that most Mg was extracted with AL, then with KCl. Values of<br />
exchangeable Mg are largely equal with the results obtained with EDTE as Mg c<strong>on</strong>tent.<br />
Result is c<strong>on</strong>vertible according to the formula:<br />
EDTE − Mg<br />
Exch.-Mg (mg/100 g) =<br />
9−K CONCLUSIONS<br />
Different soluti<strong>on</strong>s are used for extracti<strong>on</strong> to assess the Ca and Mg c<strong>on</strong>tent of soil samples.<br />
Measurements prove that most quantities were obtained with AL-soluti<strong>on</strong>, lowest quantities<br />
with water as solvent.<br />
Values of exchangeable Ca and Mg c<strong>on</strong>tent largely coincide with the results of EDTE-Ca<br />
and EDTE-Mg. The results obtained with the different methods are c<strong>on</strong>vertible to each<br />
other.<br />
REFERENCES<br />
Filep T. (2002): Characterisati<strong>on</strong> of the nutrient buffer capacity of K, Ca, Mg in two<br />
Hungarian soils. Agrokémia és Talajtan 2002. 73-78 p.<br />
Filep Gy. (1988): Talajkémia. Akadémiai Kiadó. Budapest.<br />
Németh K. (1971): Talajvizsgálatok elektromos ultraszűrővel. Agrokémia és Talajtan. 20:<br />
515 p.<br />
356<br />
A
ABSTRACT<br />
BIOLOGICAL INVESTIGATION ON AN EROSION CATENA<br />
B. Szeder 1 , B. Sim<strong>on</strong> 1 , M. Dombos 2 , T. Szegi 1<br />
1 Szent István University, Department of Soil Science and Agrochemistry,<br />
2103 Gödöllő, Páter K. u. 1. Hungary<br />
2 Research Institute of Soil Science and Agrochemistry,<br />
Budapest, Herman O. U. 15. Hungary,<br />
Biological investigati<strong>on</strong>s were carried out <strong>on</strong> an erosi<strong>on</strong> catena <strong>on</strong> Szent István<br />
University Experimental Farm, north central Hungary. Four soil profiles (Chernozems<br />
and Calcisols) were examined in detail for the general physical, chemical parameters.<br />
The aim was to find out quantitative relati<strong>on</strong>ship between the physical and the<br />
biological (and biochemical) degradati<strong>on</strong> processes. The level of physical degradati<strong>on</strong><br />
was expressed by the moisture retenti<strong>on</strong> capacity and the depth of humus rich horiz<strong>on</strong>.<br />
The degradati<strong>on</strong> of soil fauna was examined by the number and the species compositi<strong>on</strong><br />
of collembolans al<strong>on</strong>g the transect in eleven sampling points. The physical degradati<strong>on</strong>,<br />
biochemical and biological parameters showed str<strong>on</strong>g relati<strong>on</strong>ship: with increasing<br />
erosi<strong>on</strong> and decreasing depth of the humus layers most investigated parameters showed<br />
degradati<strong>on</strong>. The number and the abundance of Collembola had significant decreasing<br />
tendency with increasing level of degradati<strong>on</strong>. The results suggest that soil degradati<strong>on</strong>,<br />
decreasing moisture holding c<strong>on</strong>diti<strong>on</strong>s affect str<strong>on</strong>gly the collembolan community. The<br />
physical, chemical parameters seemed to be more resistant to changes in soils compared<br />
to the biological aspects, thus, it is assumed that the biological parameters can be used<br />
as early indicators.<br />
Keywords: degradati<strong>on</strong>, biological indicators, Collembola, erosi<strong>on</strong> catena<br />
INTRODUCTION<br />
Loss of soil organic matter is a global problem related to soils and the envir<strong>on</strong>ment.<br />
Most of the fertile Chernozems of agricultural soils in Hungary developed <strong>on</strong> calcareous<br />
loess. Chernozems have been cultivated for several centuries in Hungary (Stefanovits,<br />
1999). However, most of the Chernozem soils in the country has experienced some kind<br />
of soil degradati<strong>on</strong>, including decrease in organic matter, compacti<strong>on</strong>, erosi<strong>on</strong> and/or<br />
structural degradati<strong>on</strong>, mainly due to inappropriate land use practices. These<br />
degradati<strong>on</strong> processes are generally accompained by biological degradati<strong>on</strong>, i.e. loss of<br />
species richness. Soil fauna is essential in the decompositi<strong>on</strong> processes of biomass,<br />
synthesis of humus, mixing soil particles, development of soil structure and it influences<br />
several other soil characteristics that de<strong>term</strong>ine normal soil functi<strong>on</strong>s. Soil degradati<strong>on</strong><br />
processes influence the compositi<strong>on</strong>, populati<strong>on</strong> size and activity of the soil fauna<br />
(Larsen, 2004; Bíró, 2005; Veisz, 2004).<br />
Soil quality can be evaluated by using several soil parameters, depending <strong>on</strong> the<br />
scale and objective of the evaluati<strong>on</strong>. These parameters can be, for instance, the soil<br />
organic matter and soil aggregate stability (Gardi et al., 2002). In case of an erosi<strong>on</strong><br />
risk, soil quality can be characterized by using different parameters, such as organic<br />
matter c<strong>on</strong>tent, aggregate stability, and enzyme activities. However, these parameters<br />
357
tend not to give enough informati<strong>on</strong> about the biological state or activity and diversity<br />
of soils.<br />
Collembola (springtails), as a representative group of soil fauna, has relatively low<br />
biomass, but it is extremely important in influencing the structure of some soils. For<br />
example, “alpine pitch rendzinas” <strong>on</strong> limest<strong>on</strong>e have a black humus layer that is formed<br />
almost entirely of Collembola feces. However, most soils have milli<strong>on</strong>s of collembolan<br />
fecal pellets <strong>on</strong> each square meter (Hopkin, 2002). Springtails play important role in<br />
decompositi<strong>on</strong> and soil respirati<strong>on</strong> through feeding <strong>on</strong> fungal hyphae. These effects are<br />
beneficial in the aspect of soil structure, moisture regime, soil organic c<strong>on</strong>tent, cati<strong>on</strong><br />
exchange capacity, nutrient availability, that de<strong>term</strong>ine healthy soil functi<strong>on</strong>s (Cole,<br />
2005). It is believed that soil degradati<strong>on</strong> processes lead changes in the activity and<br />
compositi<strong>on</strong> of the soil fauna (Larsen, 2004). Soil scientists have been c<strong>on</strong>sidering a<br />
highly diverse community of soil organisms to indicate good soil quality (Brady and<br />
Weil, 1999).<br />
The objective of this presented study was to find quantitative relati<strong>on</strong>ship between<br />
the physical and the biological (and biochemical) degradati<strong>on</strong> processes al<strong>on</strong>g an<br />
erosi<strong>on</strong> catena. The degradati<strong>on</strong> of soil fauna was examined by the number and the<br />
species compositi<strong>on</strong> of collembolans. It was expected that not <strong>on</strong>ly the abiotic factors<br />
(soil-moisture, aerati<strong>on</strong>, compacti<strong>on</strong>) but also biotic factors, such as microbiological<br />
activity are important ecological factors for soil animal communities. The appropriate<br />
measurements and scaling of these effects can serve as a basis for soil biological<br />
degradati<strong>on</strong> assessments.<br />
MATERIALS AND METHODS<br />
1. Study area: The study was c<strong>on</strong>ducted <strong>on</strong> Szent István University Experimental Farm<br />
in Józsefmajor, north central Hungary. Four soil profiles were examined in detail <strong>on</strong> a<br />
topographic catena, representing different levels of erosi<strong>on</strong> (no, low, high erosi<strong>on</strong> and<br />
accumulati<strong>on</strong> profiles). The first two profiles are Chernozem soils, the third <strong>on</strong>e, which<br />
is the most eroded profile, is a Calcisol and the fourth <strong>on</strong>e at the bottom of the valley is<br />
a Cumulic Chernozem.<br />
2. Laboratory analyses: The following general laboratory analyses were carried out <strong>on</strong><br />
the four soil profiles: soil organic matter (SOM %) de<strong>term</strong>ined by Walkley–Black<br />
method (Walkley, 1947); cati<strong>on</strong> exchange capacity (CEC) by Mehlich method;<br />
pH(H2O) and pH(KCl) de<strong>term</strong>ined electrometrically (Buzás, 1988); texture; CaCO3<br />
c<strong>on</strong>tent by Scheibler method; and bulk density (BD) de<strong>term</strong>inati<strong>on</strong> (Buzás, 1988). The<br />
simplified moisture retenti<strong>on</strong> was also de<strong>term</strong>ined <strong>on</strong> the soil samples to find out the<br />
level of physical degradati<strong>on</strong>.<br />
Besides the general laboratory analyses the following biochemical properties were<br />
de<strong>term</strong>ined: water soluble carb<strong>on</strong> (WSC) and carbohydrates (WSChy) (Garcia, 1997),<br />
enzimatic characterizati<strong>on</strong> (Veisz, 2004), urease, and ß-glucosidase activity (Garcia,<br />
1997).<br />
3. Experimental design for biological aspects: Collembola was sampled by regular<br />
sampling, al<strong>on</strong>g a transect in eleven points with 20 m distances from each other. The<br />
first four points bel<strong>on</strong>ged to the n<strong>on</strong> eroded regi<strong>on</strong>, whereas the 5-8th points were low<br />
eroded and the 9th, 10th points represented the most eroded parts of the transect. Soil<br />
358
profiles were typical for catena positi<strong>on</strong>s, i.e. the plateau positi<strong>on</strong> was the n<strong>on</strong>-eroded<br />
regi<strong>on</strong>, the middle slope positi<strong>on</strong> was low eroded, the lower slope positi<strong>on</strong> was the most<br />
eroded and at the valley the accumulated soil profile was found.<br />
Four soil core samples were taken in each points al<strong>on</strong>g the transect from the upper 5 cm<br />
by using 5 cm diameter sampling pots.<br />
RESULTS AND DISCUSSIONS<br />
The results of the laboratory analyses are given in Table 1. The depth of soil organic<br />
matter was 90 cm in the n<strong>on</strong> eroded profile, whereas it was <strong>on</strong>ly 10 cm at the highly<br />
eroded site showing high physical degradati<strong>on</strong>. The soil organic carb<strong>on</strong> (SOC) c<strong>on</strong>tent<br />
and the cati<strong>on</strong> exchange capacity (CEC) of the soils decreased with increasing erosi<strong>on</strong><br />
rate. The water soluble carb<strong>on</strong> (WSC) showed similar tendency. As for the bulk density<br />
(BD), higher values were found in the upper slope positi<strong>on</strong>s. C<strong>on</strong>cerning the enzymatic<br />
characterizati<strong>on</strong>, less activity of urease (involved in N cycle) and<br />
ß-glucosidase (involved in the C cycle) were experienced in the highly degraded slope<br />
positi<strong>on</strong>s, suggesting less biological activity related to degradati<strong>on</strong>.<br />
Table 1: General laboratory parameters of the top horiz<strong>on</strong>s<br />
of the four soil profiles al<strong>on</strong>g the catena<br />
Profile DH SOC WSC WSCh CaCO3 CEC pH Ure B-Gl Texture BD<br />
(upper h.) cm % ppm ppm % cmol·kg -1<br />
μmols μmols FAO g·cm -3<br />
NE 90 2,2 106,2 2,1 0 30 6,1 0,75 21,14 CL 1,4<br />
LE 40 1,5 103,6 1,4 0 28 6,1 0,66 17,20 CL 1,5<br />
HE 10 0,5 95,5 7,6 12 24 6,1 0,49 12,44 CL 1,3<br />
AP 80 2,5 119,2 6,4 2 40 7,6 1,28 35,61 CL 1,2<br />
Profile designati<strong>on</strong>s: NE – N<strong>on</strong> Eroded, LE – Low Eroded, HE – High Eroded,<br />
AP – Accumulated Profile; DH - depth of humus layer (cm); SOC – Soil organic carb<strong>on</strong><br />
(%); WSC - Water soluble carb<strong>on</strong> (ppm); WSCh - Water soluble carbohydrates (ppm);<br />
CEC - Cati<strong>on</strong> exchange capacity (cmol·kg -1 ); Ure – Urease activity (μmols N-<br />
NH4 + /g.h); B-Gl – B-Glucosidase activity (μmols PNF/g.h); CL- clay loam; BD - Bulk<br />
density (g·cm -3 ).<br />
The degradati<strong>on</strong> of soil fauna was examined by the number and the species composti<strong>on</strong><br />
of collembolans al<strong>on</strong>g the erosi<strong>on</strong> gradient. Eleven sampling points (Figure 1) were<br />
chosen al<strong>on</strong>g the slope al<strong>on</strong>g the four soil profiles that were examined in detail. Except<br />
for the highest point (number 0 in Figure 1) in the transect, which might have been a<br />
sampling error, abundances of springtails decreased highly. C<strong>on</strong>cerning the erosi<strong>on</strong><br />
grades, the soil living springtails showed significant differences in their abundances<br />
(F(2,41)= 9.2, p
The average values of abundances, derived from the four subsamples, showed str<strong>on</strong>g<br />
correlati<strong>on</strong> with the moisture retenti<strong>on</strong> of soil (r=0.89) (Figure 4.).<br />
Figure 1: Number of Springtails<br />
(Collembola) al<strong>on</strong>g the erosi<strong>on</strong> gradient<br />
Figure 3: Simplified moisture retenti<strong>on</strong> at<br />
the erosi<strong>on</strong> levels<br />
Figure 2: Number of Springtails<br />
(Collembola) at the erosi<strong>on</strong> levels<br />
Figure 4: Correlati<strong>on</strong> between water<br />
retenti<strong>on</strong> and springtail abundance<br />
Figure 5 shows that most springtails preferred the pH 6-7. Less individuals were found<br />
above pH 7. As far as the Collembola species compositi<strong>on</strong> is c<strong>on</strong>cerned, we found much<br />
less species <strong>on</strong> the high eroded area than the reference and low eroded areas (Table 2).<br />
360
Table 2: Collembola species compositi<strong>on</strong> am<strong>on</strong>g the erosi<strong>on</strong> gradient<br />
Reference, n<strong>on</strong><br />
eroded profile<br />
Low eroded<br />
profile<br />
High<br />
eroded<br />
Collembola species<br />
ref 1 2 3 4 5 6 7 8 9 10<br />
Entomobria handschini 2 1<br />
Entomobria multifasciata 27 10 7 3 2<br />
Heteromurus nitidus 35 12 11 1 2 4 15<br />
Folsomia cf. penicula 5 2 1 29 1 1 2 1 4<br />
Lepidicyrtus cyaneus 8 7 22 1 4 1<br />
Lepidicyrtus paradoxus 1 1<br />
Ceratophysella cf. armata 3 1 1 1<br />
Orchesella cincta 5 9 8 7 6<br />
Orchesella sp. 1 1<br />
Poduromorpha sp. 1 2<br />
Sminthurus elegans 8 5 1 8 7 27 1 3<br />
Sminthurinus aureus 2 1 1<br />
Tomocerus sp. 1<br />
abundance<br />
30,00<br />
20,00<br />
10,00<br />
0,00<br />
The pH and the abundance of springtails<br />
A<br />
A<br />
6,50 7,00 7,50<br />
A<br />
pH<br />
A A<br />
A<br />
A<br />
A<br />
A<br />
A<br />
Linear Regressi<strong>on</strong><br />
abundance = 110,69 + -13,88 * pH<br />
R-Square = 0,64<br />
Figure 5: Correlati<strong>on</strong> between pH and abundance of springtails<br />
361
CONCLUSIONS<br />
The physical degradati<strong>on</strong>, biochemical and biological parameters showed str<strong>on</strong>g<br />
relati<strong>on</strong>ships <strong>on</strong> the studied erosi<strong>on</strong> catena. With erosi<strong>on</strong> and decreasing depth of the<br />
humus layers most investigated other parameters showed degradati<strong>on</strong>.<br />
The number and the abundance of Collembola were investigated in this study al<strong>on</strong>g a<br />
transect representing different levels of erosi<strong>on</strong> and soil degradati<strong>on</strong> (organic matter<br />
decrease, soil compacti<strong>on</strong>, etc.). We found that al<strong>on</strong>g a transect the abundance of<br />
springtails has significant decreasing tendency with increasing level of degradati<strong>on</strong>. The<br />
results suggest that soil degradati<strong>on</strong> affects str<strong>on</strong>gly the collembolan community. In<br />
case of the eroded area we experienced higher pH (above 7) because of the distance of<br />
the calcareous parent material from the soil surface were less than the other sites. This<br />
result suggests that due to degradati<strong>on</strong> processes soil moisture c<strong>on</strong>diti<strong>on</strong>s str<strong>on</strong>gly affect<br />
the belowground animal communities, such as springtails.<br />
The examined enzyme activities are important informati<strong>on</strong> to characterize the measure<br />
of soil degradati<strong>on</strong>. The land degradati<strong>on</strong> seems to be a key factor for variati<strong>on</strong> in the<br />
behaviour of the examined enzyme.<br />
The physical, chemical parameters compared to the biological parameters showed the<br />
following correlati<strong>on</strong>s: the physical features are more resistant to changes in soils<br />
compared to the biological aspects, thus, it is assumed that the biological parameters can<br />
be used as early indicators.<br />
ACKNOWLEDGEMENTS<br />
INDEX Nr. GOCE-CT-2003-505450<br />
Hungarian Scientific Research Fund (OTKA – Nr. D 048592)<br />
REFERENCES<br />
Bíró, B., Villányi, I., Füzy, A., Naár, Z., (2005): Baktériumok és gombák<br />
kol<strong>on</strong>izációja génmódosított (Bt-) és izogénes k<strong>on</strong>troll kukorica rizoszférájában,<br />
Agrokémia és Talajtan, 2005, vol. 54, N o s 1-2, 189-202 pp.<br />
Brady N. C., Weil R. R. (1999): The Nature and Properties of Soils. Twelfth Editi<strong>on</strong>.<br />
Prentice Hall. N. J. 404-405, 760-761.<br />
Buzás, I. (Szerk.) (1988): Talaj- és agrokémiai talajvizsgálati módszerkönyv 2.<br />
A talajok fizikai-kémiai és kémiai vizsgálati módszerei. (Methods of soil and<br />
agricultural chemistry analyses 2. Physical-chemical and chemical methods of soil<br />
analyses.) Mezőgazdasági Kiadó. Budapest. Hungary.<br />
Cole L., Buckland S. M., Bardgett R. D. (2005): Relating microarthropod community<br />
structure and diversity to soil fertility manipulati<strong>on</strong>s in temperate grassland. Soil<br />
biology and biochemistry 37:99, 1707-1717.<br />
Garcia, C., T. Hernandez, A. Roldan, J. Albaladejo (1997): Biological and<br />
biochemical quality of a semiarid soil after induced devegetati<strong>on</strong>. Journal of<br />
Envir<strong>on</strong>mental Quality 26, 1116-1122.<br />
Gardi, C., M. Tomaselli, V. Parisi, A. Petraglia, C. Santini. (2002): Soil quality<br />
indicators and biodiversity in northern Italian permanent grasslands. European<br />
Journal of Soil Biology. 38, 103-110.<br />
Hopkin, S.P. (2002): Collembola. Encyclopedia of Soil Science, p. 207-210.<br />
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Larsen, T., P. Schj<strong>on</strong>ning, J. Axelsen. (2004): The impact of soil compacti<strong>on</strong> <strong>on</strong><br />
euedaphic Collembola. Applied Soil Ecology. 26, 273-281.<br />
Stefanovits, P., Gy. Filep, Gy. Füleky (1999): Soil Science. (In Hungarian.)<br />
Mezőgazda Kiadó. Budapest. 92-123.<br />
Veisz, O, Bencze, S, Janda, T, (2004): Changes in the activity of antioxidant enzymes<br />
in cereal species during the winter CEREAL RES COMMUN 32 (4): 493-500.<br />
Walkley, A. (1947): A critical examinati<strong>on</strong> of a rapid method for de<strong>term</strong>ining organic<br />
carb<strong>on</strong> in soils: Effect of variati<strong>on</strong>s in digesti<strong>on</strong> c<strong>on</strong>diti<strong>on</strong>s and of inorganic soil<br />
c<strong>on</strong>stituents. Soil Sci. 63. 251–263.<br />
363
THE PRESENT STATE OF THE ROMANIAN AGRO-ALIMENTARY<br />
CONSUMPTION<br />
Anca M<strong>on</strong>ica Brata<br />
University of Oradea, Faculty of Envir<strong>on</strong>mental Protecti<strong>on</strong><br />
ABSTRACT<br />
The alimentary c<strong>on</strong>sumpti<strong>on</strong> is not and cannot be identical in the countries and areas of the<br />
entire world, no matter how attractive the internati<strong>on</strong>al c<strong>on</strong>sumpti<strong>on</strong> patterns would be. The<br />
specialists in alimentati<strong>on</strong> and the internati<strong>on</strong>al organisms with resp<strong>on</strong>sibilities in this field<br />
recommend the remodelling of the alimentary c<strong>on</strong>sumpti<strong>on</strong> and encourage having a healthy<br />
alimentati<strong>on</strong>, based <strong>on</strong> each nati<strong>on</strong>’s accumulati<strong>on</strong>s and showing their value as being a<br />
cultural nutriti<strong>on</strong>al thesaurus. It is c<strong>on</strong>sidered that the alimentary educati<strong>on</strong> of the populati<strong>on</strong><br />
and the improvement of the health insurance systems can c<strong>on</strong>tribute to the limitati<strong>on</strong> of<br />
hunger.<br />
Am<strong>on</strong>gst the well developed countries and the developing <strong>on</strong>es, there are great gaps<br />
regarding the average c<strong>on</strong>sumpti<strong>on</strong> per inhabitant, but the deficiencies are especially<br />
structural and qualitative.<br />
Keywords: alimentati<strong>on</strong>, annual and m<strong>on</strong>thly c<strong>on</strong>sumpti<strong>on</strong>.<br />
INTRODUCTION<br />
The humanity develops c<strong>on</strong>tinuously. This is a fact that cannot be ignored, which modifies<br />
the humans’ way of thinking and behaviour. This evoluti<strong>on</strong>al process takes place in every<br />
field of life, including the alimentary <strong>on</strong>e.<br />
The evoluti<strong>on</strong> of the human species implied permanent changes in the alimentary<br />
behaviour, in the alimentati<strong>on</strong> manner, but also in the process of obtaining alimentary<br />
products. The increase of people’s culture and civilizati<strong>on</strong>, the technical and scientific<br />
discoveries, the research in the alimentati<strong>on</strong> and alimentary field, all these have c<strong>on</strong>tributed<br />
in a great extent to these changes. The diversity of the alimentary products, the<br />
quantificati<strong>on</strong> of their metabolic effects and relating the aliments to the populati<strong>on</strong> health<br />
state, have led to a need of a careful design of alimentary products, that takes into account<br />
the nutritious value of aliments and the expected effect <strong>on</strong> alimentati<strong>on</strong>.<br />
The problem of insuring a sufficient level of the populati<strong>on</strong>’s alimentati<strong>on</strong> from the<br />
quantitative, qualitative and structural point of view has become for most countries and for<br />
the governmental and n<strong>on</strong>-governmental organisms – nati<strong>on</strong>al, regi<strong>on</strong>al and internati<strong>on</strong>al – a<br />
more powerful preoccupati<strong>on</strong> in the last half of the century. This preoccupati<strong>on</strong> is the<br />
c<strong>on</strong>sequence of the c<strong>on</strong>scious knowing of the c<strong>on</strong>necti<strong>on</strong> between health and the access to<br />
various sources of food. Because the expansi<strong>on</strong> and the limits of the access to the sources of<br />
alimentati<strong>on</strong> differ from an area to another, from a country to another and even within the<br />
same country from an area to another, the taking of some measures regarding the<br />
ameliorati<strong>on</strong> of the alimentati<strong>on</strong> matter becomes essential [Bulgaru, 2001].<br />
364
The alimentary problem has c<strong>on</strong>stituted and still c<strong>on</strong>stitutes a major preoccupati<strong>on</strong>,<br />
nati<strong>on</strong>ally and internati<strong>on</strong>ally, mainly focusing <strong>on</strong> the most efficient solving of the relati<strong>on</strong><br />
between resources and c<strong>on</strong>sumpti<strong>on</strong> necessities.<br />
At every people, al<strong>on</strong>g its existence and development, alimentary traditi<strong>on</strong>s, habits and<br />
skills have installed more or less durable, that c<strong>on</strong>tinue and will c<strong>on</strong>tinue to play a<br />
predominant role in the nutriti<strong>on</strong> of the respective populati<strong>on</strong>, even under the impact of the<br />
modern alimentati<strong>on</strong> principles, based <strong>on</strong> their scientific and rati<strong>on</strong>al character. The main<br />
factors of this phenomen<strong>on</strong> are the ambient geographical envir<strong>on</strong>ment, family and external<br />
influences [Gavrilescu, Giurca, 2000].<br />
At present, the evoluti<strong>on</strong> of alimentati<strong>on</strong> generates a rati<strong>on</strong>al alimentary behaviour,<br />
implies the knowing of the body’s needs and the efficient ways to satisfy them. The<br />
supporting of a rati<strong>on</strong>al alimentati<strong>on</strong> represents a preoccupati<strong>on</strong> of both the specialists of<br />
nutriti<strong>on</strong> and the governments of the states and of the nati<strong>on</strong>al and internati<strong>on</strong>al organisms<br />
founded and organized for this purpose.<br />
Thus, the rec<strong>on</strong>sidering of the aliments’ c<strong>on</strong>sumpti<strong>on</strong> has become necessary, its<br />
relati<strong>on</strong> with the needs of the body, its adaptati<strong>on</strong> to the climate and working c<strong>on</strong>diti<strong>on</strong>s.<br />
RESULTS AND DISCUSSIONS<br />
In Romania, in the late years, the populati<strong>on</strong>’s real incomes were c<strong>on</strong>tinuously decreasing.<br />
Under these c<strong>on</strong>diti<strong>on</strong>s, the housekeepers’ incomes were almost entirely intended for<br />
covering the daily needs – especially the alimentary needs – and the compulsory payments<br />
(taxes, fees, etc.), without being able to save significant amounts of m<strong>on</strong>ey, except for the<br />
patr<strong>on</strong>s’ families.<br />
In order to focus <strong>on</strong> the evoluti<strong>on</strong> of the goods’ c<strong>on</strong>sumpti<strong>on</strong> after 1989 we will<br />
observe table 23 where we can find the structure of the c<strong>on</strong>sumpti<strong>on</strong> expenses <strong>on</strong> groups of<br />
families as employers, peasants, pensi<strong>on</strong>ers.<br />
Table 1. The structure of the c<strong>on</strong>sumpti<strong>on</strong> expenses <strong>on</strong> groups of families, after the<br />
m<strong>on</strong>thly income per pers<strong>on</strong>, in the year 1990 comparatively to the year 2000<br />
Expenses for food Expenses for n<strong>on</strong>- Expenses for payment<br />
alimentary goods<br />
of services<br />
1990 2000 1990 2000 1990 2000<br />
Employers 46,8% 50,6% 39,2% 27,8% 14,0% 21,6%<br />
Peasants 63,7% 57,4% 28,3% 34,8% 8,0% 7,8%<br />
Pensi<strong>on</strong>ers 59,2% 54,3% 22,7% 29,1% 18,1% 16,6%<br />
Source: Anuarul Statistic al Romaniei, (The Statistic Annual of Romania), 2001<br />
The following aspects can be noticed [Anuarul Statistic, 1991-2001]:<br />
In the expenses for alimentary products there can be noticed an increase with 3.8% at<br />
employers, and at peasants and pensi<strong>on</strong>ers a decrease with 6.3%, respectively 4.9%.<br />
365
In the expenses for the n<strong>on</strong>-alimentary goods there can be noticed a decrease with 11.4% at<br />
employers, and at peasants and pensi<strong>on</strong>ers a decrease with 6.5%, respectively 6.4%.<br />
The expenses for duties increase with 7.6% at employers and at peasants and<br />
pensi<strong>on</strong>ers they decrease with 0.2%, respectively with 1.5%.<br />
On resistance media, the alimentary expenses represent, in the total of the c<strong>on</strong>sumpti<strong>on</strong><br />
expenses, 56% in urban envir<strong>on</strong>ment and 58% in rural envir<strong>on</strong>ment. The above menti<strong>on</strong>ed<br />
data indicate the fact that, in our country, not even the families with the highest income have<br />
a structure of the c<strong>on</strong>sumpti<strong>on</strong> expenses comparable with that of the western-European<br />
countries, where the alimentary expenses do not exceed 20% of the total expenses for<br />
housekeeping.<br />
As regards the c<strong>on</strong>sumpti<strong>on</strong> <strong>on</strong> populati<strong>on</strong> categories in the post-December period, it<br />
has been achieved in different c<strong>on</strong>diti<strong>on</strong>s, following the decline line of the nati<strong>on</strong>al<br />
ec<strong>on</strong>omy, the significant decrease of the agricultural and industrial producti<strong>on</strong>, the<br />
agriculture system, as well as the new system of distributi<strong>on</strong> under the c<strong>on</strong>diti<strong>on</strong>s of private<br />
acti<strong>on</strong>s. One thing that can be said is that the average annual level of the peasants’<br />
c<strong>on</strong>sumpti<strong>on</strong> showed a tendency of increase, overtaking the c<strong>on</strong>sumpti<strong>on</strong> in the employers’<br />
families that recorded significant decrease. This fact can be explained by the fact that the<br />
main source of forming the alimentary c<strong>on</strong>sumpti<strong>on</strong> is the own producti<strong>on</strong>.<br />
The alimentary c<strong>on</strong>sumpti<strong>on</strong> in Romania during the analysed period is the following:<br />
Table 2. The average annual c<strong>on</strong>sumpti<strong>on</strong> per inhabitant, for the main alimentary<br />
products<br />
1990 1995 2000 2005<br />
Cereals 213,6 215,8 219,7 219,8<br />
Potatoes 59,4 71 86,5 97,9<br />
Vegetables 127 140,4 134,3 183,3<br />
Fruit 59,5 45,8 44,5 77,4<br />
Sugar 27,3 23,5 23 25,6<br />
Milk 140,1 188,6 193 238,9<br />
Eggs 246 197 206 289<br />
Meat 61 51,2 48,3 65,5<br />
Source: Anuarul Statistic al Romaniei, (The Statistic Annual of Romania), 1990-2006<br />
There can be seen an increase of the potatoes, vegetables, cereals and milk c<strong>on</strong>sumpti<strong>on</strong>.<br />
The meat c<strong>on</strong>sumpti<strong>on</strong> is low enough as compared with other countries [Anuarul statistic,<br />
1991-2006].<br />
The presentati<strong>on</strong> of the average annual c<strong>on</strong>sumpti<strong>on</strong> of the main alimentary products<br />
<strong>on</strong> countries is also very interesting.<br />
366
Table 3. The average annual c<strong>on</strong>sumpti<strong>on</strong> per inhabitant, for the main alimentary<br />
products, <strong>on</strong> countries, kg/inhabitant in 2002<br />
Country Vege- Potato Fruit Meat Alcoholic Sugar Milk Cereals<br />
ables<br />
drinks<br />
Romania 137,32 89,75 46,07 42,27 53,27 22,73 123,17 218,33<br />
Poland 124,48 134,77 62,4 67,63 78,5 35,27 181,65 154,18<br />
Slovakia 80,58 77,53 62,93 69,57 79,25 36,07 188,87 131,68<br />
Hungary 106,67 67,62 63,01 72,28 83,8 37,77 190,12 125,32<br />
Check<br />
Republic<br />
76,15 79,6 86,08 90,31 110,39 42,25 211,54 105,56<br />
Bulgaria _ 32,16 126,58 106,22 171,92 48,48 239,77 104,41<br />
Source: Anuarul Statistic al Romaniei, (The Statistic Annual of Romania), 2003<br />
As regards the vegetables c<strong>on</strong>sumpti<strong>on</strong> we are situated <strong>on</strong> advanced positi<strong>on</strong>s, the average<br />
annual c<strong>on</strong>sumpti<strong>on</strong> is 137.3 kg/inhabitant, over the average of the European Uni<strong>on</strong> (122.57<br />
kg), or of the US (125.8 kg/inhabitant), while in other developing countries, vegetables are<br />
c<strong>on</strong>sumed in an annual average of 98.8 kg/inhabitant [Anuarul statistic 2003].<br />
In the developing countries, the potatoes c<strong>on</strong>sumpti<strong>on</strong> is over the average of the<br />
developed countries. The average of the potatoes c<strong>on</strong>sumpti<strong>on</strong> is 78.65 kg/inhabitant<br />
annually, and in the US there is an average of about 65.27 kg/inhabitant. A Romanian<br />
c<strong>on</strong>sumes annually an average of 89.75 kg of potatoes (in 2002).<br />
Romania is much under the average of the developed countries and developing<br />
countries at the fruit meat, milk and sugar c<strong>on</strong>sumpti<strong>on</strong>. The cereals c<strong>on</strong>sumpti<strong>on</strong> in our<br />
country has the highest c<strong>on</strong>sumpti<strong>on</strong> in comparis<strong>on</strong> with the countries analysed in the above<br />
table.<br />
A basic aliment in the human diet is milk. From milk we can prepare various products,<br />
which de<strong>term</strong>ined the occurrence of a real alimentary industry with influence <strong>on</strong> human’s<br />
feeding manner. By introducing the technology of transforming the liquid milk in powder<br />
milk, it can be transported at l<strong>on</strong>g distances and can be stored for l<strong>on</strong>g periods of time. This<br />
thing allowed that in the periods when the producti<strong>on</strong> is higher than c<strong>on</strong>sumpti<strong>on</strong>, to create<br />
stocks that can be used in the moment when the producti<strong>on</strong> is lower than the c<strong>on</strong>sumpti<strong>on</strong>.<br />
Table 4. The average milk c<strong>on</strong>sumpti<strong>on</strong> <strong>on</strong> countries, 2003 (l/inhabitant)<br />
Country Annual average milk c<strong>on</strong>sumpti<strong>on</strong> (l/inhabitant)<br />
Bulgaria 163,8<br />
Hungary 176,71<br />
Romania 188,87<br />
Poland 190,12<br />
Check Republic 203,57<br />
Source: Anuarul Statistic al Romaniei, (The Statistic Annual of Romania), 2004<br />
367
Am<strong>on</strong>g the developing countries, Romania is <strong>on</strong> an average positi<strong>on</strong> as regards the annual<br />
average c<strong>on</strong>sumpti<strong>on</strong> of milk, but in comparis<strong>on</strong> to other countries in the European Uni<strong>on</strong> or<br />
USA, Romania is <strong>on</strong> an inferior positi<strong>on</strong> than these countries [Anuarul statistic, 2004].<br />
For an extended presentati<strong>on</strong> of the alimentary c<strong>on</strong>sumpti<strong>on</strong> in Romania, the<br />
presentati<strong>on</strong> of the daily average calories c<strong>on</strong>sumpti<strong>on</strong> is necessary.<br />
Table 5. Alimentary c<strong>on</strong>sumpti<strong>on</strong> expressed in calories and nutritious factors, in a daily<br />
average, per inhabitant<br />
Calories Protides Lipids Fats<br />
Year animals total animals total animals total<br />
1990 711 3038 42,8 96,7 49,8 93,4 433,3<br />
1992 671 2758 38,3 88,3 46,2 80,2 403,2<br />
1994 690 2872 39 92,9 47,1 78 431,7<br />
Source: Anuarul Statistic al Romaniei, (The Statistic Annual of Romania), 1991-1995<br />
As <strong>on</strong>e can notice, the calories c<strong>on</strong>sumpti<strong>on</strong> has decreased starting from 1990. In 1994 it<br />
was ameliorated. It is c<strong>on</strong>sidered that in Romania it would be necessary a daily average<br />
c<strong>on</strong>sumpti<strong>on</strong> of 3400-3500 calories per inhabitant with a structure of 1/3 calories of animal<br />
origin and a c<strong>on</strong>sumpti<strong>on</strong> of 100 grams proteins, 60% of animal origin. It would mean a<br />
decent average c<strong>on</strong>sumpti<strong>on</strong>, close to the balanced <strong>on</strong>e. At present we have a deficit of<br />
about 20-25% at the calories number and of 35-40% at the proteins of animal origin. There<br />
is, as it can be explained, a great difference of the alimentary c<strong>on</strong>sumpti<strong>on</strong> according to the<br />
income, fact that leads to more decreased levels, that is at surviving limits, of absolute<br />
hunger but of specific hunger too, due to malnutriti<strong>on</strong>, to the lack of some important<br />
nutritious elements in the populati<strong>on</strong>’s c<strong>on</strong>sumpti<strong>on</strong> ratio as the protides, vitamins and<br />
mineral salts [Anuarul statistic,1991-1995].<br />
Examining the low level of the alimentary c<strong>on</strong>sumpti<strong>on</strong> in Romania, it has to be taken<br />
into account the important and well-known fact that it has a significant impact <strong>on</strong> the<br />
populati<strong>on</strong>’s general behaviour, of the labour’s productivity, of health state, of demographic<br />
indices, the level of general and infantile mortality. We have to note the fact that the<br />
infantile mortality in our country is 2-3 times higher than in other western-European<br />
countries, and the average life time is situated with 6 years in minus than in most European<br />
countries, and the mortality affects more and more the poor populati<strong>on</strong>, and the less fed<br />
people.<br />
REFERENCES<br />
1. Mircea Bulgaru – Dreptul de a minca, Editura Ec<strong>on</strong>omica, 2001<br />
2, Dinu Gavrilescu, Daniela Giurca – Ec<strong>on</strong>omie Agroalimentara, Editura Expert, Bucuresti,<br />
2000<br />
3. Anuarele statistice ale României -1991-2006<br />
368
FROM FARM TO FORK – EVALUATION OF CROPPING FIELDS WITH<br />
REGARD TO FOOD AND FEED SAFETY AFTER LONG-STANDING<br />
CULTIVATION<br />
Péter Sipos, Zsuzsanna Szathmáry, Árpád Tóth, Diána Ungai,<br />
Dóra Hovánszki, Zoltán Győri<br />
University of Debrecen, Centre of Agricultural Sciences, Institute of Food Science,<br />
Quality Assurance and Microbiology<br />
138 Böszörményi Street, Debrecen, H-4032, Hungary<br />
ABSTRACT<br />
From the aspects of quality c<strong>on</strong>trol system in accordance to food safety, the pesticide<br />
residues and toxic or potentially toxic elements are important parameters to examine not<br />
<strong>on</strong>ly from the products and its raw materials point of view, but from the external<br />
resources (as soil and applied matters, e.g. manure) as well. The sensitivity and<br />
accumulati<strong>on</strong> properties of the different plants and cultures is different according to the<br />
above menti<strong>on</strong>ed factors, and the aim of producti<strong>on</strong> also de<strong>term</strong>ines the upper level of<br />
limit values. On the other hand, in regulati<strong>on</strong>s of several countries like Hungary there is<br />
no differentati<strong>on</strong> as regards utilizati<strong>on</strong> and accumulati<strong>on</strong> properties.<br />
In a case study we have examined the soil parameters of food safety of differently<br />
utilised (intensive cereal producti<strong>on</strong>, extensive meadow and bio producti<strong>on</strong>) cropping<br />
fields which were cultivated for a l<strong>on</strong>g time, so the presence of c<strong>on</strong>taminants in the soil<br />
profile was assumed. We found that the examined fields are suitable for producing safe<br />
foods, but in the case of intensive cultivati<strong>on</strong> some accumulati<strong>on</strong> of pesticides and<br />
heavy metals is observable.<br />
Keywords: food safety, pesticide remains, toxic elements, different levels of land use<br />
INTRODUCTION<br />
In latest decades there has been an increasing demand to extend the more or less<br />
developed quality c<strong>on</strong>trol system of food product processing to the raw material<br />
producti<strong>on</strong> and supply. This would lead to getting closer to the fully introduced food<br />
and feed safety in the whole plant-animal processing line. Beside this, it was obligatory<br />
to apply the HACCP system by food processors and traders. The picture of applied<br />
quality c<strong>on</strong>trol systems is very diverse nowadays; strict applicati<strong>on</strong> and m<strong>on</strong>itoring is<br />
used in the processing of horticultural crops, but in the case of field crops, especially<br />
cereals and forage crops it is far from being effective.<br />
To examine the possibilities of the development of a quality c<strong>on</strong>trol system in the<br />
whole plant-feed-animal-food chain, i.e. “from farm to fork” we have started a research<br />
program with the c<strong>on</strong>tributi<strong>on</strong> of industrial partners. Our aim is to create a base<br />
specificati<strong>on</strong> that helps to apply the ISO 22000 standard and to develop a quality c<strong>on</strong>trol<br />
and tracing system specified for Hungarian circumstances. To examine the process we<br />
made a case study for each c<strong>on</strong>tributor. To de<strong>term</strong>ine whether the fields taken into these<br />
examinati<strong>on</strong>s are suitable for safe producti<strong>on</strong>, we analysed their pesticide remains and<br />
heavy metal c<strong>on</strong>tent. On these sites the crop producti<strong>on</strong> activity has been lasting for a<br />
l<strong>on</strong>g time, so these examinati<strong>on</strong>s seemed necessary, because of the applied<br />
369
agrochemicals as fertilizers and pesticides, the probability of c<strong>on</strong>taminati<strong>on</strong> or<br />
accumulati<strong>on</strong> was c<strong>on</strong>siderable.<br />
The group of organochlorines are widely used pesticides and are important in the<br />
aspects of food safety (Kan and Meijer, 2007). The peak of their applicati<strong>on</strong> was in the<br />
’50ies and ‘60ies. Many of them were banned later in the most countries in Europe and<br />
North America, but in developing countries large amounts have been applied in the<br />
‘90ies (Kannan et al., 1997) and are still in use nowadays in several countries. The<br />
informati<strong>on</strong> about organochlorine c<strong>on</strong>taminati<strong>on</strong> is mostly limited to DDTs and lindane,<br />
but it is easy to accept that other dangerous pesticide remains are still in our soils<br />
(Shegunova, 2007). It may be also proved by the fact that both Hungarian and European<br />
Uni<strong>on</strong> soil and water quality standards c<strong>on</strong>tain target and interventi<strong>on</strong> values for DDTs,<br />
drins as aldrin and endrin, and HCH and HCB (Oldal et al., 2006). Besides, earlier<br />
studies <strong>on</strong> Hungarian surface water m<strong>on</strong>itoring reports that 8 out of the examined 21<br />
pesticide residues exceeded the 0,1 μg/l limit c<strong>on</strong>centrati<strong>on</strong> required by EU directives<br />
(Kárpáti et al., 1998).<br />
From the aspect of food safety another important soil property is its element<br />
c<strong>on</strong>tent, primarily the heavy metal c<strong>on</strong>tent. Excessive amounts of metals originated<br />
from agricultural or industrial producti<strong>on</strong> often cause human acti<strong>on</strong>s (Biehl and Buck,<br />
1987). It is important to m<strong>on</strong>itor the cadmium, lead, mercury and arsenic c<strong>on</strong>tents in<br />
both soils and the products from different locati<strong>on</strong>s, although the latter is not heavy<br />
metal by original definiti<strong>on</strong>, but in several cases it poses similar risk. Besides limit<br />
values in different nati<strong>on</strong>al and internati<strong>on</strong>al standards, the Ministry of Agriculture in<br />
Netherlands published the so-called LAC values, which define the limit values that are<br />
“the lowest level in soil that, when exceeded, may give rise to the emergence of<br />
negative impacts <strong>on</strong> the yield and the quality of agricultural products, and with the<br />
health of human beings and animals” (Brus et al., 2005). This recommendati<strong>on</strong> gives<br />
different limit values according to both soil type (as sand or clay) and grown plant (as<br />
grassland, arable land with flowers or arable land with food for human c<strong>on</strong>sumpti<strong>on</strong>). In<br />
Hungary the current limits are declared in 10/2000. (VI.2.) KöM-EüM-FVM-KHVM<br />
ministry order in which the background (A) levels, c<strong>on</strong>taminati<strong>on</strong> limit (B) levels and<br />
envir<strong>on</strong>mental remediati<strong>on</strong> c<strong>on</strong>taminati<strong>on</strong> limit (D) levels are de<strong>term</strong>ined. Naturally, the<br />
aim of producti<strong>on</strong> often raises a claim to decrease the maximum heavy metal level in<br />
soils, e.g. in the case of producing drug or medicine raw materials.<br />
In our case study we had an opportunity to evaluate and estimate three kinds of<br />
cropping sites in the aspects of food safety; an intensively used (for wheat and maize<br />
producti<strong>on</strong>), an extensively used (for meadow) and <strong>on</strong>e used in bio producti<strong>on</strong>.<br />
MATERIALS AND METHODS<br />
Soil samples: Examined sites were the followings: two winter wheat producti<strong>on</strong> sites<br />
near to Szentes and Sárrétudvari, two maize producti<strong>on</strong> sites by Nádudvar and<br />
Hajdúböszörmény, a meadow near to Bak<strong>on</strong>szeg and a bio producti<strong>on</strong> area at the<br />
Látókép Experimental Stati<strong>on</strong> of University of Debrecen, Centre of Agricultural<br />
Sciences. Site samplings were made in the summer of 2005 from the upper 20<br />
centimetres of soil and from the 100-130 cm depth and 200-235 depth as well. In 2006<br />
370
we made a deeper vertical sampling to 300 centimetres depth of soil and we separated<br />
10-12 layers in the profiles based <strong>on</strong> their physical properties.<br />
Pesticide residue analysis: Pesticide residue analysis was made in the Soil and Plant<br />
C<strong>on</strong>servati<strong>on</strong> Service of Miskolc in 2005 and in the Institute of Food Science, Quality<br />
Assurance and Microbiology in 2006. Sample preparati<strong>on</strong> was carried out as follows:<br />
after milling and drying, 50 g of the soil was accurately weighed and added 7 ml of 0.2<br />
M amm<strong>on</strong>ium-chloride (Sigma). After resting for approximately 15 minutes nhexane:acet<strong>on</strong>e<br />
(1:1) mixture was added, and the soluti<strong>on</strong> was shaked for 2 hours in<br />
laboratory shaking machine. The mixture was filtered <strong>on</strong> Büchner funnel and the<br />
acet<strong>on</strong>e was separated with 700 ml of distilled water. The phase of n-hexane was dried<br />
<strong>on</strong> sodium-sulphate and c<strong>on</strong>centrated with rotati<strong>on</strong> evaporating machine. The residue<br />
was dissolved in 5 ml n-hexane. The extract was cleaned by column-chromatography,<br />
the column filled with 8 g of Alumina (activity: V.). The evaluti<strong>on</strong> was carried out with<br />
n-hexane, finally the soluti<strong>on</strong> was evaporated again and dissolved in 1 ml of n-hexane.<br />
During the sample preparati<strong>on</strong> we used organic solvents in the quality of GC-residue<br />
analysis (Scharlau). The detecti<strong>on</strong> of the analytes was performed <strong>on</strong> a Varian CP-3800<br />
gas chromatograph with ECD detector (Nederland). Operating c<strong>on</strong>diti<strong>on</strong>s: CP 8907<br />
Factor Four Capillary Column, 15 m * 0.25 mm, temperature program: 50 °C, 1 min;<br />
50-180 °C, 25 °C min-1; 180-250 °C, 4°C min-1; 250°C, 1 min, helium carrier gas (1.0<br />
ml min-1, 6.0 Linde), injected volume: 1 µl, temperature of the detector: 250 °C. The<br />
standard materials and internal standards were prepared from MERCK Chlorinated<br />
Pesticide Mix 40.<br />
Heavy metal c<strong>on</strong>tent analysis: De<strong>term</strong>inati<strong>on</strong> of heavy metal c<strong>on</strong>centrati<strong>on</strong>s was made<br />
in the Institute of Food Science, Quality Assurance and Microbiology. Samples were<br />
dried at 60 o C, ground, sieved through a 2 mm mesh sieve and digested in HNO3-H2O2<br />
according to Hungarian methodological standard procedure: MSZ 21470-50:1998<br />
(Kovács et al., 2000). Merck and BDH standard soluti<strong>on</strong>s were used to prepare the<br />
stock-soluti<strong>on</strong>s, and REANAL (Budapest, Hungary) solid chemicals were also used.<br />
Ultrapure water was used to prepare the soluti<strong>on</strong>s (Millipore, Paris, France). The<br />
de<strong>term</strong>inati<strong>on</strong>s were made by a Perkin-Elmer Optima 3300 DV Inductively Coupled<br />
Plasma - Optical Emissi<strong>on</strong> Spectrometry (ICP-OES) equipment.<br />
RESULTS AND DISCUSSIONS<br />
In 2005 by the analysis of element compositi<strong>on</strong> we found that the examined fields show<br />
significant differences in macro nutrient c<strong>on</strong>centrati<strong>on</strong>s (N, P, K, S), but show<br />
surprisingly homogenous distributi<strong>on</strong> in the case of several micro nutrients and toxic<br />
elements (Table 1). We found no significant amount of pesticide residues in the<br />
ploughed upper soil layer (0-20 cm layer), but in the case of maize and winter wheat<br />
fields a small degree of accumulati<strong>on</strong> was observed (Table 2). These results from the<br />
pesticide residue analysis induced us to c<strong>on</strong>tinue the sampling in wider soil profile.<br />
We found as a result of analysis in 2006 that the bio producti<strong>on</strong> field at Látókép<br />
did not c<strong>on</strong>tain detectable amounts of pesticide residues. In the case of other sites the<br />
detected amount of pesticide residues was diverse in the case of different chemicals, but<br />
in all case it was under the c<strong>on</strong>taminati<strong>on</strong> limit value (A value) of 10/2000. (VI.2.)<br />
KöM-EüM-FVM-KHVM ministry order in the whole soil profile (Figure 1). The peaks<br />
371
visible at lower soil layers does not mean risks in producti<strong>on</strong> and in food safety in the<br />
case of the examined cultivated plants, partly because of the relatively low<br />
c<strong>on</strong>centrati<strong>on</strong>s, partly because we made additi<strong>on</strong>al product (grain and tissues of animals<br />
fed <strong>on</strong> these forages) analysis <strong>on</strong> residues and we have not found detectable amount of<br />
pesticide residues in this products.<br />
Table 1: Element c<strong>on</strong>tent of upper ploughed soil layers of examined fields (mg/kg)<br />
nati<strong>on</strong>wide<br />
element Nádudvar Hajdúböszörmény Bak<strong>on</strong>szeg Sárrétudvari Szentes average*<br />
P 1032 887 881 1009 918 535<br />
K 4443 2858 3265 3145 3658 2952<br />
S 123 118 138 152 159 128<br />
Zn 76,4 76,4 72 73 70 47<br />
Cu 18 14 12 15 12 18<br />
Ni 20 19 19 20 19 14<br />
Mn 762 704 608 536 643 457<br />
Cd 0,957 0,92 0,94 0,93 0,96 0,81<br />
Pb 24 22 22 21 20 14<br />
* average of TIM (Hungarian Soil Informati<strong>on</strong> and M<strong>on</strong>itoring System) samples<br />
Table 2: Pesticide residues of different soil levels of examined fields, 2005 (μg/kg)<br />
Depth Sum of HCH Sum of drins<br />
Sum of DDT /<br />
DDD / DDE<br />
Hajdúböszörmény<br />
(maize)<br />
0-20<br />
100-130<br />
200-235<br />
0, 024397<br />
35, 75<br />
24, 45<br />
0, 002113<br />
0, 72<br />
0, 84<br />
0, 071583<br />
11, 77<br />
8, 97<br />
0-20 0, 025776 0, 00248 0, 009218<br />
Nádudvar (maize) 100-130 32, 95 4, 94 2, 41<br />
200-235 30, 69 3, 6 1, 92<br />
0-20 0, 00764 0, 00017 0, 01774<br />
Szentes (wheat) 100-130 10, 26 0, 24 0, 58<br />
Sárrétudvari<br />
(wheat)<br />
Bak<strong>on</strong>szeg<br />
(meadow)<br />
Látókép (bio)<br />
200-235 10, 12
Although the toxic element c<strong>on</strong>tent of upper soil levels showed no high values (and<br />
therefore risk in food safety) again we analysed the element compositi<strong>on</strong> of soil samples<br />
besides the pesticide residues. In the case of the bio field of Látókép we measured lower<br />
values than the limit value by Hungarian orders, in homogenous distributi<strong>on</strong> in the more<br />
or less deep soil layers. On the other hand, in the case of intensively cultivated fields the<br />
cadmium c<strong>on</strong>centrati<strong>on</strong> is near to the B value, suggesting c<strong>on</strong>taminati<strong>on</strong> in certain<br />
layers (Figure 2). Although the measured values are lower than the c<strong>on</strong>taminati<strong>on</strong> level<br />
(C value) currently not in arrangement and the envir<strong>on</strong>mental remediati<strong>on</strong><br />
c<strong>on</strong>taminati<strong>on</strong> limit (D value), it is recommended to analyse this parameter in the case<br />
of those sensitive plants that are susceptible to accumulati<strong>on</strong> of cadmium. The little<br />
higher c<strong>on</strong>centrati<strong>on</strong> (~10-15%) than the natural average value of copper in the case of<br />
some samples is a c<strong>on</strong>sequence of intensive agr<strong>on</strong>omy. From the results the near to limit<br />
value of zinc c<strong>on</strong>centrati<strong>on</strong> at Szentes in lower soil layers is also remarkable.<br />
Nádudvar<br />
(maize)<br />
Szentes<br />
(wheat)<br />
Depth (cm)<br />
Depth (cm)<br />
0<br />
50<br />
100<br />
150<br />
200<br />
250<br />
300<br />
0<br />
50<br />
100<br />
150<br />
200<br />
250<br />
300<br />
0 5 10 μg/kg 15 20 25<br />
sum of HCHs<br />
sum of drins<br />
sum of DDTs<br />
0 5 10 μg/kg 15 20 25<br />
sum of HCHa<br />
sum of drins<br />
sum od DDTs<br />
Figure 1: Pesticide residues in the examined soil profiles, 2006 (μg/kg)<br />
373
Látókép<br />
Nádudvar<br />
Szentes<br />
Depth (cm)<br />
Depth (cm)<br />
Depth (cm)<br />
0 20 40 60 80 100 120<br />
0<br />
50<br />
100<br />
150<br />
200<br />
250<br />
300<br />
(mg/kg)<br />
0 20 40 60 80 100 120<br />
0<br />
50<br />
100<br />
150<br />
200<br />
250<br />
300<br />
(mg/kg)<br />
0 20 40 60 80 100 120<br />
0<br />
50<br />
100<br />
150<br />
200<br />
250<br />
300<br />
(mg/kg)<br />
Figure 2: Toxic element c<strong>on</strong>tent in the examined soil profiles, 2006 (mg/kg)<br />
374<br />
Cd<br />
Cu<br />
Pb<br />
Zn<br />
Cd<br />
Cu<br />
Pb<br />
Zn<br />
Cd<br />
Cu<br />
Pb<br />
Zn
ACKNOWLEDGEMENTS<br />
This work was supported by ALAP1-00058/2004 GAK: “Farm to fork” for cereals for<br />
animal food (2005-) and GVOP-3.1.1.-2004-05-0416/3.0: Applicati<strong>on</strong> of modern bio<br />
and food analytical methods in food raw material processing (2005-) research projects<br />
REFERENCES<br />
10/2000. (VI.2.) KöM-EüM-FVM-KHVM együttes rendelete. Magyar Közlöny, 53:<br />
3156-3167<br />
Biehl, M.L., Buck, W.J., (1987): Chemical c<strong>on</strong>taminants: their metabolism and their<br />
residues. J. Food Protect. 50. 12, 1058–1073.<br />
Brus, D.J., de Gruijter, J. J., Römkens, P.F.A.M. (2005): Probabilistic quality<br />
standards for heavy metals in soil derived from quality standards in crops.<br />
Geoderma 128 (2005) 301– 311.<br />
Kan, C.A., Meijer, G.A.L. (2007): The risk of c<strong>on</strong>taminati<strong>on</strong> of food with toxic<br />
substances present in animal feed. Animal Feed Science and Technology, 133, 84–<br />
108.<br />
Kannan, K., Tanabe, S., Giesy, J.P., Tatsukawa, R. (1997): Organochlorine<br />
pesticides and polychlorinated biphenyls in foodstuffs from Asian and Oceanic<br />
countries. Reviews in Envir<strong>on</strong>mental C<strong>on</strong>taminati<strong>on</strong> and Toxicology 152, 1-55.<br />
Kárpáti, Z., Győrfi, L., Csanády, M., Károly, G., Krómer, I. (1998): Ivóvizek<br />
növényvédő szer szennyezettsége. Egészségtudomány, 42, 143–152.<br />
Kovács, B., Prokisch, J., Győri, Z., Balla Kovács, A., Palencsár, A.J. (2000): Studies<br />
<strong>on</strong> Soil Sample Preparati<strong>on</strong> for Inductively Coupled Plasma Atomic Emissi<strong>on</strong><br />
Spectrometry Analysis. Commun. Soil. Sci. Plant Anal., 31.11-14, 1949-1963.<br />
Oldal, B., Maloschik, E., Uzinger, N., Ant<strong>on</strong>, A., Székács, A. (2006): Pesticide<br />
residues in Hungarian soils Geoderma 135. 163–178.<br />
Shegunova, P., Klánová, J., Holoubek, I. (2007): Residues of organochlorinated<br />
pesticides in soils from the Czech Republic. Envir<strong>on</strong>mental Polluti<strong>on</strong> 146. 257-<br />
261.<br />
375
MICROCLIMATIC STUDY OF THE MAIN WINE-GROWING<br />
REGIONS IN ROMANIA DURING 2006<br />
DRD. Dorin POPA<br />
* Universitatea Oradea, Facultatea de Protectia Mediului<br />
ABSTRACT:<br />
From a climatic point of view, Romania differs from other geographical regi<strong>on</strong>s, the result<br />
being specific microclimates favourable for vine cultivati<strong>on</strong>. Our microclimate influences<br />
cultivated vine sorts and the wine quality.<br />
Analyzing climatic particularities of five wine-growing regi<strong>on</strong>s during 2006, from a<br />
point of view of thermic resources, pluviometric c<strong>on</strong>diti<strong>on</strong>s and climatic events, we can<br />
c<strong>on</strong>clude that, from a climatic point of view, 2006 was a favourable year for vine culture,<br />
securing premises for a good producti<strong>on</strong>, in quantity as well as in quality.<br />
Key-words: climate, vine, wine-growing regi<strong>on</strong>s, quality.<br />
1.1 CLIMATIC CHARACTERIZATION<br />
1.1.1. Wine-growing regi<strong>on</strong> Moldavian Hills (Dealurile Moldovei)<br />
Climate particularities during 2006 and their c<strong>on</strong>sequences over vine plantati<strong>on</strong>s will be<br />
presented based <strong>on</strong> specific time periods, analyzing critical climate factors having impact<br />
over vegetati<strong>on</strong> status and producti<strong>on</strong> potential.<br />
From a thermic point of view, 2006 was a year warmer than usual, the medium<br />
temperature oscillating between 8,6 and 11,4 0 C. 2006 started with a cold winter, even frosty<br />
at night and in the morning. In January, the snow was set <strong>on</strong> most of the land, except the<br />
south of the regi<strong>on</strong>, where its absence resulted in damages. The lowest temperature in<br />
January was between –19,9 0 C and –26,8 0 C. As for the highest temperature, this was<br />
between 31,4 0 C and 36,1 0 C, during August.<br />
Analysing the rainfall amount, there has been c<strong>on</strong>cluded that, overall, in Moldavia was<br />
recorded in 2006 a surplus of 7,3 l/m 2 than the average multi-annual amount. The highest<br />
values were recorded in the north of the regi<strong>on</strong>, where the rainfall amount had a surplus of<br />
19,2 % than the average multi-annual amount. At the other side is the Central-South of the<br />
regi<strong>on</strong>, where the deficit was of 10%.<br />
On a m<strong>on</strong>thly basis, the rainfall amount during 2006 was higher from January to May,<br />
except for February and June. The highest amount was in August, the surplus being of 99 %<br />
over the average amount. The daily water quantities were in most of the places over 30<br />
l/m 2 . The deficit of buds due to frost was of 11 – 15 % in the South and 20 – 35 % in the<br />
North-Central regi<strong>on</strong>.<br />
Over January, February and the beginning of March, the soil stayed frozen, with a<br />
depth of 1-10 cm <strong>on</strong> most of the agricultural regi<strong>on</strong>, even 20-35 cm <strong>on</strong> some parts.<br />
Summer was cold at the beginning, with high thermic amplitudes and a high haziness,<br />
delaying the growth and development processes at all the sorts. All al<strong>on</strong>g the summer there<br />
were reported rainfalls, mainly sudden showers, but also torrential. All over the regi<strong>on</strong>, <strong>on</strong><br />
small areas, stormy winds and hail storms partially affected the vine plantati<strong>on</strong>s.<br />
Autumn started with a nice and warm weather, with few rainfalls. The end of autumn<br />
and the beginning of winter (December, 2006) was marked by a high thermic regime and<br />
few rainfalls. It is important to point out that in December 2006 there was no snow layers in<br />
the regi<strong>on</strong>.<br />
376
l/mp<br />
120<br />
100<br />
80<br />
60<br />
40<br />
20<br />
0<br />
Fig. 3 M<strong>on</strong>thly evoluti<strong>on</strong> of rainfalls compared by average multi-annual amount<br />
in Moldavia (situated East from Siret)<br />
28,8 25,9<br />
10<br />
24,2<br />
67,7<br />
26,4<br />
53,9<br />
42,9<br />
64,6<br />
58,7<br />
85,8<br />
77,3<br />
I II III IV V VI VII VIII IX X XI XII<br />
58<br />
66,6<br />
105,4<br />
52,8<br />
19<br />
43,4<br />
2006 normal<br />
1.1.2 Wine-growing regi<strong>on</strong> Podişul Transilvaniei<br />
2006 was a year of quite reduced thermic resources, with an average annual temperature of<br />
9.9°C and a pluviometric regime of 589.4 mm close to normal (644.6 mm) (Tabel 1).<br />
During winter there were low temperature values, the lowest being -20.3°C (in January),<br />
partially affecting buds viability of the sorts sensitive to frosty weather, requiring cutting<br />
compensati<strong>on</strong>s in spring.<br />
Vine started vegetati<strong>on</strong> later, but the richer thermic resources during summer led to<br />
recovery of this delay, the vegetati<strong>on</strong> development being normal. During August the<br />
haziness was high and there were massive rainfalls (169,4 mm), leading to a delay of grape<br />
maturati<strong>on</strong> and creating favourable c<strong>on</strong>diti<strong>on</strong>s of Botrytis cinerea attack. The first frost was<br />
recorded <strong>on</strong> 17.10 (-2.8°C).<br />
Tabel 1: Main climate data for podgoria Tarnave Vineyard – Viticultural Center Blaj<br />
Climatic Average m<strong>on</strong>thly Extreme Extreme Total rainfalls ( mm)<br />
indicator temperature temp. temp.<br />
M<strong>on</strong>th ( o C) lowest<br />
( o highest<br />
C) ( o realized normal diff.<br />
C)<br />
+/ -<br />
January -5,6 -20,3 7,1 9,2 26,3 -17,1<br />
February -0,4 -18,0 16,0 11,1 21,2 -10,1<br />
March 4,4 -12,5 21,5 63,9 28,9 +35,0<br />
April 12,1 -0,5 23,5 80,1 68,3 +11,8<br />
May 15,5 4,4 31,5 54,3 80,2 -25,9<br />
June 18,6 5,5 32,5 88,4 93,6 -5,2<br />
July 21,5 9,5 33,2 17,0 99,0 -82,0<br />
August 19,5 9,6 33,2 169,4 64,0 +105,4<br />
September 16,6 4,5 27,1 27,8 56,7 -28,9<br />
October 10,9 -3,5 28,0 31,7 36,6 -4,9<br />
November 5,2 -5,5 17,1 12,8 36,5 -23,7<br />
December 0,5 -11,5<br />
377<br />
9,1 23,7 33,3 -9,6<br />
14,8<br />
34<br />
7,3<br />
32,6<br />
7<br />
29
1.1.3 Viticultural regi<strong>on</strong> Banat Crisana<br />
Annual thermic regime was slowly in excess (an annual average temperature of 13,0 o C) and<br />
slowly reduced during vine vegetati<strong>on</strong> period (April – September average values of<br />
19,1 o C). Haighest thermic value was recorded in July ( 39,1 o C), from June till September<br />
the temperature values being over 30 o C, slowly affecting the photosynthetical process at a<br />
foliar level. The lowest thermic value was recorded in January ( –15,1 o C), negative<br />
temperature being recorded from October till March, but the fruit buds were not affected.<br />
Buds viability was slowly affected in depressi<strong>on</strong>ary areas, without significant losses.<br />
Thermic accumulati<strong>on</strong> from the standard vegetati<strong>on</strong> period (April – September) favourised<br />
in a certain amount sugars accumulati<strong>on</strong>, stopped by the highest temperature values from<br />
the vegetati<strong>on</strong> period.<br />
Insolati<strong>on</strong> value was close to normal, the photosynthetic process and the crop quality<br />
not being disturbed. From a hydrological point of view, there was a surplus of moisture<br />
c<strong>on</strong>tent, favourising cryptogamic diseases. Normal humidity values were recorded in July-<br />
August, favourising grape maturati<strong>on</strong>.<br />
As for the rainfall regime, according to the real evaporati<strong>on</strong>, 2006 had higher amounts,<br />
a hydrological deficit being recorded in July, September and October.<br />
The dangerous winter c<strong>on</strong>diti<strong>on</strong>s recorded were hoar-frost, block frost and white frost.<br />
Hoar-frosts were recorded from October (4 days) to March (1 day), block and white frost<br />
being recorded in December (1 day) and February (3 days). Am<strong>on</strong>g the worm seas<strong>on</strong><br />
c<strong>on</strong>diti<strong>on</strong>s there were recorded 2 cases of hail-storm in June and str<strong>on</strong>g wind weather, even<br />
stormy weather in April and June.<br />
1.1.4 Viticultural regi<strong>on</strong> Colinele Dobrogei<br />
Examining the thermic regime of 2005-2006 viticultural year, we can say that its values<br />
were higher than the average multi-annual values. Thus, during the vegetative rest period,<br />
November, December and January had an average m<strong>on</strong>thly temperature higher than<br />
normal, and during the vegetati<strong>on</strong> period, from April to January, all m<strong>on</strong>ths had higher<br />
temperature values than normal, the difference ranging from 0,7 0 C in April (lowest) to<br />
3,7 0 C in August (highest).<br />
During summer the highest temperature recorded was between 33,0 0 C and 36,0 0 C,<br />
stimulating the vegetative raise of the vines, <strong>on</strong> the background of the massive rainfalls<br />
from the sec<strong>on</strong>d part of summer.<br />
We can c<strong>on</strong>clude that the 2005-2006 viticultural year, vine took advantage from a<br />
more effective thermic regime than the average, leading to a superior viticultural<br />
producti<strong>on</strong>, mostly in quantity and less in quality, due to the reduced number of days with<br />
sunshine. Regarding the pluviometric regime, we can menti<strong>on</strong> the lower rainfall amounts in<br />
June, a situati<strong>on</strong> that created a str<strong>on</strong>g hydrological pressure in the plantati<strong>on</strong>, irrigati<strong>on</strong>s<br />
being mandatory. But the amount of rainfalls in July, August and September solved the<br />
problem. The relative air humidity was of 85%, higher than the average 73%, influencing<br />
and favourising cryptogamic disease attacks <strong>on</strong> vine, especially mildew <strong>on</strong> sensitive sorts,<br />
but also blast and botrytis <strong>on</strong> the other sorts.<br />
1.1.5 Viticultural regi<strong>on</strong> of Muntenian and Oltenian Hills<br />
Analysing viticultural year 2005-2006 (October 2005- September 2006) we find average<br />
temperature values higher than normal and less rainfalls, leading to an extensi<strong>on</strong> of vine<br />
shoot maturati<strong>on</strong> and a delay of the relative rest of the vine (fig. 1).<br />
The reduced amount of rainfalls in May (16,4 mm, compared to the average of 60,1 mm)<br />
and the lower thermic regime c<strong>on</strong>tinued to have a negative influence <strong>on</strong> plant development.<br />
Blooming started late (16 th - 19 th of June) and lasted till the end of the m<strong>on</strong>th. The rains<br />
378
during this period, though not so significant in quantity (43,6 mm), created most favourable<br />
c<strong>on</strong>diti<strong>on</strong>s for the development of pathogenic fungus.<br />
Thermic balance during July (Σ°738,0) and August (Σ°710) and the normal level of<br />
rainfalls led to a recovery of growing differences and a normal development of the<br />
vegetative system and of the grapes.<br />
Climatic c<strong>on</strong>diti<strong>on</strong>s in September, total hours of insolati<strong>on</strong>, rainfalls and the active<br />
thermic balance favoured the accumulati<strong>on</strong> of sugars, of coloured substances and of<br />
specific fragrance of every sort.<br />
ToC/ mm<br />
60.0<br />
50.0<br />
40.0<br />
30.0<br />
20.0<br />
10.0<br />
0.0<br />
Fig. 1 Evoluti<strong>on</strong> of the average daily temperature ( o C) and of rainfalls (mm),<br />
during 01.04 -30.09.06<br />
April<br />
ToC Σmm<br />
May<br />
June<br />
1 11 21 1 11 21 31 10 20 30 10 20 30 9 19 29 8 18 28<br />
If we analyse the climatic profile (for the vegetati<strong>on</strong> period) for 2006 (tabel 4) and we<br />
compare it with the normal values of the viticultural center Valea Calugareasca and with the<br />
optimal values for obtaining quality crops, we can see that all the indicators had optimal or<br />
almost optimal values in order to obtain quality white and red wines, having c<strong>on</strong>trolled origin<br />
denominati<strong>on</strong> (DCO).<br />
379<br />
July<br />
August<br />
September
Tabel 4 Climatic profile during vine vegetati<strong>on</strong> period in 2006 Valea Calugareasca Viticultural<br />
Center<br />
Indicator name<br />
Normal<br />
values<br />
Values<br />
recorded<br />
in 2006<br />
Optimal<br />
period<br />
Observati<strong>on</strong>s<br />
Global thermic balance ( o C) 3635 3562<br />
2700-<br />
3500<br />
Useful thermic balance ( o C) 1801 1716<br />
1000-<br />
1700<br />
> 17 o 23,8<br />
C table wines<br />
> 19 o Average temperature in July (<br />
C high quality wines<br />
o C) 25,2<br />
18.5-23.2 o<br />
> 21 C quality wines having<br />
c<strong>on</strong>troled<br />
denominati<strong>on</strong><br />
origin<br />
Highest average temperature in<br />
August ( o C)<br />
29,0<br />
26,8<br />
24.5-30.0<br />
Values higher than 34 o C are<br />
restrictive, beacuse they lead<br />
to physiological process<br />
blocking.<br />
Highest temperature in August<br />
( o 34,4<br />
C)<br />
34,2 2300 favourability for<br />
aromatic sorts.<br />
Indicator of coolness at night IF 19,8 13,6<br />
>18<br />
< 12<br />
Worm nights, favourable to<br />
maturati<strong>on</strong><br />
Cold nights, unfavourable to<br />
maturati<strong>on</strong>.<br />
1.2. CONCLUSIONS<br />
Each wine-growing regi<strong>on</strong> has distinctive features from a microclimatic point of view, but<br />
we can make a general analysis, taking into account the comm<strong>on</strong> climatic elements during<br />
the analysed period, as it follows:<br />
• thermical resources were high, a little over the multinati<strong>on</strong>al average values;<br />
380
• the pluviometric regime was also higher than the average multinati<strong>on</strong>al values, being<br />
quite uniformly distributed throughout the whole year;<br />
• climatic accidents as the hail-storms in Moldavia and Dobrogea did not severely affect<br />
vine plantati<strong>on</strong>s.<br />
Losses of buds due to low temperatures during winter (-18 0 C) were of 0 and 35%, a<br />
normal situati<strong>on</strong>, solved by compensati<strong>on</strong> cuttings.<br />
Autumn was l<strong>on</strong>g and warm, favourising sugars accumulati<strong>on</strong> and colour for the red<br />
grape sorts.<br />
An excepti<strong>on</strong> from a microclimatic point of view for 2006 was Transylvannia Plateau,<br />
where thermic resources were quite low, the annual average temperature being 9,9 0 C, the<br />
pluviometric regime 589,4 mm, the average multiannual value being of 644,6 mm. Also,<br />
the lowest temperature (-20,3 0 C) affected frost-sensitive buds. Haziness and massive<br />
rainfalls during the ripening period favourised mouldiness <strong>on</strong> grapes.<br />
On the whole, we can say that 2006 was a microcimatic faourable year for most of the<br />
wine-growing regi<strong>on</strong>s, this leading to good producti<strong>on</strong>s, in quantity as well as in quality.<br />
BIBLIOGRAPHY<br />
* Ministerul Mediului şi Gospodăririi apelor<br />
* Administraţia Naţi<strong>on</strong>ală de Meteorologie www.inmk.ro<br />
* Raport 2006 ICDVV Valea Călugărească<br />
* Raport 2006 SCDVV Odobeşti<br />
* Raport 2006 SCDVV Bujoru<br />
* Raport 2006 SCDVV Murfatlar<br />
* Raport 2006 SCDVV Blaj<br />
* Raport 2006 SCDVV Iaşi<br />
* Raport 2006 SCDVV Miniş Măderat<br />
381
WHEAT GRAINS PROCESSING INFLUENCE AT QUALLITY PARAMETERS<br />
Timar Adrian<br />
University of Oradea, faculty for Envir<strong>on</strong>mental Protecti<strong>on</strong><br />
ABSTRACT<br />
This study try to find if processing of wheat grains is important. Althrough we try to<br />
find how parameters of grains is changed during proccesing and if this parameters are<br />
significant improuved. Paper is a part of my Phd. Thesis and will be coroborate with<br />
parameters evoluti<strong>on</strong>s during storage time in thre diferent systems.<br />
Key words : wheat grains, gluten, starch, rheologichall parameters, wheat grains purity.<br />
1. INTRODUCTION<br />
For evaluati<strong>on</strong> the importance of processing we use data imputs for following<br />
parameters: Organolepticall analysis, Physical analysis (Purity, Hectolitric weight, 1000<br />
grains weight, Absolut weight, Specific weight, Grains humidity), Chemical analysis<br />
(Mineral c<strong>on</strong>tents, Ph, Proteins c<strong>on</strong>tent, Gluten c<strong>on</strong>tent, Starch c<strong>on</strong>tent), Rheological<br />
analysis (Elasticity of gluten, Extensibillity of gluten, Degradati<strong>on</strong> of gluten). Methods<br />
used for analysis are according with romanian standards and are quottati<strong>on</strong> in latest<br />
studys.<br />
2. MATERIALS AND METHODS<br />
Taking samples: We use to take samples cilindrical and electromechanical probes. From<br />
serface and upper layers samples was taken with cilindrical probes. From deep layers<br />
samples was taken with electromechanical probes. Procedure was according to Thierer<br />
L.V. 1976 and Duda M. 2003.<br />
Obteining working samples :We form successively elementar, brutto, homogenized,<br />
laboratory and work samples according with Mureşan et al. 1986.<br />
2.1.Organoleptical analysis<br />
Was study colour, aspect, smell and tste of grains according Thierer L.V. 1966. If this<br />
parameters was out of normal range grains was c<strong>on</strong>sidered out of standards and study of<br />
those samples was ended.<br />
2.2.Physical analysis<br />
We study follow parameters : Purity, Hectolitric weight, 1000 grains weight, Absolut<br />
weight, Specific weight, Grains humidity.<br />
1./ Purity was study according romanian method, Muresan T. 1986 and Duda M. 2003.<br />
P% = mx100/M %<br />
m – weight of elements in g;<br />
M – initial weight of sample in g.<br />
2./ Hectolitric weight was study with hectolitrical 20 l ballance according Muresan T.<br />
1986 and Duda M. 2003.<br />
3./ 1000 grains weight was study with analitical ballance 0,01 g precisi<strong>on</strong> range<br />
accooording Muresan T. 1986 and Duda M. 2003.<br />
4./ Absolut weight was study according Muresan T. 1986 and Duda M. 2003, using<br />
follow formula :G = (100 – U)G/100<br />
U - humidity of grains, %; G – 1000 grains weight, g.<br />
382
1. Specific weight was study according Muresan T. 1986 and Duda M. 2003,<br />
using follow formula: m = G/V<br />
G – 1000 grains weight, g; V - volumme of 1 000 grains, cm 3 .<br />
2.2.Chemical analysis<br />
We study following parameters : Mineral c<strong>on</strong>tents, Ph, Proteins c<strong>on</strong>tent, Gluten c<strong>on</strong>tent,<br />
Starch c<strong>on</strong>tent.<br />
1. Mineral c<strong>on</strong>tents was made according Le<strong>on</strong>te M. 2006 by calcinati<strong>on</strong> at<br />
600 °C.<br />
2. Ph was measured by titrating with NaOH according Thierer L.V. 1966.<br />
3. Proteins c<strong>on</strong>tent was study by mineralyzade and titrating this extract<br />
according Le<strong>on</strong>te M. 2006. Results was using in following foemula :<br />
P%=[(V1 – V)x0,007x5,70]x100/m<br />
V1 - H2SO4 0,5 n volume in ml;<br />
V - NaOH 0,5 n volume in ml;<br />
m – samples weight, în g;<br />
0,007 – N, g equivalent in 1 ml H2SO4 0,5 n;<br />
5,70 - proteic coeficient of wheat, g for 1 g N.<br />
4. Gluten c<strong>on</strong>tent was study according Thiere L.V. 1966 and Le<strong>on</strong>te M.<br />
2003.<br />
5. Starch c<strong>on</strong>tent was study by Ewers method according Thiere L.V. 1966<br />
and Le<strong>on</strong>te M. 2003.<br />
2.3. Rheological analysis<br />
We study following parameters : Elasticity of gluten, Extensibillity of gluten,<br />
Degradati<strong>on</strong> of gluten.<br />
1. Elasticity of gluten measured lenght until epruvets was tear according Thiere<br />
L.V. 1966 and Le<strong>on</strong>te M. 2003.<br />
2. Extensibillity of gluten was study according Thiere L.V. 1966 and Le<strong>on</strong>te M.<br />
2003, and measured capacity of gluten sphere to regain initial shape after<br />
ending deformati<strong>on</strong>. Results was expressed in c<strong>on</strong>venti<strong>on</strong>al value from 1 for<br />
very elastic and 5 for unelastic sample.<br />
3. Degradati<strong>on</strong> of gluten was study according Auerman method quotatti<strong>on</strong> by<br />
Thiere L.V. 1966 and Le<strong>on</strong>te M. 2003.<br />
2.4. Experimental Methodic<br />
Samples was study according following schema :<br />
V1 – raw materials;<br />
V2 – processing with fanning maschine type TA 1215 according Costin I. 1983 and<br />
Bucurescu N. 1992;<br />
V3 – processing with fanning maschine type TA 1215 and selector type MCS 5<br />
according Costin I. 1983 and Bucurescu N. 1992;<br />
V4 – processing with fanning maschine type TA 1215, selector type MCS 5 and<br />
gravitati<strong>on</strong>al typ G 3,3 equipment according Costin I. 1983 and Bucurescu N.<br />
1992<br />
2.5. Biological material<br />
We study wheat, Triticum aestivum L. ssp. vulgare Host McKey, kind named „Dropia”.<br />
Dropia, was created by I.C.C.P.T. Fundulea, in 1992 and registered in 1993.<br />
383
Grain is big, ovoid, and colour is red.. 1000 grains weight is around 44 şi 48 g. and<br />
hectolitrical weight is around 73 şi 74,5 g.<br />
Dropia is in B1-A2 quallity groupe, protein c<strong>on</strong>tent is 13 – 14,8 %, wet gluten 23,3<br />
– 38,8 % and dry gluten 9,2 – 15,4 %.<br />
This study has been d<strong>on</strong>e in 15 july 2005 at harvest moment.<br />
2.6. Statistics methodic<br />
We use Polifaact statistic processing software.<br />
3. RESULTS AND DISCUSSION<br />
3.1. Physical analysis<br />
Table 1. Purity, P % results<br />
Nr. crt. Variant P, (%) % Diference Semnificati<strong>on</strong><br />
1 V1 81,00 100,00 - mt.<br />
2 V2 81,80 101,00 0,80 ***<br />
3 V3 82,60 102,00 1,60 ***<br />
4 V4 84,30 104,10 3,30 ***<br />
DL (p 5%)= 0,01; DL (p 1%)=0,02; DL (p 0,1%)=0,02<br />
After every kind of processing of grains purity grows very significant so in V4 we have<br />
the best results.<br />
Table2. Hectolitric weight MH % results<br />
Nr. crt. Variant MH, (kg) % Diference Semnificati<strong>on</strong><br />
1 V1 77,47 100,00 - mt.<br />
2 V2 77,87 100,50 0,40 ***<br />
3 V3 78,17 100,90 0,70 ***<br />
4 V4 78,47 101,30 1,00 ***<br />
DL (p 5%)=0,10; DL (p 1%)=0,15 ; DL (p 0,1%)=0,24<br />
After every kind of processing of grains Hectolitric weight grows very significant so in<br />
V4 we have the best results.<br />
Table.3. 1000 grains weight, MMB g results<br />
Nr. crt. Variant MMB, (g) % Diference Semnificati<strong>on</strong><br />
1 V1 47,00 100,00 - mt.<br />
2 V2 47,80 101,70 0,80 ***<br />
3 V3 47,27 100,60 0,27 ***<br />
4 V4 47,60 101,30 0,60 ***<br />
DL (p 5%)=0,06; DL (p 1%)=0,09; DL (p 0,1%)=0,14<br />
After every kind of processing of grains 1000 grains weight grows very significant so in<br />
V4 we have the best results.<br />
384
Table 4. Absolut weight MA g results<br />
Nr. crt. Variant MA, (g) % Diference Semnificati<strong>on</strong><br />
1 V1 33,12 100,00 - mt.<br />
2 V2 33,96 102,50 0,84 ***<br />
3 V3 34,22 103,30 1,10 ***<br />
4 V4 35,35 106,70 2,23 ***<br />
DL (p 5%)=0,01; DL (p 1%)=0,02; DL (p 0,1%)=0,02<br />
After every kind of processing of grains Absolute weight grows very significant so in<br />
V4 we have the best results.<br />
Table 5. Specific weight MS g results<br />
Nr. crt. Variant MS, (g) % Diference Semnificati<strong>on</strong><br />
1 V1 1,40 100,00 - mt.<br />
2 V2 1,44 103,10 0,04 ***<br />
3 V3 1,49 106,20 0,09 ***<br />
4 V4 1,50 107,10 0,10 ***<br />
DL (p 5%)=0,01; DL (p 1%)=0,01, DL (p 0,1%)=0,02<br />
After every kind of processing of grains Specific weight grows very significant so in V4<br />
we have the best results.<br />
3.2. Chemical analysis<br />
Table 6. Mineral c<strong>on</strong>tents % results<br />
Nr. crt. Varianta Mineral c<strong>on</strong>tents, (%) % Diference Semnificati<strong>on</strong><br />
1 V1 1,82 100,00 - mt.<br />
2 V2 1,82 100,00 - -<br />
3 V3 1,82 100,00 - -<br />
4 V4 1,81 99,40 - 0,01 ooo<br />
DL (p 5%)= - ; DL (p 1%)= - , DL (p 0,1%)= -<br />
After every kind of processing of mineral c<strong>on</strong>tent decrease very significant in V4<br />
especialy but because DL is nule this is a insignificant variance.<br />
Table7. Ph results<br />
Nr. crt. Variant Ph<br />
1 V1 2,5<br />
2 V2 2,5<br />
3 V3 2,5<br />
4 V4 2,5<br />
Is no variati<strong>on</strong> in course of processing.<br />
385
Table 8. Proteins c<strong>on</strong>tent % results<br />
Nr. crt. Variant Proteins c<strong>on</strong>tent, (%) % Diference Semnificati<strong>on</strong><br />
1 V1 13,80 100,00 - mt.<br />
2 V2 13,50 97,80 - 0,30 ooo<br />
3 V3 13,45 97,50 - 0,35 ooo<br />
4 V4 13,21 95,70 - 0,59 ooo<br />
DL (p 5%)=0,01; DL (p 1%)=0,02, DL (p 0,1%)=0,02<br />
Because external comp<strong>on</strong>ents are eliminated during processing after every stage protein<br />
level is decreasing. Rate of decreasing is very significant.<br />
Table 9. Gluten c<strong>on</strong>tent % results<br />
Nr. crt. Variant Gluten c<strong>on</strong>tent, (%) % Diference Semnificati<strong>on</strong><br />
1 V1 28,80 100,00 - mt.<br />
2 V2 28,87 100,20 0,07 mt.<br />
3 V3 28,90 100,30 0,10 *<br />
4 V4 29,27 101,60 0,47 ***<br />
DL (p 5%)=0,09, DL (p 1%)=0,13; DL (p 0,1%)=0,21<br />
Variant V4 is the first what improuve very significant gluten c<strong>on</strong>tent. V2 is almoust the<br />
same lice witness sample and V3 is a little improuvment just significant.<br />
Table 10. Starch c<strong>on</strong>tent % results<br />
Nr. crt. Variant Starch c<strong>on</strong>tent, (%) % Diference Semnificati<strong>on</strong><br />
1 V1 59,37 100,00 - mt.<br />
2 V2 59,83 100,80 0,46 ***<br />
3 V3 59,94 101,00 0,57 ***<br />
4 V4 60,70 102,20 1,33 ***<br />
DL (p 5%)=0,06, DL (p 1%)=0,09, DL (p 0,1%)=0,14<br />
After every kind of processing of grains Specific weight grows very significant so in V4<br />
we have the best results.<br />
3.3.Rheological analysis<br />
Table11. Elasticity of gluten % results<br />
Nr. crt. Variant Elasticity of gluten % % Diference Semnificati<strong>on</strong><br />
1 V1 2,00 100,00 - mt.<br />
2 V2 2,00 100,00 - mt.<br />
3 V3 2,00 100,00 - mt.<br />
4 V4 1,00 50,00 - 1,00 ooo<br />
DL (p 5%)=0,10; DL (p 1%)=0,15; DL (p 0,1%)=0,24<br />
386
Very significant decrease are in V4 because this stage reduce also proteins c<strong>on</strong>tent. The<br />
rest of variants are like witness.<br />
Table 12. Extensibillity of gluten mm results<br />
Nr. crt. Variant Extensibillity of gluten, (mm) % Diference Semnificati<strong>on</strong><br />
1 V1 15,30 100,00 - mt.<br />
2 V2 15,30 100,00 - mt.<br />
3 V3 15,32 100,00 0,02 -<br />
4 V4 15,80 50,00 0,50 ***<br />
DL (p 5%)=0,03; DL(p 1%)=0,04; DL(p 0,1%)=0,07<br />
Just in V4 we find very significant value, but because DL are very small this is<br />
especialy in V3 just theoreticaly.<br />
Table 13. Degradati<strong>on</strong> of gluten mm results<br />
Nr. crt. Variant Degradati<strong>on</strong> of gluten, (mm) % Diference Semnificati<strong>on</strong><br />
1 V1 42,03 100,00 - mt.<br />
2 V2 41,43 98,60 - 0,60 ooo<br />
3 V3 40,73 96,90 - 1,30 ooo<br />
4 V4 40,50 96,40 - 1,53 ooo<br />
DL (p 5%)=0,15, DL(p 1%)=0,23; DL (p 0,1%)=0,37<br />
Decreasing very significant of gluten degradati<strong>on</strong> in every variants is obvious and show<br />
in V4 variant the best result.<br />
4.CONCLUSIONS<br />
After this study we find very significant improuvments for almoust all parameters and<br />
is obvious why is necessary to process the wheat with complete technological line.<br />
The improuvments are so important to justify the best and complex producti<strong>on</strong> line, full<br />
scale plant.<br />
REFERENCES<br />
1. Ardelean M., Sestraş R., Cordea Mirela, Tehnică experimentală horticolă, Edit.<br />
Academicpres, Cluj – Napoca, 2005;<br />
2. Bucurescu N. ş.a. : Sămânţa şi pregătirea acesteia pentru însămânţări, Edit. Ceres,<br />
Bucureşti, 1992;<br />
3. Costin, I., Tehnologii de prelucrare a cerealelor în industria morăritului, Ed.<br />
Tehnica, Bucureşti, 1983;<br />
4. Duda M. M., Vârban D., Muntenu S., Fitotehnie, Îndrumător de lucrări practice,<br />
partea I, Edit. AcademicPress, Cluj – Napoca, 2003;<br />
5. Le<strong>on</strong>te M., Tehnologii şi utilaje în industria morăritului, Edit. Millenium, Piatra<br />
Neamţ, 2003;<br />
6. Mureşan T., Pană N.P., Cseresnyes Z, Producerea şi c<strong>on</strong>trolul calităţii seminţelor<br />
agrocole, Edit. Ceres, Bucureşti, 1986;<br />
387
7. Thierer L V. Tehnologia recepţi<strong>on</strong>ării, depozitării, c<strong>on</strong>diţi<strong>on</strong>ării şi c<strong>on</strong>servării<br />
produselor agricole, Edit. Ceres, 1976;<br />
8. Thierer L.V., Dumitrescu M., Huştiu I., Oprescu I., Tehnologia recepţi<strong>on</strong>ării,<br />
depozitării, c<strong>on</strong>diţ<strong>on</strong>ării şi c<strong>on</strong>servării produselor agricole, Ed. Ceres, Bucureşti,<br />
1971;<br />
9. Thierer, L.V., De<strong>term</strong>inarea calităţii produselor agricole vegetale, Bucureşti, Edit.<br />
Agro- Silvică, 1966;<br />
388
OCEAN DYNAMICS: AIR-SEA INTERFACE GASES EXCHANGE 1<br />
Speranta Coldea,<br />
Faculty of Envir<strong>on</strong>ment Protecti<strong>on</strong>, University of Oradea,<br />
Oradea, Str. Gen. Magheru Nr.26, Romania<br />
ABSTRACT<br />
Having known the tracer c<strong>on</strong>servati<strong>on</strong> equati<strong>on</strong> and discussed ocean transport, we will<br />
c<strong>on</strong>sider now the processes at the air-sea interface that need to be specified in order to<br />
solve the tracer c<strong>on</strong>tinuity equati<strong>on</strong>. The most important process that we must c<strong>on</strong>sider<br />
here there is the gas exchange. This process has a major impact <strong>on</strong> the distributi<strong>on</strong> of<br />
gases within both the ocean as well as within the atmosphere. We will introduce an<br />
overview of the major gases in the atmosphere and ocean and some important processes<br />
in which they are involved. Further we will discuss the solubility of gases in seawater<br />
and the processes at the air-sea interface that c<strong>on</strong>trol the exchange between the<br />
atmosphere and ocean.<br />
Keywords: atmosphere – ocean interacti<strong>on</strong>, gases exchange.<br />
1. INTRODUCTION<br />
The terrestrial atmosphere compositi<strong>on</strong> is a unique <strong>on</strong>e in our solar system due to the<br />
influence <strong>on</strong> l<strong>on</strong>g <strong>term</strong>s of biological processes up<strong>on</strong> al the basic comp<strong>on</strong>ents, with the<br />
excepti<strong>on</strong> of noble gases (see Table.nr.1). The noble gases are near the equilibrium state<br />
between ocean and atmosphere (with the excepti<strong>on</strong> of a supersaturati<strong>on</strong> due to air<br />
bubbles solving, bubbles that are injected in ocean by the braking tides, and by the<br />
fluctuati<strong>on</strong>s resulted from mixing, heating and from the water-air fluxes). The<br />
distributi<strong>on</strong> of ocean noble gases is de<strong>term</strong>ined especially by their solubility. The<br />
equilibrium distributi<strong>on</strong> between the atmosphere and ocean of the basic gases is<br />
de<strong>term</strong>ined by the chemical and biological processes that produce the gas c<strong>on</strong>sumpti<strong>on</strong><br />
in the ocean. From the eight reactive gases given in the Table nr.1, three of them, e.g.<br />
O2, CO2 and N2O, are produced and c<strong>on</strong>sumed in great quantities by the oceanic<br />
processes. The Ocean doesn’t produce methane, but the flux of this gas to the<br />
atmosphere is negligible compared to that from terrestrial sources (∼ 2%). The ocean<br />
produces and c<strong>on</strong>sumes also N2, but we d<strong>on</strong>’t know if the ocean is a source or a sink, or<br />
it is neuter for this gas. Some gases have an impact <strong>on</strong> the thermal state of our planet by<br />
capti<strong>on</strong> of l<strong>on</strong>g wave radiati<strong>on</strong> emitted by the Earth, from so called greenhouse effect.<br />
We must observe that CO2 and N2O are in bigger quantities compared with the<br />
beginning of the industrial era and c<strong>on</strong>tribute semnificatively to the greenhouse effect.<br />
We will explore partially the vertical and horiz<strong>on</strong>tal distributi<strong>on</strong> of the gases, and we<br />
will give a special attenti<strong>on</strong> to the c<strong>on</strong>centrati<strong>on</strong>s at the ocean surface in c<strong>on</strong>tact with the<br />
atmosphere. This distributi<strong>on</strong> has some unusual particularities for O2, CO2, and N2O<br />
that we will introduce in our analysis. The n<strong>on</strong>quilibrium state between ocean and<br />
atmosphere of CO2 is larger than that of O2. For CO2 the curve seems to begin to be<br />
larger and it is opposite to that of O2. The same biological processes which decay O2 in<br />
the deep waters will enlarge the c<strong>on</strong>centrati<strong>on</strong> of the total carb<strong>on</strong>. This fact explains<br />
why the curve for CO2 at the equator is opposite to that of O2, but doesn’t explain why<br />
389
the maximum of CO2 have the tendency to be larger than that for O2. We introduce the<br />
following questi<strong>on</strong> at which we must give a resp<strong>on</strong>se: which are the relative<br />
c<strong>on</strong>tributi<strong>on</strong>s of the biological and physical processes to this distributi<strong>on</strong> of gases.<br />
From the measurements it can be seen that the N2O distributi<strong>on</strong> is near to the<br />
atmospheric equilibrium <strong>on</strong> the most latitudes, with the excepti<strong>on</strong> of those closed to the<br />
equator, where N2O is in a big quantity that would imply the existence of an oceanic<br />
source. We need to see which this source is. To answer such questi<strong>on</strong>s we need to know<br />
the gases solubilities, that tell us at which c<strong>on</strong>centrati<strong>on</strong>s are the gases. We must also<br />
discuss about the gases exchange rates that de<strong>term</strong>ines how rapidly a corps immersed in<br />
water will be in equilibrium with the atmosphere at the ocean surface. In a future paper<br />
we will discuss also about the biological processes that take place inside the ocean,<br />
processes that together with the physical <strong>on</strong>es force the gases to go out from the<br />
equilibrium state with atmospheric air.<br />
2. THE GASES SOLUBILITIES<br />
We need to estimate the gas c<strong>on</strong>centrati<strong>on</strong> in mmol ⁄ m 3 in sea water, which is not in<br />
equilibrium with the gas from wet air above of ocean. The gases atmospheric<br />
c<strong>on</strong>centrati<strong>on</strong>s are given as partial pressures in physical atmosphere (atm) defined by<br />
the relati<strong>on</strong>:<br />
pA = χA ⋅P (1)<br />
where the molar fracti<strong>on</strong> χA is the moles number of the comp<strong>on</strong>ent A in the unity of air,<br />
P being the total atmospheric pressure in atm. The most suitable definiti<strong>on</strong> for the gases<br />
solubility is given as:<br />
CA = SA ⋅pA = SA⋅χA⋅P (2)<br />
where CA is the soluti<strong>on</strong> c<strong>on</strong>centrati<strong>on</strong> of the comp<strong>on</strong>ent A in mmol⋅m -3 , equal with 1<br />
μmol⋅l -1 ; SA is the solubility in mmol⋅m -3 ⋅atm -1 . This parameter is related to Bunsen<br />
coefficient β or to the parameter that is functi<strong>on</strong> of solubility, F. These parameters could<br />
be related. In the Table nr.2 the equati<strong>on</strong> and the coefficient for calculati<strong>on</strong> of Bunsen<br />
coefficients and of solubility fracti<strong>on</strong>s F for some gases are given. The solubility decays<br />
with the increasing of the temperature for all gases.<br />
3. THE GAS EXCHANGE<br />
The vertical moti<strong>on</strong>s in the atmosphere and in the ocean are intensively suppressed near<br />
the air-ocean water interface. There were been proposed a variety of models to explain<br />
the effect of this phenomen<strong>on</strong> <strong>on</strong> the gas transfer transverse to the air-water interrface.<br />
We will take into c<strong>on</strong>siderati<strong>on</strong> the stagnant film model. This model supposes that the<br />
turbulence suppressi<strong>on</strong> at the sea-air interface could be represented by molecular<br />
diffusi<strong>on</strong> inside a stable film of finite depth, both in ocean and in the atmosphere. This<br />
model could be easier visualized and gives us the possibility of a direct discussi<strong>on</strong> for<br />
the relative importance of the atmospheric and ocean boundary layers. The model of the<br />
stagnant film represents the transiti<strong>on</strong> from the turbulence inside the free surface fluid to<br />
the molecular diffusi<strong>on</strong> from inside the stable film by a simple structure made from two<br />
layers, illustrated in the Fig.1. If we take into account the fact that doesn’t exist nor<br />
sources nor sinks inside the film, it may be affirmed that fluxes in these films will be<br />
given by the first Fick law:<br />
390
∂ C<br />
Φ = - ε<br />
(3)<br />
∂z<br />
where Φ is in mmol⋅m -2 ⋅ s - 1 . Gas flux inside the atmospheric film is represented by the<br />
expressi<strong>on</strong>:<br />
)<br />
0<br />
0 k<br />
Φ = − k ( p − p ) = − a ( C − C<br />
(4)<br />
a a a a S a a<br />
and inside the ocean film we have the relati<strong>on</strong>:<br />
Φ = − k ( C<br />
0<br />
− C )<br />
(5)<br />
o o o o<br />
Table 1: The atmospheric compositi<strong>on</strong> influenced by chemical and biological<br />
processes<br />
________________________________________________________________<br />
(a)The atmospheric compositi<strong>on</strong> (b) Influenced by chemical<br />
(molar fracti<strong>on</strong> in dry air) and biological processes<br />
___________________________ppm___________________Yes___________ No___<br />
N2 0,78084 ± 0,00004 CO2 358 Ar N2<br />
O2 0, 20946 ± 0,00002 Ne 18,18 ± 0,04 Ne O2<br />
Ar 0,00934 ± 0,00001 He 5,24 ± 0,004 He CO2<br />
CH4 1,72 Kr CH4<br />
Kr 1,14 ± 0,01 Xe H2<br />
H2 0,55 N2O<br />
N2O 0,312 CO<br />
Xe 0,087 ± 0,001 O3<br />
CO 0,05 - 0,20<br />
O3 0,01 - 0,50<br />
See West and East, 1982, for all gases, with excepti<strong>on</strong> of CO2, CH4 and N2O, that are<br />
given after Hought<strong>on</strong>, 1996.<br />
The index 0 indicates the c<strong>on</strong>centrati<strong>on</strong> and the partial pressure at the air-water<br />
interface, where z = 0.<br />
This partial atmospheric pressure of a gas was c<strong>on</strong>verted in the c<strong>on</strong>centrati<strong>on</strong><br />
equivalent in water, C, by using the solubility S. In these equati<strong>on</strong>s ka and ko are the gas<br />
exchange coefficients, having the units of velocity and respectively of solubility. ko is<br />
also named transfer velocity. ka ⁄ S and ko have values of ε ⁄ Δz in the model of stagnant<br />
film, where Δz is the depth of this film.<br />
The measurements of pressures and of partial c<strong>on</strong>centrati<strong>on</strong>s are made above the<br />
air-water interface, never <strong>on</strong> this ocean surface. We will c<strong>on</strong>sider from the c<strong>on</strong>tinuity at<br />
this interface that Φa = Φo = Φ and Ca 0 = Co 0 , with the aim to eliminate the surface<br />
c<strong>on</strong>centrati<strong>on</strong>s from eq.(4) and eq.(5) . It will be obtained that:<br />
Φ = − K( C − C )<br />
(6)<br />
a o<br />
391
with<br />
= +<br />
K k<br />
o<br />
1 1<br />
ka ⁄ S is much larger than 1000 cm ⁄ hour. As compared with the values of order of 10<br />
cm⁄hour for ko, that permits us to ignore the stable film from the atmosphere, this will<br />
give K = kO, and<br />
Φ = − k ( C − C )<br />
(8)<br />
O a O<br />
or, as a functi<strong>on</strong> of partial pressures, we have:<br />
Φ = − k ( p − p )<br />
(9)<br />
s<br />
with ks = kOS.<br />
The measurements of the gas exchange c<strong>on</strong>stants were made in wind tunnels and<br />
using in situ markers for a variety of gases, under various wind velocities and<br />
temperature c<strong>on</strong>diti<strong>on</strong>s. The measurements indicate a dependency of fricti<strong>on</strong> in water,<br />
u*O, for kO of and also of Schmidt number, defined as<br />
Sc = ν ⁄ ε (10)<br />
where ν is the kinematic viscosity, ko of the form<br />
kO = γ -1 Sc -n u*O (11)<br />
and γ is a n<strong>on</strong>-dimensi<strong>on</strong>al c<strong>on</strong>stant. The exp<strong>on</strong>ent n at the number Sc is included to<br />
permit that the gas exchange do not comports as when it should suffer a transversal<br />
transiti<strong>on</strong> to a stable film with kO = ε ⁄ Δz (e.g. with n = 1). n could be de<strong>term</strong>ined by<br />
gases exchange measurements with more that a single gas and a heat exchange in the<br />
same c<strong>on</strong>diti<strong>on</strong>s:<br />
log( k 1 / k 2)<br />
n = − O O<br />
(12)<br />
log( Sc1<br />
/ Sc2)<br />
In Table nr.2 are given different models for the gases exchange coefficient. These have<br />
a difference between them of more than a factor of 2.<br />
4. CONCLUSIONS<br />
There were made laboratory studies and also in situ experiments. The laboratory<br />
studies were made in hydrodynamic tunnels, with which are analyzed the gases<br />
exchanges and the exchange coefficients are measured by measurement of the<br />
equilibrium for a marker mass in water. The exchange <strong>on</strong> a given period of time is the<br />
flux. The exchange coefficient is obtained making the rapport between the flux and the<br />
difference of c<strong>on</strong>centrati<strong>on</strong> air-water, measured with eq.(5). Such experiments have<br />
indicated that the gases exchange coefficients at the air-water interface varies as<br />
functi<strong>on</strong>s of wind velocity, U, giving us the idea that of existence of three linear regimes<br />
separated by the transiti<strong>on</strong> from a plate water surface to <strong>on</strong>e with <strong>on</strong>e with rugosities,<br />
and from a rugous surface to <strong>on</strong>e for which the tides begin to break up. The theory of<br />
the micrometeorological boundary layer, that tell us that n = - 2 ⁄ 3 [1]-[4], seems to<br />
be good when the water surface is plate. The Model of the substituti<strong>on</strong> film, which<br />
proposes the existence of a film that is periodicaly substituted by the fluid volume, gives<br />
392<br />
a<br />
S<br />
k<br />
a<br />
O<br />
(7)
n = - 1 ⁄ 2 [1]-[4]. The later model is applied when the water surface is rugous (with<br />
tides).<br />
Table nr.2<br />
___________________________________________________________Solubility<br />
Symbol C<strong>on</strong>versi<strong>on</strong> to SA<br />
___________________________________________________________<br />
For using with pA humid = pA dry (1 - pH2O/P)<br />
−3<br />
β A 6 mmol m<br />
Bunsen coefficient βA SA = × 10 −1<br />
VA mol l<br />
Solubility functi<strong>on</strong> F<br />
F<br />
3<br />
6 mmol m<br />
−<br />
SA =<br />
× 10<br />
( P − p ) −1<br />
H2O<br />
mol l<br />
______________________________________________________________________<br />
For using with pA dry<br />
β<br />
−3<br />
6<br />
Bunsen coefficient βA SA =<br />
A mmol m<br />
× 10 −1<br />
V mol l<br />
A<br />
−3<br />
F 6 mmol m<br />
Solubility functi<strong>on</strong> F SA = × 10 −1<br />
p mol l<br />
It can be observed that a not so soluble gas as O2 has an exchange rate larger than a<br />
more soluble gas as CO2. This fact is perfectly compatible with the important role of the<br />
bubbles in the gas exchange process previously discussed. The not so soluble gases<br />
exchange is increased by a bigger fracti<strong>on</strong>al quantity when the bubbles are developed,<br />
due to the fact that the gases with a lower solubility have a lower transfer coefficient<br />
transversal to the air-sea interface than the gases with bigger solubility. The bubbles<br />
transfer could be separately discussed. A result of the laboratory studies is: it<br />
dem<strong>on</strong>strates that the sensibility of gases exchange for a variety of c<strong>on</strong>diti<strong>on</strong>s, that are<br />
difficult to reproduce in a laboratory situati<strong>on</strong>.<br />
A series of techniques for studying the gases exchange in a more realistic mode<br />
were developed, in the field c<strong>on</strong>diti<strong>on</strong>s. There are three such techniques:<br />
• The method of the rad<strong>on</strong> deficiency<br />
• The method of the oceanic radiocarb<strong>on</strong> inventory, and<br />
• The dual method of a tracer.<br />
We will refer to in situ (in field) studies, describing them and presenting here <strong>on</strong>ly the<br />
method of the rad<strong>on</strong> deficiency.<br />
(a) The method of the rad<strong>on</strong> deficiency is based <strong>on</strong> the Rad<strong>on</strong>-222 distributi<strong>on</strong> at<br />
the ocean surface. This radioisotope, with the de half value life time of 3,825 days, is<br />
formed by the decay of Radiu-226 at oceanic water surface. We know from<br />
393
c<strong>on</strong>centrati<strong>on</strong> measurements of the radium how much rad<strong>on</strong>-222 is formed in a unit of<br />
time. The measurements effectuated in the mixed layer present a deficiency relative<br />
to the quantity at which we should expect. The flux to the atmosphere can be estimated<br />
from this deficiency and the gas exchange coefficient is found from the rapport of the<br />
flux to the air–water c<strong>on</strong>centrati<strong>on</strong> gradient. We will present a simple estimati<strong>on</strong>, but we<br />
will firstly c<strong>on</strong>sider the process of a series of decays, which imply radium-226 and<br />
rad<strong>on</strong>-222. Radiu-226 has a half value life time much l<strong>on</strong>ger than rad<strong>on</strong>-222. These two<br />
isotopes are put in isolati<strong>on</strong> until when they reaches equilibrium, where the exchange<br />
rates for which the two isotopes are decayed are equal. It is better to give the<br />
c<strong>on</strong>centrati<strong>on</strong> of these radioisotopes by the activity, noted as A, which is equal with the<br />
product between the decay c<strong>on</strong>stant λ and the atoms number that are present in a<br />
volume unit:<br />
A Rn-222 = λ⋅CRn-222 (13)<br />
where CRn-222 represents the atom c<strong>on</strong>centrati<strong>on</strong> per volume unit. In the case of our<br />
c<strong>on</strong>sidered isotopes the c<strong>on</strong>centrati<strong>on</strong> is given in dpm ⁄100 Kg, where dpm is the<br />
number of decays in a minute. The decay c<strong>on</strong>stant is given as a rapport between ln(2)<br />
and the mean half value life time. The equilibrium activity of Rn-222 is of the form A Rn-<br />
222 Ra-226<br />
= A .<br />
The rad<strong>on</strong> deficiency due to the gas exchange is given by:<br />
Φ = h(A Ra-226 - A Rn-222 ) (14)<br />
where h is the Rad<strong>on</strong>-222 layer depth and the activities are the values <strong>on</strong> this layer. The<br />
flux will be of the form:<br />
Φ = - kapă(CRn-222)aer - (CRn-222)water =<br />
k<br />
= −<br />
water<br />
[ A ) − ( A ) ]<br />
(15)<br />
λ Rn−222<br />
air Rn−222<br />
water<br />
The c<strong>on</strong>centrati<strong>on</strong> of the atmospheric rad<strong>on</strong> Rn-222 is much lower than that from the<br />
ocean. Then we can ignore the activity of Rn-222air and we make equal the last two<br />
equati<strong>on</strong>s, obtaining the following result:<br />
⎛ A ⎞<br />
k = λ h⎜<br />
Ra − 226<br />
− 1⎟<br />
(16)<br />
water ⎜ A ⎟<br />
⎝ Rn − 222 ⎠<br />
From the made measurements it was obtained (in a mixed layer of 40 m in depth) that<br />
the rapport of Ra-226 to Rn-222 is 1,6, giving a pushing velocity kappa of the form:<br />
ln( 2)<br />
−1<br />
k<br />
water<br />
= 40 m(<br />
1,<br />
6 − 1)<br />
= 4,<br />
4 m d = 10 cm / hour (17)<br />
3,<br />
85d<br />
(c) Applicati<strong>on</strong>s<br />
One of the advantages of in situ researches is that they have permitted a better<br />
understanding of the role played by organic films for de<strong>term</strong>inati<strong>on</strong> of the gases<br />
exchanges. After obtaining such results more types of models for this phenomen<strong>on</strong> of<br />
the gas exchange were been given. It is not the aim of this paper to develope models,<br />
which were <strong>on</strong>ly used in the next discussi<strong>on</strong> and could be given in the Table nr.3. We<br />
wish that, <strong>on</strong> these models base to answer at some questi<strong>on</strong>s that were been not<br />
394
elucidated yet. We must also imply the supersaturati<strong>on</strong> of 3% oxygen observed any<br />
place in the oceans., but also we must c<strong>on</strong>sider a lot of other processes: from the<br />
sensibility to temperature of the oxygen solubility it can be observed that a water<br />
heating, water that is in equilibrium with the atmosphere leads to a supersaturati<strong>on</strong> in O;<br />
an other mechanism which must be c<strong>on</strong>sidered is the mixing.- a mixing of waters at two<br />
different temperatures will also give a supersaturati<strong>on</strong>. The third mechanism which<br />
must be c<strong>on</strong>sidered as c<strong>on</strong>tributing to the supersaturati<strong>on</strong> is bules forming.<br />
One of the known model for bule forming was d<strong>on</strong>e by Woolf and Thorpe (1991);<br />
At a wind velocity of 7,5 m⁄s this model gives a supersaturati<strong>on</strong> in oxygen of 0,1 %. The<br />
theory of bule injecti<strong>on</strong> is probably the cause of a quart from the observed<br />
supersaturati<strong>on</strong>. The remaining suprasaturati<strong>on</strong> in O may be given by some processes as<br />
are the oxygen producti<strong>on</strong> by photosynthesis in the ocean surface layer. The<br />
suprasaturati<strong>on</strong> in O leads at a leak of oxygen to the atmosphere gas flux. Another<br />
mechanism that must be c<strong>on</strong>sidered is the oxygen leak to the ocean <strong>term</strong>ocline by a<br />
vertical exchange. The oxygen producti<strong>on</strong> by photosynthesis will equilibrate the leak<br />
rate by these two mechanisms. We can now estimate the oxygen leak by gas exchange.<br />
We will also include the bule injecti<strong>on</strong> for the temperature correcti<strong>on</strong>s and it is<br />
obtained:<br />
Φ = - kapă[(1 + ζ +Δe)⋅(CO2)aer – (CO2)apă] (18)<br />
The oxygen c<strong>on</strong>centrati<strong>on</strong> in water is suprasaturated by 3 % related to the oxygen<br />
c<strong>on</strong>centrati<strong>on</strong> in the air, e.g. (CO2)water = 1,03 (CO2)air , where (CO2)air = 0,241 molm -3 is<br />
the saturati<strong>on</strong> c<strong>on</strong>centrati<strong>on</strong>, at the surface global mean temperature of 17,64 0 C and<br />
with a water salinity of 24,78 %. The correcti<strong>on</strong> given by the process of bule injecti<strong>on</strong> is<br />
of 0,007, at a wind velocity of 7,5 ms -1 .<br />
From the oxygen solubility we can estimate a surface temperature lower with<br />
0,3 0 C, that will give ζ = 0,004. For the gas exchange the coefficient obtained from the<br />
model of Liss and Merlival is given in the table nr.3. By using the mean global<br />
temperature it is obtained for the Schmidt number ScO = 663. This value gives a gas<br />
exchange coefficient of 17,4 cm⁄hour from the model of Wanninkhof, and a value of<br />
10,9 cm ⁄ hour from the model of Liss and Merlival. C<strong>on</strong>verting these values in units of<br />
m⁄hour and substituting all the obtained values the gas exchange flux will be<br />
Φ = - 4,4 la - 7,0 m -2 yard -1 (19)<br />
The oxygen producti<strong>on</strong> estimati<strong>on</strong> is superimposed <strong>on</strong>to the typical estimati<strong>on</strong>s for the<br />
oxygen biological producti<strong>on</strong> in the surface ocean layer that is of the order of 5 mol m-2<br />
yard-1.<br />
On the basis of the previously made analysis, which combines different<br />
expressi<strong>on</strong>s from various model for the oxygen superasaturati<strong>on</strong> with some in situ<br />
obtained empirical data, we could give the c<strong>on</strong>clusi<strong>on</strong> that the superasaturati<strong>on</strong> in O,<br />
globally observed at the ocean surface could be explained by a combinati<strong>on</strong> of the<br />
following processes: the bule injecti<strong>on</strong>, the effect of a lower temperature of the surface<br />
water, and, the most important mechanism – the producti<strong>on</strong> of oxygen by<br />
photosynthesis. The phenomen<strong>on</strong> of oxygen O2 suprasaturati<strong>on</strong> c<strong>on</strong>ducted by the<br />
biological processes and the net oxygen flux to the atmosphere which this implies must<br />
be equilibrated in the subsaturati<strong>on</strong> regi<strong>on</strong>s by a net flux of oxygen to the ocean. Other<br />
way the level of O2 should decrease has not been observed yet. A z<strong>on</strong>e where there is<br />
395
oxygen subsaturati<strong>on</strong> is that of equator, and we d<strong>on</strong>’t discuss here this particular regi<strong>on</strong>.<br />
As a c<strong>on</strong>clusi<strong>on</strong> the results obtained for the mean gases exchanges could be given in kg<br />
and estimati<strong>on</strong>s for the increasing in radiocarb<strong>on</strong> obtained from different threedimensi<strong>on</strong>al<br />
models are given in the Table nr.4. The last column in Table nr.4 gives the<br />
radiocarb<strong>on</strong> increasing normalized to the global inventory estimated at 7,67 x 10 9 atoms<br />
cm -2 .<br />
Table 3: The expressi<strong>on</strong>s for the exchange coefficients kwater given as a c<strong>on</strong>venti<strong>on</strong> in<br />
cm⁄hour, the wind velocity U is in ms-1, obtained by the various models proposed for<br />
the gas exchange<br />
(1) Liss si Merlival (1986)<br />
kwater = 0,17U(Sc ⁄ 600) -0,5 for U ≤ 3,6 ms -1<br />
kwater = (U – 3,4)2,8(Sc ⁄ 600) -0,5 for 3,6 < U < 13 ms -1<br />
kwater = (U – 8,4)5,9 (Sc ⁄ 600) -0,5 for U > 13 ms -1<br />
(2) Wanninkhov (1992)<br />
kwater = 0,39Umean 2 (Sc ⁄ 600) -0, 5 to be used with the mean wind velocity<br />
kwater = 0,31U 2 (Sc ⁄ 600) -0,5 to be used with the mean wind velocity <strong>on</strong> short <strong>term</strong><br />
(3) Butin and Etcheto (1995)<br />
kwater = 0,34(U 2 + σu 2 (Sc ⁄ 600) -0,5<br />
(4) Broecker (1985)<br />
kg = 0,0118 (U – 2)<br />
(5) Maier-Reimer and<br />
Hasselmann (1987)<br />
kg = 0,050<br />
Table 4:. The mean gases exchanges and the estimated in radiocarb<strong>on</strong> increasing<br />
Formulati<strong>on</strong> for kg global mean of radiocarb<strong>on</strong> increasing<br />
the gas exchange (mol m -2 yard -1 ppm -1 )<br />
The models results<br />
(a) Brocker (1985) 0,061 1,000<br />
(b) Liss and Merlival (1986) 0,029 0,607<br />
( c) Maier-Reimer and Hasselmann 0,050 -<br />
(1987)<br />
(d) Broecker (1985 and 1995) 1,070 and 1,074<br />
REFERENCES<br />
396
RESEARCHES ON THE QUICK METHOD OF SUGAR PRODUCTIVITY<br />
EVALUATION<br />
Gheorghe Sarca<br />
University of Oradea, Faculty for Envir<strong>on</strong>mental Protecti<strong>on</strong><br />
ABSTRACT<br />
In a sugar factory it is necessary to be able to calculate very quickly the sugar<br />
productivity possible to obtain by processing the beet having a certain technological<br />
quality. That is the reas<strong>on</strong> why, in the following lines, it is presented a very quick<br />
method used by the Institute of Alimentary Chemistry, the laboratory for sugar research<br />
and engineering, for such kind of quick calculati<strong>on</strong>s, but with a high theoretical and<br />
practical precisi<strong>on</strong>.<br />
INTRODUCTION<br />
Table 1 is completed after the calculati<strong>on</strong>s.<br />
Table1. 1 The quick calculati<strong>on</strong> of sugar productivity<br />
No Name of product<br />
Quantity of: Purity C<strong>on</strong>tent of:<br />
Product Dry Sugar % Dry Sugar<br />
substance<br />
substance<br />
Kg/100 kg Kg/100 kg Kg/100 kg % ˚Brix %<br />
beet beet beet<br />
C SU Z O B P<br />
0 1 2 3 4 5 6 7<br />
1 Beet noodles 100,00 - 16,00 - - 16,00<br />
2 Unde<strong>term</strong>inable<br />
sugar losses, at<br />
diffusi<strong>on</strong><br />
- - 0,20 - - -<br />
3 Moist beet pulp 80,00 - - - - -<br />
4 Pressed beet pulp 31,10 - 0,28 - 18,00 0,9<br />
5 Water from the<br />
beet pulp’s<br />
pressing<br />
48,90 - - - - -<br />
6 Total water for<br />
diffusi<strong>on</strong><br />
103,70 - - - - -<br />
7 Fresh water for<br />
diffusi<strong>on</strong><br />
54,80 - - - - -<br />
8 Diffusi<strong>on</strong> juice 123,70 18,19 15,52 85,30 14,70 12,50<br />
9 Mud from<br />
carb<strong>on</strong>ati<strong>on</strong><br />
10,00 - 0,07 - - 0,7<br />
10 Unde<strong>term</strong>inable<br />
sugar losses, at<br />
purificati<strong>on</strong><br />
- - 0,05 - - -<br />
397
11 Thin juice 124,70 17,11 15,40 90,00 13,70 12,35<br />
12 Thick juice 26,30 17,11 15,40 90,00 65,00 58,50<br />
13 Unde<strong>term</strong>inable<br />
sugar losses, at<br />
steaming, boiling<br />
and<br />
crystallizati<strong>on</strong> in<br />
three products<br />
- - 0,30 - - -<br />
14 Total products - 17,02 15,10 - - -<br />
15 Molasses 5,66 4,80 2,88 60,00 82,00 50,85<br />
16 Molasses type<br />
50%<br />
5,76 4,80 2,88 60,00 83,33 50,00<br />
17 Moist crystal<br />
sugar<br />
12,300 12,24 12,22 99,85 99,5 99,35<br />
18 Crystal sugar 12,246 12,24 12,22 99,85 99,95 99,80<br />
19 Water<br />
evaporated at the<br />
sugar drying<br />
0,054 - - - - -<br />
For calculating this total the following method is recommended:<br />
C1 has the value of 100.00 because the total is calculated for 100 kg beet noodles;<br />
P1 is de<strong>term</strong>ined with the laboratory analysis. In the given example it was admitted the<br />
value of 16.00%;<br />
Z1=P1;<br />
Z2 – the value recognized by the extractor’s supplier is accepted. In the given example<br />
the value of 0.20% was admitted, specific to the extractor type DDS;<br />
SU2 is c<strong>on</strong>sidered 30% from Z2, because, by fermentati<strong>on</strong> or enzymatic processes, the<br />
sugar is not wholly changed into volatile substances and water, but it is particularly<br />
changed in lactic acid. The recommendati<strong>on</strong> is given by The Laboratory for sugar<br />
research and engineering at the Institute of Alimentary Chemistry, Bucharest;<br />
C3 is c<strong>on</strong>sidered 80% of C1;<br />
P4 is de<strong>term</strong>ined by laboratory analyses. In the given example the value of 0.9% is<br />
accepted;<br />
B4 is de<strong>term</strong>ined by laboratory analyses. In the given example the value of 18% is<br />
accepted;<br />
C4 is calculated from Muck’s diagram, according to the known methodology;<br />
Z4 = (C4+P4)/100;<br />
C5 = C3 – C4;<br />
Z8 = Z1 – Z2 – Z3<br />
P8 is de<strong>term</strong>ined by laboratory analyses. In the given example the value of 12.50% is<br />
accepted;<br />
B8 is de<strong>term</strong>ined by laboratory analyses. In the given example the value of 14.70˚Brix<br />
is accepted;<br />
Q8 = (P8/B8) – 100;<br />
Z8 = ((Z1 – Z2 – Z4)/P8) – 100;<br />
398
SU8 = (Za/QB) – 100;<br />
C8 = (Z8/P8) – 100;<br />
C6 = C8 + C3 – C1;<br />
C7 = C6 – C5;<br />
P9 is de<strong>term</strong>ined by laboratory analyses. In the given example the value of 0.70% is<br />
accepted;<br />
C9 is accepted as being equal with 10.00 kg / 100 kg beet;<br />
Z9 = (C9 x P9)/100;<br />
Z10 = 0.05;<br />
B11 is de<strong>term</strong>ined by laboratory analyses. In the given example the value of 13.70˚Brix<br />
is accepted;<br />
P11 is de<strong>term</strong>ined by laboratory analyses. In the given example the value of 12.35% is<br />
accepted;<br />
Z11 = Z8 – Z9 – Z10;<br />
SU11 = SU8 – SU12;<br />
Q11 = (Z11/SU11) x 100;<br />
C11 = (Z11/P11) x 100;<br />
B12 is de<strong>term</strong>ined by laboratory analyses. In the given example the value of 65.00˚Brix<br />
is accepted;<br />
P12 is de<strong>term</strong>ined by laboratory analyses. In the given example the value of 58.50% is<br />
accepted;<br />
Q12 = (P12/B12) – 100;<br />
SU12 = SU11;<br />
Z3 is accepted with the value of 0.3 kg/100 kg beet;<br />
Z14 = Z12 – Z13;<br />
B15 is de<strong>term</strong>ined by laboratory analyses. In the given example the value of 82.00˚Brix<br />
is accepted;<br />
P15 is de<strong>term</strong>ined by laboratory analyses. In the given example the value of 50.85% is<br />
accepted;<br />
Q15 = (P15/B15) – 100;<br />
Z15 = (SU11-Z11) x (Q15/(100-Q15));<br />
C15 = (Z15/P15) x 100;<br />
SU15 = (Z15/P15) - 100;<br />
Q16 = Q15<br />
P16 = 50;<br />
B16 = (P16/Q16) x 100;<br />
SU16 = SU15;<br />
C16 = (Z16/P16) – 100;<br />
Z17 = Z14 – Z15;<br />
B17 is de<strong>term</strong>ined by laboratory analyses. In the given example the value of 99.50% is<br />
accepted;<br />
P17 is de<strong>term</strong>ined by laboratory analyses. In the given example the value of 99.35% is<br />
accepted;<br />
Q17 = (P17/B17) x 100;<br />
SU17 = (Z17-Q17) x 100;<br />
C17 = (Z17/p17) x 100;<br />
399
SU18 = SU17;<br />
Q18 = (Z18/SU18) – 100;<br />
B18 is de<strong>term</strong>ined by laboratory analyses. In the given example the value of 99.95% is<br />
accepted;<br />
P18 is de<strong>term</strong>ined by laboratory analyses. In the given example the value of 99.80% is<br />
accepted;<br />
C18 = (Z18/P18) – 100;<br />
C19 = C17 – C18.<br />
CALCULATION FORMULAS OF SOME TECHNOLOGICAL PARAMETERS<br />
Calculati<strong>on</strong> of the water quantity for diluting the thick mass final product<br />
The following relati<strong>on</strong> is used in order to calculate the water quantity necessary for the<br />
diluting of the thick mass final product before the additi<strong>on</strong>al crystallizati<strong>on</strong> through<br />
cooling:<br />
[1 l water/100 kg thick mass final product], in which:<br />
B is the c<strong>on</strong>tent of dry substance in the thick mass final product while discharging it<br />
from the vacuum apparatus, in ˚Brix:<br />
NZ1 – c<strong>on</strong>tent of n<strong>on</strong>-sugar in the thick mass final product while discharging it from the<br />
vacuum apparatus, in kg/100 kg thick mass;<br />
A1 – c<strong>on</strong>tent of water in the thick mass final product while discharging it from the<br />
vacuum apparatus, in kg/100 kg thick mass;<br />
NZ2 – c<strong>on</strong>tent of n<strong>on</strong>-sugar expected to be produced by diluting the thick mass final<br />
product, in kg/100 kg thick mass;<br />
A2 – c<strong>on</strong>tent of water in the thick mass final product after diluti<strong>on</strong>, in kg/100 kg thick<br />
mass<br />
Relati<strong>on</strong>s between the values of different indicators de<strong>term</strong>ined in laboratory analyses<br />
The following indicators are:<br />
O – purity;<br />
Z – c<strong>on</strong>tent of saccharose polar-metrical de<strong>term</strong>ined from a product, expressed in %;<br />
S – c<strong>on</strong>tent of apparent dry substance of a product refractory-metrical de<strong>term</strong>ined,<br />
expressed in ˚Brix;<br />
A – c<strong>on</strong>tent of water of a product, expressed in kg/100 kg product;<br />
NZ – c<strong>on</strong>tent of n<strong>on</strong>-sugar of a product, representing the sum of the c<strong>on</strong>tent of organic<br />
substances and mineral substances in a product, expressed in kg/100 kg product;<br />
Z/A – ratio n<strong>on</strong>-sugar / water or solubility of sugar in water;<br />
C – c<strong>on</strong>tent of c<strong>on</strong>ductor-metric ash of a product, expressed in %;<br />
SO – c<strong>on</strong>tent of organic substances in a product, expressed in %;<br />
RO – organic ratio that represents the ratio between the c<strong>on</strong>tent of organic substances<br />
and c<strong>on</strong>ductor-metric ash in a product;<br />
CS – saline coefficient, expressed in the ratio between the c<strong>on</strong>tent of sugar and<br />
c<strong>on</strong>ductor-metric ash in a product<br />
400
There are the following calculati<strong>on</strong> relati<strong>on</strong>s between these indicators:<br />
Z<br />
Q = * 100 =<br />
B<br />
A = 100 – B [kg/100 kg product];<br />
S / A*<br />
( 100 − B)<br />
B<br />
Q*<br />
B<br />
Z =<br />
100<br />
Z<br />
B = * 100 =<br />
Q<br />
=<br />
= Z/A * (100 - B ) [%]<br />
100*<br />
S / A<br />
S / A + NZ / A<br />
Z / A*<br />
( 100 − B)<br />
* 100 [%]<br />
Q<br />
Z Q*<br />
B<br />
NZ = B – Z = * 100 - [%]<br />
Q 100<br />
Z<br />
Z/A =<br />
100 − B<br />
=<br />
Q*<br />
B<br />
[%]<br />
100*<br />
( 100 − B)<br />
B − Z Z / Q − Z Z ⎛ 1 ⎞<br />
RO = - 1 =<br />
- 1 = * ⎜<br />
C C<br />
⎜<br />
−1⎟<br />
[%]<br />
C ⎝ Q ⎠<br />
But Z/C is the saline coefficient, noted CS. It results:<br />
⎛1<br />
− Q ⎞<br />
RO = CS * ⎜<br />
⎟ - 1,<br />
⎝ Q ⎠<br />
From where results:<br />
Q<br />
CS = (1 – RO) * .<br />
1−<br />
Q<br />
REFERENCES<br />
1. Banu C., 1992 – „Progrese tehnice, tehnologice şi ştiinţifice în industria<br />
alimentară”, Ed. Tehnică, Bucureşti.<br />
2. Beceanu Dumitru, Chira Adrian, 2003 – „Tehnologia produselor agricole”, Ed.<br />
Ec<strong>on</strong>omică, Bucureşti.<br />
3. Sarca Gheorghe, 2004 – „Materii prime vegetale”, Ed. Universităţii din Oradea.<br />
4. <str<strong>on</strong>g>Internati<strong>on</strong>al</str<strong>on</strong>g> Sugar Journal, 2006 december.<br />
5. Popescu V., Stroia A., 1998 – „Tehnologii moderne de industrializarea sfeclei de<br />
zahăr”, Ed. Ceres, Bucureşti.<br />
401<br />
[%]
RESEARCH REGARDING THE CONCENTRATION OF THIN JUICE AND<br />
THICK JUICE<br />
Gheorghe Sarca<br />
University of Oradea, Faculty for Envir<strong>on</strong>mental Protecti<strong>on</strong><br />
ABSTRACT<br />
The thin juice boils off in an installati<strong>on</strong> working <strong>on</strong> the multiple effects principle. The<br />
process the c<strong>on</strong>centrati<strong>on</strong> is based <strong>on</strong> is the vaporizati<strong>on</strong> through boiling the thin juice at<br />
boiling temperature.<br />
INTRODUCTION<br />
The highest temperature sugar is submitted to during the technological process of cane<br />
processing is in the first body of the installati<strong>on</strong> and it has values of 126-129 o C. Due to<br />
the high temperature and the time these temperatures are maintained, losses of sugar<br />
occur and they are presented in table 1.<br />
Table 1. Quantity of decomposed sugar in the vaporizati<strong>on</strong> stati<strong>on</strong><br />
with multiple effect in mg/hour<br />
Temperature Apparent c<strong>on</strong>tent of dry substance of juices that boil off in the<br />
vaporizati<strong>on</strong> stati<strong>on</strong>, in °BRIX<br />
o C 15 25 35 45 55 65<br />
80 0,0443 0,0300 0,0156 0,0178 0,0199 0,0232<br />
85 0,0614 0,0420 0,0222 0,0261 0,0295 0,0330<br />
90 0,0789 0,0540 0,0289 0,0343 0,0391 0,0450<br />
95 0,0964 0,0660 0,0356 0,0426 0,0487 0,0555<br />
100 0,1139 0,0780 0,0422 0,0507 0,0583 0,0675<br />
105 0,1384 0,0936 0,0489 0,0587 0,0679 0,0792<br />
110 0,1629 0,1092 0,0556 0,0666 0,0775 0,0891<br />
115 0,1748 0,1186 0,0622 0,0747 0,0861 0,0991<br />
120 0,2822 0,2340 0,1856 0,2268 0,2677 -<br />
125 0,5329 0,5081 0,4838 0,5938 0,7043 -<br />
130 2,0552 1,4609 0,8666 1,0234 1,1799 -<br />
135 3,5775 - - - - -<br />
140 5,0999 - - - - -<br />
In figure 1 is presented the chart of a modern installati<strong>on</strong> of thin juice c<strong>on</strong>centrati<strong>on</strong> in a<br />
sugar plant. This installati<strong>on</strong> reduces to minimum the time for heating the juice,<br />
ensuring the decrease of thick juice dyeing. The capacity of such an installati<strong>on</strong>,<br />
manufactured by the French company MAGUIN, is of 14000-20000 t cane/24 hours.<br />
- entry of thin juice<br />
- high pressure steam<br />
- steams 124 o C<br />
402
- preheaters<br />
- towards c<strong>on</strong>densati<strong>on</strong><br />
- thick juice<br />
- heating<br />
- general services<br />
Fig. 1 - Thin juice c<strong>on</strong>centrati<strong>on</strong> installati<strong>on</strong><br />
MATERIALS AND METHODS<br />
Another c<strong>on</strong>centrati<strong>on</strong> installati<strong>on</strong> used in sugar plants is that of four effects and “0<br />
body” whose thermal regime is presented in table 2. This installati<strong>on</strong> is profitable<br />
especially in the c<strong>on</strong>diti<strong>on</strong>s of juice processing with no thermal stability, as well as in<br />
the case of plants that d<strong>on</strong>’t have electric vortex wheels.<br />
Table 2. Research methods regarding the temperature regime at the stati<strong>on</strong> of<br />
vaporizati<strong>on</strong> c<strong>on</strong>centrati<strong>on</strong> with four effects and “0 body”<br />
Basic indicators<br />
Number of vaporizati<strong>on</strong> effect<br />
0 1 2 3 4<br />
Heating steam temperature 141 114 106 97 84<br />
Heating steam pressure, ATA 3,89 1,71 1,91 0,95 0,56<br />
Useful temperature difference, °C 25,5 6,2 6,4 10,4 22,4<br />
Boiling temperature for juice, °C 116,3 108,6 100,4 87,4 61,4<br />
Temperature loss due to<br />
hydrostatic pressure °C<br />
0,3 0,6 1,4 2,4 3,4<br />
Sec<strong>on</strong>dary steam temperature, °C 115 107 98 84 57<br />
Sec<strong>on</strong>dary steam pressure, ATA 1,77 1,35 0,99 0,58 0,184<br />
Temperature losses in the steam<br />
pipes °C<br />
1 1 1 1 1<br />
C<strong>on</strong>densate temperature, o C 139 112 104 95 81<br />
403
The main technological and energetical tasks of a thin juice c<strong>on</strong>centrati<strong>on</strong> installati<strong>on</strong><br />
are the following:<br />
• c<strong>on</strong>centrati<strong>on</strong> of thin juice up to the limit of avoiding the danger of accidental or<br />
unc<strong>on</strong>trolled crystallizati<strong>on</strong> of saccharose before vacuum apparatus. Generally, in<br />
the c<strong>on</strong>diti<strong>on</strong>s of Romania, the thick juice can c<strong>on</strong>centrate up to 69,5-70,00 o Brix<br />
• recovering the evaporated water from the thin juice and ensuring the requierement<br />
of sec<strong>on</strong>dary steam necessary for technological purposes specific to the plant<br />
• c<strong>on</strong>densati<strong>on</strong> of the steam received from the vortex wheel under the shape of<br />
c<strong>on</strong>densate, that returns to the Thermoelectric Power Plant of the plant.<br />
The correct operati<strong>on</strong> of the thin juice c<strong>on</strong>centrati<strong>on</strong> stati<strong>on</strong> is ensured by the operating<br />
way of the main users of sec<strong>on</strong>dary steam produced as an effect of juice c<strong>on</strong>centrati<strong>on</strong>.<br />
Am<strong>on</strong>g the main users, the preheaters of juices and syrups have an important place.<br />
In tables 3, 4 and 5 are present the total coefficients of thermal transfer, in<br />
W/(m 2 .grade), of diffusi<strong>on</strong> juice, clear juice I, thin juice and thick juice or syrups.<br />
Table 3.Total coefficients of thermal transfer, in W/(m 2 .grade) at diffusi<strong>on</strong> juice<br />
preheaters<br />
Temperature of heating Total coefficients of thermal transfer at circulati<strong>on</strong><br />
agent, speed of juice, in m/s of:<br />
o C<br />
0,1 0,2 0,3 0,4 0,5<br />
Under 100 375 655 836 1017 1220<br />
Over 100 466 815 1045 1275 1513<br />
Table 4. Total coefficients of thermal transfer, in W/(m 2 .grade) at diffusi<strong>on</strong> juice,<br />
clear juice I and thin juice preheaters<br />
Circulati<strong>on</strong> speed<br />
of juice through the<br />
preheaters pipes<br />
Temperature difference, Δt, o C<br />
m/s 10 15 20 40 50<br />
I. For diffusi<strong>on</strong> juice (raw juice)<br />
1,0 710 676 648 627 606 578<br />
1,5 941 901 857 815 780 731<br />
2,0 1163 1164 1101 1045 976 920<br />
12,3 1303 1234 1164 1101 1045 976<br />
2. For clear juice I and thin juice<br />
1,0 2071 1931 1833 1743 1659 1589<br />
1,5 2231 2126 2022 1917 1645 1722<br />
2,0 2440 2280 2182 2043 1952 1645<br />
2,5 2580 2931 2301 2182 2022 1882<br />
404
Circulati<strong>on</strong> speed of<br />
the c<strong>on</strong>centrated<br />
syrup<br />
Table 5. Total coefficients of thermal transfer, in W/(m 2 .grade)<br />
at thick juice preheaters (c<strong>on</strong>centrated syrup)<br />
Thermal transfer<br />
coefficients for c<strong>on</strong>tent of<br />
dry substance in °BRIX,<br />
DE<br />
m/s 60 65<br />
0,3 404 299<br />
0,5 676 466<br />
0,7 752 543<br />
0,9 927 676<br />
Observati<strong>on</strong>s<br />
In the case of calculati<strong>on</strong> of<br />
newly assembled preheaters we<br />
will use values of juice or syrup<br />
speed:for juices 1,5 m/s<br />
- for syrups 0,7 m/s<br />
RESULTS AND DISCUSSIONS<br />
Results obtained in the principles of using the thin juice c<strong>on</strong>centrati<strong>on</strong> installati<strong>on</strong><br />
In the thin juice tank, placed in fr<strong>on</strong>t of the c<strong>on</strong>centrati<strong>on</strong> installati<strong>on</strong>, there must be a<br />
permanent stock of juice that ensures the feeding of the c<strong>on</strong>centrati<strong>on</strong> installati<strong>on</strong> for<br />
10-15 min . If this stock of thin juice is lacking we have to introduce in the tank<br />
c<strong>on</strong>densate from the third level of the c<strong>on</strong>centrati<strong>on</strong> stati<strong>on</strong> with multiple effect.<br />
The functi<strong>on</strong>ing of the c<strong>on</strong>centrati<strong>on</strong> stati<strong>on</strong> is based <strong>on</strong> the circulati<strong>on</strong> of the juice<br />
and of the forward flow steam, with the gradual reducti<strong>on</strong> of pressure and temperature.<br />
From every body of the c<strong>on</strong>centrati<strong>on</strong> stati<strong>on</strong>, sec<strong>on</strong>dary steams are taken over and<br />
directed for heating in various technological points of the plant, at various equipments<br />
and installati<strong>on</strong>s. Generally, the last c<strong>on</strong>centrati<strong>on</strong> effect of a c<strong>on</strong>centrati<strong>on</strong> installati<strong>on</strong><br />
works under vacuum, the equipment that represent this effect being c<strong>on</strong>nected to its<br />
own barometric c<strong>on</strong>denser.<br />
The quantity of vaporized and removed water from the juice in the c<strong>on</strong>centrati<strong>on</strong> stati<strong>on</strong><br />
depends <strong>on</strong> the following factors:<br />
• c<strong>on</strong>tent of sugar of the processed cane and the sutirage made at the sugar extracti<strong>on</strong><br />
installati<strong>on</strong><br />
• diluti<strong>on</strong> grade of juice during the calco-carb<strong>on</strong>ic purificati<strong>on</strong> process<br />
• final c<strong>on</strong>centrati<strong>on</strong> of thick juice that is evacuated from the last level of the<br />
c<strong>on</strong>centrati<strong>on</strong> installati<strong>on</strong>.<br />
The physical and chemical phenomena that occur during the thin juice c<strong>on</strong>centrati<strong>on</strong> are<br />
the following:<br />
• saccharose decompositi<strong>on</strong> under the influence of high temperature<br />
• formati<strong>on</strong> of coloring substances that de<strong>term</strong>ine the increase of the dyeing with 5<br />
units ICUMSA/min – 100 g saccharose. Generally, during the c<strong>on</strong>centrati<strong>on</strong> of the<br />
juice, the dyeing increases with 50-100%<br />
• decompositi<strong>on</strong> of groups of n<strong>on</strong>-saccharates, for example, amides and inverted<br />
sugar existing in the juice, firstly of those quantities that have not been destroyed<br />
in the specific phases of calco-carb<strong>on</strong>ic purificati<strong>on</strong>. As an effect of amides<br />
decompositi<strong>on</strong> amm<strong>on</strong>ia is given off;<br />
• precipitati<strong>on</strong> of insoluble salts, firstly carb<strong>on</strong>ate, sulphite, oxalate and aluminiumcalcium<br />
silicates. The phenomen<strong>on</strong> is due to the increase of the juice c<strong>on</strong>centrati<strong>on</strong>,<br />
405
ecoming an oversaturated soluti<strong>on</strong> for many calcium salts. Also, by the<br />
decompositi<strong>on</strong> of groups of n<strong>on</strong>-saccharates, oxalic acid is formed and it produces,<br />
through the reacti<strong>on</strong> with calcium i<strong>on</strong>s, insoluble salts or hardly soluble calcium<br />
<strong>on</strong>es. The sediments produced in the c<strong>on</strong>centrati<strong>on</strong> stati<strong>on</strong>, partially deposit<br />
themselves <strong>on</strong> the thermal exchange surfaces and partially leave the installati<strong>on</strong>,<br />
being carried al<strong>on</strong>g by the thick juice flow;<br />
• decrease in the alkalinity of the juice as compared to the dry substance c<strong>on</strong>tent, as a<br />
result of the carb<strong>on</strong>ate neutralizati<strong>on</strong> by the formed acids, the juice giving off<br />
carb<strong>on</strong> dioxide;<br />
• in the case of juice overcarb<strong>on</strong>izati<strong>on</strong> at the sec<strong>on</strong>d carb<strong>on</strong>izati<strong>on</strong>, in the steamers<br />
of the c<strong>on</strong>centrati<strong>on</strong> stati<strong>on</strong>, the decompositi<strong>on</strong> of the calcium hydrocarb<strong>on</strong>ate in<br />
calcium carb<strong>on</strong>ate hardly soluble and in CO2 occurs.<br />
The thick juice resulted from the c<strong>on</strong>centrati<strong>on</strong> of the thick juice in the c<strong>on</strong>centrati<strong>on</strong><br />
stati<strong>on</strong> must have the following technical features:<br />
1. the c<strong>on</strong>tent of dry substance: 62-70 o Brix, according to its purity and the fulfilment<br />
of the thermostability criteria<br />
2. dyeing: 1500-2500 units ICUMSA/100 o Brix<br />
3. c<strong>on</strong>tent of calcium salts in the case of technological mature and healthy cane: 50-60<br />
mg CaO/100 o Brix<br />
4. alkalinity to phenolphthalein: 0,005-0,06 g CaO/100 ml.<br />
5. In these c<strong>on</strong>diti<strong>on</strong>s, the quantity of thick juice that results from 100 kg cane is 23-<br />
28 kg.<br />
REFERENCES<br />
1. Banu C., 1992 – „Progrese tehnice, tehnologice şi ştiinţifice în industria<br />
alimentară”, Ed. Tehnică, Bucureşti.<br />
2. Beceanu Dumitru, Chira Adrian, 2003 – „Tehnologia produselor agricole”, Ed.<br />
Ec<strong>on</strong>omică, Bucureşti.<br />
3. Sarca Gheorghe, 2004 – „Materii prime vegetale”, Ed. Universităţii din Oradea.<br />
4. <str<strong>on</strong>g>Internati<strong>on</strong>al</str<strong>on</strong>g> Sugar Journal, 2006 december.<br />
5. Popescu V., Stroia A., 1998 – „Tehnologii moderne de industrializarea sfeclei de<br />
zahăr”, Ed. Ceres, Bucureşti.<br />
406
RESEARCH ON SEEDING AGENTS TO EFFECT THE SUGAR<br />
CRYSTALLIZATION<br />
Gheorghe Sarca<br />
Oradea University, Faculty of Envir<strong>on</strong>mental Protecti<strong>on</strong><br />
ABSTRACT<br />
The quality of sugar is the decisive factor in ensuring the sugar commercializing<br />
through the c<strong>on</strong>servati<strong>on</strong> of old markets and expansi<strong>on</strong> towards new markets. The sugar<br />
quality is also the key element which de<strong>term</strong>ines the price maintenance in according<br />
with the expenditures performed or even the diminuti<strong>on</strong> of the price as a c<strong>on</strong>sequence of<br />
the quality lessening.<br />
INTRODUCTION<br />
Securing the technological process efficiency is mainly d<strong>on</strong>e through the following:<br />
• Decreasing to the minimum level the sugar amount re-circulated from the 1 st step<br />
of crystallizati<strong>on</strong> to the 2 nd <strong>on</strong>e;<br />
• Decreasing to the minimum level the sugar amount re-circulated from the 2 nd step<br />
of crystallizati<strong>on</strong> to the 3 rd <strong>on</strong>e;<br />
• Permanent preoccupati<strong>on</strong> to avoid the drawing in the molasses of those small<br />
crystals that can pas through the loops of the centrifugal sieves;<br />
• Obtaining the top quality sugar mainly defined by:<br />
- soluti<strong>on</strong> colouring, sugar-in-water soluti<strong>on</strong>s transparency<br />
- polarisati<strong>on</strong>, which shows the amount of saccharose c<strong>on</strong>tent out of the crystal<br />
sugar;<br />
- humidity and the c<strong>on</strong>tent of c<strong>on</strong>ductor-metrical ashes;<br />
- the size the shape and the crystal homogeneity;<br />
all these are being worldwide achieved by the way in which the crystallizing and<br />
shaping technology regarding the emerged crystals is c<strong>on</strong>ducted.<br />
MATERIALS AND METHODS<br />
Methods for starting the saccharose crystallizati<strong>on</strong><br />
There are well-know 3 methods for saccharose crystallizati<strong>on</strong> starting:<br />
• seeding in the field of supersaturating metastable by inserting desired quantity of<br />
crystallizing germs in the c<strong>on</strong>centrated syrup;<br />
• crystallizati<strong>on</strong> centres formati<strong>on</strong> by a thermal shock effect in the super-saturati<strong>on</strong><br />
in<strong>term</strong>ediate level;<br />
• sp<strong>on</strong>taneous or natural crystallizati<strong>on</strong> in the unstable field.<br />
The main method for starting the saccharose crystallizati<strong>on</strong> out of its super-saturated<br />
soluti<strong>on</strong>s takes place through the seeding method. The seeding is the preliminary<br />
c<strong>on</strong>diti<strong>on</strong> for the transiti<strong>on</strong> to the boiling and to the c<strong>on</strong>trolled and automatic<br />
crystallizati<strong>on</strong> of the sugar as <strong>on</strong>ly in this way <strong>on</strong>e can reach to the process<br />
reproducibility to the<br />
407
Seeding with powder sugar<br />
The quality of the obtain sugar relies, and especially the homogeneity of the crystals<br />
obtained, the shape of the crystals, the occurrence and the c<strong>on</strong>glomerates c<strong>on</strong>tent, all<br />
depend <strong>on</strong> the way the seeding is d<strong>on</strong>e. In Romania, powder sugar has been used for a<br />
very l<strong>on</strong>g time and it is still being used as seeding agent. The use of powder sugar in<br />
alcoholic suspensi<strong>on</strong> presents certain advantages as against the use of <strong>on</strong>ly powder<br />
sugar as such. The advantages of the seeding with powder sugar are the following:<br />
• <strong>on</strong>e can accurately establish the number of germs c<strong>on</strong>tained in the seeding soluti<strong>on</strong><br />
which leads us to the awareness of the total number of germs introduced in the<br />
vacuum apparatus;<br />
• <strong>on</strong>e can ensure the uniformity of the germs from their size point of view;<br />
• <strong>on</strong>e can achieve the advanced dispersal of the sugar particles out of the alcoholic<br />
suspensi<strong>on</strong>. This dispersal attracts itself an improvement of the sugar’ quality both<br />
from its crystal aspect point of view and from chemical point of view. Due to the<br />
advanced disposal degree, the c<strong>on</strong>glomerate formati<strong>on</strong> is avoided. The crystal<br />
sugar c<strong>on</strong>glomerates are the formati<strong>on</strong>s resulted from the sugar crystals b<strong>on</strong>ding.<br />
During their shaping these c<strong>on</strong>glomerates include “mother” syrup. This fact leads<br />
to the augmentati<strong>on</strong> of the c<strong>on</strong>ductor-metrical and carb<strong>on</strong>ic ashes in the crystal<br />
sugar;<br />
• achievement of a better homogenising of the crystallizati<strong>on</strong> germs within the<br />
supersaturated syrup mass;<br />
• the opportunity for preparing a higher amount of powder sugar alcoholic<br />
suspensi<strong>on</strong> due to its l<strong>on</strong>g-lasting stability;<br />
• replacing the sugar powder, c<strong>on</strong>sider “classical agent for seeding” which is due<br />
also to the humidity c<strong>on</strong>tained in the sugar crystals which is freed through the<br />
crystals’ milling which enhance the crystal sugar tendency to form in the lob run<br />
of crystal lumps.<br />
A method for sugar powder suspensi<strong>on</strong> preparati<strong>on</strong> in isopropyl alcohol<br />
In the specialised literature several method for the preparing the alcoholic suspensi<strong>on</strong><br />
for sugar crystallisati<strong>on</strong> seeding are menti<strong>on</strong>ed. These methods have been studied<br />
within the Sugar Research and Engineering Laboratory from the Food Chemistry<br />
Institute Bucharest, but <strong>on</strong>e can adapt them to our working methods. The method<br />
adopted c<strong>on</strong>sists in sugar milling directly in isopropyl alcohol.<br />
The method is two-phased <strong>on</strong>e, namely:<br />
• obtaining the base suspensi<strong>on</strong>;<br />
• the base suspensi<strong>on</strong> diluti<strong>on</strong> and obtaining the seeding suspensi<strong>on</strong>.<br />
On the basis of the experimental results obtained in the laboratory, which c<strong>on</strong>sist in<br />
sugar milling in isopropyl alcohol in a 5.0 l milling machine <strong>on</strong> balls working <strong>on</strong> rolling<br />
and having the a 46 rotati<strong>on</strong>s/min spin several working c<strong>on</strong>diti<strong>on</strong>s have been established<br />
which are presented bellow.<br />
Obtaining the base suspensi<strong>on</strong><br />
• for a charge <strong>on</strong> employ:<br />
• 100 grams of crystal sugar having a granulati<strong>on</strong> of 0.2-0.8 mm;<br />
• 200 ml. of isopropyl alcohol;<br />
408
• about 160 porcelain balls having a diameter of 30 mm with a total mass of 1.0 kg;<br />
• about 600 porcelain balls having the diameter of 12 mm with a total mass of 1.o<br />
kg;<br />
• the milling period is 1 hour.<br />
Obtaining the seeding suspensi<strong>on</strong> Out of the base suspensi<strong>on</strong> thus obtained, the seeding<br />
suspensi<strong>on</strong> is prepared through its diluti<strong>on</strong> in isopropyl alcohol in a 1:200 ratio.<br />
Maturing the seeding suspensi<strong>on</strong> The thus prepared powder sugar suspensi<strong>on</strong> in<br />
isopropyl alcohol, called seeding suspensi<strong>on</strong>, is let to repose for a maturing period of 4<br />
weeks. In the meanwhile, the germs smaller than 1 mm are being dissolved while the<br />
sugar dissolved crystallizes <strong>on</strong> larger particles. Subsequent to this maturing period<br />
equilibrium is established while the number of sugar particles from the seeding<br />
suspensi<strong>on</strong> actually remains c<strong>on</strong>stant.<br />
Criteria for ensuring the reproducibility of the seeding suspensi<strong>on</strong> quality In order to<br />
always obtain a suspensi<strong>on</strong> with appreciatively the same number of particles <strong>on</strong> the<br />
mass unit, <strong>on</strong>e recommends the observance of the following measures:<br />
• using the sugar with the same granulati<strong>on</strong> for milling namely the sugar of 0.2-0.8<br />
mm, deprived of powder or c<strong>on</strong>glomerates;<br />
• maintaining the ratio “sugar parts” against the “isopropanol parts”. As smaller<br />
quantity of sugar is used <strong>on</strong> a isopropanol unit as brutish the sugar from the<br />
obtained suspensi<strong>on</strong> will be. The best results are obtained using for a part of<br />
granulati<strong>on</strong> sugar of 0.2 mm-0.8 mm or a ratio of 2.0-2.2 parts of isopanorol;<br />
• observing the milling period for the same mill machine functi<strong>on</strong>ing c<strong>on</strong>diti<strong>on</strong>s<br />
(criteria). The milling period prol<strong>on</strong>ging does not imply a better milling.<br />
A method for powder sugar preparati<strong>on</strong> in ethyl alcohol<br />
The permanent tendency to improve the preparati<strong>on</strong> process of powder sugar suspensi<strong>on</strong><br />
preparati<strong>on</strong> during the saccharose crystallizati<strong>on</strong> striking, in the sense to improve the<br />
dispersal technique in solvent, led to a new preparati<strong>on</strong> method of the seeding<br />
suspensi<strong>on</strong> which is based <strong>on</strong> saccharose reduced solubility in ethyl alcohol. This<br />
method c<strong>on</strong>sists in sugar precipitati<strong>on</strong> from a saturated soluti<strong>on</strong> in ethyl alcohol. The<br />
specialised literature recommends a similar method which c<strong>on</strong>sists in precipitating of a<br />
21 grams of sugar which is found in a saturated soluti<strong>on</strong> at 55ºC, a soluti<strong>on</strong> of 28ºBrix<br />
c<strong>on</strong>centrati<strong>on</strong> or 28% in ethyl alcohol.<br />
Preparing the base suspensi<strong>on</strong> The laboratory experiments accomplished <strong>on</strong> the milling<br />
machine based <strong>on</strong> balls allowed the elaborating of the following working method:<br />
• <strong>on</strong>e introduce 21 grams ethyl alcohol in the milling machine <strong>on</strong> which <strong>on</strong>e pour, in<br />
a thin stripe, a 28% sugar soluti<strong>on</strong> which c<strong>on</strong>tains 420 grams of powder sugar;<br />
• <strong>on</strong>e activate the milling machine in rotating (spin) moti<strong>on</strong>. The milling functi<strong>on</strong>ing<br />
period is of 30 minutes during which the sugar crystallizing under the suspensi<strong>on</strong><br />
form;<br />
• after the sugar crystallized under suspensi<strong>on</strong>, the suspensi<strong>on</strong> is pulled out of the<br />
milling machines <strong>on</strong> balls and it is let for reposing for 10 up to15 minutes;<br />
• <strong>on</strong>e replace the supernatant liquid with a new fresh film (layer) of ethyl alcohol;<br />
• the mixture of ethyl alcohol and sugar is again introduced in the milling machine<br />
<strong>on</strong> balls which is activated for another 10 minutes.<br />
409
• The sugar suspensi<strong>on</strong> thus prepared can be used after a minimum period of two<br />
days, as n the meanwhile it becomes stable. In order to be preserved for a l<strong>on</strong>g<br />
period of time, the film of supernatant alcohol is replaced <strong>on</strong>e more time with fresh<br />
ethylic alcohol.<br />
Form the point of view of shape and size, the sugar crystals of the suspensi<strong>on</strong> thus<br />
prepared are very homogeneous.<br />
Preparing the seeding substance Out of the base suspensi<strong>on</strong> <strong>on</strong>e prepare the seeding<br />
suspensi<strong>on</strong> through the diluti<strong>on</strong> of 1,200 in isopropyl alcohol. In comparis<strong>on</strong> with the<br />
method described, the current method which employs ethyl alcohol for base suspensi<strong>on</strong><br />
preparati<strong>on</strong> is easier to be executed.<br />
Establishing the number of germs introduced within a “decocti<strong>on</strong>” The crystal sugar<br />
obtaining of a certain granulati<strong>on</strong> depends <strong>on</strong> the number of germs introduced at a<br />
single seeding in the vacuum apparatus. Before reaching to this, <strong>on</strong>e establish the<br />
number of germs from the seeding suspensi<strong>on</strong> and then <strong>on</strong>e calculate the required<br />
quantity (amount) functi<strong>on</strong> of the size of the sugar crystals which is sought to be<br />
obtained. For establishing the number of germs in the sugar suspensi<strong>on</strong> there are used<br />
the following methods:<br />
- the microscopically method;<br />
- the sedimentati<strong>on</strong> method;<br />
- the sieving (sifting) method<br />
In this purpose <strong>on</strong>e can employ; the sedimentati<strong>on</strong> balance, the Andreasen dropper or<br />
the Coulter counter. Bearing in mind the number of germs which are c<strong>on</strong>tained in the<br />
seeding suspensi<strong>on</strong>, we calculate the necessary amount to effect crystallizati<strong>on</strong>. In<br />
practice, a surplus of germens is employed in experiment due to the different supersaturati<strong>on</strong><br />
in the comp<strong>on</strong>ent parts of the vacuum apparatus, especially in the case in<br />
which this is not endowed with a mechanical system for stirring up. The exceeding<br />
quantity of crystallizing germs depends <strong>on</strong> other factors such as, foe example:<br />
- the crystallizing step at which the striking (effecting) is been d<strong>on</strong>e;<br />
- the c<strong>on</strong>structive type (model) and the capacity of the vacuum apparatus;<br />
- the pressure of the heating steam.<br />
In the case of the A product the specialised literature recommends the use of a<br />
crystallizati<strong>on</strong> germs surplus of 30-50% in comparis<strong>on</strong> with the theoretical necessary.<br />
The crystallizing efficaciousness (rated capacity) from a gross mass is theoretically<br />
know as it can be calculated using the calculati<strong>on</strong> formula based <strong>on</strong> gross mass purity<br />
and of the inter-crystalline syrup and <strong>on</strong> the dry-mass c<strong>on</strong>tent of the gross mass. The<br />
calculati<strong>on</strong> formula for the crystals extracted of gross mass’ efficaciousness (rated<br />
capacity) is the following:<br />
where:<br />
SUmg is the c<strong>on</strong>tent of dry-mass of the gross-mass în ºBrix:<br />
Qmgg – purity of the gross mass in %<br />
Qsi – purity of the inter-crystalline syrup, meaning the purity of the green syrup l in %<br />
410
For the crystal mass the following formula is used:<br />
p = d 3 X 1.1 [mg],<br />
where:<br />
p=is the desired individual mass of a single sugar crystal, in mg:<br />
d – is the web dimensi<strong>on</strong> of the whole corresp<strong>on</strong>ding to the desired size of the sugar<br />
crystal, in mm;<br />
The specific mass of the crystal sugar in n<strong>on</strong>-settled way<br />
For example if we want to obtain sugar crystals with the web dimensi<strong>on</strong> of the whole<br />
sized of 0,7mm, the calculati<strong>on</strong> is d<strong>on</strong>e as following:<br />
- the desired individual mass of a single sugar crystal which will be obtained<br />
after crystallizati<strong>on</strong> is calculated using the following formula:<br />
0.7 3 X 1.1 = 0.3773 mg;<br />
a single t<strong>on</strong>e of crystal sugar obtained through crystallizati<strong>on</strong>, formed through<br />
crystallizati<strong>on</strong> and c<strong>on</strong>stituted from crystals with an individual mass of 0.3773 mg, will<br />
c<strong>on</strong>tain:<br />
1 000 000 000 : 0.3773 = 28 X 10 8 crystals<br />
This number represents also the number of crystallizati<strong>on</strong> germs of which must be,<br />
theoretically, introduced in the vacuum apparatus for crystallizati<strong>on</strong> striking (initiating)<br />
and to obtaining of the t<strong>on</strong>e of crystal sugar.<br />
If we take into c<strong>on</strong>siderati<strong>on</strong>:<br />
- the vacuum apparatus capacity 50l;<br />
- the crystal efficaciousness (output rate) 45% we can calculate:<br />
- the amount of crystal sugar which must be obtained at a single decocti<strong>on</strong><br />
namely: 50 X 0.45 = 22.5 t<strong>on</strong>es<br />
- the number of crystals c<strong>on</strong>tained by the decocti<strong>on</strong> thus obtained is:<br />
22.5 X (28 – 10 8 ) = 630 X 10 8 crystals<br />
RESULTS AND DISCUSSIONS<br />
This number, namely 630 X 10 8 represents also the number of germs which must be<br />
introduced in the vacuum apparatus at crystal initiati<strong>on</strong> (striking).<br />
If we c<strong>on</strong>sider the fact that through the previous measurements is has been established<br />
that in 100 ml of seeding suspensi<strong>on</strong> there are 325 X 10 9 crystallizati<strong>on</strong> germs, an<br />
amount of 20 ml seeding suspensi<strong>on</strong> calculated according to the following formula<br />
(630-10 8 ) : (325-10 9 ) = 20 ml will be required.<br />
Taking into account the exceeding amount of crystallizing germs, 30 – 50% sugar the<br />
real amount of seeding suspensi<strong>on</strong>, introduced in the vacuum apparatus at a single<br />
decocti<strong>on</strong> with 0.7 mm crystals, is calculated using the following formula:<br />
20 X (1.30 – 1.50) – 20 ml<br />
This is the way of seeding suspensi<strong>on</strong> preparati<strong>on</strong> and also the way to calculate the<br />
seeding suspensi<strong>on</strong> volume required to initiate (strike) the crystal sugar in the above<br />
menti<strong>on</strong>ed c<strong>on</strong>diti<strong>on</strong>s and data.<br />
411
REFERENCES<br />
1. Banu C., 1992 – „Progrese tehnice, tehnologice şi ştiinţifice în industria<br />
alimentară”, Ed. Tehnică, Bucureşti.<br />
2. Beceanu Dumitru, Chira Adrian, 2003 – „Tehnologia produselor agricole”, Ed.<br />
Ec<strong>on</strong>omică, Bucureşti.<br />
3. Sarca Gheorghe, 2004 – „Materii prime vegetale”, Ed. Universităţii din Oradea.<br />
4. <str<strong>on</strong>g>Internati<strong>on</strong>al</str<strong>on</strong>g> Sugar Journal, 2006 december.<br />
412
WATER DIFFUSION IN BREAD DURING BAKING<br />
Ruska L.*, Chereji Rodica**, Purcărea Cornelia**, Timar A.**<br />
* Bakery Bicaciu<br />
** University of Oradea, Faculty of Envir<strong>on</strong>mental Protecti<strong>on</strong><br />
ABSTRACT<br />
A study of heat and water transport in bread during baking was performed. Loaves of bread<br />
were fermented twice and baked in a c<strong>on</strong>venti<strong>on</strong>al ven at 225 0 C, with no forced c<strong>on</strong>vecti<strong>on</strong>,<br />
for 35 min. The local water c<strong>on</strong>tent and two or three different temperatures were measured<br />
inside the bread during the baking process. Bread baked from wheat flour was used to<br />
investigate the mechanisms of water transport inside a loaf during baking. The water<br />
c<strong>on</strong>tent was measured in the centre, 1 cm from the base, in the bottom crust, 1 cm under the<br />
top surface and in the top crust. The temperature was measured <strong>on</strong> the bottom surface, in<br />
the centre, and 1 mm from the optic fibre. The results indicate that up to 70±5 0 C, where a<br />
structural change is taking place, the water c<strong>on</strong>tent in the centre changes little. However,<br />
after reaching this temperature the water c<strong>on</strong>tent in the centre of the loaf rises due to<br />
vapour transport from the warmer regi<strong>on</strong>s. The water appears to be moving towards the<br />
coldest regi<strong>on</strong>, and not towards the geometrical centre.<br />
Keywords: water; diffusi<strong>on</strong>; transport; baking; bread<br />
INTRODUCTION<br />
During baking, starch is gelatinised and proteins are denaturated. The denaturati<strong>on</strong> of<br />
proteins releases water while the gelatinisati<strong>on</strong> of starch absorbs water. These two<br />
phenomena occur during the same temperature interval of 60-85 0 C, and c<strong>on</strong>tribute to the<br />
change from dough to crumb (Marst<strong>on</strong>, P. E. and Wannan, 1976). The temperature-rise at<br />
the surface and heat transport towards the centre also involves water diffusi<strong>on</strong><br />
(Thorvaldss<strong>on</strong>, K. and Skjoldebrand, 1996) and therefore a change in the water c<strong>on</strong>tent<br />
distributi<strong>on</strong>.<br />
The water c<strong>on</strong>tent affects the gelatinisati<strong>on</strong> of the starch Kokini, J. L., Lai, L. S. and<br />
Chedid, L., 1992) and the final quality of the bread. During baking, the water c<strong>on</strong>tent <strong>on</strong> the<br />
surface of the loaf becomes lower than in the centre. This, in combinati<strong>on</strong> with the high<br />
temperature, is <strong>on</strong>e of the factors that makes the crust different from the crumb. An example<br />
of a reacti<strong>on</strong> that is affected by the water c<strong>on</strong>tent and c<strong>on</strong>tributes to the crust formati<strong>on</strong> is<br />
the Maillard reacti<strong>on</strong>, which results in the browning of the crust and the development of<br />
bread aromas (Skjoldebrand C., 1986). C<strong>on</strong>versely, the starch gelatinisati<strong>on</strong> and the protein<br />
denaturati<strong>on</strong> affect the diffusi<strong>on</strong> of water by absorbing and releasing water. This makes the<br />
baking process very complex. In 1972 Auerman suspected that the water c<strong>on</strong>tent rose in the<br />
centre of a loaf during baking, due to evaporati<strong>on</strong> and c<strong>on</strong>densati<strong>on</strong>. Both Sluimer and<br />
Krist-Spit, 1978 and de Vries et al. 1998 found an increase in the water c<strong>on</strong>tent, with 3.5 g<br />
water 100 g bread in the centre of the loaf immediately after baking.<br />
413
The objective of this study was to investigate whether the transport of water towards the<br />
centre actually occurs during the baking process and to establish when the water transport<br />
starts, ends, and what directi<strong>on</strong> it takes. A sec<strong>on</strong>d objective was to investigate whether the<br />
rise in water c<strong>on</strong>tent inside the loaf is centred at the coldest regi<strong>on</strong> or whether it is<br />
geometrically centred.<br />
MATERIALS AND METHODS<br />
Bread recipe: A standard recipe for white French bread was used in this study with the<br />
following quantities and ingredients for <strong>on</strong>e loaf: wheat flour 370 g, water 200 g, salt 6 g,<br />
sugar 6 g, oil 6 g, dry yeast 4.5 g.<br />
Water c<strong>on</strong>tent and temperature measurements: During the baking processes the optic fibre<br />
was placed at five different sites in the loaf, in the centre, 1 cm from the base, in the bottom<br />
crust, 1 cm under the top surface and in the top crust. The temperatures in the oven and in<br />
the loaf were measured with copper-c<strong>on</strong>stantan thermocouples type T. The thermocouples in<br />
the loaf were placed <strong>on</strong> the bottom surface, in the centre or 1 mm from the optic fibre. In<br />
two of the loaves the temperature was measured at seven different places during the baking<br />
to locate the coldest regi<strong>on</strong> and study the temperature gradient. The thermocouples were<br />
placed, (a) <strong>on</strong> the top surface, (b) <strong>on</strong> the bottom surface, (c) <strong>on</strong> the side surface, (d) a third<br />
of the way to the centre from the bottom surface, (e) half way to the centre from the top<br />
surface, (f) in the centre, (g) two thirds of the way to the centre from the bottom surface.<br />
Oven: The study used a c<strong>on</strong>venti<strong>on</strong>al oven with a good temperature stability ±3 0 C and no<br />
forced c<strong>on</strong>vecti<strong>on</strong>.<br />
Baking procedure: The ingredients were mixed in a dough processor at 80 rpm for 2 min<br />
and at 185 rpm for 3 min. The dough was left to rise for 35 min and mixed again at 185 rpm<br />
for 2 min. It was then divided into three pieces which were rolled out into rectangular slabs.<br />
The rectangular slabs were placed <strong>on</strong> top of each other in the baking tin, with the<br />
thermocouples and the optic fibre between. The rectangular, 1.5 L baking tin had<br />
dimensi<strong>on</strong>s of 9x25x6 cm. At <strong>on</strong>e end of the tin there was an open slot, from the base to the<br />
top, where the optic fibre and the thermocouples could protrude. The dough was allowed to<br />
rise for 45 min in the tin before being placed into the oven, where it was baked for 35 min at<br />
225 0 C.<br />
Separati<strong>on</strong> of crust and crumb: In <strong>on</strong>e loaf the crust was cut off immediately after baking.<br />
The crust pieces were separated into crust in direct c<strong>on</strong>tact with the baking tin, and crust in<br />
c<strong>on</strong>tact with the air. The crust in c<strong>on</strong>tact with the baking tin was separated into side and<br />
bottom crust. The crust was dried and weighed to see how much of the dough had become<br />
crust and how much water had been present as dough. The water c<strong>on</strong>tent of the crust in<br />
c<strong>on</strong>tact with the air was compared with the total water loss of the loaf directly after baking.<br />
The water c<strong>on</strong>tent of the crust in c<strong>on</strong>tact with the baking tin was compared with the rise in<br />
measured water c<strong>on</strong>tent in the centre of the loaf.<br />
414
RESULTS<br />
Temperature distributi<strong>on</strong>: When the loaf is placed in the oven the temperature rises <strong>on</strong> the<br />
surfaces Figure 1. The temperature is approximately the same <strong>on</strong> all of the surfaces which<br />
are in c<strong>on</strong>tact with the baking tin (i.e. <strong>on</strong> the base and sides of the loaf). The temperature of<br />
the base rises slightly faster than the side temperatures due to heat c<strong>on</strong>ducti<strong>on</strong> through the<br />
base from the ceramic plate in the oven. At the top surface the temperature is slightly lower,<br />
0-10 0 C, than the base temperature, up to approximately 80 0 C. The top surface temperature is<br />
higher than the bottom temperature from 0 0 C to approximately 30 0 C, when removed from<br />
the oven. Throughout the loaf there is a temperature gradient, with the coldest regi<strong>on</strong> a little<br />
below the centre Figure 1. When the temperature reaches 100 0 C there is a plateau in the<br />
temperature rise, until all of the water has evaporated. On the surface there is no plateau<br />
whereas behind the crust, in the crumb of the loaf, the temperature remains at the plateau.<br />
Water c<strong>on</strong>tent distributi<strong>on</strong>: The water c<strong>on</strong>tent measured in the centre decreases when the<br />
loaf is placed in the oven, until the temperature reaches approximately 70"5 8C. There is<br />
then a change in the measured water c<strong>on</strong>tent, which begins to increase. Figure 2 shows a<br />
typical graph of the changes in temperature and water c<strong>on</strong>tent in the centre during the<br />
baking. At 1 cm underneath the top surface the changes in the measured water c<strong>on</strong>tent are<br />
identical, although the initial rise is approximately 3 g water 100 g bread and the decrease is<br />
approximately 13 g water 100 g bread Figure 3. At the top and absolute bottom the decrease<br />
in water c<strong>on</strong>tent commences immediately and a crust is formed.<br />
Figure 1: Changes in temperature inside the loaf during baking (a) top surface; (b)<br />
bottom surface; (c) side surface; (d) 1 cm from the bottom surface; (e) 6.8 cm from the<br />
bottom surface; (f) centre, 4.6 cm from the bottom surface; (g) 3.5 cm from the bottom<br />
surface. Total heights 9.0 cm<br />
DISCUSSION<br />
Temperature distributi<strong>on</strong>: During the fermentati<strong>on</strong> of the dough the temperature distributi<strong>on</strong><br />
in the loaf is fairly c<strong>on</strong>stant. The surface temperature of the loaf rises quickly during baking.<br />
415
The high temperature <strong>on</strong> the surface leads to heat transfer into the loaf. The water in the<br />
pores inside the loaf c<strong>on</strong>tributes to the transfer of heat through the material. This occurs due<br />
to the evaporati<strong>on</strong> of the water at the warm end of a pore and by c<strong>on</strong>densati<strong>on</strong> of the water<br />
at the cold end, known as the heat pipe effect or the Watt principle. This causes a<br />
temperature gradient inside the loaf. The partial water vapour pressure is directly related to<br />
the temperature, and a vapour pressure gradient is built up from the surface to the centre of<br />
the loaf. The results show that the temperature of the surfaces _ in direct c<strong>on</strong>tact with the<br />
baking tin Figure 1, curves b. and c is generally lower than the temperature of the top<br />
surface Figure 1, curve a . This is probably why the coldest regi<strong>on</strong> is centred slightly below<br />
the geometrical centre of the loaf Figure 1, curves e, f and g.<br />
Water c<strong>on</strong>tent distributi<strong>on</strong>: From the results it can be seen that the measured water c<strong>on</strong>tent<br />
in the centre of the loaf decreased up to a centre temperature of approximately 70±5 0 C. This<br />
is probably due to the quantity of dough in the measuring volume in fr<strong>on</strong>t of the optic fibre<br />
decreasing as the total volume of the loaf increases. Earlier measurements have shown that<br />
there is a good correlati<strong>on</strong> between the rise of the loaf and the decrease in the measured<br />
water c<strong>on</strong>tent. The true water c<strong>on</strong>tent is probably c<strong>on</strong>stant during this period as the pore<br />
system is not c<strong>on</strong>tinuous in the dough, which prevents the water from moving. When the<br />
temperature reaches approximately 70 0 C the chemical changes commence, which make the<br />
dough change into bread. The pore structure previously c<strong>on</strong>sists of discrete pores in which<br />
the water vapour is captured.<br />
Figure 2: Typical diagram of the changes in water c<strong>on</strong>tent in the centre of the loaf. The<br />
decrease in water c<strong>on</strong>tent during the first period in the oven is a measured decrease <strong>on</strong>ly<br />
and not an actual decrease. The water c<strong>on</strong>tent starts to rise in the centre, when the<br />
temperature has reached 70±5 0 C<br />
416
During the chemical changes the pore structure is opened up and becomes c<strong>on</strong>tinuous. In<br />
this new pore system the water is more free to move. As can be seen from the results, the<br />
measured water c<strong>on</strong>tent in the centre starts to increase at approximately 70±5 0 C, which<br />
c<strong>on</strong>firms the theory that the water is not free to move until the structural change occurred.<br />
The partial water vapour pressure gradient caused by the temperature gradient attempts to<br />
reduce itself.<br />
Figure 3: Typical diagram of the changes in water c<strong>on</strong>tent under the top crust of the loaf.<br />
The decrease in water c<strong>on</strong>tent during the first period in the oven is a measured decrease<br />
<strong>on</strong>ly, not an actual decrease. When the temperature has reached 70±5 0 C the water<br />
c<strong>on</strong>tent increases with approximately 3 g water/100 g bread and then slowly decreases<br />
with approximately 13 g water/100 g bread<br />
The water vapour therefore starts to move towards the centre and towards the surface. As<br />
can be seen in Figure 2 the water c<strong>on</strong>tent appears to increase in the centre when the<br />
temperature has reached 70±5 0 C, and appears to decrease 1 cm from the bottom surface. At<br />
1 cm under the top crust, Figure 3, the results show an increase in the measured water<br />
c<strong>on</strong>tent immediately after the temperature has reached 75 0 C. This is probably due to the<br />
transport of water vapour towards the colder parts being prevented by the dough. Only when<br />
the interior has changed to bread can the water move freely. Part of the rise could also be<br />
due to compressi<strong>on</strong> of the crumb near the surface, as the crust has started to form and the<br />
interior is still rising. Part of the rise could therefore be a measured rise <strong>on</strong>ly and not an<br />
actual rise. Figure 4 shows the cross secti<strong>on</strong> of a loaf with a rough divisi<strong>on</strong> into regi<strong>on</strong>s of<br />
increasing and decreasing water c<strong>on</strong>tent, at the time point when the loaf is removed from the<br />
oven. The water c<strong>on</strong>tents of the regi<strong>on</strong>s have been estimated as a mean value from the<br />
measured water c<strong>on</strong>tents. The water c<strong>on</strong>tent of the dough was 41.8 g waterr100 g dough.<br />
The crust is set at 0 g water/100 g bread, the regi<strong>on</strong> below the top crust, DV10 decreasing<br />
417
volume 10 g water/100 g bread is set to a decrease of 10 g water/100 g bread, the regi<strong>on</strong><br />
around the centre, DV2 decreasing volume 2 g water/100 g bread is set to a decrease of 2 g<br />
water /100 g bread and the centre volume, IV increasing volume, is increasing. In the crust<br />
separati<strong>on</strong> investigati<strong>on</strong>, the crust weighed 80.9 g which corresp<strong>on</strong>ds to 139 g of the dough.<br />
The remainder of the dough, 441 g, would then have become crumb. From Figure 4 the<br />
volume of the crumb regi<strong>on</strong>s has been estimated as a percentage of the total crumb volume;<br />
DV10=18.5 cm 3 /100 cm 3 , DV2=30.6 cm 3 /100 cm 3 and IVs50.9 cm 3 /100 cm 3 .<br />
Figure 4: Cross secti<strong>on</strong> of a loaf with a rough divisi<strong>on</strong> into regi<strong>on</strong>s of increasing and<br />
decreasing water c<strong>on</strong>tent, immediately after the loaf is removed from the oven. All figures<br />
are in mm. The water c<strong>on</strong>tent of the regi<strong>on</strong>s has been estimated to a mean value out of the<br />
measured water c<strong>on</strong>tents. The crust is set to 0 g water/100 g, the regi<strong>on</strong> just below the top<br />
crust, DV10, is set to a decrease of 10 g water/100 g bread, the regi<strong>on</strong> around the centre,<br />
DV2, is set to a decrease of 2 g water/100 g bread and the centre, IV, is increasing.<br />
On the surface in direct c<strong>on</strong>tact with the baking tin the water cannot diffuse into the air, so<br />
all the water that evaporates in the warm parts must move towards the colder parts in the<br />
centre of the loaf. If the water from DV2, the bottom crust and the side crust, except for the<br />
12 g of water, moved towards the centre, this would lead to a mean value of the water<br />
c<strong>on</strong>tent in IV of 51.4 g water/100 g bread.<br />
418
CONCLUSIONS<br />
The water c<strong>on</strong>tent in the centre of the loaf rises during the baking process. The water moves<br />
towards the centre by the evaporati<strong>on</strong> of water vapour near the surface, where the<br />
temperature is high, and c<strong>on</strong>densati<strong>on</strong> closer to the centre, where the temperature is lower.<br />
There is a change in the measured water c<strong>on</strong>tent when the temperature reaches 70±5 0 C. This<br />
implies that the chemical changes that cause the bread to change from dough to crumb and<br />
open up the pore structure occur just prior to this temperature interval. The transport<br />
towards the centre starts when the temperature has reached 70±5 0 C and stops when the<br />
temperature reaches 100 0 C and there is no temperature gradient left. The coldest regi<strong>on</strong><br />
occurs just below the geometrical centre and the water vapour moves towards that regi<strong>on</strong>.<br />
As can be seen in the objectives, the c<strong>on</strong>clusi<strong>on</strong>s fulfil the aims of this research work.<br />
REFERENCES<br />
MARSTON, P. E. AND WANNAN, T. L. Bread baking the transformati<strong>on</strong> from dough to<br />
bread. The Bakers Digest, 50( 4 ), 24-49 ,1976.<br />
THORVALDSSON, K. AND SKJOLDEBRAND, C. Water Transport in Meat during<br />
Reheating. Journal of Food Engineering, 29, 13-21, 1996.<br />
KOKINI, J. L., LAI, L. S. AND CHEDID, L. L. Effect of Starch Structure <strong>on</strong> Starch<br />
Rheological Properties. Food. Technology, 46 (6) , 124-139, 1992.<br />
SKJOLDEBRAND, C. Heat and Mass Transfer in Baking of Bread. SIK Service Series No.<br />
752, 1986.<br />
AUERMAN, L. J. Backprozess. In: Technologie der Brother stellung. Basel: VEB<br />
Fachbuchverlag, pp. 237-278, 1977.<br />
SLUIMER, P. AND KRIST-SPIT, C. E. Heat transport in dough during the baking of bread.<br />
In: MORTON, I. D. Ed. Cereals in a European C<strong>on</strong>text. Basel: Ellis Horwood Ltd., pp.<br />
355-363, 1987.<br />
DE VRIES, U., SLUIMER, P. AND BLOCKSMA, A. H. A quantitative model for heat<br />
transport in dough and crumb during baking. In: ASP N. G. Ed. Cereal Science and<br />
Technology in Sweden. Basel: Lund University Chemical Centre, pp. 174-188, 1988.<br />
419
BREAD PROPERTIES AND CRUMB STRUCTURE<br />
Ruska L.*, Chereji Rodica**, Purcărea Cornelia**, Timar A.**<br />
* Bakery Bicaciu<br />
** University of Oradea, Faculty of Envir<strong>on</strong>mental Protecti<strong>on</strong><br />
ABSTRACT<br />
The relati<strong>on</strong>ship between bread-crumb cellular structure and many aspects of quality in a<br />
loaf of white bread justifies investigati<strong>on</strong>s of how the structure arises during processing of<br />
the dough. Following a brief overview of the development of bread cellular structure in the<br />
dough, three parts of the literature pertaining to crumb appearance (visual texture) and<br />
bread quality are reviewed, with emphasis <strong>on</strong> the mechanical properties (physical texture)<br />
of the crumb. The importance of an objective segmentati<strong>on</strong> of the two macroscopic phases<br />
(crumb cells and cell walls solids) is emphasised in digital image analysis studies of breadcrumb<br />
structure. A review of studies where mechanical properties have been measured in<br />
fundamental units has secti<strong>on</strong>s <strong>on</strong> the mechanical properties of the composite structure and<br />
<strong>on</strong> recent analyses of the mechanical properties of the solid phase. Finally, models which<br />
have been used to relate structure to mechanical properties. Compared with the rule of<br />
mixtures, these bounds represent a good (52%) improvement in the ability to predict values<br />
for bread crumb moduli (crumb firmness).<br />
Keywords: Bread;Crumb; Mechanical properties; Structure; Processing. 1<br />
INTRODUCTION<br />
The relati<strong>on</strong>ship between crumb structure and crumb appearance may be self-evident, but<br />
this same crumb structure is also a de<strong>term</strong>inant of loaf volume (Zghal, Scanl<strong>on</strong>, &<br />
Sapirstein, 1999), the resilience of the loaf [the typical c<strong>on</strong>sumer purchasing decisi<strong>on</strong><br />
assessment test (P<strong>on</strong>te & Ovadia, 1996)], the texture during eating and even the taste<br />
(Baker, 1939). Therefore, some knowledge of the structure that defines crumb appearance<br />
will permit us to predict many of the quality attributes of bread from knowledge of the<br />
ingredients and how they are processed into the cellular structure <strong>on</strong> which bread quality is<br />
established. The review addresses three parts of the literature pertaining to crumb<br />
appearance, bread crumb physical texture and bread quality.<br />
MATERIALS AND METHODS<br />
1. Formati<strong>on</strong> of the bread crumb<br />
Bread: Bread as a solid is ‘‘soft’’ (Scanl<strong>on</strong>, Sapirstein, & Fahloul, 2000), and, like many<br />
other foodstuffs (Campbell & Mougeot, 1999), is comprised, at a macroscopic level, of two<br />
phases- a fluid (air) and a solid (cell wall material). When viewing a cross-secti<strong>on</strong> of bread<br />
crumb (Fig.1), it is apparent that the solid phase is entirely c<strong>on</strong>nected (Torquato, 2000);<br />
420
those porti<strong>on</strong>s that are not, are not bread crumb, but merely bread crumbs. In Fig. 1 it can be<br />
seen that the air cells are isolated. However, in three dimensi<strong>on</strong>s, at least a porti<strong>on</strong> (and<br />
perhaps most) of the cells are c<strong>on</strong>nected. The volume fracti<strong>on</strong> of the phases and the nature<br />
of their c<strong>on</strong>nectivity (Warren & Kraynik, 1997) de<strong>term</strong>ines the structure, and c<strong>on</strong>sequently<br />
the mechanical properties of the bread.<br />
Fig. 1. Digital image of bread crumb.<br />
Breadmaking: The mix ingredients that are used to create a basic formula dough are flour,<br />
water, leavening agent (yeast or chemicals) and sodium chloride. In order to c<strong>on</strong>vert these<br />
mix ingredients into the structure shown in Fig. 1 a number of processing operati<strong>on</strong>s are<br />
performed. The operati<strong>on</strong>s are carried out in such a way that the dough can possess the<br />
appropriate mechanical properties which will permit it to retain gas and thus produce a wellexpanded<br />
loaf of bread with an even crumb structure. Three objectives are sought in the<br />
processing operati<strong>on</strong>s :<br />
1. mixing and development of the dough (mixing and fermenting);<br />
2. formati<strong>on</strong> of a foam structure in the dough (moulding, proofing and baking);<br />
3. stabilizati<strong>on</strong> of a porous structure by altering the molecular c<strong>on</strong>figurati<strong>on</strong> of the<br />
polymeric comp<strong>on</strong>ents in the cell walls through the applicati<strong>on</strong> of heat (baking).<br />
2. Evaluating bread crumb cellular structure<br />
Crumb cellular structure (or its grain) is an important quality criteri<strong>on</strong> used in commercial<br />
baking and research laboratories to judge bread quality al<strong>on</strong>gside taste, crumb colour and<br />
crumb physical texture. Bread crumb visual texture accounts for approximately 20% of the<br />
weighting used in judging bread quality. Regardless of the weight assigned to it, crumb<br />
grain is believed to have c<strong>on</strong>siderable importance in defining bread quality since the<br />
accuracy in scoring other quality attributes in bread (e.g. loaf volume, loaf symmetry)<br />
depends <strong>on</strong> the underlying crumb grain characteristics. In bread crumb scoring, the<br />
examined parameters are crumb fineness (open versus closed cells), uniformity, cell shape,<br />
and cell wall thickness (Pyler, 1988).<br />
Image acquisiti<strong>on</strong>: In examining the visual texture of bread, particularly if <strong>on</strong>e is trying to<br />
obtain quantitative informati<strong>on</strong> <strong>on</strong> its structural organisati<strong>on</strong> to de<strong>term</strong>ine the influence of<br />
formula or processing c<strong>on</strong>diti<strong>on</strong>s, the goal is to enhance the c<strong>on</strong>trast between the two phases<br />
421
in the image. In examining the structure of other cellular products, a number of image<br />
acquisiti<strong>on</strong> methods have been used including:<br />
1. transmissi<strong>on</strong> of light through thin secti<strong>on</strong>s, where the solid phase appears at the low end<br />
of the grey level scale;<br />
2. reflecti<strong>on</strong> of light from the surface of the specimen , where the shadows cast by cut cell<br />
walls attenuate the light intensity that is reflected from the recessed cell walls, so that<br />
the solid phase appears at the high end of the grey level scale;<br />
3. x-ray tomography, where differences in absorpti<strong>on</strong> between the two phases permits<br />
n<strong>on</strong>destructive evaluati<strong>on</strong> of the internal structure.<br />
3. Mechanical properties of bread crumb<br />
3.1. Measuring crumb mechanicalp roperties<br />
Compressi<strong>on</strong>: The most comm<strong>on</strong>ly used method to measure crumb physical texture is<br />
indentati<strong>on</strong> (AACC, 1983), a compressive loading test from which ‘‘modulus’’ values can<br />
be obtained. Deformati<strong>on</strong> of a crumb specimen between parallel plates in a uniaxial<br />
compressi<strong>on</strong> test can also be used to measure the mechanical properties of bread crumb.<br />
Fig. 2 shows a typical stress–strain curve for bread crumb under compressi<strong>on</strong>, to<br />
indicate two important parameters used for characterizing the properties of the bread crumb:<br />
Young’s modulus and critical stress. Bread crumb stressed parallel to the loaf’s l<strong>on</strong>gest axis<br />
had values for Young’s modulus 2–3 times those of specimens compressed in the other two<br />
directi<strong>on</strong>s, and values for critical stress 2–2.5 times those in the other directi<strong>on</strong>s. Uniaxial<br />
compressi<strong>on</strong> of wheat starch bread (also tested fresh) gave a value of 18 kN m_2 for the<br />
compressive modulus of crumb specimens compressed parallel to the l<strong>on</strong>g axis. Fresh<br />
specimens of a soft wheat flour bread, compressed parallel to the l<strong>on</strong>g axis of the loaf,<br />
yielded rather low values for the compressive modulus at 300 N m_2 (Piazza & Masi,<br />
1995).<br />
Fig. 2. Typical stress strain curves for bread crumb under tensile and<br />
compressive loading. Young’s modulus (E), critical stress (_c), and failure stress (_f)<br />
denoted.<br />
422
Tensi<strong>on</strong>: For bread prepared under laboratory baking c<strong>on</strong>diti<strong>on</strong>s from the same wheats, but<br />
loaded in tensi<strong>on</strong> parallel to the vertical axis of the loaf, values for Young’s modulus were<br />
8.0 and 7.0 kN m_2 C<strong>on</strong>siderably larger values have been obtained when very small strain<br />
(0.02%) dynamic compressive/ tensile tests have been used to de<strong>term</strong>ine the mechanical<br />
properties of bread crumb: a value of approximately 50 kN m_2 having been obtained for<br />
the elastic part of Young’s modulus. The ability to propagate a crack in bread crumb<br />
specimens with a l<strong>on</strong>g notch means that it is possible to derive a fracture toughness value<br />
for bread crumb.<br />
3.2. Measuring mechanical properties of cell walls<br />
Although the mechanical properties of the bread are a functi<strong>on</strong> of the crumb cell wall<br />
materials and the structure created by processing, changes in bread crumb physical texture<br />
with shelf life (staling) are ascribed solely to cell wall property changes. A number of<br />
factors account for this. Of the two phases in bread crumb, <strong>on</strong>ly the physical properties of<br />
the solid phase will change. Most studies indicate that changes in the starch polymers in the<br />
crumb walls are resp<strong>on</strong>sible for the firming of bread crumb over time (Zobel & Kulp, 1996),<br />
although loss of moisture will evidently c<strong>on</strong>tribute to a change in the properties of the cell<br />
walls which would be manifest as a firming of the crumb (Piazza & Masi, 1995). Changes<br />
in bread crumb structure are not usually discussed in relati<strong>on</strong> to staling (Zobel & Kulp,<br />
1996), even though image analysis studies (Zghal et al., 1999) indicated substantial<br />
structural changes arising from crumb moisture losses, with the magnitude of these changes<br />
dependent <strong>on</strong> flour type used to make the crumb. Therefore, to eliminate structural<br />
rearrangements within the crumb from affecting studies of firming mechanisms within<br />
crumb wall comp<strong>on</strong>ents, direct measurement of the properties of the solid phase is desirable.<br />
The most comm<strong>on</strong> method of measuring the mechanical properties of the solid phase is to<br />
perform mechanical measurements <strong>on</strong> the compressed crumb (Scanl<strong>on</strong> et al., 2000).<br />
RESULTS AND DISCUSSION<br />
4. Relati<strong>on</strong>ship between structure and physical texture of cellular solids<br />
In order to predict mechanical parameters that can be used to characterize bread physical<br />
texture, physical models must be employed to relate the mechanical properties to the<br />
structure quantified by imaging techniques. The degree of sophisticati<strong>on</strong> of the model is to a<br />
certain extent de<strong>term</strong>ined by the amount and type of informati<strong>on</strong> provided by the imaging<br />
system. Therefore, a number of factors that influence the mechanical properties of cellular<br />
solids will be described al<strong>on</strong>g with descripti<strong>on</strong>s of specific models and/or experiments that<br />
have linked these structural properties to mechanical properties.<br />
4.1. Measurements of structure related to volume fracti<strong>on</strong><br />
Bulk density: Bulk density, like density, is defined as mass per unit volume (kg m_3). Bulk<br />
density has been used to describe the density of the cellular solid. However, bulk density is<br />
usually used to describe the packing density of commodities such as grains, where the space<br />
423
etween discrete particles c<strong>on</strong>tributes to a reducti<strong>on</strong> in density of the bulk collecti<strong>on</strong><br />
compared to the density of the individual grains. Given the discussi<strong>on</strong> in Secti<strong>on</strong> 2.1 <strong>on</strong> the<br />
nature of c<strong>on</strong>nectivity of bread crumb, bulk density will not be used to describe the density<br />
of bread and other cellular solids. Where the <strong>term</strong> appears to have been used to describe the<br />
density of the cellular solid, it will be discussed simply as density (_), as distinct from solid<br />
or cell wall density (_s) which is used to define the density of the materials comprising the<br />
solid phase.<br />
Density: As for other foamed polymeric materials, density (or its inverse, specific volume,<br />
m3 kg_1) has an enormous effect <strong>on</strong> the mechanical behaviour of bread crumb (Fig. 3).<br />
Because of the large c<strong>on</strong>trast in densities between the two macroscopic phases of bread<br />
crumb, a simple measurement of bread density permits the volume fracti<strong>on</strong>s to be calculated<br />
without recourse to image analysis for de<strong>term</strong>ining the distributi<strong>on</strong> of the two phases in the<br />
bread. Str<strong>on</strong>g relati<strong>on</strong>ships between density and various mechanical properties have been<br />
reported for bread and other cellular food products. For bread crumb differing in<br />
compositi<strong>on</strong> (flours of 100% rye, 50% blend of wheat and rye, and<br />
Fig. 3. Typical compressive stress strain curves for bread crumb created by proofing for<br />
various times to generate different densities<br />
100% wheat), crumb firmness linearly increased while crumb elasticity linearly decreased<br />
with increasing density.<br />
The rate of change in mechanical properties of bread crumb as a functi<strong>on</strong> of density was<br />
dependent <strong>on</strong> bread compositi<strong>on</strong>, with rye bread having the highest slope.<br />
Relative density: Because there is variati<strong>on</strong> in cell wall density with changes in formulati<strong>on</strong><br />
or processing c<strong>on</strong>diti<strong>on</strong>s, then the density of the cellular solid would be expected to change,<br />
and therefore the effect of the structure of bread <strong>on</strong> its physical texture would be partially<br />
424
obscured. It has been stated that relative density is the dominant physical character<br />
representing the three-dimensi<strong>on</strong>al structure of cellular solids, and it has been used in many<br />
studies, including those of biological materials(Warbut<strong>on</strong> et al., 1990), to quantify the<br />
dependence of mechanical properties <strong>on</strong> structure.<br />
4.2. Predicting crumb properties with little or no imaging informati<strong>on</strong><br />
Informati<strong>on</strong> <strong>on</strong> the volume fracti<strong>on</strong>s of the individual phases can be obtained from imaging<br />
data, but it is more readily gained from density measurements. The distributi<strong>on</strong> of phases<br />
within the crumb may be irregular (Fig. 1), but statistical homogeneity<br />
Fig. 4. Illustrati<strong>on</strong> of statistical homogeneity in bread crumb. Thresholded<br />
images are <strong>on</strong>e-eighth secti<strong>on</strong>s taken from different regi<strong>on</strong>s of Fig. 1. Exact<br />
structures are different, but statistical homogeneity is<br />
apparent.<br />
within the bread crumb is assumed. This refers to instances where a subregi<strong>on</strong> of crumb can<br />
be selected whose volume fracti<strong>on</strong>s and properties are essentially the same as those of the<br />
bread crumb specimen as a whole, even though the actual c<strong>on</strong>nectivities may be different.<br />
An illustrati<strong>on</strong> of statistical homogeneity in bread crumb is given in Fig. 4.<br />
CONCLUSIONS<br />
As for other composite cellular solids, density exerts a str<strong>on</strong>g influence <strong>on</strong> the mechanical<br />
properties of bread crumb. With informati<strong>on</strong> <strong>on</strong> the physical properties of the solid phase of<br />
bread crumb, it is possible from a knowledge of bread crumb density to define a range of<br />
modulus values which the bread crumb will definitely possess. Quantificati<strong>on</strong> of additi<strong>on</strong>al<br />
structural informati<strong>on</strong> by image analysis will permit improvements in predictive capacity.<br />
To take better advantage of models used for predicting the physical properties of industrial<br />
and biomedical cellular solids from their structure, precise and systematic measurements of<br />
the mechanical properties of breads created from various raw materials and processing<br />
c<strong>on</strong>diti<strong>on</strong>s is advocated.<br />
425
ACKNOWLEDGEMENTS<br />
We are grateful for research funding from The Backery Bicaciu.<br />
REFERENCES<br />
AACC. (1983). AACC method 74-09: Bread firmness by universal testing machine. In<br />
Approved methods of the American Associati<strong>on</strong> of CerealChemists Vol II (8th ed.). St.<br />
Paul, MN: American Associati<strong>on</strong> of Cereal Chemists, Inc. BAKER, J. C. (1939). The<br />
permeability of bread by air. CerealChemistry, 16, 730–733.<br />
CAMPBELL, G. M., & MOUGEOT, E. (1999). Creati<strong>on</strong> and characterizati<strong>on</strong> of aerated<br />
food products. Trends in Food Science & Technology, 10, 283–296.<br />
PIAZZA, L., & MASI, P. (1995). Moisture redistributi<strong>on</strong> throughout the bread loaf during<br />
staling and its effect <strong>on</strong> mechanical properties. CerealChemistry , 72, 320–325.<br />
PONTE, J. G. Jr., & OVADIA, D. Z. (1996). Instrumental methods. In R. E. Hebeda, & H.<br />
F. Zobel (Eds.), Baked goods freshness (pp. 151– 170). New York: Marcel Dekker, Inc.<br />
PYLER, E. J. (1988). Baking science and technology, volume II (pp. 850– 910). Merriam,<br />
KS: Sosland.<br />
SCANLON, M. G., SAPIRSTEIN, H. D., & FAHLOUL, D. (2000). Mechanical properties<br />
of bread crumb prepared from flours of different dough strength. Journalof<br />
CerealScience , 32, 235–243.<br />
TORQUATO, S. (2000). Modeling of physical properties of composite materials.<br />
<str<strong>on</strong>g>Internati<strong>on</strong>al</str<strong>on</strong>g>Journalof Solids and Structures, 37, 411–422.<br />
WARBURTON, S. C., D<strong>on</strong>ald, A. M., & Smith, A. C. (1990). The deformati<strong>on</strong> of brittle<br />
starch foams. Journalof Materials Science, 25, 4001–4007.<br />
WARREN, W. E., & KRAYNIK, A. M. (1997). Linear elastic behavior of a low-density<br />
Kelvin foam with open cells. Journalof Applied Mechanics, 64, 787–794.<br />
ZGHAL, M. C., SCANLON, M. G., & SAPIRSTEIN, H. D. (1999). Predicti<strong>on</strong> of bread<br />
crumb density by digital image analysis. Cereal Chemistry, 76, 734–742.<br />
ZOBEL, H. F., & KULP, K. (1996). The staling mechanism. In R. E. Hebeda, & H. F.<br />
Zobel (Eds.), Baked goods freshness (pp. 1–64). New York: Marcel Dekker, Inc.<br />
426
THE MATHEMATICAL MODELLING OF THE PRESSURE REGULATOR<br />
AND OF THE ELECTROMAGNETIC INJECTION AT ENGINES WITH THE<br />
REVOLUTION AND THE TEMPERATURE OF THE ENVIRONMENT<br />
SURROUNDINGS<br />
Vasile Blaga 1 , Nicolae Chioreanu 2 , Adriana Cătaş 3<br />
1 University of Oradea, Faculty of Envir<strong>on</strong>ment Protecti<strong>on</strong>,<br />
2 University of Oradea, Faculty of Envir<strong>on</strong>ment Protecti<strong>on</strong><br />
3 University of Oradea, Faculty of Science, Department of Mathematics,<br />
acatas@uoradea.ro<br />
ABSTRACT<br />
The author proposes a pers<strong>on</strong>al model for the calculati<strong>on</strong> of pressure regulator and<br />
electromagnetic injecti<strong>on</strong>, the volume of fuel injected in cycle and the durati<strong>on</strong> of the<br />
injecti<strong>on</strong> with the number of rotati<strong>on</strong> at the total full charge load. This kind of mode can<br />
be used in the modeling system of electr<strong>on</strong>ic gasoline injecti<strong>on</strong> mo<strong>on</strong>lit or multiunit. To<br />
carry out this model it is necessary modeling engines with spark lighting cycle with<br />
gasoline injecti<strong>on</strong> with a model helping cycle proposed by the author.<br />
Keywords: pressure regulator, valve, pulverizati<strong>on</strong>, durati<strong>on</strong> of injecti<strong>on</strong>.<br />
THE OPTIMUM CALCULATION OF THE PRESSURE REGULATOR<br />
Pressure regulator maintains c<strong>on</strong>stant pressure injecti<strong>on</strong> in supplying installati<strong>on</strong>. [3; 5].<br />
Calculati<strong>on</strong> scheme of the pressure regulator is presented in figure 1.<br />
Figure 1: Scheme of pressure regulator: 1-membrane; 2- springs; 3- valve; 4-<br />
c<strong>on</strong>necti<strong>on</strong> to the tank; 5- entrance c<strong>on</strong>necti<strong>on</strong>; 6- exit c<strong>on</strong>necti<strong>on</strong>; 7- housing.<br />
427
Static balance equati<strong>on</strong> of regulator membrane is given by the following relati<strong>on</strong>:<br />
F = F + F ,<br />
(1)<br />
a<br />
ga<br />
b<br />
where:<br />
2<br />
2<br />
π ⋅ Dr<br />
π ⋅ Dr<br />
Fa = Ka ⋅ f; Fga = ⋅ pga<br />
; Fb<br />
= ⋅ pb<br />
;<br />
4<br />
4<br />
and Ka is elastic c<strong>on</strong>stant springs; f- spring arrow; D-diameter of the regulator<br />
membrane; Fa-power pressure of spring; Fga –pressure power from manifold; pga –<br />
pressure from the manifold; pb –gasoline pressure in masterly of the injecti<strong>on</strong>; Fb –the<br />
gasoline injecti<strong>on</strong> power.<br />
π ⋅ D<br />
4<br />
π ⋅ D<br />
4<br />
2<br />
2<br />
r ⋅ pb<br />
= Ka⋅<br />
f +<br />
r ⋅ pga<br />
After the changing, relati<strong>on</strong> (1) become :<br />
4<br />
p b = ⋅ Ka<br />
f pga<br />
;<br />
2 ⋅ +<br />
π ⋅ Dr<br />
or:<br />
p b = K r + pga<br />
;<br />
(2)<br />
where:<br />
Kr – is the c<strong>on</strong>stant regulator.<br />
4 ⋅ Ka<br />
⋅ f<br />
= 2<br />
π ⋅ D<br />
;<br />
K<br />
Kr r<br />
r<br />
=<br />
1..<br />
4<br />
THE OPTIMUM CALCULATION OF THE ELECTROMAGNETIC INJECTION<br />
The injector proposed by the author are with electr<strong>on</strong>ic command Renix type, with<br />
c<strong>on</strong>ic top needle with four holes of pulverizati<strong>on</strong> or M<strong>on</strong>o-Motr<strong>on</strong>ic with c<strong>on</strong>ic top of<br />
needle with three holes of pulverizati<strong>on</strong>.<br />
The volume of gasoline injected in cycle is proporti<strong>on</strong>al with the injecti<strong>on</strong> pressure<br />
and the durati<strong>on</strong> of injecti<strong>on</strong>. [3;6].<br />
The secti<strong>on</strong> of the passing pulverizati<strong>on</strong> hole is de<strong>term</strong>ined by the following relati<strong>on</strong> :<br />
[6]<br />
__<br />
__<br />
d d '<br />
d 1<br />
Aa<br />
Ac<br />
Ac<br />
sa<br />
sin<br />
;<br />
2 2<br />
2 sa<br />
sin ⎟ ⎡ ⎛ ⎞⎤<br />
⎛ β ⎞⎛<br />
− ⎞<br />
= π ⋅ ⎢ + ⎜ − ⎟<br />
⎟⎥<br />
= π ⋅ ⎜ ⎟ ⎜<br />
(3)<br />
⎣ ⎝ ⎠⎦<br />
⎝ ⎠⎝<br />
⋅ β ⎠<br />
where: AI-passing secti<strong>on</strong> offered by c<strong>on</strong>ic top needle; sa-raising up needle; dv-diameter<br />
of the needle in top z<strong>on</strong>e; β-c<strong>on</strong>e angle tight; dp-sack diameter.<br />
Raising up needle sa is c<strong>on</strong>sidered to be c<strong>on</strong>stant.<br />
428<br />
,
It was marked with AI the area of the leaking secti<strong>on</strong> near the needle top of injector with<br />
c<strong>on</strong>ic top.<br />
Ai = f(sa, β, d) = ct;<br />
Ai = π(dp - 0,5 sa sinβ)sa sin β/2;<br />
sa = 0,15 mm; dp = 1…1,2 mm; β = 60 0 .<br />
Figure 2: The calculati<strong>on</strong> scheme of electromagnetic injector<br />
Discharge of gasoline which passes the injector leaking secti<strong>on</strong> is calculated by the<br />
relati<strong>on</strong>:<br />
2(<br />
pb<br />
− pga<br />
)<br />
Q = μ ⋅ A<br />
; (4)<br />
b i i<br />
ρb<br />
where μI coefficient of discharge in the secti<strong>on</strong> offered by the needle; μi = 0,8 - 0,93 ;<br />
Taking in c<strong>on</strong>siderati<strong>on</strong> relati<strong>on</strong> (2):<br />
Q<br />
2K<br />
= μ ⋅ A<br />
(5)<br />
r<br />
b i i<br />
ρb<br />
where: Ai is the leaking secti<strong>on</strong> at the injector; pb – gasoline pressure at the entrance of<br />
injector; pga – air pressure from the manifold; ρb – gasoline density; Kr - c<strong>on</strong>stant<br />
regulator pressure; Qb- discharge of the gasoline.<br />
In the other hands it is lanown from the relati<strong>on</strong> of the discharge definiti<strong>on</strong>, that is<br />
the leaking fuel volume during a time unit.<br />
We obtain<br />
Q<br />
b<br />
V<br />
=<br />
t<br />
b<br />
i<br />
mcb<br />
=<br />
ρ ⋅t<br />
b<br />
i<br />
From the equality of relati<strong>on</strong> (5) and (6) results un durati<strong>on</strong> of the injecti<strong>on</strong>, ti:<br />
mcb<br />
1<br />
ti<br />
= ⋅<br />
ρb<br />
μ ⋅ A ⋅<br />
2K<br />
r<br />
mcb<br />
=<br />
;<br />
μi<br />
⋅ Ai<br />
⋅ 2K<br />
r ⋅ ρb<br />
1<br />
d =<br />
λ ⋅ Lo<br />
m<br />
=<br />
m<br />
i<br />
i<br />
ρ<br />
b<br />
429<br />
(6)<br />
cb<br />
aer<br />
;
m<br />
cb<br />
maer<br />
maer<br />
mad<br />
m<br />
= = ⋅ = ξ ⋅d<br />
⋅mad<br />
; ξ =<br />
λ ⋅ L m λ ⋅ L<br />
m<br />
o<br />
ad<br />
o<br />
where ξ is the coefficient which represents the ratio between necessary air quantity for<br />
burning moor and the mixed quantity fuel accepted mad. Results:<br />
t<br />
ξ ⋅ d ⋅m<br />
ad<br />
i = [s], [7]<br />
μi<br />
⋅ Ai<br />
⋅ 2K<br />
r ⋅ ρb<br />
where: mcb is the volume of the fuel; maer – the volume of the air aspirated by the<br />
engine; mad – quantity of mixed fuel admitted; d – proporti<strong>on</strong>ing.<br />
In figure 3 is represented the variati<strong>on</strong> of the volume of the fuel mcb with the revoluti<strong>on</strong><br />
and the temperature of the envir<strong>on</strong>ment surroundings. In figure 4 is represented the<br />
variati<strong>on</strong> of the durati<strong>on</strong> of injecti<strong>on</strong> ti with the revoluti<strong>on</strong> and the temperature of the<br />
envir<strong>on</strong>ment surroundings.<br />
The working of the curled engine with lambda transmitter and catalyst, makes<br />
coefficient λ to be menti<strong>on</strong>ed as close as possible to λ=1 (stoechiometric dosage).<br />
On the base of a pers<strong>on</strong>al model the author realized analytic calculati<strong>on</strong> of pressure<br />
in the manifold pga and the admissi<strong>on</strong> pressure pa, the calculati<strong>on</strong> of engine pressure<br />
regulator and the durati<strong>on</strong> ti of the electromagnetic injector.<br />
The equati<strong>on</strong> of working engine by spark equipped with electr<strong>on</strong>ic engine injecti<strong>on</strong><br />
were de<strong>term</strong>ined and introduced in the calculator.<br />
For the modeling cycle engines with spark lighting with engine injecti<strong>on</strong> are noticed<br />
the initial dates, pressure expressi<strong>on</strong> from un manifold pga and the pressure of air at the<br />
end of the admissi<strong>on</strong> pa, the thermal volume charging qcb, the filling efficiency ηv, the<br />
raising ratio of the pressure in α, at volum c<strong>on</strong>stant burning, raising ratio of the after<br />
burning volume δ, the temperature of the evacuated gasoline Te, the coefficient of the<br />
residual burning gasoline γr, the temperature at the end of the admissi<strong>on</strong> Ta.<br />
All these expressi<strong>on</strong> were analytic calculated and correlated am<strong>on</strong>g them in order<br />
to be introduced in the calculator. The calculator gets the command to repeat this<br />
operati<strong>on</strong> till the getting of the imposed error of the engines with spark lighting<br />
parameters.<br />
It is calculated the mechanical theoretic work proposed Ltp, the average pressure<br />
proposed ptp, theoretical proposed efficiency and the mechanical loosing of the cycle.<br />
430<br />
aer<br />
ad<br />
;
mcb<br />
[g]<br />
mcb<br />
[g]<br />
mcb<br />
[g]<br />
0,040<br />
0,030<br />
0,020<br />
0,010<br />
0,000<br />
0,70<br />
0,800,90<br />
1,00<br />
lambda 1,10<br />
1,20<br />
500<br />
1625<br />
2750<br />
3875<br />
5000<br />
Speed [1/min]<br />
0<br />
0<br />
0<br />
0<br />
0<br />
0<br />
0<br />
0<br />
0<br />
0<br />
500 1000 1500 2000 2500 3000 3500 4000 4500 5000 5500<br />
0<br />
0<br />
0<br />
0<br />
0<br />
0<br />
0<br />
0<br />
0<br />
Speed [1/min]<br />
0,70 0,80 0,90 1,00 1,10 1,20 1,30<br />
lambda<br />
mcb<br />
0,030-0,040<br />
0,020-0,030<br />
0,010-0,020<br />
0,000-0,010<br />
mcb,<br />
lamda = 1<br />
Series1<br />
mcb<br />
[g],<br />
n = 5250<br />
Series1<br />
Figure 3: The variati<strong>on</strong> of the volume of the fuel mcb with the revoluti<strong>on</strong> and the<br />
temperature of the envir<strong>on</strong>ment surroundings<br />
431
ti<br />
[ms]<br />
ti<br />
[ms]<br />
ti<br />
[ms]<br />
6<br />
5<br />
4<br />
3<br />
2<br />
1<br />
8,00<br />
6,00<br />
4,00<br />
2,00<br />
0,00<br />
0,70<br />
0,800,90<br />
1,00<br />
lambda 1,10<br />
1,20<br />
5000<br />
500<br />
1625<br />
2750<br />
3875<br />
Speed<br />
[1/min]<br />
0<br />
500 1000 1500 2000 2500 3000 3500 4000 4500 5000 5500<br />
7<br />
6<br />
5<br />
4<br />
3<br />
2<br />
1<br />
0<br />
Speed [1/min]<br />
0,70 0,80 0,90 1,00 1,10 1,20 1,30<br />
lambda<br />
ti<br />
6,00-8,00<br />
4,00-6,00<br />
2,00-4,00<br />
0,00-2,00<br />
ti,<br />
lamda = 1<br />
Series1<br />
ti,<br />
n = 5250<br />
Series1<br />
Figure 4: The variati<strong>on</strong> of the durati<strong>on</strong> of injecti<strong>on</strong> ti with the revoluti<strong>on</strong> and the<br />
temperature of the envir<strong>on</strong>ment surroundings<br />
432
The calculati<strong>on</strong> of engines with spark lighting parameter, was secti<strong>on</strong>ed in 10<br />
proceedings and functi<strong>on</strong>s. Near the declared c<strong>on</strong>stant values at the beginning of the<br />
program, it is c<strong>on</strong>sidered as initial dates, presumed to be lanown choosing arbitrary<br />
from the statistical dates of engines with spark lighting cycle: Tao=322 K; Tzo=2530 K;<br />
Tuo =2660 K and kco =1,3; kvo =1,3; kuo =1,2; kdo=1,3; keo=1,3; without them are not<br />
possible to calculate in general way all others coefficients and the other temperature and<br />
the physical seize which characterize the cycle. In this way the seize above will play the<br />
parameter role, variable will the temperature in which thermal process evaluate. These<br />
temperatures depend <strong>on</strong> the adiabatic coefficients which in fact are stability by going<br />
though many times of the cycle until these coefficients become c<strong>on</strong>stant with 0,000009<br />
error.<br />
This exit decisi<strong>on</strong> from the cycle for a certain revoluti<strong>on</strong> is given by the diminish of the<br />
c<strong>on</strong>stant error of temperature Ta under 1,5 K.<br />
The calculati<strong>on</strong> of algorithm is made up of the following steps:<br />
1) The initial parameter relati<strong>on</strong> Tao, Tzo, Tuo şi kco, kvo, kuo, kdo, keo for <strong>on</strong>e revoluti<strong>on</strong>.<br />
2) The cycle is going through, obtaining new values for k coefficients and Ta, Tz, Tu<br />
temperatures.<br />
3) It is made differences between old and new calculated values (∆k, ∆T).<br />
4) If the differences between (∆k, ∆T) are lower than the imposed error, then the new<br />
calculated values are valid and they are going to the next revoluti<strong>on</strong> value and 1-4<br />
points are repeated.<br />
5) If the differences (∆k, ∆T) are above the imposed errors then the cycle is going<br />
through by the recalculati<strong>on</strong> of the adiabatic coefficient values and temperature,<br />
c<strong>on</strong>sidered new value as initial parameter. This re-going through is realized until<br />
the descend of the differences (∆k, ∆T) under the imposed errors.<br />
This program calculate the characteristic seize of <strong>on</strong>e thermodynamic cycle in general<br />
way <strong>on</strong>ly with the restricti<strong>on</strong> of the energetic efficiency of burning ηar and the using<br />
coefficient of warmth during the burning stage at a c<strong>on</strong>stant volume ξo, which define as<br />
a performant middle of study.<br />
CONCLUSIONS<br />
The engines with spark lighting modeling cycle with the gasoline injecti<strong>on</strong> proposed by<br />
the author c<strong>on</strong>sist of the presentati<strong>on</strong> of the initial dates of the calculati<strong>on</strong> program,<br />
calculati<strong>on</strong> and the correlati<strong>on</strong> between the engines with spark lighting parameter<br />
expressi<strong>on</strong> with the gasoline injecti<strong>on</strong> for the achievement of the program calculati<strong>on</strong>.<br />
The engines with spark lighting cycle calculati<strong>on</strong> proposed, with gasoline injecti<strong>on</strong> is an<br />
helping cycle for the simulati<strong>on</strong> <strong>on</strong> the calculator of the gasoline injecti<strong>on</strong>. The<br />
simulati<strong>on</strong> <strong>on</strong> the calculator permit the de<strong>term</strong>inati<strong>on</strong> of the theoretical ec<strong>on</strong>omicaltehnical<br />
parameters proposed; mechanical work theoretical proposed corresp<strong>on</strong>ding to<br />
the diagram, theoretical pressure proposed, theoretical efficiency, theoretical specific<br />
c<strong>on</strong>sumpti<strong>on</strong> proposed. The loosing mechanical pressure and the pumping are<br />
calculated, the theoretical ec<strong>on</strong>omical-tehnical parameters of the engine.<br />
Using the proposed model by the author the volume of the fuel is calculated which<br />
reset of the engine cycle in functi<strong>on</strong> with the revoluti<strong>on</strong> of the total load, the debit of the<br />
433
gasoline which goes through the electromagnetic secti<strong>on</strong> of the injecti<strong>on</strong>, reaching the<br />
aim in view, the de<strong>term</strong>inati<strong>on</strong> of the injecti<strong>on</strong> durati<strong>on</strong> with the revoluti<strong>on</strong> in total load.<br />
It is c<strong>on</strong>ceived the calculati<strong>on</strong> of the pressure regulator and of the electromagnetic<br />
injector. It was made the calculati<strong>on</strong> of the injecti<strong>on</strong> durati<strong>on</strong> ti with the revoluti<strong>on</strong> and<br />
load which represents the model proposed by the author.<br />
It was effectuated a engines with spark lighting program for the calculati<strong>on</strong> of<br />
parameters with the gasoline injecti<strong>on</strong> with the under- program:<br />
• the calculati<strong>on</strong> program of engines with spark lighting parameters (depending <strong>on</strong> n<br />
and λ at to=-35...+45ºC and po=1·10 2 kPa) [3].<br />
• the calculati<strong>on</strong> program of engines with spark lighting parameters (depending <strong>on</strong> n<br />
and to at λ=1 and po=1·10 2 kPa) presented in annex B;[3].<br />
REFERENCES<br />
1) Apostolescu, N. and Chiriac, R. , Process of burning in driving with inward<br />
alight.The Technical Editi<strong>on</strong>, Bucharest 1998.<br />
2) Băţagă, N. , Driving with inward alight.The Didactic and Pedagogical Editi<strong>on</strong>,<br />
Bucharest 1996.<br />
3) Blaga, V. , The c<strong>on</strong>tributi<strong>on</strong> at modeling of gasoline injecti<strong>on</strong> at M.A.S. Work of<br />
doctor`s. The Technical University of Cluj-Napoca 2000.<br />
4) Blaga, V., The dynamic of car, The University Editi<strong>on</strong> Oradea,2005.<br />
5) Blaga, V.,Engine with gasoline injecti<strong>on</strong>, The University Editi<strong>on</strong> Oradea,2005.<br />
6) Delanette, M., Les Motors a injecti<strong>on</strong>. Editi<strong>on</strong> Tehniques pour L’automobil et<br />
L’industrie, iunie, 1989.<br />
7) Negrea, V. D., Venetia Sandu The combating of medium polutti<strong>on</strong> in motor vehicle.<br />
The Technical Editi<strong>on</strong>, Bucharest, 2000<br />
8) Turcoiu, T., B<strong>on</strong>coi, J. şi Time, Al, Equipment’s from injecti<strong>on</strong> for engine with<br />
internal burning. The Technical Editi<strong>on</strong>, Bucharest 1987.<br />
434
THE VARIATION OF EFFECTIVE POWER AND SPECIFIC EFFECTIVE<br />
FUEL CONSUME OF 106-20 ENGINE WITH THE REVOLUTION AND THE<br />
TEMPERATURE OF THE ENVIRONMENT SURROUNDINGS<br />
Vasile Blaga 1 , Nicolae Chioreanu 2 , Adriana Cătaş 3<br />
1 University of Oradea, Faculty of Envir<strong>on</strong>ment Protecti<strong>on</strong>,<br />
2 University of Oradea, Faculty of Envir<strong>on</strong>ment Protecti<strong>on</strong><br />
3 University of Oradea, Faculty of Science, Department of Mathematics,<br />
acatas@uoradea.ro<br />
ABSTRACT<br />
Working system of engine is definite by rotati<strong>on</strong> and load, but besides those, it requires<br />
knowledge about thermal system, that is machine part temperature situati<strong>on</strong>, the<br />
temperature of cooling fluid, admitted air temperature, evacuati<strong>on</strong> gases temperature<br />
height correcti<strong>on</strong>s.<br />
Engine load suits effective power Pe delivered by engine to a certain rotati<strong>on</strong>. The<br />
load is regulated through speed pedal, c<strong>on</strong>trolling M<strong>on</strong>o-Motr<strong>on</strong>ic mixture quality<br />
system and air quality of other systems. Outside engine acti<strong>on</strong> applied of c<strong>on</strong>sumer<br />
(motor vehicle or brake trying <strong>on</strong> trial engine stand) represent engine attempt when<br />
thermal system of the engine doesn’t vary in time, working system is stabilized, c<strong>on</strong>trary<br />
the engine is working in provisi<strong>on</strong>al system.<br />
Keywords: gasoline, injecti<strong>on</strong>, lambda, effective power, revoluti<strong>on</strong>.<br />
THE GENERAL MODEL FUEL INJECTION MODEL<br />
Load remark may be realized through comparis<strong>on</strong> between effective power Pe and<br />
c<strong>on</strong>tinued effective power Pec developed <strong>on</strong> the some rotati<strong>on</strong>, named load coefficient<br />
χ= Pe/Pec [2];[3].<br />
Through c<strong>on</strong>tinued effective power it can be understand maximum value of power,<br />
which can be developed by an engine permanently, at a named rotati<strong>on</strong>, without<br />
modificati<strong>on</strong> of engine indexes and without appearance of some wear defecti<strong>on</strong>s. For<br />
short lapse of time (for example 15 min. for <strong>on</strong>e hour interval) the engine can develop a<br />
superior power, named in<strong>term</strong>ittent superior power Pei. Maximum c<strong>on</strong>tinuously power<br />
settled by industrial unit engineering is the nominal power Pen, and adequate/suitable<br />
rotati<strong>on</strong>, nominal rotati<strong>on</strong>.<br />
The load according power Pec (χ=1) is named complete load the <strong>on</strong>e according Pei<br />
power absolute load and according idly running Pe=0 (theoretical χ=0) null load.<br />
Between null and complete load (0
Mixture quality estimate is d<strong>on</strong>e frequently by air excess coefficient x, definite by report<br />
χ=L/Lo, where L means air quantity which is available for burning <strong>on</strong>e kilo fuel, and Lo<br />
is minimum quantity of air available to burn <strong>on</strong>e kilo fuel.<br />
For theoretic dosage χ=1, for poorest <strong>on</strong>e χ>1 and for the richest χ=1.<br />
Block scheme of general model used modulate gasoline injecti<strong>on</strong> system of M.A.S. is<br />
presented by figure 1 [3].<br />
Figure 1: Block scheme of general model used modelling gasoline injecti<strong>on</strong><br />
system of engines spark igniti<strong>on</strong><br />
It includes correlati<strong>on</strong> system of aspirated air by engine with a quantity of gasoline<br />
injected <strong>on</strong> cycle.<br />
Entering magnitudes into the system are x load, n rotati<strong>on</strong>, angular speed, positi<strong>on</strong><br />
of obdurate and the emergence <strong>on</strong>e are effective moment, effective efficiency, effective<br />
c<strong>on</strong>summati<strong>on</strong>, polluting emissi<strong>on</strong>. Engine process is influenced by thermal system of T<br />
engine.<br />
Following the study made for many different fuel injecti<strong>on</strong> systems it can be<br />
noticed that every system no matter that it is a mechanic or electric <strong>on</strong>e, they have basic<br />
scheme, based <strong>on</strong> internal burning engine working. Mechanic systems have <strong>on</strong> their<br />
principle scheme base two circuits: fuel circuit, air circuit and: suitable correcti<strong>on</strong>.<br />
Electr<strong>on</strong>ic systems includes, beside that, the electric circuit and firing circuit for<br />
Motr<strong>on</strong>ic system [4]. The main injecti<strong>on</strong> equipment load is correlating air quantity<br />
aspirated by the engine with quantity of injected fuel <strong>on</strong> the cycle, such that may result<br />
the best dosage every engine working system. That can be possible with correlati<strong>on</strong><br />
between air and fuel quantities and a series of engine working parameters (air, pressure,<br />
into the collecting input obturater positi<strong>on</strong>), engine rotati<strong>on</strong> [1]; [5].<br />
Figure 2 represents three dimensi<strong>on</strong>al variati<strong>on</strong>s effective power of 106-20 engine<br />
with Bosh M<strong>on</strong>o-Motr<strong>on</strong>ic MA 1.7 gasoline injecti<strong>on</strong> and, figure 3 represents three<br />
dimensi<strong>on</strong>al variati<strong>on</strong> of specific effective fuel c<strong>on</strong>sume of the same typo of engine, fit<br />
with the same injecti<strong>on</strong> system.<br />
436
437
Pe<br />
[kW]<br />
60<br />
50<br />
40<br />
30<br />
20<br />
10<br />
0<br />
n = 5250[rot/min]<br />
lambda = 1.0<br />
-40 -30 -20 -10 0 10 20 30 40 50<br />
to<br />
Figure 2: Variati<strong>on</strong>s effective power of 106-20 engine with the revoluti<strong>on</strong> and the<br />
temperature of the envir<strong>on</strong>ment surroundings<br />
438
ce<br />
[g/kW h]<br />
400.019<br />
350.019<br />
300.019<br />
250.019<br />
200.019<br />
150.019<br />
100.019<br />
50.019<br />
0.019<br />
319.83<br />
n = 5250[rot/min]<br />
lambda = 1.0<br />
-40 -30 -20 -10 0 10 20 30 40 50<br />
to<br />
Figure 3: Variati<strong>on</strong> of specific effective fuel c<strong>on</strong>sume of 106-20 engine with the<br />
revoluti<strong>on</strong> and the temperature of the envir<strong>on</strong>ment surroundings<br />
Quantity of air aspirated by engine can be expressed after enumerated functi<strong>on</strong>al<br />
parameters must to count <strong>on</strong> injecti<strong>on</strong> pressure, flowing secti<strong>on</strong> through injector and<br />
injecti<strong>on</strong> length.<br />
Often it is prefer that opening length of electromagnetic injector to depend <strong>on</strong><br />
depressi<strong>on</strong> from the input collector, because quantity of injecti<strong>on</strong> is correlating with<br />
quantity of air aspirated <strong>on</strong> the cycle with, as it is known from injecti<strong>on</strong> mechanic.<br />
Equipments are after depressi<strong>on</strong> from admitted collector dependence of rotati<strong>on</strong> is<br />
smaller in this case following to be d<strong>on</strong>e rotati<strong>on</strong> correcti<strong>on</strong>s for working systems witch<br />
demand such correcti<strong>on</strong>s. Flowing correcti<strong>on</strong>s of injected gasoline into cylinder are<br />
439
demanded for a successi<strong>on</strong> of transitory system of enfin working, cooling start, cooling<br />
liquid temperature, lubricati<strong>on</strong> oil temperature atmosphere pressure.<br />
Accounting all those, to realize injecti<strong>on</strong> equipment’s it must be applied the<br />
fundamental principle, graphically the gasoline from the tank and represses it to<br />
electromagnet injectors. gasoline pressure upstream injectors are c<strong>on</strong>stantly menti<strong>on</strong>ed<br />
with a pressure regulator, which allows the return to reservoir of gasoline excess<br />
suppressed by supply pump.<br />
Electromagnetic injectors associated to each engine cylinder, are opened <strong>on</strong>ce a<br />
cycle (of <strong>on</strong>e cameo shaft rotati<strong>on</strong>) by stream impulses proceeded from a computer.<br />
C<strong>on</strong>trol impulse length depends <strong>on</strong> depressi<strong>on</strong> from impute collector, engine rotati<strong>on</strong><br />
and a series of other correcti<strong>on</strong> dimensi<strong>on</strong>s [5].<br />
CONCLUSIONS<br />
Those dimensi<strong>on</strong>s are perceived with electro-mechanical translators’ help, being<br />
transmitted to the computer like electrical dimensi<strong>on</strong>s.<br />
On this basically principle can be modelled an important variety of resp<strong>on</strong>d to<br />
demanding of performance increase of engines, dominating affective c<strong>on</strong>sumpti<strong>on</strong> of<br />
fuel and polluting emissi<strong>on</strong> factors from evacuati<strong>on</strong> gases as much as necessity<br />
encumbering and adaptability.<br />
REFERENCES<br />
1. Apostolescu, N. and Chiriac, R. , Process of burning in driving with inward<br />
alight.The Technical Editi<strong>on</strong>, Bucharest 1998.<br />
2. Băţagă, N. , Driving with inward alight. The Didactic and Pedagogical Editi<strong>on</strong>,<br />
Bucharest 1996.<br />
3. Blaga, V. , The c<strong>on</strong>tributi<strong>on</strong> at modeling of gasoline injecti<strong>on</strong> at M.A.S. Work of<br />
doctor`s. The Technical University of Cluj-Napoca 2000.<br />
4. Blaga, V.,Engine with gasoline injecti<strong>on</strong>, The University Editi<strong>on</strong> of Oradea,2005.<br />
5. Delanette, M., Les Motors a injecti<strong>on</strong>. Editi<strong>on</strong> Tehniques pour L’automobil et<br />
L’industrie, iunie, 1989.<br />
6. Negrea, V. D., Venetia Sandu, The combating of medium polutti<strong>on</strong> in motor<br />
vehicle. The Technical Editi<strong>on</strong>, Bucharest, 2000.<br />
440
THE EFFECT OF THE GENOTYPE AND EXPLANT UPON THE SOMATIC<br />
EMBRYO INDUCTION IN THREE QUERCUS SPECIES<br />
AI. Timofte*, M. Palada-Nicolau**, M. Ardelean***, D. Pamfil***<br />
*University of Oradea, Romania<br />
**Forest Research and Management Institute, Research Center Simeria, Romania<br />
***University of Agriculture and Veterinary Medicine Cluj-Napoca, Romania<br />
ABSTRACT<br />
The somatic embryogenesis is an advanced method for cl<strong>on</strong>al propagati<strong>on</strong> and a useful<br />
tool for ex situ c<strong>on</strong>servati<strong>on</strong> of genetic resources.<br />
Three oak species were tested for the somatic embryogenesis ability: Quercus robur<br />
(three provenances), Q.petraea (<strong>on</strong>e provenance), Q.frainetto (two provenances).<br />
The most important factor affecting the efficiency of somatic embryogenesis was<br />
found to be the developmental stage of the zygotic embryos used as juvenile explants.<br />
The high embryogenic ability of young zygotic embryos and low embryogenic ability of<br />
maturing zygotic <strong>on</strong>es showed the str<strong>on</strong>g positive relati<strong>on</strong>ship between somatic embryo<br />
inducti<strong>on</strong> and accumulati<strong>on</strong> of reserve nutrients in cotyled<strong>on</strong>s.<br />
Two nutritive culture media and two growth horm<strong>on</strong>e combinati<strong>on</strong>s were tested.<br />
The effect nutritive media was insignificant, but the effect of horm<strong>on</strong>e combinati<strong>on</strong> was<br />
either week, or masked by the effect of explant type or that of the genotype. As for<br />
genotype, some differences were found am<strong>on</strong>g species, as well as am<strong>on</strong>g the<br />
provenances of the same species, regardless the explants type. These differences were<br />
analysed statistically and some week correlati<strong>on</strong>s were found.<br />
The subsequent phases of the somatic embryogenesis process (multiplicati<strong>on</strong>,<br />
germinati<strong>on</strong> and plant regenerati<strong>on</strong>) were not related to the initial embryo efficiency,<br />
but <strong>on</strong>ly to the quality of stabilized embryogenic lines.<br />
Key words: oak species, somatic embryogenesis, explants, genotype, statistics.<br />
INTRODUCTION<br />
The in vitro cl<strong>on</strong>al propagati<strong>on</strong> in oaks is an important step in the breeding activity<br />
using cl<strong>on</strong>al strategies, and in the same time a tool for the applicati<strong>on</strong> of somacl<strong>on</strong>al<br />
selecti<strong>on</strong> procedures in the improvement of adaptability traits.<br />
The use of somatic embryogenesis in oak cl<strong>on</strong>al micropropagati<strong>on</strong> is preferable in<br />
the applicati<strong>on</strong>s c<strong>on</strong>cerning somacl<strong>on</strong>al variati<strong>on</strong> and selecti<strong>on</strong>, because the plants<br />
regenerated from somatic embryos usually have m<strong>on</strong>ocellular origin and c<strong>on</strong>sist in<br />
cl<strong>on</strong>es of l<strong>on</strong>g-<strong>term</strong> culture, submitted of in vitro selecti<strong>on</strong> procedures.<br />
Generally, the somatic embryogenesis can be c<strong>on</strong>sidered an efficient mean of<br />
cl<strong>on</strong>al propagati<strong>on</strong>, when the following c<strong>on</strong>diti<strong>on</strong>s are accomplished:<br />
• the stable embryogenic cultures c<strong>on</strong>stantly produce a big amount of somatic<br />
embryos<br />
• the embryogenic ability of such cultures can be maintained for l<strong>on</strong>g time by serial<br />
adventitious embryogenesis<br />
• the somatic embryos can be efficiently c<strong>on</strong>verted into acclimatable plants.<br />
• Starting with the first successful embryogenic inducti<strong>on</strong>s (Chalupa, 1987, 1990,<br />
441
Gingas and Lineberger, 1989; Jorgensen, 1988, 1993), the method was<br />
c<strong>on</strong>tinuously improved. As a result of many experiments, nowadays is rather easy<br />
to obtain a big amount of somatic embryos out of juvenile explants, and also to<br />
c<strong>on</strong>serve the embryogenic ability of the selected somacl<strong>on</strong>es for l<strong>on</strong>g periods<br />
(Wilhelm et al., 1996). The embryogenesis occurs mostly direct <strong>on</strong> the explant,<br />
without a callus phase, and the stabilisati<strong>on</strong> of embryogenic cultures is based <strong>on</strong><br />
the serial adventitious embryogenesis.<br />
This advanced method of plant propagati<strong>on</strong> was also applied in the ex situ c<strong>on</strong>servati<strong>on</strong><br />
of genetic resources, using the cryopreservati<strong>on</strong> techniques (Jorgensen, 1991; Tutkova<br />
and Wilhelm, 1999).<br />
The somatic embryogenesis in oak, as an alternative to propagati<strong>on</strong> by cuttings,<br />
provides the possibility of mass producti<strong>on</strong> of cotyled<strong>on</strong>ary embryos, that can be either<br />
cryopreserved, or maintained by l<strong>on</strong>g-<strong>term</strong> culture as stable embryogenic somacl<strong>on</strong>es<br />
(Wilhelm et. al., 1996, 2000).<br />
The aim of this paper is the improvement of somatic embryogenesis technique in<br />
different oak genotypes, analysing the relati<strong>on</strong>ship am<strong>on</strong>g diverse factors affecting the<br />
embryo inducti<strong>on</strong> efficiency<br />
MATERIAL AND METHODS<br />
Plant material: As a genetical background, the plant material was represented by three<br />
oak species, in each of them <strong>on</strong>e or more provenances being investigated: Quercus<br />
robur (three provenances), Q. petraea (<strong>on</strong>e provenance) and Q. frainetto (two<br />
provenances).<br />
The immature acorns harvested at 4 different dates were dissected and used as<br />
sources of explants. The explants were represented by immature zygotic embryos in<br />
different developmental stages or fragments of more advanced embryos. The<br />
developmental stages of zygotic embryos have been defined and correlated with the<br />
morphological characteristics of acorns. (Palada-Nicolau, Hausman, 2001)<br />
Culture medium: Four media were tested for the inducti<strong>on</strong> of embryogenesis,<br />
representing combinati<strong>on</strong>s between two nutrient recipes (1/2 MS – Murashige, Skoog,<br />
1962) and DCR (Gupta, Durzan, 1985) and two combinati<strong>on</strong>s of growth regulators:<br />
QE1: 1/2MS with 10μM/l 2,4-D and 2μM/l BAP<br />
QE2: 1/2MS with 5μM/l NAA - 5μM/l BAP<br />
QE3: DCR with 10μM/l 2,4-D and 2μM/l BAP<br />
QE4: DCR with 5μM/l NAA - 5μM/l BAP<br />
After two subcultures <strong>on</strong> this medium, a horm<strong>on</strong>e-free ½ MS medium was used for the<br />
proliferati<strong>on</strong> of embryogenic structures<br />
The protocol: Sterilisati<strong>on</strong> of immature acorns by immersi<strong>on</strong> for 10 min. in soluti<strong>on</strong> of<br />
HgCl2 0,2 %, followed by abundant rinsing in sterile distilled water;<br />
a) Dissecti<strong>on</strong> of acorns in aseptic c<strong>on</strong>diti<strong>on</strong>s, isolati<strong>on</strong> of embryos and inoculati<strong>on</strong> <strong>on</strong><br />
agar solidified culture medium. Petri dishes (∅ = 10 cm) c<strong>on</strong>taining cca. 25 ml<br />
medium were used. Five explants were deposed in each Petri dish. In the case of<br />
the more advanced embryos (stages 3b and 4) around ¾ of the cotyled<strong>on</strong>s were<br />
removed, and the embryo with the basal part of cotyled<strong>on</strong>s was planted <strong>on</strong> the<br />
culture medium.<br />
442
) The cultures were incubated at cca. 22 o C in darkness. The durati<strong>on</strong> of subcultures<br />
ranged between 21 and 30 days.<br />
c) The observati<strong>on</strong>s were made at the end of first two subcultures. All reacti<strong>on</strong>s of the<br />
explant to the culture c<strong>on</strong>diti<strong>on</strong>s (embryogenesis, callogenesis, rhyzogenesis and<br />
germinati<strong>on</strong>) were recorded. If necessary, a binocular stereomicroscope was used.<br />
Structure of the experiment<br />
3 oak species: Quercus robur, Quercus petraea, Quercus frainetto<br />
6 provenances: 3 of Quercus robur, <strong>on</strong>e of Q. petraea and two of Q. frainetto.<br />
4 developmental stages of explants (4 explant types): stages 1, 2, 3 and 4<br />
4 culture media, represented by:<br />
2 variants of nutritive medium : ½ MS (Murashige-Skoog, 1962) and DCR (Gupta,<br />
Durzan, 1985)<br />
2 variants of growth regulator combinati<strong>on</strong>s, for each nutritive medium<br />
In order to establish the effect of culture medium up<strong>on</strong> the efficiency of somatic embryo<br />
inducti<strong>on</strong>, comparis<strong>on</strong>s were made between nutritive media (QE1 + QE2, versus QE3 +<br />
QE4), <strong>on</strong> <strong>on</strong>e hand, and between the two combinati<strong>on</strong>s of growth regulators (QE1 +<br />
QE3, versus QE2 + QE4), <strong>on</strong> the other hand. Three replicati<strong>on</strong>s c<strong>on</strong>sisting of five to ten<br />
explants have been counted for each parameter.<br />
Statistical analyses<br />
In order to validate the results of the oak somatic embryogenesis experiments, in which<br />
the main parameters were genotypes, explant stages and culture media, statistical<br />
analyses were made, using three ways ANOVA, lay/out with incomplete blocks, for the<br />
character „embryogenic inducti<strong>on</strong>” to the species Q. robur and Q. frainetto. The results<br />
have been presented as bilateral analysis tables, exclusively for the experimental factors<br />
and interacti<strong>on</strong>s found in ANOVA table having the test F significant.<br />
For the species Q. petraea, <strong>on</strong>ly two ways ANOVA in randomised blocks design was<br />
used, for experiments in three replicati<strong>on</strong>s.<br />
'<br />
Because some experimental data had values “0”, the transformati<strong>on</strong> x = x + 1 was<br />
used, all calculati<strong>on</strong>s being performed with transformed values (Ardelean et al., 2005).<br />
All the results were reported to the number of viable explants.<br />
The statistical validati<strong>on</strong> of the differences am<strong>on</strong>g the experimental variants was made<br />
by the multiple range test (Duncan or Turkey). These tests allowed the objective<br />
comparis<strong>on</strong> of all differences between every two variants.<br />
RESULTS AND DISCUSSION<br />
1. Somatic embryogenesis experiments<br />
The somatic embryo inducti<strong>on</strong> was mainly direct. The embryos and embryo clusters<br />
were visible <strong>on</strong> the explant surface after 30 to 45 days (Fig. 1).<br />
They were detached from the explant tissue and cultivated <strong>on</strong> media with the same<br />
compositi<strong>on</strong> to stabilisati<strong>on</strong>.<br />
The stable embryogenic cultures were cultivated <strong>on</strong> media with small<br />
c<strong>on</strong>centrati<strong>on</strong> of cytokinines or <strong>on</strong> horm<strong>on</strong>e-free medium, in order to obtain<br />
proliferati<strong>on</strong> by serial adventitious embryogenesis (Fig. 2).<br />
443
Fig. 1 – Formati<strong>on</strong> of somatic embryos and<br />
embryo clusters <strong>on</strong> the cotyled<strong>on</strong>nary explant<br />
Fig. 2 – Stable oak embryogenic<br />
culture<br />
Besides embryogenesis, other reacti<strong>on</strong>s of the explants to the culture c<strong>on</strong>diti<strong>on</strong>s were<br />
observed: callogenesis, rhyzogenesis and germinati<strong>on</strong>.<br />
The viability of explants after two subcultures ranged between 55 to 85% at the young<br />
explants (stage and 2), and 71 to 100% in the case of more developed explants (stages 3<br />
and 4). The differences between species and provenances, c<strong>on</strong>cerning the viability were<br />
due mostly to technical reas<strong>on</strong>s (distance to the field, number of days in c<strong>on</strong>servati<strong>on</strong><br />
before dissecti<strong>on</strong>, etc.).<br />
All records c<strong>on</strong>cerning the efficiency of somatic embryo inducti<strong>on</strong> and the<br />
frequency of other reacti<strong>on</strong> of the explants were reported <strong>on</strong>ly to the viable explants.<br />
It was found that the most important factor affecting the somatic embryo inducti<strong>on</strong><br />
efficiency was the developmental stage of the explant. In all oak species, the<br />
embryogenic ability decreased c<strong>on</strong>stantly from the youngest to the more advanced<br />
zygotic embryos used as explants (Fig. 3). The same trend was recognised in the case of<br />
the provenances inside the species, regardless the culture medium used for the inducti<strong>on</strong><br />
(Fig. 4 and 5).<br />
% embryogenic inducti<strong>on</strong><br />
50<br />
45<br />
40<br />
35<br />
30<br />
25<br />
20<br />
15<br />
10<br />
5<br />
0<br />
Stage 1 Stage 2 Stage 3 Stage 4<br />
explant type (developmental stage of zygotic embryo)<br />
Q. robur<br />
Q. petraea<br />
Q. frainetto<br />
Fig. 3 - The effect of explant type (developmental stage of zygotic embryo)<br />
up<strong>on</strong> the embryogenic ability, in three Quercus species<br />
444
% embryogenic inductio<br />
60<br />
50<br />
40<br />
30<br />
20<br />
10<br />
0<br />
Stage 1 Stage 2 Stage 3 Stage 4<br />
explant type (developmental stage of zygotic embryo)<br />
Târgu-Mureş<br />
Dumbrava<br />
Stefanesti<br />
mean<br />
Fig. 4 - The effect of explant type (developmental stage of zygotic embryo)<br />
up<strong>on</strong> the embryogenic ability, in three provenances of Quercus robur<br />
As for the genotype, there were differences am<strong>on</strong>g both species (Fig. 3) and<br />
provenances inside species (Fig. 4, 5, 6).<br />
% embryogenic inducti<strong>on</strong><br />
40<br />
35<br />
30<br />
25<br />
20<br />
15<br />
10<br />
5<br />
0<br />
Stage 1 Stage 2 Stage 3 Stage 4<br />
explant type (developmental stage of zygotic embryo)<br />
Griva<br />
Pad.Sarului<br />
mean<br />
Fig. 5 - The effect of explant type (developmental stage of zygotic embryo)<br />
up<strong>on</strong> the embryogenic ability, in two provenances of Quercus frainetto<br />
445
Regarding the species, the best embryogenic ability was found in Q. petraea and the<br />
worst, in Q. frainetto. The embryogenic ability of Q. robur (average, 17,8%) and Q.<br />
petraea (av. 18,5%) was similar, but that of Q. frainetto was significantly inferior<br />
(average 10,4%).<br />
C<strong>on</strong>cerning provenances, in Q. robur, the best results were obtained in the<br />
provenance Tg. Mures (av. 22,9%) and the worst, in Dumbrava Sibiului (av. 12,9).<br />
Between the Q. frainetto provenances, Griva was superior (av. 12,5%), and Padurea<br />
sarului was the worst (average 8,3%).<br />
Statistical analyses in two ways Anova system were made in order to validate the<br />
correlati<strong>on</strong>s between the effect of genotype and developmental stage.<br />
% embryogenic inducti<strong>on</strong><br />
25<br />
20<br />
15<br />
10<br />
5<br />
0<br />
genotypes<br />
average<br />
Tg.Mureş Dumbrava Sib. Stefanesti Mihaesti Griva Pad.Sarului<br />
Q. robur Q. petraea<br />
Q. frainetto<br />
genotypes<br />
Fig. 6 - The effect of the genotype (species, provenances) up<strong>on</strong> the<br />
embryogenic ability in oak<br />
The effect of culture medium proved to be totally insignificant, besides the differences<br />
observed am<strong>on</strong>g the variants cultivated <strong>on</strong> different media. The situati<strong>on</strong> was the same<br />
as well for the nutrients as for growth regulators<br />
In order to discriminate the role of each factor up<strong>on</strong> the efficiency of embryogenic<br />
inducti<strong>on</strong>, three-ways Anova statistical analyses were performed.<br />
2. STATISTICS<br />
2.1. Trifactorial experiments<br />
2.1.a. The effect of provenance, stage of explant and culture medium up<strong>on</strong> the embryo<br />
inducti<strong>on</strong> efficiency in Quercus robur<br />
The three way Anova for the effect of provenance, stage of explant and culture medium<br />
up<strong>on</strong> the embryo inducti<strong>on</strong> efficiency in Quercus robur is presented in the Table 1.<br />
446
Table 1 The three way Anova type 3x4x4 for the effect of (provenance x stage of<br />
explant x culture medium) up<strong>on</strong> the embryo inducti<strong>on</strong> efficiency in Quercus robur<br />
Source of variati<strong>on</strong> MSD DF MS F-test<br />
TOTAL 4.569 143<br />
Big plots 0.198 8<br />
Replicati<strong>on</strong>s 0.003 2<br />
Factor A (PROVENANCE) 0.164 2 0.082 10.52* > 6.94; 18.00<br />
Error (a) 0.031 4 0.008<br />
Mean plots 2.895 27<br />
Factor B (STAGE) 2.453 3 0.818 44.75** > 3.16; 5.09<br />
Interacti<strong>on</strong> A x B 0.112 6 0.019 1.02 < 2.66; 4.01<br />
Error (b) 0.329 18 0.018<br />
Small plots 1.476 108<br />
Factor C (MEDIUM) 0.037 3 0.012 0.97 < 2.74; 4.07<br />
Interacti<strong>on</strong> A x C 0.045 6 0.008 0.59 < 2.23; 3.06<br />
Interacti<strong>on</strong> B x C 0.058 9 0.006 0.50 < 2.01; 2.66<br />
Interacti<strong>on</strong> A x B x C 0.419 18 0.023 1.83* > 1.74; 2.21<br />
Error (c) 0.917 72 0.013<br />
Looking in the table, <strong>on</strong>e can notice that the calculated F is bigger than the theoretical F<br />
for the transgressi<strong>on</strong> probability of 5% and 1% <strong>on</strong>ly in the cases of provenance, explant<br />
stage and the threefold interacti<strong>on</strong> provenance x stage x culture medium, that means the<br />
variants variability is due to real causes, true differences existing am<strong>on</strong>g the<br />
experimental variants.<br />
The results showed a significant acti<strong>on</strong> of the provenance and a highly significant<br />
acti<strong>on</strong> of the stage of explant up<strong>on</strong> the embryogenesis. The threefold interacti<strong>on</strong> of<br />
above-menti<strong>on</strong>ed factors was also significant.<br />
The existence of the significant triple interacti<strong>on</strong> shoved the utility to study the averages<br />
of all combinati<strong>on</strong> of parameters (Săulescu et al.., 1967), presented in the following<br />
three tables of test Duncan.<br />
The data from the table 2 present the effects of provenance and stage of explant<br />
up<strong>on</strong> the embryogenic ability in Q. robur.<br />
Looking in the table 2, <strong>on</strong>e can notice that the provenance Dumbrava Sibiului<br />
presented a rate of embryo inducti<strong>on</strong> significantly inferior to the other two provenances<br />
that are similar and that the provenance Tg. Mures was the best. As for the explant<br />
types, the early developmental stages of the zygotic embryos 1 and 2 had a significant<br />
positive effect up<strong>on</strong> the embryo inducti<strong>on</strong> efficiency, but the more advanced stages (3<br />
and 4) had a negative effect, comparing to the average.<br />
447
Table 2 The effect of provenance and developmental stage of explant up<strong>on</strong> the rate of<br />
embryogenic inducti<strong>on</strong> in Q. robur (test Duncan)<br />
Nr.<br />
crt.<br />
Stage of explants<br />
(zygotic<br />
embryos)<br />
Provenance<br />
Embryogenic inducti<strong>on</strong> and its significance*<br />
S1 S2 S3 S4<br />
Average <strong>on</strong><br />
provenances<br />
1 Tg. Mureş 0.88a 0.66abc 0.58bc 0.47c 0.65 A<br />
2 Ştefăneşti 0.78ab 0.66abc 0.61bc 0.45c 0.63 A<br />
3 Dumbrava Sibiului 0.73abc 0.64abc 0.48c 0.42c 0.57 B<br />
Average <strong>on</strong> stages 0.80M 0.65N 0.56NP 0.44P<br />
*Differences am<strong>on</strong>g any variants followed by <strong>on</strong>e comm<strong>on</strong> letter are not significant.<br />
DS 5% for the interacti<strong>on</strong> between provenance and stage: 0,225-0,261.<br />
DS 5% for provenance: 0,050-0,051.<br />
DS 5% for the stage of explants: 0,134-0,145.<br />
We can c<strong>on</strong>clude that it is very important to harvest the acorns for somatic<br />
embryogenesis experiments in early stages, and when it is not possible, to plant <strong>on</strong><br />
culture medium a big number of explants (5 fold more than usually), in order to ensure<br />
the somatic embryo inducti<strong>on</strong>, despite of the very low efficiency.<br />
It is also useful to preview a big number of explants when new provenances are tested,<br />
because of their effect up<strong>on</strong> the embryogenesis.<br />
Table 3 The effect of provenance and culture medium up<strong>on</strong> the rate of embryogenic<br />
inducti<strong>on</strong> in Q. robur (test Duncan)<br />
Nr.<br />
crt.<br />
Provenance<br />
Culture<br />
medium<br />
Rate of embryo inducti<strong>on</strong>(%) and significance*<br />
QE1 QE2 QE3 QE4<br />
Average <strong>on</strong><br />
provenances<br />
1 Tg. Mureş 0.64ab 0.65ab 0.65ab 0.65ab 0.65 A<br />
2 Ştefăneşti 0.60ab 0.59ab 0.62ab 0.69a 0.63 A<br />
3 Dumbrava Sibiului 0.57b 0.57b 0.55b 0.58b 0.57 B<br />
Average <strong>on</strong> culture media 0,61M 0,60M 0,61M 0,64M<br />
*Differences am<strong>on</strong>g any variants followed by <strong>on</strong>e comm<strong>on</strong> letter are not significant.<br />
DS 5% for the interacti<strong>on</strong> between provenance and medium: 0,092-0,110.<br />
DS 5% for provenance: 0,050-0,051.<br />
DS 5% for culture medium: 0,184-0,200.<br />
As it can be seen in the table 3, the effect of culture medium up<strong>on</strong> the rate of embryo<br />
inducti<strong>on</strong> was insignificant, despite of some differences am<strong>on</strong>g provenances.<br />
We can c<strong>on</strong>clude that the compositi<strong>on</strong> of culture medium was not a limitative<br />
factor, media with different nutritive compositi<strong>on</strong>s and auxin/cytokinin ratios of 5/1<br />
(QE1, QE3) and 1/1 (QE2, QE4) being equally suitable for the inducti<strong>on</strong> of somatic<br />
embryogenesis.<br />
448
Table 4 The effect of developmental stage of explant and culture medium up<strong>on</strong> the<br />
rate of embryogenic inducti<strong>on</strong> in Q. robur (test Duncan)<br />
Nr.<br />
crt.<br />
Stage of explants<br />
(zygotic<br />
embryos)<br />
Culture medium<br />
Rate of embryo inducti<strong>on</strong>(%) and significance*<br />
Average<br />
S1 S2 S3 S4<br />
<strong>on</strong><br />
culture<br />
media<br />
1 QE1 0.76abc 0.65cde 0.57defg 0.45ghi 0.61 A<br />
2 QE2 0.79ab 0.67cde 0.51fghi 0.43i 0.60 A<br />
3 QE3 0.82a 0.61def 0.56efghi 0.44hi 0.61 A<br />
4 QE4 0.83a 0.69bcd 0.59def 0.46ghi 0,64 A<br />
Average <strong>on</strong> stages 0.80M 0.65N 0.56NP 0.44P<br />
*Differences am<strong>on</strong>g any variants followed by <strong>on</strong>e comm<strong>on</strong> letter are not significant.<br />
DS 5% for the interacti<strong>on</strong> between culture medium and stage: 0,106-0,129.<br />
DS 5% for culture medium: 0,184-0,200.<br />
DS 5% for the stage of explants: 0,134-0,145.<br />
In the table 4 it is shown that the best results were obtained in the stage 1, <strong>on</strong> the media<br />
QE3 and QE4, and the worst, in the stage 4 <strong>on</strong> the medium QE 2.<br />
2.1.b. The effect of provenance, stage of explant and culture medium up<strong>on</strong> the embryo<br />
inducti<strong>on</strong> efficiency in Quercus frainetto<br />
The three way Anova for the effect of provenance, stage of explant and culture medium<br />
up<strong>on</strong> the embryo inducti<strong>on</strong> efficiency in Q.s frainetto is presented in the Table 5.<br />
Table 5 The three way Anova type 3x4x4 for the effect of (provenance x stage of<br />
explant x culture medium) up<strong>on</strong> the embryo inducti<strong>on</strong> efficiency in Quercus frainetto<br />
Source of variati<strong>on</strong> MSD DF MS<br />
F-test<br />
TOTAL 2.524 95<br />
Big plots 0.081 5<br />
Replicati<strong>on</strong>s 0.010 2<br />
Factor A (PROVENANCE) 0.002 1 0.002 0,053.49; 5.95<br />
Interacti<strong>on</strong> A x B 0.045 3 0.015 1,95
The results showed a highly significant effect of the stage of explant. All the other<br />
acti<strong>on</strong>s are insignificant (table 6).<br />
Table 6 The effect of developmental stage of explant up<strong>on</strong> the rate of embryogenic<br />
inducti<strong>on</strong> in Q. frainetto (test Duncan)<br />
Stage of explants (zygotic Average <strong>on</strong> stages Significance of the<br />
embryos)<br />
differences*<br />
S1 0.746 A<br />
S2 0.548 B<br />
S3 0.488 BC<br />
S4 0.424 C<br />
*Differences am<strong>on</strong>g any variants followed by <strong>on</strong>e comm<strong>on</strong> letter are not significant.<br />
DS 5% for stage: 0,110-0,118.<br />
The high embryo inducti<strong>on</strong> rate in the stage 1 proved to be highly significant, as well as<br />
the very low embryo inducti<strong>on</strong> efficiency obtained in the stage 4.<br />
2.2 Bifactorial experiments in Quercus petraea: the effect of stage of explant and<br />
culture medium up<strong>on</strong> the embryo inducti<strong>on</strong> efficiency<br />
(4 stages x 4 culture media)<br />
All differences between the results obtained <strong>on</strong> different culture media and also the<br />
interacti<strong>on</strong>s stage x medium were insignificant, but the differences am<strong>on</strong>g stages were<br />
found to be highly significant (Table 7)<br />
Table 7 The effect of developmental stage of explant up<strong>on</strong> the rate of embryogenic<br />
inducti<strong>on</strong> in Q. petraea (test Duncan)<br />
Stage of explants (zygotic Average <strong>on</strong> stages Significance of the<br />
embryos)<br />
differences*<br />
S1 0.814 M<br />
S2 0.590 N<br />
S3 0.422 P<br />
S4 0.311 Q<br />
*Differences am<strong>on</strong>g any variants followed by <strong>on</strong>e comm<strong>on</strong> letter are not significant.<br />
DS 5% for stage: 0,086-0,093.<br />
CONCLUSION<br />
1. All oak species tested Quercus robur (three provenances), Q. petraea (<strong>on</strong>e<br />
provenance) and Q. frainetto (two provenances) and all provenances tested inside<br />
each species showed embryogenic ability: the possibility of somatic embryo<br />
inducti<strong>on</strong> out of very juvenile explants (cotyled<strong>on</strong>s of immature zygotic embryos).<br />
2. The efficiency of somatic embryo inducti<strong>on</strong> varied between the variants of the<br />
experiment from 50 % (Q. robur, provenance Tg. Mures, stage 1) to 1,7 % (Q.<br />
frainetto, stage 4), excluding the variants giving negative results, especially those<br />
with explants in the stage 4.<br />
3. The most important factor affecting the efficiency of somatic embryo inducti<strong>on</strong><br />
450
was the explant type (the developmental stage of the zygotic embryo used as<br />
explant); the early developmental stages of the zygotic embryos 1 and 2 had a<br />
significant positive effect up<strong>on</strong> the embryo inducti<strong>on</strong> efficiency, but the more<br />
advanced stages (3 and 4) had a negative effect, comparing to the average.<br />
4. We can c<strong>on</strong>clude that it is very important to harvest the acorns for somatic<br />
embryogenesis experiments in early stages, and when it is not possible, to plant <strong>on</strong><br />
culture medium a big number of explants (5 fold more than usually), in order to<br />
ensure the somatic embryo inducti<strong>on</strong>, despite of the very low efficiency.<br />
5. The sec<strong>on</strong>d factor affecting the embryogenic ability, as importance, was the<br />
genotype. The best embryogenic ability was found in Q. petraea and the worst, in<br />
Q. frainetto. The embryogenic ability of Q. robur (average, 17,8%) and Q. petraea<br />
(av. 18,5%) was similar, but that of Q. frainetto was significantly inferior (average<br />
10,4%).<br />
6. C<strong>on</strong>cerning provenances, in Q. robur, the best results were obtained in the<br />
provenance Tg. Mures (av. 22,9%) and the worst, in Dumbrava Sibiului (av. 12,9).<br />
Between the Q. frainetto provenances, Griva was superior (av. 12,5%), and<br />
Padurea sarului was the worst (average 8,3%).<br />
7. The effect of culture medium proved to be totally insignificant, besides the<br />
differences observed am<strong>on</strong>g the variants cultivated <strong>on</strong> different media. The<br />
situati<strong>on</strong> was the same as well for the nutrients as for growth regulators<br />
8. The compositi<strong>on</strong> of culture medium was not a limitative factor, media with<br />
different nutritive compositi<strong>on</strong>s and auxin/cytokinin ratios of 5/1 (QE1, QE3) and<br />
1/1 (QE2, QE4) being equally suitable for the inducti<strong>on</strong> of somatic embryogenesis.<br />
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Fagus sylavtica and somatic embryos of Aesculus hippocastanum" - In: Ahuja<br />
(Ed.), Woody Plant Biotechnology, Plenum press: 355-356.<br />
8. Jorgensen, J., 1993 - "Embryogenesis in Quercus petraea" - Ann. Sci. For. 50,<br />
Suppl.1: 344s-350s.<br />
451
9. Murashige, T., Skoog, F., 1962 - "A revised medium for rapid growth and<br />
bioassays with tobacco tissue cultures" - Physiol. Plant. 15: 473-497.<br />
10. Palada-Nicolau, M., Hausman, J-F, 2001 -Comparis<strong>on</strong> between somatic and<br />
zygotic embryo development in Quercus robur L. - Plant Biosystems, 135 (1) x-xx,<br />
2001, pg. 1-9.<br />
11. Săulescu, N. P., Săulescu, N.N., 1967 – „Câmpul de experienţă”, ed. a II-a- -<br />
Editura Agro-Silvică, Bucureşti<br />
12. Tutkova, M., Wilhelm, E., 1999 - "Cryopreservati<strong>on</strong> of Quercus robur somatic<br />
embryos" In vitro Cell. Dev. Biol. Plant, 35: 180<br />
13. Wilhelm, E., Burg, A., Berenyi, M., Endelmann, M., Rodler, R. 1996 - "Plantlet<br />
regenerati<strong>on</strong> via somatic embryogenesis and investigati<strong>on</strong>s <strong>on</strong> Agrobacterium<br />
tumefaciens mediated transformati<strong>on</strong> of oak (Quercus robur) - In: Ahuja et al.<br />
(Eds.), Somatic Cell Genetics and Molecular Genetics of Trees, Kluwer press: 125-<br />
130<br />
14. Wilhelm, E, 2000 - Somatic embryogenesis in oak (Quercus spp.) In Vitro Cellular<br />
& Developmental Biology – Plant series, Volume 36: Issue 5<br />
452
ABSTRACT<br />
COMPARATIVE TESTS FOR SEPARATION METHODS OF<br />
ORGANOCHLORINE PESTICIDES IN MILK<br />
Adriana Chiş, Vasile Bara<br />
University from Oradea, Faculty for Envir<strong>on</strong>mental Protecti<strong>on</strong><br />
Oradea, Romania<br />
In the case of food that c<strong>on</strong>tains more than 2%, OCP, is de<strong>term</strong>ined through separati<strong>on</strong>,<br />
together with fat. This is why the comparis<strong>on</strong> of various methods presented by the<br />
specialised literature through which it can be separated from milk fat, is very useful.<br />
The validity of the methods has been tested through the comparis<strong>on</strong> of the de<strong>term</strong>ined<br />
values with the <strong>on</strong>es de<strong>term</strong>ined through the reference method indicated for the<br />
de<strong>term</strong>inati<strong>on</strong> of milky fat. The applicati<strong>on</strong> of various methods shows that there are not<br />
major quantitative differences; however, some of them are closet to the reference<br />
values. This thing is good to know in order to be able to apply in safety c<strong>on</strong>diti<strong>on</strong>s, the<br />
fastest method, the less expensive or, simply, the most accessible in the laboratory that<br />
performs the de<strong>term</strong>inati<strong>on</strong>.<br />
Keywords: organochlorine pesticides (OCP), fat, milk<br />
1. INTRODUCTION<br />
Fat separati<strong>on</strong> from milk is the first phase in OCP de<strong>term</strong>inati<strong>on</strong>, from milk. Despite the<br />
fact that OCP usage has been limited due to the toxic effects and bioaccumulati<strong>on</strong>,<br />
research <strong>on</strong> this kind of food c<strong>on</strong>taminant is necessary because they occur under<br />
sanitary - veterinary surveillance (Savu C-tin and Georgescu Narcisa, 2004). The<br />
m<strong>on</strong>itoring of food chemical c<strong>on</strong>taminati<strong>on</strong>, is an essential compound of food safety<br />
and, according to Codex Alimentarius, it supplies valuable informati<strong>on</strong> <strong>on</strong> the identity<br />
of chemical c<strong>on</strong>taminants for the selecti<strong>on</strong> of priorities in what regards their follow-up<br />
and limits. The program GEMS/FOOD (Global Envir<strong>on</strong>ment M<strong>on</strong>itoring System/Food<br />
C<strong>on</strong>taminati<strong>on</strong> M<strong>on</strong>itoring and Assessment Program) of evaluati<strong>on</strong> of the chemical<br />
c<strong>on</strong>taminants level in foods, is essential to reach this purpose. According to this<br />
program we can observe that the de<strong>term</strong>inati<strong>on</strong>s do not refer <strong>on</strong>ly to the primary foods<br />
such as milk and diary but also to the <strong>on</strong>es al<strong>on</strong>g the food chain, that can be resp<strong>on</strong>sible<br />
of c<strong>on</strong>taminati<strong>on</strong>. In this regard we can state exemplify the cereals. The organochlorine<br />
compounds are followed to the total diets, as shown in Table 1<br />
Table 1: Chemical c<strong>on</strong>taminants under sanitary – veterinary surveillance –<br />
extract (after Hura Carmen, 2005 )<br />
C<strong>on</strong>taminants Food<br />
Chlorine compounds: DDT (metabolites),<br />
HCH (isomers); polychlorine bi-phenyls,<br />
dioxins, etc.<br />
453<br />
Milk, powder milk, butter, eggs, animal<br />
and vegetal fat, fish, cereals, oil, mother<br />
milk, total diets (menu)
2. MATERIALS AND METHODS<br />
OCP are liposoluble substances, which leads to their accumulati<strong>on</strong> in fats. For this<br />
reas<strong>on</strong>, in the qualitative and quantitative de<strong>term</strong>inati<strong>on</strong> of this kind of toxic the first<br />
phase is the separati<strong>on</strong> of fat from the matrix elements, in this case, cow milk. (Hura<br />
Carmen, 2006) However, for this kind of food product the OCP-kind residues c<strong>on</strong>tent,<br />
is expresses in mg/kg fat. So in order to be sure of an accurate extracti<strong>on</strong> of the toxic for<br />
the purpose of its quantitative de<strong>term</strong>inati<strong>on</strong> is useful to know the efficiency of fat<br />
separati<strong>on</strong> methods. This is the preliminary phase of quantitative de<strong>term</strong>inati<strong>on</strong> of<br />
pesticides. (Guidelines <strong>on</strong> Good Laboratory Practice in Pesticide residue Analysis,<br />
1993)<br />
2.1 De<strong>term</strong>inati<strong>on</strong> of fat c<strong>on</strong>tent<br />
2.2.1– Reference method<br />
Fat c<strong>on</strong>tent of tested probes has been de<strong>term</strong>ined using as reference method, the etherchlorhydric<br />
method (Weinbull method), applied to 100g of test, working with two types<br />
of whole milk. We have worked with a Soxhlet extractor with four posts. After the<br />
mineralizati<strong>on</strong>, the fat was extracted in p.a. oil ether, which has been divided in sand<br />
bath, under the niche. For each test there were three de<strong>term</strong>inati<strong>on</strong>s plus the witness test.<br />
2.2.2 Applicati<strong>on</strong> of alternative methods of milk fat separati<strong>on</strong><br />
For the extracti<strong>on</strong> of fat from milk, we used the following methods:<br />
- Extracti<strong>on</strong> with AOAC method<br />
- Column extracti<strong>on</strong><br />
- Partiti<strong>on</strong> extracti<strong>on</strong><br />
Extracti<strong>on</strong> through the AOAC method modified:<br />
(Associati<strong>on</strong> of Analytical Communities)<br />
The method has been modified (Sprecht, W. in 1987 and Cuniff, P. in 1997) in the<br />
proporti<strong>on</strong>al diminuti<strong>on</strong> (1:10) of all reactive quantities used, due to the volume of the<br />
test tubes for centrifugati<strong>on</strong> at our disposal. So, in a 50 ml centrifugati<strong>on</strong> test tube, put<br />
10 lm ethanol, 0.1 g potassium oxalate and 10 ml milk. Mix it very well. Add 5 ml<br />
ethylic ether and agitate it thoroughly for 1 minute, then we add 5 ml oil ether and<br />
agitate it thoroughly for 1 minute. Afterwards, centrifuge it for 5 minutes at 1500<br />
rot/min; afterwards transfer the solvent layer in a 100 ml separati<strong>on</strong> funnel with 50 ml<br />
water and 3 ml soluti<strong>on</strong> of saturated sodium chlorine. Extract the residue twice agitating<br />
thoroughly with 5 ml ethylic ether: oil ether 1 : 1 (v/v) Centrifuge it and transfer the<br />
solvent layer in the separati<strong>on</strong> funnel after each extracti<strong>on</strong>. Carefully mix combined<br />
extracts with water. Drain and remove water layer. Wash the solvent layer, twice, with<br />
two porti<strong>on</strong>s of water, of 100 ml each, remove water each time. For dehydrati<strong>on</strong>, pass<br />
the solvent soluti<strong>on</strong> through a column of anhydrous sodium sulphate of 50 mm and<br />
exterior diameter of 25 mm; collect the eluate in a Berzelius glass. Wash the column<br />
with small porti<strong>on</strong>s of ether (three times, 5 ml each time). Gather all etheric extracts in a<br />
weighted vase and evaporate the solvent from the combined extracts at steam bath<br />
temperature under a current of air, to obtain the fat.<br />
454
Column extracti<strong>on</strong> (Beck, H. and Mathar W. 1985)<br />
Mix 10 ml of liquid milk sample in a mortar with see sand and sodium sulphate (1 + 1<br />
mix) to obtain a dry friable product. Transfer the mix in the extracti<strong>on</strong> glass-tube<br />
(interior diameter 12 mm and 300 mm length). Before this, introduce in the tube a tap of<br />
“glass cott<strong>on</strong>” and a layer of 2 cm of sodium sulphate, for dehydrati<strong>on</strong>. Eluate the<br />
column with a mix 2:1 (v/v) n-hexane and acet<strong>on</strong>e. Collect the eluate and evaporate it in<br />
a spinning evaporator at approximately 50 ºC under reduced pressure, using ballo<strong>on</strong>s of<br />
c<strong>on</strong>stant weight to de<strong>term</strong>ine fat.<br />
Partiti<strong>on</strong> extracti<strong>on</strong> (SR EN 1528-2, 2004)<br />
In a 1000 ml glass, add 100 g milk, 500 ml 2:1 (V/V) n-hexane and acet<strong>on</strong>e mix and<br />
homogenize for 4 minutes. Allow phases to separate. Decant the superior organic layer<br />
in a separati<strong>on</strong> funnel which c<strong>on</strong>tains 500 ml sodium sulphate soluti<strong>on</strong>. Add in the glass<br />
50 ml 2:1 n-hexane-acet<strong>on</strong>e mix and decant in the separati<strong>on</strong> funnel to ensure the<br />
qualitative transfer of the organic phase. Agitate the separati<strong>on</strong> funnel for 30 sec<strong>on</strong>ds.<br />
Allow phases to separate and remove inferior, watery phase.<br />
Agitate the organic layer with another 500 ml sodium sulphate soluti<strong>on</strong>. Remove<br />
inferior layer as before but keep about 2 ml in the funnel. Spin the separati<strong>on</strong> funnel<br />
around its axis to remove all the water from the vase’s walls. When all water settled,<br />
drain the remaining watery phase and remove it. Put approximately 20 g sodium<br />
sulphate with frit glass and pass the organic phase through the sodium sulphate in a<br />
round bottom ballo<strong>on</strong> which has been weighted first. Evaporate the soluti<strong>on</strong> in a<br />
spinning rotator at approximately 50 ºC under reduced pressure.<br />
RESULTS AND DISCUSSIONS<br />
For each separati<strong>on</strong> method, the final phase is the <strong>on</strong>e when the solvent is removed<br />
through evaporati<strong>on</strong> and the fat is de<strong>term</strong>ined gravimetrically.<br />
MG = (M2 –M1) / M1<br />
% GR = MG /M X 100<br />
MG - FAT MASS<br />
% GR – FAT PERCENTAGE<br />
M – MASS OF THE WORKING TEST<br />
M1 – mass of the recipient where extracts are collected, empty<br />
M2 – mass of the recipient where the extracts are collected, after removing the solvent<br />
Note : for the column extracti<strong>on</strong> method we de<strong>term</strong>ined the density of the test because<br />
the methods implies the introducti<strong>on</strong> of a certain volume, while the calculus refer to the<br />
masses.<br />
The results obtained are presented in the tables 2 and 3 for the two types of test in work.<br />
455
Table 2: De<strong>term</strong>inati<strong>on</strong> results for test L1<br />
Fat c<strong>on</strong>tent %<br />
Methods<br />
Reference<br />
method<br />
AOAC<br />
Column<br />
extracti<strong>on</strong><br />
Partiti<strong>on</strong><br />
extracti<strong>on</strong><br />
De<strong>term</strong>inati<strong>on</strong> 1 4.02 3.87 3.99 4.04<br />
De<strong>term</strong>inati<strong>on</strong> 2 4.22 4.08 4.15 4.28<br />
De<strong>term</strong>inati<strong>on</strong> 3 4.15 3.93 4.12 4.33<br />
Nr 3 3 3 3<br />
Average 4.130 3.960 4.087 4.217<br />
Standard deviati<strong>on</strong> 0.101 0.108 0.085 0.155<br />
t 1.9913 0.5642 -0.8155<br />
Table 3: De<strong>term</strong>inati<strong>on</strong> results for test L2<br />
Fat c<strong>on</strong>tent %<br />
Methods<br />
Reference<br />
method<br />
AOAC<br />
Column<br />
extracti<strong>on</strong><br />
Partiti<strong>on</strong><br />
extracti<strong>on</strong><br />
De<strong>term</strong>inati<strong>on</strong> 1 3.9 3.73 3.81 4.02<br />
De<strong>term</strong>inati<strong>on</strong> 2 3.85 3.62 3.98 4.09<br />
De<strong>term</strong>inati<strong>on</strong> 3 4.07 3.89 4.12 4.15<br />
Nr 3 3 3 3<br />
Average 3.940 3.747 3.970 4.087<br />
Standard deviati<strong>on</strong> 0.115 0.136 0.155 0.065<br />
t 1.8414 -0.2692 -1.9274<br />
The statistic analysis was performed through the t – student test (Ardelean M. 2005) and<br />
it shows the fact that the differences are not significant between the reference method<br />
and the three applied methods, which are used for pesticide separati<strong>on</strong> with fat in milk<br />
tests that are supposed to be c<strong>on</strong>taminated.<br />
However the AOAC method values are the most far away from the reference <strong>on</strong>e<br />
and the “t” value for 4 degrees of freedom are very close to the value that make the<br />
distincti<strong>on</strong> from un-significant values to significant values. But, since we worked at this<br />
case <strong>on</strong> a reduced scale, as shown from the descripti<strong>on</strong> of the work methods, it is<br />
possible that the deviati<strong>on</strong> is influenced by this aspect.<br />
However, the extracti<strong>on</strong> method and the partiti<strong>on</strong> method have results very close to<br />
<strong>on</strong>e another in respect to the reference method and can be used for the fat separati<strong>on</strong> in<br />
the case of pesticides de<strong>term</strong>inati<strong>on</strong> from milk tests, which can be seen in the graphics<br />
from figure 1 and 2. Later, the separated fracti<strong>on</strong> is submitted to qualitative and<br />
quantitative purificati<strong>on</strong> and de<strong>term</strong>inati<strong>on</strong> through instrumental methods.<br />
456
Fat Percentage<br />
Fat percentage<br />
4.4<br />
4.3<br />
4.2<br />
4.1<br />
4<br />
3.9<br />
3.8<br />
3.7<br />
3.6<br />
3.5<br />
4.2<br />
4.1<br />
4<br />
3.9<br />
3.8<br />
3.7<br />
3.6<br />
3.5<br />
4.22<br />
4.15<br />
4.02<br />
4.08<br />
3.93<br />
3.87<br />
4.15<br />
4.12<br />
3.99<br />
1 2 3 4<br />
Ref AOAC Extr Part<br />
Figure 1: Variati<strong>on</strong> %Gr for L1<br />
4.07<br />
3.9<br />
3.85<br />
3.89<br />
3.73<br />
3.62<br />
4.12<br />
3.98<br />
3.81<br />
1 2 3 4<br />
Ref AOAC Extr Part<br />
Figure 2: Variati<strong>on</strong> %Gr for L2<br />
457<br />
4.33<br />
4.28<br />
4.04<br />
4.15<br />
4.09<br />
4.02<br />
Series1<br />
Series2<br />
Series3<br />
Series1<br />
Series2<br />
Series3
6. REFERENCES<br />
1. Beck H, Mathar W, 1985, Bundesgessundheitsdl. 28, pag 1 -12<br />
2. Cunniff, P. (Ed): Official Methods of Analysis of the AOAC INTERNATIONAL<br />
[Metode Oficiale de Analiză AOAC Internaţi<strong>on</strong>al], 16 !h editi<strong>on</strong>, Arlingt<strong>on</strong> VA USA<br />
1995, Voi 1, Chapter 10 pp 1-10, Method No 970.52<br />
3. Guidelines <strong>on</strong> Good Laboratory Practice in Pesticide residue Analysis [Ghid de<br />
Bună Practică de Laborator la analiza reziduurilor de pesticide], Codex alimentarius<br />
commissi<strong>on</strong>. In Codex Alimentarius Volume Two, Pesticide residues in food -<br />
Rome; Food and Agriculture Organizati<strong>on</strong> of the United Nati<strong>on</strong>s (FAO); World<br />
Healh Organizati<strong>on</strong> (WHO) 1993 Part 4.3, pp 417-455<br />
4. Hura Carmen, 2005, Chemical c<strong>on</strong>taminati<strong>on</strong> of food in Romania (in orig.<br />
C<strong>on</strong>taminarea chimică a alimentelor în România) 2005, Editura Cermi, Iaşi<br />
5. Hura Carmen, 2006, Lab guide – Analysis methods for food products (in orig. Ghid<br />
de laborator – Metode de analiză pentru produse alimentare), Editura Cermi, Iaşi<br />
6. Savu C-tin, Georgescu Narcisa, 2004, Food safety (in orig. Siguranţa alimentelor),<br />
Editura Semne, Bucureşti<br />
7. Specht, W.1987,:Organochlorine and organophosphorus pesticides [Pesticide<br />
organoclorurate şi organofosforice]. In: Deutsche Forschungsgemeinschaft, Manual<br />
of Pesticide Residue Analysis, VCH Verlagsgesellschaft Weinheim, Voi. 1, pp 309-<br />
319, Method S 10.<br />
8. *** SR EN 1528-2, octomber 2004, Fat food products. De<strong>term</strong>inati<strong>on</strong> of pestidices<br />
and polychlorbiphenyls (in orig. Produse alimentare grase. De<strong>term</strong>inarea<br />
pesticidelor şi policlorbifenililor (PCB) ), Part 2 : Extracti<strong>on</strong> of fat and PCB and<br />
de<strong>term</strong>inati<strong>on</strong> of fat c<strong>on</strong>tent (in orig. Extracţia grăsimii, pesticidelor şi PCB-urilor<br />
şi de<strong>term</strong>inarea c<strong>on</strong>ţinutului în grăsime)<br />
458
HIGHLIGHTING THE STATE OF FRESHNESS OF THE FRESH RAW<br />
MATERIAL FISH ON THE BASIS OF THE PHYSICAL-CHEMICAL<br />
PROPERTIES<br />
Purgea Ram<strong>on</strong>a<br />
University of Oradea,Faculty for Envir<strong>on</strong>mental Protecti<strong>on</strong><br />
ABSTRACT<br />
The assessment of the state of freshness of the raw material fish may be achieved beside<br />
the sensorial analyses (smell, colour, c<strong>on</strong>sistency, etc.) also based <strong>on</strong> the physicalchemical<br />
<strong>on</strong>es: the biological particularities of the fish, the structure and chemical<br />
compositi<strong>on</strong> of the tissues, the technology of capturing and life suppressi<strong>on</strong> impart<br />
certain characteristics to the biochemical processes in the fish flesh (Eftimie V.M.,<br />
2001).<br />
Keywords: fish, easily hydrolysable nitrogen, amm<strong>on</strong>iac, H2S and pH.<br />
1./ INTRODUCTION<br />
After the death of the fish, the biochemical processes from the fish flesh are carried out<br />
with an increased dynamics that leads to the faster installati<strong>on</strong> of the specific<br />
modificati<strong>on</strong>s. The fish flesh is rich in protein and n<strong>on</strong>-protein nitrogen and poor in<br />
carb<strong>on</strong> hydrates (Ward, D.R. et al., 1991, C<strong>on</strong>stantiniu S., 1999).<br />
2./ MATERIALS AND METHODS<br />
178 samples of whole fresh fish were drawn, from the two units, A and B, seeking the<br />
verificati<strong>on</strong> of 16 batches (8 batches from unit A and 8 batches from unit B). The draw<br />
of samples was performed after classifying the fish raw material in quality classes<br />
(based <strong>on</strong> species, size, corporal weight, state of freshness, etc.) from the batches of<br />
indigenous fish, which were verified from sanitary-veterinary point of view at the<br />
recepti<strong>on</strong> in the two fish processing units.<br />
91 samples from 8 batches from unit A and 87 samples from 8 batches of unit B<br />
were drawn. After collecti<strong>on</strong>, the fresh fish samples were introduced in sterile packages<br />
(polyethylene <strong>on</strong>e time use bags), in order to prevent the subsequent c<strong>on</strong>taminati<strong>on</strong> and<br />
were sent to the laboratory with a view to performing the physical-chemical<br />
examinati<strong>on</strong> for de<strong>term</strong>ining: the easily hydrolysable nitrogen, the amm<strong>on</strong>iac in freestate,<br />
the sulphurated hydrogen and the pH. The whole fresh raw material fish must<br />
corresp<strong>on</strong>d from physical-biochemical point of view to the c<strong>on</strong>diti<strong>on</strong>s of quality<br />
foreseen in STAS 5386-86 (Zaika,L.L. et al., 2000).<br />
According to STAS 5386-86 (Romanian standardisati<strong>on</strong>) , the c<strong>on</strong>diti<strong>on</strong>s of<br />
admissibility for the fresh fish are:<br />
• easily hydrolysable nitrogen, mg NH3/100 g = 30<br />
• amm<strong>on</strong>iac in free-state (Nessler reacti<strong>on</strong>) = absent<br />
• H2S = absent<br />
• pH = maximum 6,2.<br />
459
3./ RESULTS AND DISCUSSIONS<br />
From the 16 analyzed batches (8 in unit A and 8 in unit B), in 8 of them (3 batches from<br />
unit A and 5 batches from unit B) <strong>on</strong>e highlighted exceedings of the maximum admitted<br />
limits of the physical-chemical parameters, according to the stipulati<strong>on</strong>s of STAS<br />
5386/86.<br />
From the 178 draws performed in the 2 units, a number of 37 of them resp<strong>on</strong>ded<br />
positively as a result of the physical-chemical examinati<strong>on</strong> to the following parameters:<br />
the easily hydrolysable nitrogen, the amm<strong>on</strong>iac in free-state, the sulphurated hydrogen<br />
and the pH.<br />
In table 1 the results obtained as a result of performing the physical-chemical<br />
examinati<strong>on</strong> of the samples of whole fresh raw material fish.<br />
As a result of the performed physical-chemical examinati<strong>on</strong>s <strong>on</strong>e obtained 37<br />
positive samples from the 178 analyzed (20.78% in the two units with a c<strong>on</strong>taminati<strong>on</strong><br />
rate of 17.58% in 16/91 samples) in unit A and 24, 13% (21/87 samples in unit B (Table<br />
1).<br />
Predominant were exceedings of the maximum limits for the easily hydrolysable<br />
nitrogen, which were recorded in 12 of the 178 analyzed samples (6, 74%) in the two<br />
units, followed by values above the allowed limits of the pH in 9 of the analyzed<br />
samples (5, 05%) and positive results of the Nessler reacti<strong>on</strong> in 10 samples (5, 61%) and<br />
of the reacti<strong>on</strong> for highlighting H2S in 5 samples (2,8%).<br />
In 6 of the 16 batches <strong>on</strong>e did not record exceedings of the maximum allowed<br />
limits for the physical-chemical parameters, according to the stipulati<strong>on</strong>s of STAS<br />
5386/86 (batches 1A, 4A, 6A, 7A, 4B and 6 B).<br />
The inadequate results of the physical-chemical parameters of freshness obtained<br />
as a result of performing the laboratory examinati<strong>on</strong>s corroborate with the results of the<br />
organoleptic examinati<strong>on</strong> of the batches of fresh raw material fish.<br />
Table 1: The results of the physical-chemical examinati<strong>on</strong> of the batches of fresh raw<br />
material fish<br />
Crt. Batch 2A Batch 3A Batch 5A Batch 1B Batch 2B Batch 5B<br />
No. NH3 pH NH3 pH NH3 pH NH3 pH NH3 pH NH3 pH<br />
1. 39 7,1 41,9 7,3 51,9 7,6 52,3 7,7 52,0 7,6 51,4 7,5<br />
(the values above the maximum allowed limits of the NH3 and pH parameters)<br />
4./ CONCLUSIONS<br />
As a result of the physical-chemical de<strong>term</strong>inati<strong>on</strong>s performed for the samples of fresh<br />
raw material fish <strong>on</strong>e recorded exceedings of the maximum allowed limits for the easily<br />
hydrolysable nitrogen, pH, positive results of the Nessler reacti<strong>on</strong> and of the reacti<strong>on</strong> for<br />
highlighting H2S (sulphurated hydrogen).<br />
460
REFERENCES<br />
1. Eftimie V.M. Particularitatile morfo-structurale, c<strong>on</strong>sideratii biochimice,<br />
microbiologice si parazitologice ale pestelui materie prima, destinat procesarii.<br />
Referat 2 in cadrul tezei de doctorat, Facultatea de Medicina Veterinara Bucuresti,<br />
iunie 2001.<br />
2. Zaika,L.L., J.G.Phillips,J.S.Fanellli and O.J.Scullen. 2000. Revised model for<br />
aerobic growth of Shigella flexneri to extend the validity of predicti<strong>on</strong> at<br />
temperatures between 10 and 19ºC.Intl. J. Food Microbiol. 41; 9-19.<br />
3. Ward, D.R. and C.R. Hackney (eds). 1991. Microbiology of Marine Food<br />
Products. Van. Nostrand Reinhold, New York.<br />
4. C<strong>on</strong>stantiniu S.-Yersinii. Biologie si diagnostic de laborator, Ed. Cors<strong>on</strong>, Iasi, 1999<br />
461
ASSESSMENT OF THE HYGIENIC QUALITY OF THE FISH FRESH RAW<br />
MATERIAL BASED ON THE BACTERIOLOGICAL EXAMINATIONS<br />
Purgea Ram<strong>on</strong>a<br />
University of Oradea, Faculty for Envir<strong>on</strong>mental Protecti<strong>on</strong><br />
ABSTRACT<br />
Immediately after catching it, the flesh and some internal organs of fish are normally<br />
sterile. There are however, numerous microorganisms, especially bacteria, <strong>on</strong> the skin<br />
(103-109/g), <strong>on</strong> the gills(103-107/g) and in the digestive tube(103-108/g). The number<br />
of bacteria from different parts of the organism vary very much and reflect, mostly, the<br />
degree of c<strong>on</strong>taminati<strong>on</strong> (Barzoi D.,Apostu S., 2002, Yasumoto, T. et al., 1993).<br />
Keywords: fish, microorganisms: sporulated gram-positive bacteria and gram-negative<br />
bacteria<br />
1./ INTRODUCTION:<br />
The pathogenic germs present in fish may multiply, reaching c<strong>on</strong>taminati<strong>on</strong> levels<br />
capable of de<strong>term</strong>ining the occurrence of illnesses in man.<br />
2./ MATERIALS AND METHODS<br />
With a view to highlighting the microbial c<strong>on</strong>taminati<strong>on</strong>, 178 samples of whole fresh<br />
fish were drawn, from 2 units, A and B, <strong>on</strong>e m<strong>on</strong>itored the verificati<strong>on</strong> of 16 batches (8<br />
batches from unit A and 8 batches from unit B). The draw of samples was performed<br />
after classifying the fish raw material in quality classes (based <strong>on</strong> species, size, corporal<br />
weight, state of freshness, etc.) from the batches of indigenous fish, which were verified<br />
from sanitary-veterinary point of view at the recepti<strong>on</strong> in the two fish processing units.<br />
91 samples from 8 batches from unit A and 87 samples from 8 batches of unit B were<br />
drawn. After collecti<strong>on</strong>, the fresh fish samples were introduced in sterile packages<br />
(polyethylene <strong>on</strong>e time use bags), in order to prevent the subsequent c<strong>on</strong>taminati<strong>on</strong> and<br />
were sent to the laboratory with a view to performing the microbiological examinati<strong>on</strong>.<br />
The whole fresh raw material fish must corresp<strong>on</strong>d from bacteriological point of view to<br />
the c<strong>on</strong>diti<strong>on</strong>s of quality foreseen in STAS 5386-86 (Barzoi D. et al., 1999, Huss H.H.,<br />
2004).<br />
According to STAS 5386-86 (Popa G. et al., 1991) the whole fresh fish must not<br />
c<strong>on</strong>tain bacteria in the depths of the muscular mass; gram positive bacteria are allowed<br />
(cocci, bacilli), which do not produce in cultures gases, indol, sulphurated hydrogen<br />
under the c<strong>on</strong>diti<strong>on</strong> that the fish does not present organoleptic and physical-chemical<br />
modificati<strong>on</strong>s.<br />
3./ RESULTS<br />
From the 16 analyzed batches (8 in unit A and 8 in unit B), in 15 of them <strong>on</strong>e<br />
highlighted the presence of bacteria in the depths of the muscular masses.<br />
From the 178 draws performed in the 2 units, a number of 124 of them resp<strong>on</strong>ded<br />
positively as a result of the bacteriologic examinati<strong>on</strong> for the presence of the bacterian<br />
flora in the depths of the muscular masses.<br />
462
In table 1 the results obtained as a result of the direct isolati<strong>on</strong> <strong>on</strong> the 3 culture<br />
envir<strong>on</strong>ments used in parallel for the highlighting of the mesophile aerobe bacteria and<br />
respectively of the anaerobe bacteria or opti<strong>on</strong>ally anaerobe from the samples of whole<br />
fresh raw material fish are shown.<br />
Table 1: Evaluati<strong>on</strong> of the bacterian c<strong>on</strong>taminati<strong>on</strong> of the fresh raw material fish at<br />
recepti<strong>on</strong> in the two processing units<br />
Crt. Unit/batch<br />
Isolati<strong>on</strong> envir<strong>on</strong>ments Total positive<br />
No.<br />
M1 M2 M3 samples/ batch<br />
1. 1A 10/10 10/10 9/10 10/10(100%)<br />
2. 2A 10/12 10/12 9/12 10/12(83,33%)<br />
3. 1B 8/10 8/10 7/10 8/10(80%)<br />
4. 2B 12/13 12/13 11/13 12/13(92,3%)<br />
5. 3B 7/10 7/10 7/10 7/10(70%)<br />
6. 4B 10/11 9/11 9/11 10/11(90,9%)<br />
7. 5B 4/10 2/10 2/10 4/10(40%)<br />
8. 6B 10/12 10/12 9/10 10/12(83,33%)<br />
9. 7B 11/11 9/11 8/11 11/11(100%)<br />
10. 8B 2/10 0/10 0/10 2/10(20%)<br />
11. 3A 0/11 0/11 0/11 0/11(0%)<br />
12. 4A 9/12 9/12 9/12 9/12(75%)<br />
13. 5A 4/13 4/13 3/13 4/13(30,76%)<br />
14. 6A 9/10 9/10 9/10 9/10(90%)<br />
15. 7A 8/12 8/12 8/12 8/12(66,66%)<br />
16. 8A 10/11 9/11 9/11 10/11(90,9%)<br />
T1<br />
15 c<strong>on</strong>taminated 124/134 115/134 109/134 124/134<br />
T2<br />
batches<br />
16 Analyzed<br />
batches<br />
(92,53%)<br />
124/178<br />
(69,66%)<br />
(85,82%)<br />
115/178<br />
(64,60%)<br />
(81,34%)<br />
109/178<br />
(61,23%)<br />
(92,53%)<br />
124/178<br />
(69,66%)<br />
M1=Nutritive glucosed bulli<strong>on</strong>. M2=Nutritive glucosed agar, M3=Nutritive glucosed<br />
bulli<strong>on</strong> with liver, T1= Total samples + from the c<strong>on</strong>taminated batches,<br />
T2=Total samples + from the draws of the two units<br />
As a result of the performed bacteriologic examinati<strong>on</strong>s <strong>on</strong>e obtained 124 positive<br />
samples from the 134 analyzed (92.53% in the two units with a c<strong>on</strong>taminati<strong>on</strong> rate of<br />
65.93% in 60/91 samples) in unit A and 73,56% (64/87 samples) in unit B<br />
The anaerobe bacteria of type Bacillus were predominant, these being isolated in<br />
85 samples from the 178 samples analyzed in the two units(47.75%), followed by the<br />
type Clostridium, found in 25 samples of the 178 samples analyzed(14,04%); the aerobe<br />
bacteria(cocci an bacilli) were isolated in proporti<strong>on</strong> of 7,86% of the total of<br />
c<strong>on</strong>taminated samples.<br />
In 1 of the 16 batches the presence of the bacterian flora was not c<strong>on</strong>firmed in the<br />
depths of the muscular masses(batch 3A).<br />
463
For the culture envir<strong>on</strong>ments that allow the growth and development of the aerobe<br />
mesophile bacteria(glucosed bulli<strong>on</strong> respectively glucosed nutritive agar) <strong>on</strong>e obtained<br />
rates of positivity of 92,53% in the c<strong>on</strong>taminated batches and respectively 85,82% and<br />
<strong>on</strong> the envir<strong>on</strong>ment of culture used for cultivating anaerobe bacteria(the glucosed<br />
nutritive bulli<strong>on</strong> with liver), the positivity rate was of 81,34%.<br />
One of the analyzed batches proved to be c<strong>on</strong>taminated in proporti<strong>on</strong> of 100% with<br />
aerobe mesophile bacteria: batch 1A; other batches presented a c<strong>on</strong>taminati<strong>on</strong> of over<br />
90%; the batch 2B(92,3%); the batches 8A and 7B(90.9%); the batch 6A (90%),<br />
according to the table 1. In the other batches, the c<strong>on</strong>taminati<strong>on</strong> rate observed was more<br />
reduced: 66,66% for batch 7A, 70% for batch 3B, 75% for batch 4A, 81,8% for batch<br />
7B etc.<br />
Table 2: Distributi<strong>on</strong> of the bacterian stems isolated as a result of the analysis of the<br />
batches of whole fresh fish in the units A and B<br />
Crt. No. Unit Sporulated gram-positive bacteria Gram-negative<br />
Bacillus Type Clostridium Type bacteria<br />
1. A 44/91 (48,35%) 10/91 (10,98%) 6/91 (6,59%)<br />
2. B 41/87 (47,12%) 15/87 (17,24%) 8/87 (9,19%)<br />
3. A+B 85/178 (47,75%) 25/178 (14,04%) 14/178 (7,86%)<br />
4./ CONCLUSIONS<br />
1. The fresh raw material fish presented as a result of the evaluati<strong>on</strong> of the bacterian<br />
c<strong>on</strong>taminati<strong>on</strong> at the recepti<strong>on</strong> in the two units A and B a c<strong>on</strong>taminati<strong>on</strong> of 65,93%<br />
in unit A and of 73,65% in unit B.<br />
2. The results obtained from the samples of fresh raw material fish drawn from the<br />
two units A and B reveal a level of c<strong>on</strong>taminati<strong>on</strong> with sporulated gram positive<br />
bacteria of type Bacillus of 47, 75% followed by the type Clostridium 14,04%; the<br />
mixed gram negative microbial flora (cocci and bacilli) represented a level of<br />
c<strong>on</strong>taminati<strong>on</strong> of 7,86% from the total of analyzed samples.<br />
REFERENCES<br />
1. Barzoi D.,Negut M., Meica S.Toxiinfectiile alimentare. Ed.Diac<strong>on</strong><br />
Coresi,Bucuresti,1999.<br />
2. Barzoi D.,Apostu S. Microbiologia produselor alimentare. Ed. Risoprint, Cluj-<br />
Napoca, 2002<br />
3. Huss H.H. Ababouch L., Gram L. 2004. Assesment and management of seafood<br />
safety and quality. FAO Fischeries Technical Paper, Rome,444.<br />
4. Popa G.,Stanescu V. C<strong>on</strong>trolul sanitar veterinar al produselor de origine animala.<br />
Ed. Didactica si Pedagogica, Bucuresti, 1991<br />
5. Yasumoto, T.. and M.Murata. 1993. Marine toxins. Chem. Rev. 93;1897-1909<br />
464
FORENSIC ASPECTS REGARDING DISASTER MANAGEMENT<br />
Carmen Radu*, Camelia Buhaş*,Nicolae Csep**, Radu Gabriela* Andrei Csep*<br />
* University of Oradea – Facultyof Medicine and Pharmacy<br />
** University of Oradea – Faculty for Envir<strong>on</strong>mental Protecti<strong>on</strong><br />
ABSTRACT<br />
Recent events, especially internati<strong>on</strong>al, but also some that happened in our country,<br />
such as the Mihăileşti amm<strong>on</strong>ium nitrate truck explosi<strong>on</strong>, led us to believe that we have<br />
to be prepared for a rapid, correct and professi<strong>on</strong>al reacti<strong>on</strong>. Therefore, establishing the<br />
correct identity of each body represents an extremely important stage within the disaster<br />
approach. Key words: disaster, identificati<strong>on</strong> center, identifying the bodies, morgue,<br />
autopsy.<br />
INTRODUCTION<br />
The <strong>term</strong> disaster (catastrophe) can be approached according to the initial event:<br />
• the accident is a sudden and unpredictable event, causing in the organism a injury<br />
or a functi<strong>on</strong>al disorder, permanent or temporary (Robert)<br />
• the catastrophe is a phenomen<strong>on</strong> of a greater extensi<strong>on</strong> producing a complete<br />
disorder or even a violent end in the existence of an individual. It is syn<strong>on</strong>ymous to<br />
the calamity, cataclysm, disaster or drama.<br />
• the calamity is a public misfortune, including starvati<strong>on</strong>, war, and epidemic.<br />
• the word cataclysm comes from the Greek „kataklusmos” which means deluge and<br />
can disturb the Earth’s crust through earthquakes, flooding, etc.<br />
• the disaster comes from „disastro” – astrological <strong>term</strong> meaning a fatal event, a<br />
serious misfortune and therefore destructi<strong>on</strong> and ruins. It signifies the influence of a<br />
star ceasing to be favorable, <strong>on</strong> the other side it is a misfortune produced by fate.<br />
• the scourge represents all the big calamities to fall up<strong>on</strong> the populati<strong>on</strong>: avalanches,<br />
flooding, huge tides.<br />
• the sinister c<strong>on</strong>stitutes a natural catastrophical event, caused by bad weather and<br />
which produces destructi<strong>on</strong>s and losses (fires, flooding, shipwrecks, earthquakes,<br />
etc.)<br />
As a c<strong>on</strong>clusi<strong>on</strong>, a catastrophe may be defined as a sudden and unusual, of great<br />
ampleness, hitting a community and causing important damages both as c<strong>on</strong>cerns the<br />
humans (great number of victims), and as c<strong>on</strong>cerns the materials (deteriorati<strong>on</strong>s and<br />
destructi<strong>on</strong>s of the envir<strong>on</strong>ment).<br />
According to art. 2 of the Ordinance 47/1994 (approved by Law 124/1995) in Romania<br />
disaster means:<br />
• natural destructive phenomen<strong>on</strong> of geological or meteorological origin or the<br />
sickening a great number of pers<strong>on</strong>s or animals suddenly occurred as a mass<br />
phenomen<strong>on</strong>.<br />
• event with extremely serious effects <strong>on</strong> the envir<strong>on</strong>ment, caused by an accident<br />
The mass disaster represents an event which is not in a direct relati<strong>on</strong>ship with an act<br />
of war and which produces the loss of at least 10 human lives or when the number of<br />
bodies is bigger they could investigate by means of local resources.<br />
465
From their origin’s point of view, the disasters are divided in:<br />
• natural, being represented by massive ecological destructi<strong>on</strong>s, which can<br />
overwhelm a community needing help from outside. E.g. earthquakes, flooding,<br />
hurricanes (typho<strong>on</strong>s) and tornadoes, land sliding, volcanic erupti<strong>on</strong>s.<br />
• artificial, which are also divided in: technological (events caused by human errors<br />
or omissi<strong>on</strong>s such as: inadequate transport of dangerous substances, toxic gas<br />
leaks), social (represented by deliberate human acts, such as wars, revoluti<strong>on</strong>s,<br />
terrorists’ acts, events followed by huge life and material losses c<strong>on</strong>stituting the<br />
social disasters) and ecological (caused especially by people and which affects the<br />
Earth, the flora and fauna <strong>on</strong> multiple levels. E.g. destroying the planetary forest<br />
and some animal species.<br />
The disasters can have the following c<strong>on</strong>sequences:<br />
• injured people needing treatment and urgent transportati<strong>on</strong> to the hospital;<br />
• deceased who need an identificati<strong>on</strong> as accurate as possible;<br />
• survivors who had not been injured and do not need medical care;<br />
• victims of disasters who lost their houses and urgently need shelter, hence the<br />
importance of creating a center for receiving such victims,<br />
• relatives or friends who ask for informati<strong>on</strong> about the pers<strong>on</strong>s involved in the<br />
accident;<br />
The sanitary infrastructure is composed of: local medical dispensaries, regi<strong>on</strong>al<br />
hospitals, reference hospitals, specialized clinics. All public resources must be<br />
integrated in this system so that they could meet the nati<strong>on</strong>al plan’s objectives.<br />
MATERIALS AND METHODS<br />
The forensic’s attributi<strong>on</strong>s in case of disasters:<br />
• to search the place, to record the deaths, to assist the body packing and labeling<br />
together with the Police and the Prosecutor (a map of he place will be drawn <strong>on</strong><br />
which each body or segments of the body will be marked)<br />
• when in situati<strong>on</strong>s involving c<strong>on</strong>taminati<strong>on</strong> risks when researching the place<br />
(chemical, biological accidents) the forensic will instruct the team involved in<br />
recuperating the bodies and they will wear protecti<strong>on</strong> suits.<br />
• he has a very important role in supervising the operati<strong>on</strong>s at the identificati<strong>on</strong><br />
center and especially in the actual identificati<strong>on</strong> of the bodies (a mixed team<br />
composed of prosecutors, police officers, emergency ambulance representatives,<br />
firemen, forensics, dentists, anthropologists, technical experts, etc.)<br />
• will supervise the pers<strong>on</strong>s involved in the incident, who have no injuries, but who<br />
can develop the post-traumatic stress disorder.<br />
The operati<strong>on</strong>s at the Identificati<strong>on</strong> Center (the center will be organized at the nearest<br />
morgue, but a temporary morgue can be also arranged in a warehouse, hangar, military<br />
base, etc.)<br />
I will now menti<strong>on</strong> the necessary steps for a good functi<strong>on</strong>ing of the center:<br />
• the recepti<strong>on</strong> of the bodies at the morgue (by a team formed of forensics, police<br />
officers, medical students);<br />
466
• depositing the bodies and the evidences (which will be packed, labeled and sealed);<br />
• the identificati<strong>on</strong> center’s structure (the center’s leader, usually a prosecutor, will<br />
collaborate with other instituti<strong>on</strong>s in order to obtain the necessary equipment,<br />
scientists);<br />
• identifying the bodies;<br />
• the autopsy;<br />
• the relatives’ recepti<strong>on</strong>;<br />
• the cooperati<strong>on</strong> with other agencies (essential for a correct investigati<strong>on</strong>).<br />
Methods of investigati<strong>on</strong>:<br />
• the visual investigati<strong>on</strong> is relative method due to the circumstances of the<br />
identificati<strong>on</strong> d<strong>on</strong>e by a family member of a close friend of the victim. The<br />
forensic takes part at the accomplishment of the spoken portrait based <strong>on</strong> the<br />
Bertill<strong>on</strong> method; a modern way of identifying is represented by the robot portrait.<br />
• the palmar or plantar prints: are based <strong>on</strong> the existence of the ante mortem pints<br />
for comparis<strong>on</strong> or they can be collected <strong>on</strong> the pers<strong>on</strong>al items.<br />
• the od<strong>on</strong>tological identificati<strong>on</strong> – the dental structures are very resistant, even in<br />
case of severe mutilati<strong>on</strong>s, advanced decompositi<strong>on</strong>s or even burns, but this<br />
method also is based <strong>on</strong> the existence of a ante mortem material for comparis<strong>on</strong><br />
(declarati<strong>on</strong>s, dental files, dental radiographies)<br />
• the genetic identificati<strong>on</strong> – a sample of spleen or muscle is collected and<br />
represents the source for the DNA procedure. In this case too <strong>on</strong>e needs ante<br />
mortem data or the genetic profile can be de<strong>term</strong>ined using biological samples<br />
from the close relatives.<br />
• For identificati<strong>on</strong> the classical methods can be used, represented by the serological<br />
methods (blood categories, Rh) or modern techniques of genotypicity: the DNA<br />
analysis by the methods RFLP and STR. (the most comm<strong>on</strong> <strong>on</strong>e)<br />
• the anthropological identificati<strong>on</strong> by comparing the anthropological characters of<br />
the group or individuals (note the importance of comparing the ante mortem<br />
radiographies with the post mortem <strong>on</strong>es.)<br />
• another way of identificati<strong>on</strong> is the facial rec<strong>on</strong>structi<strong>on</strong> by special techniques.<br />
RESULTS AND DISCUSSION<br />
Identificati<strong>on</strong> c<strong>on</strong>clusi<strong>on</strong>s, may be placed within <strong>on</strong>e of the following categories:<br />
• positive identificati<strong>on</strong>;<br />
• probable identificati<strong>on</strong>;<br />
• unidentified ( situati<strong>on</strong> when the gathered informati<strong>on</strong> regarding the body will be<br />
kept in case of a future identificat<strong>on</strong>).<br />
The forensic has to make sure of the sustained efforts of the team in order to establish<br />
the correct identificati<strong>on</strong> of each body. The difficulties in identifying the bodies are<br />
according to the nature of the disaster, with special difficulties in case of body<br />
carb<strong>on</strong>izati<strong>on</strong>, decompositi<strong>on</strong>, and disintegrati<strong>on</strong> (plane accidents, explosi<strong>on</strong>s).<br />
Unfortunately in some cases it is impossible to do the identificati<strong>on</strong>, in which case the<br />
body will remain unidentified. In an investigati<strong>on</strong>, the decisi<strong>on</strong> to aband<strong>on</strong> the<br />
467
identificati<strong>on</strong> does not have to be taken quickly, but when taken the informati<strong>on</strong> about<br />
the body will be kept for a possible future identificati<strong>on</strong>.<br />
ACKNOWLEDGEMENTS<br />
In order for the identificati<strong>on</strong> team’s activity to develop in time and systematically, both<br />
<strong>on</strong> site and at the Identificati<strong>on</strong> Center I propose the introducti<strong>on</strong> of some forms to be<br />
filled in by the cor<strong>on</strong>er. Each cor<strong>on</strong>er will follow 10 cases at most, thus receiving 10<br />
forms numbered from 1 to 10; the sec<strong>on</strong>d cor<strong>on</strong>er gets the forms from 11 to 20, etc.<br />
According to the number of available cor<strong>on</strong>ers and according to the collective<br />
accident’s complexity (number of victims) the repartiti<strong>on</strong> of the cases / cor<strong>on</strong>er is<br />
variable, but not more than 10 cases / cor<strong>on</strong>er.<br />
THE PROCESSING FORM: attached to each case, filled in and signed by the chief of<br />
each compartment.<br />
FORM NO. 1 IDENTIFICATION PROTOCOL<br />
Recepti<strong>on</strong><br />
-body<br />
-body fragment<br />
Pictures<br />
Pers<strong>on</strong>al items<br />
Fingerprints<br />
Radiological<br />
investigati<strong>on</strong>s<br />
Dental<br />
investigati<strong>on</strong>s<br />
Laboratory<br />
Mortuary<br />
services<br />
Identificati<strong>on</strong><br />
number<br />
Place<br />
<strong>on</strong> the map<br />
DATE SIGNATURE<br />
FORM NO. 2 BODY FRAGMENTS IDENTIFICATION<br />
Body fragment<br />
(identificati<strong>on</strong> number)<br />
1.<br />
2.<br />
Place<br />
<strong>on</strong> the<br />
map<br />
Type of<br />
tissue or<br />
organ<br />
Type of<br />
lesi<strong>on</strong><br />
DATE SIGNATURE<br />
468<br />
Signature Date<br />
Biological traces and<br />
biological c<strong>on</strong>diti<strong>on</strong>
FORM NO. 3 ITEM IDENTIFICATION<br />
Pers<strong>on</strong>al objects<br />
(identificati<strong>on</strong> number)<br />
Place<br />
<strong>on</strong> the map<br />
DATE SIGNATURE<br />
Type C<strong>on</strong>tent Traces<br />
REFERENCES<br />
1. Matei V., Actualităţi în medicina legală- Note de curs , Timişoara ,2001;<br />
2. Beliş V., Tratat de medicină legală, vol. I şi II, Ed. Medicală, Bucureşti, 1995;<br />
3. Proceedings of the 1st Asian-Pacific C<strong>on</strong>gress <strong>on</strong> Legal Medicine and Forensic<br />
Sciences, Singapore, September, 1983;<br />
4. Dogaroiu C., Drăghici V., Investigaţia dezastrelor cu multiple victime, Revista<br />
română de medicină legală, vol.X, nr.1, martie 2002;<br />
5. Şteiner N., Mănăstireanu D., Curs practic de urgenţe medico chirurgicale –<br />
Introducere în medicina de dezastre, Ed. Didactică şi Pedagogică, Bucureşti 1996;<br />
6. Radu C. C., Rolul medicului legist în accidente colective, Editura Universităţii<br />
Oradea,2006.<br />
469
ENERGY RECOVERY FROM ANIMAL PRODUCTS<br />
László Nagypál<br />
Tessedik Sámuel College Faculty of Agricultural Water and Envir<strong>on</strong>mental<br />
Management Institute of Envir<strong>on</strong>mental Sciences<br />
5540 Szarvas, P.O. Box 3, Hungary<br />
ABSTRACT<br />
In the 21 st century Hungary there will not be sufficient traditi<strong>on</strong>al sources of energy so<br />
that new kinds of energy will be required. The main aim of the research is to identify<br />
renewable sources of energy which cannot be utilized in the places where they become<br />
are available. Possible sources are the waste products of some industries which could<br />
be recycled in the ec<strong>on</strong>omy. For example, waste products which cannot be processed or<br />
sold since are unwanted by-products which typically the owners seek to dispose of. The<br />
utilizati<strong>on</strong> of renewing energy sources as an alternative to the fossil fuels is <strong>on</strong>e of the<br />
main efforts. (Barna 2002.)<br />
In agriculture various types of product and waste are produced which could be a<br />
source of energy. For example, plant residues, (energy) grass, animal manure and the<br />
carcasses of dead animals. One possibility for cheap energy is the burning of the meat<br />
meal and fat produced in the animal protein processing plants. The results of this<br />
research suggest that these materials could be used as a supplementary energy source in<br />
a cement works or coal-fed power-stati<strong>on</strong>. (Boros, Csok<strong>on</strong>ay 2002.)<br />
INTRODUCTION<br />
The animal carcasses from farms are transported in a special lorry to the Animal Protein<br />
Processing Plants, where they are treated. The process involves grinding the carcasses<br />
into granular form which are then sterilised at 133 °C.<br />
The 3 categories of animal wastes<br />
EU regulati<strong>on</strong>s identify three categories for animal carcasses.<br />
Category 1: carcasses with BSE pri<strong>on</strong> or other diseases which can be transferred by<br />
cross c<strong>on</strong>taminati<strong>on</strong>;<br />
Category 2: carcasses without risk of BSE or other diseases - manure, blood, bowels,<br />
mud of sewage of slaughter-house;<br />
Category 3: carcasses without risk of BSE or other diseases skin, blood (but not bovine<br />
blood) b<strong>on</strong>es, fur, hooves.<br />
Possibility of treatment or utilizati<strong>on</strong> of animal carcasses<br />
Category 1:<br />
– burning in an incinerati<strong>on</strong> plant,<br />
– burning with another material (e.g. with coal-dust in a coal-fed power-stati<strong>on</strong>, or<br />
cement in the cement works) (These materials can not be used in animal feed.)<br />
Category 2:<br />
– used for manure or a soil enriching material,<br />
470
– utilised in a biogas processing plant,<br />
– utilised in a composting prism,<br />
– burned in an incinerati<strong>on</strong> plant,<br />
Category 3:<br />
– utilised in the animal feed industry,<br />
– utilised in biogas processing plants,<br />
– utilised in a composting prism,<br />
– burned in an incinerati<strong>on</strong> plants,<br />
In Hungary there are six agro-processing plants which produce b<strong>on</strong>e and meat meal<br />
from different types of carcass (e.g. cat, dog, cow, pig, chicken), and there are a further<br />
six plants which produce meat meal from the wastes of nearby slaughter-house. The<br />
first group of plants incinerates the carcasses of cows infected by BSE. The pathogenic<br />
pri<strong>on</strong> is harmful in the meat meal, so must be burned. EU regulati<strong>on</strong>s d<strong>on</strong>’t allow the<br />
use of meat meal from cattle. (C<strong>on</strong>nett 2005.)<br />
The twelve plants manufacture 300.000 t<strong>on</strong>s of products and burn about 60.000 t<strong>on</strong>s<br />
meat meal and fat. The European Waste Catalogue numbers are (EWC) 020102 or<br />
020202 or 020203.<br />
MATERIAL AND METHODS<br />
Samples were taken form the meat meal and the comp<strong>on</strong>ents important from point of<br />
view of burning were analyzed. The chemical compositi<strong>on</strong> of flue gas produced in the<br />
course of burning was calculated by computer modelling and using the method of<br />
approximati<strong>on</strong>. Any parameter of the flue gas has to be lower than the legally approved<br />
values. Based <strong>on</strong> the identified compositi<strong>on</strong> of the flue gas the optimum technology of<br />
burning as well as the optimum technical parameters could be de<strong>term</strong>ined.<br />
Measurements<br />
Samples weighing a total of four thousand grams were collected from industrial fat and<br />
meat meal. The chemical compositi<strong>on</strong> of the samples was analysed in two independent<br />
laboratories (a/1-a/3, b/1-b/3) and the parameters are presented in the tables below.<br />
Table1: Analyses of a wet meat meal sample<br />
Samples Carb<strong>on</strong> (%) Hydrogen (%)<br />
a/1 42,0 4,5<br />
a/2 41,8 2,7<br />
a/3 40,6 2,3<br />
Table 2: Analyses of a dry meat meal sample<br />
Samples Carb<strong>on</strong> (%) Hydrogen (%)<br />
b/1 42,2 1,6<br />
b/2 38,8 1,9<br />
b/3 41,3 3,3<br />
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After the laboratory analyses the heat energy produced during the burning of a 100 g<br />
sample was calculated in the meat meal using the two main comp<strong>on</strong>ents (neglecting<br />
heat loss and the creati<strong>on</strong> heat of pollutants).<br />
The calculati<strong>on</strong><br />
The enthalpy of reacti<strong>on</strong> was calculated from the measured parameters.<br />
ΔH – enthalpy from reacti<strong>on</strong> (thermal energy will rise from burning of meat meal)<br />
ΣΔHkproduct-ΣΔHkstart= ΔH<br />
I. C + O2 = CO2<br />
II. 2H2 + O2 = 2 H2O<br />
For 1 mol:<br />
I. ΔH = ΔHCO2 – (ΔHC +ΔHO2) = -394 kJ<br />
II. ΔH = 2ΔHH2Ogaz– (ΔHO2 +2ΔHH2) = -484 kJ<br />
For 100 g dry sample:<br />
The arithmetical middle of b/1-b/3 samples:<br />
C<strong>on</strong>tent of carb<strong>on</strong>: 41,3 %, the c<strong>on</strong>tent of hydrogen 2,27 %.<br />
The heat from the reacti<strong>on</strong>: ΔH = 1335,66 + 274,42 =1610,08 kJ.<br />
For 1000 g sample:<br />
Later the quantity of heat for 1000 g of material was calculated at 16,1 MJ/kg.<br />
The viscosity of fat and water was analysed and the results at different temperatures are<br />
presented in the next table.<br />
Viscosity of<br />
fat (°E)<br />
Viscosity of<br />
water (°E)<br />
Table 3: Viscosity and temperature of fat and water<br />
23,4 40<br />
Temperature (°C)<br />
60 80 98,3<br />
28,15 3,89 2,35 1,78 0,51<br />
1,00 0,65 0,61 0,35 0,21<br />
Diagram 1 shows the rising viscosity of fat and water at different temperature.<br />
472
Viscosity [◦E]<br />
14<br />
12<br />
10<br />
8<br />
6<br />
4<br />
2<br />
0<br />
0 50 100 150 200 250<br />
Temperature [◦C]<br />
Diagram 1. The viscosity and temperature of fat ♦ viscosity of fat ■ water of fat<br />
RESULTS AND DISCUSSIONS<br />
• It is possible to store the fat before incinerati<strong>on</strong> <strong>on</strong>ly above the solidificati<strong>on</strong> point.<br />
To ensure its proper state of aggregati<strong>on</strong> at the time of spraying into the burner, the<br />
temperature must be maintained at 90 ºC.<br />
• Before beginning the industrial use of meat meal, the compositi<strong>on</strong> ash c<strong>on</strong>tent of<br />
the meat meal needs to be analysed to prevent corrosi<strong>on</strong>. According to the<br />
laboratory analyses, the ash c<strong>on</strong>tent, should be 3% (m/m) residual, or less than that<br />
after the burning, is 2.02 % (m/m).<br />
• The meat meal should be burned together with another material, or by pelleting.<br />
• In the 2 nd laboratory the meat meal after burning at 500 ºC was light grey which<br />
indicates that the carb<strong>on</strong> c<strong>on</strong>tent was destroyed at this temperature. In Hungary the<br />
permissible burning temperature is 850ºC for wastes free of halogens.<br />
• Based <strong>on</strong> the calculati<strong>on</strong>s a good fuel material is gained if the meat meal is burned<br />
as released energy during the reacti<strong>on</strong> is ~16 MJ/kg.<br />
REFERENCES<br />
Barna, Gy. (2002): Energy recovery from animals carcass in the rendering plants,<br />
Recycling of wastes and sec<strong>on</strong>d raw materials, 2: 53-61 p.<br />
Boros, T. (2002): The BSE and energy recovery of animal waste, Recycling of wastes<br />
and sec<strong>on</strong>d raw materials, 4: 61-64 p.<br />
Csok<strong>on</strong>ay, J. (2002): The German industry of meat meals, Recycling of wastes and<br />
sec<strong>on</strong>d raw materials, 7: 49-59 p.<br />
C<strong>on</strong>nett, P. (2005): Zero Waste 2020 versus incinerati<strong>on</strong>, St. Lawrence University, 1-22<br />
p.<br />
473
THE IMPLICATIONS OF MATERNAL TOXOPLASMOSIS INFECTION IN<br />
REPRODUCTION<br />
S<strong>on</strong>ia-Maria Drăghici, A. Csep, Ildiko Lenard, Adriana Jarca, Nicoleta Negruţ<br />
University of Oradea,Faculty of Medicine and Pharmacy<br />
ABSTRACT<br />
Toxoplasmosis is a zo<strong>on</strong>osis of parasitic etiology produced by the protozoan<br />
Toxoplasma g<strong>on</strong>dii that accidentally affects the human, most often it is latent in the<br />
immunocompetent adult and child, but turns into a severe and redoubtable disease, with<br />
grave complicati<strong>on</strong>s in fetus, newborn, and immunodepressed pers<strong>on</strong>s.Owing to these<br />
complicati<strong>on</strong>s, sustained and c<strong>on</strong>sistent prophylactic measures are necessary.<br />
Key words: toxoplasmosis, biological cycle, immunodepressi<strong>on</strong>, chr<strong>on</strong>ic endometritis,<br />
chorioretinitis, c<strong>on</strong>genital, prophylaxis.<br />
1./ Epidemiology<br />
The biological cycle of the parasite comprises the passage from the definitive host (in<br />
most of the cases the cat, but also other mammals, especially felines) -where the<br />
intestinal cycle takes place with sexed and gametog<strong>on</strong>ic multiplicati<strong>on</strong>- to the<br />
in<strong>term</strong>ediary host (human, mouse, pig, sheep, etc.) -where the extraintestinal, asexual<br />
cycle takes place.<br />
The intestinal cycle in the definitive host results in the producti<strong>on</strong> of oocysts,<br />
which are eliminated with the feces in the external envir<strong>on</strong>ment, where, through<br />
maturati<strong>on</strong>, sporocysts and sporozoites result, capable of infesting a new host.<br />
The extraintestinal cycle in the in<strong>term</strong>ediary host c<strong>on</strong>sists of the transformati<strong>on</strong> of<br />
the sporozoites in tachyzoites, of which some can persist in the organs insufficient in<br />
antibodies (brain, marrow, skeletal muscles), others are spread in lymph and blood,<br />
being destroyed in immunocompetent organisms. The critical state of balance between<br />
the host and the parasite can modify in the immunosupressed. Immunosupressi<strong>on</strong> can<br />
occur during lifetime through the appearance of either external causal factors (physical,<br />
chemical, medicinal, cortis<strong>on</strong>ic or cancer therapy), or internal (chr<strong>on</strong>ic<br />
immunodepressive diseases, immunologic, neoplasic, dysmetabolic, endocrinologic,<br />
etc.)<br />
The ways of transmissi<strong>on</strong> of toxoplasmosis from animals to human are multiple,<br />
sometimes complex, human intertransmissi<strong>on</strong> is possible parenterally (transplant of<br />
infected organs or transfusi<strong>on</strong> of infected blood), or vertically from mother to fetus (by<br />
the passage of sporozoites through the placenta directly into the fetal blood). Enteral<br />
transmissi<strong>on</strong> by the c<strong>on</strong>sumpti<strong>on</strong> of aliments c<strong>on</strong>taminated with oocysts and<br />
insufficiently or unprepared thermally, or by sporozoites present in the envir<strong>on</strong>ment<br />
surrounding the source (sick animal) is the most frequent means of transmissi<strong>on</strong> of<br />
disease from animals to human.<br />
2./ Clinical features of acquired toxoplasmosis in immunocompetent pateints<br />
Immunocompetent pers<strong>on</strong>s can be infested as early as childhood and adolescence.<br />
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As in any other cyclic, infectious, self-limiting disease, there can be observed a period<br />
of incubati<strong>on</strong> of 10-21 days, after c<strong>on</strong>sumpti<strong>on</strong> of infested meat, or even shorter, 5-18<br />
days, after ingesti<strong>on</strong> of oocysts.<br />
The state period comprises the acute infecti<strong>on</strong>, which in 80-90% of cases in<br />
asymptomatic. Thus can be explained the de<strong>term</strong>inati<strong>on</strong> in populati<strong>on</strong>al studies from<br />
many European countries (e.g. France) of an 80% of IgG antitoxoplasma antibodies<br />
possessors, indicators of them having previously acquired the infecti<strong>on</strong>.<br />
In 10-20% of cases, even in immunocompetents, the acute infecti<strong>on</strong> can manifest<br />
symptomatically, with lymphogangli<strong>on</strong>ic, febrile or m<strong>on</strong><strong>on</strong>ucleosic syndrome<br />
(adenopathy, fever, asthenia, myalgia, fugitive exanthema, hepatosplenomegaly without<br />
angina, lymphocytosis persistent for days or weeks). During this stage of acute<br />
symptomatic disease, clinical c<strong>on</strong>fusi<strong>on</strong>s can be made with other diseases that develop<br />
the so-called „m<strong>on</strong><strong>on</strong>ucleosis-like syndrome” (rubella, HIV infecti<strong>on</strong>, Epstein-Barr<br />
infecti<strong>on</strong>). Extremely rarely, immunocompetents develop myocarditis, pericarditis,<br />
hepatitis, encephalitis or meningoencephalitis. The presence of acute infecti<strong>on</strong>, either<br />
asymptomatic or symptomatic is proven by the IgM antitoxoplasma antibodies.<br />
Chr<strong>on</strong>ic (or latent) infecti<strong>on</strong> exists in all infested pers<strong>on</strong>s after the resoluti<strong>on</strong> of<br />
acute phase, for a shorter or l<strong>on</strong>ger period of time (weeks-years), sometimes generating<br />
ocular manifestati<strong>on</strong>s of recidivating chorioretinitis, at other times chr<strong>on</strong>ic toxoplasmic<br />
endometritis. The appearance probability of clinical symptomatology of chr<strong>on</strong>ic<br />
infecti<strong>on</strong> in immunocompetents is also owing to external or internal factors which<br />
generate a degree of immunodepressi<strong>on</strong>. Acutizati<strong>on</strong> of chr<strong>on</strong>ic infecti<strong>on</strong> is marked by<br />
the presence of IgM antitoxoplasma, together with elevated titers of IgG<br />
antitoxoplasma.<br />
The producti<strong>on</strong> mechanism of ocular toxoplasmosis in immunocompetents is the<br />
reactivati<strong>on</strong> of c<strong>on</strong>genital infecti<strong>on</strong> or acquired acute infecti<strong>on</strong> which eventually<br />
cr<strong>on</strong>icizes. The forms of manifestati<strong>on</strong> can be: chorioretinitis or focal necrotic retinitis<br />
(visi<strong>on</strong> disorders, scotomas, local pains, photophobia, epiphora), glaucoma with visi<strong>on</strong><br />
loss, posterior uveitis or panuveitis, papilitis with optic atrophy.<br />
Endometrial toxoplasmosis resulted after chr<strong>on</strong>icized acute infecti<strong>on</strong>, can<br />
generate sterility, aborti<strong>on</strong>, premature birth and infecti<strong>on</strong> transmissi<strong>on</strong> to the c<strong>on</strong>cepti<strong>on</strong><br />
product, resulting in c<strong>on</strong>genital malformati<strong>on</strong>s compatible or incompatible with life.<br />
Fig. No.1 – Toxoplasmic Encephalitis Fig. No.2 – Toxoplasmic Chorioretinitis<br />
475
Fig. No.3 – Acute<br />
Toxoplasmic Endometritis<br />
Fig. No.4 − Chr<strong>on</strong>ic<br />
Toxoplasmic Endometritis<br />
3./ Clinical manifestati<strong>on</strong>s of toxoplasmosis in the immuno-depressed patients<br />
The producti<strong>on</strong> mechanisms are the same as in immunocompetents: acute infecti<strong>on</strong> with<br />
all its c<strong>on</strong>sequences and the reactivati<strong>on</strong> of chr<strong>on</strong>ic infecti<strong>on</strong>, in the latter the<br />
appearance rate of chr<strong>on</strong>ic forms of disease being much higher than in<br />
immunocompetents.<br />
The categories of patients exposed to the infecti<strong>on</strong> due to a higher or lower degree<br />
of immunodepressi<strong>on</strong> are: homosexuals, users of intravenous drugs, patients with<br />
chr<strong>on</strong>ic diseases, diabetics, alcoholics, and patients treated with immunosuppressing<br />
agents (neoplasm, lymphoma, leukemia, collagenosis, organ transplant).<br />
HIV infected patients are the category with maximum risk of toxoplasmosis with<br />
fulminant and severe evoluti<strong>on</strong>, neurological, pulm<strong>on</strong>ary, ocular affecti<strong>on</strong>, sometimes<br />
with infectious shock.<br />
Cerebral toxoplasmosis may evolve with diffuse encephalopathy,<br />
meningoencephalitis with disorders of the c<strong>on</strong>science state, headache, c<strong>on</strong>vulsi<strong>on</strong>s,<br />
focal neurological lesi<strong>on</strong>s or myelopathy with motor and/or sensorial disorders of limbs,<br />
urinary bladder and intestine. Chorioretinitis is usually c<strong>on</strong>comitant with CNS<br />
infecti<strong>on</strong>.<br />
Pulm<strong>on</strong>ary affecti<strong>on</strong> comprises severe interstitial pneum<strong>on</strong>ia with acute<br />
respiratory insufficiency and decease in case of unrecognized etiology or absence of<br />
specific treatment.<br />
Toxoplasmic myocarditis is manifested with cardiac rhythm disorders, cardiac<br />
failure, at times in associati<strong>on</strong> with hemorrhagic pericarditis which poses great problems<br />
of differential diagnosis.<br />
Immunodepressed pers<strong>on</strong>s may develop less comm<strong>on</strong> clinical manifestati<strong>on</strong>s:<br />
panpituitarism, insipid diabetes, inadequate secreti<strong>on</strong> of antidiuretic horm<strong>on</strong>e, orchitis,<br />
gastrointestinal, musculoskeletal and hepatic affecti<strong>on</strong>, chr<strong>on</strong>ic endometritis and<br />
sec<strong>on</strong>dary sterility.<br />
4./ Clinical aspects of c<strong>on</strong>genital toxoplasmosis<br />
Vertical transmissi<strong>on</strong> of toxoplasmic infecti<strong>on</strong> from the mother with acute infecti<strong>on</strong> or<br />
chr<strong>on</strong>ic acutized toxoplasmosis, generate a severe disease of the embryo or fetus,<br />
476
usually incompatible with life or c<strong>on</strong>ducive to grave c<strong>on</strong>genital malformati<strong>on</strong>s and<br />
ulterior infantile mortality.<br />
Transplacental transmissi<strong>on</strong> of mother’s acute infecti<strong>on</strong> has a high incidence up to<br />
50% of cases. Chr<strong>on</strong>ic maternal infecti<strong>on</strong> may generate a lower rate of transplacental<br />
transmissi<strong>on</strong> (up to 25% of cases).<br />
In the transplacental transmissi<strong>on</strong> during acute maternal infecti<strong>on</strong>, it is very<br />
important to de<strong>term</strong>ine the moment of infecti<strong>on</strong> related to the stage of pregnancy:<br />
• In the first trimester the disease seldom occurs (10-25% of cases) but is severe,<br />
leading to sp<strong>on</strong>taneous aborti<strong>on</strong>, stillbirth or newborn with severe forms of disease.<br />
• In the sec<strong>on</strong>d trimester of pregnancy, toxoplasmosis occurs in 30-45% of cases,<br />
with in<strong>term</strong>ediary graveness<br />
• In the third trimester, the disease is more frequent (60-65% of cases), but often<br />
benign, with functi<strong>on</strong>al manifestati<strong>on</strong>s, without macroscopic organic<br />
malformati<strong>on</strong>s. During the life of the newborn, he can manifest disorders of<br />
somatic, psychological, intellectual development or functi<strong>on</strong>al-enzymatic defects<br />
There are no certain data referring to the toxoplasmosis manifestati<strong>on</strong> of the c<strong>on</strong>cepti<strong>on</strong><br />
product according to the stage of pregnancy, in the case of mothers with chr<strong>on</strong>ic<br />
reactivated toxoplasmosis by the aforementi<strong>on</strong>ed immunodeppressive factors.<br />
The risk of fetal affecti<strong>on</strong> is c<strong>on</strong>diti<strong>on</strong>ed by the immune resp<strong>on</strong>se of the mother at<br />
the moment of infecti<strong>on</strong>, the number of parasites transmitted to the fetus, and the age of<br />
the fetus at the moment of mother’s sickening. Hence, the lower the gestati<strong>on</strong>al age, the<br />
graver the fetal affecti<strong>on</strong>, but the risk of transmissi<strong>on</strong> is higher.<br />
In the untreated mother the transplacental transmissi<strong>on</strong> rate is 50%, and in the<br />
treated mother 25%. The period with a high risk of developing c<strong>on</strong>genital toxoplasmosis<br />
is c<strong>on</strong>sidered to be between the 10 th and 24 th weeks of pregnancy, and low risk between<br />
the 26 th and 40 th weeks of pregnancy.<br />
If toxoplasmosis occurs in mother in the first trimester of pregnancy, it mostly<br />
results in the death of the fetus and sp<strong>on</strong>taneous aborti<strong>on</strong>, and if it occurs later, it will<br />
manifest in the infant as symptomatic c<strong>on</strong>genital toxoplasmosis either since birth (1 in<br />
10 cases), or <strong>on</strong>ly after m<strong>on</strong>ths/years (9 in 10 cases).<br />
Fig. No.5 – Chr<strong>on</strong>ic Toxoplasmic Encephalopathy<br />
Clinical forms of disease are:<br />
• Severe forms manifested as grave ne<strong>on</strong>atal infecti<strong>on</strong>s with multivisceral affecti<strong>on</strong><br />
(fever or hypothermia, icterus and hepatosplenomegaly, hydro- and/or<br />
microcephaly, intracerebral calcificati<strong>on</strong>s, micro-ophtalmy, nystagmus, strabismus,<br />
477
glaucoma, cataract, chorioretinitis, optic atrophy with cecity, deafness, encephalitis,<br />
pneum<strong>on</strong>ia, myocarditis, hemorrhagic syndrome). These forms are incompatible<br />
with life; newborns survive, but exhibit severe sequelae: blindness, epilepsy,<br />
psycho-motor retardati<strong>on</strong> (they may appear within weeks, m<strong>on</strong>ths or years since<br />
birth)<br />
• Moderate forms manifested as hepatosplenomegaly and lymphoadenopathy, in the<br />
first two m<strong>on</strong>ths of life, then with CNS and ocular affecti<strong>on</strong> (after m<strong>on</strong>ths or years<br />
since birth)<br />
• Light forms manifested as ocular affecti<strong>on</strong> (isolated micro-ophtalmy, strabismus,<br />
chorioretinitis), neurological affecti<strong>on</strong> (hypot<strong>on</strong>ia, transient somnolence), or hepatic<br />
affecti<strong>on</strong> (persistent icterus for several m<strong>on</strong>ths afterbirth)<br />
Children with c<strong>on</strong>genital toxoplasmosis may exhibit at birth: prematurity, low weight,<br />
icterus, petechiae, ecchymoses by thrombocytopenia, anemia, hepatomegaly,<br />
splenomegaly, pneum<strong>on</strong>ia, different degrees of ocular affecti<strong>on</strong>, mental retardati<strong>on</strong>,<br />
micro- or macrocephaly, hydrocephaly, cerebral calcificati<strong>on</strong>s, epilepsy, encephalitis.<br />
Newborns without clinical signs of toxoplasmic infecti<strong>on</strong>s may ulteriorly exhibit<br />
ocular affecti<strong>on</strong> (chorioretinitis, strabismus, blindness), CNS affecti<strong>on</strong> (neurological and<br />
psychological deficiencies, mental retardati<strong>on</strong>, c<strong>on</strong>vulsi<strong>on</strong>s, learning disability),<br />
p<strong>on</strong>deral deficit. The risk of ocular affecti<strong>on</strong> in these children is 1 in 3 cases.<br />
5./ General prophylactic measures of toxoplasmosis<br />
The acute toxoplasmic infecti<strong>on</strong> is difficultly prevented in the circumstances of the<br />
existence of extrahuman natural focality (sick felines). A proof of this fact is the high<br />
number of pers<strong>on</strong>s found positive at the IgG antitoxoplasma test. The importance of<br />
acute infecti<strong>on</strong> prophylaxis is not evident at first glance, since it <strong>on</strong>ly leads to<br />
symptomatic sickness or chr<strong>on</strong>ic infecti<strong>on</strong> in a small percent of cases. However, the<br />
epidemiologic research dem<strong>on</strong>strates the severity of pathological c<strong>on</strong>sequences of<br />
infecti<strong>on</strong> in the circumstances of immune resp<strong>on</strong>se modificati<strong>on</strong> of the in<strong>term</strong>ediary<br />
host, due to the numerous immunodeppressive factors existing in the envir<strong>on</strong>ment<br />
which intervene at unexpected times.<br />
Therefore, the following general measures are necessary:<br />
• Neutralizati<strong>on</strong> of infective sources (sanitary-veterinary measures, with periodic<br />
parasitological surveillance of domestic cats, hygienizati<strong>on</strong> of animals’ food, oocyst<br />
polluti<strong>on</strong> preventi<strong>on</strong>)<br />
• Measures of alimentary hygiene: c<strong>on</strong>sumpti<strong>on</strong> of aliments (meat, milk, etc.) <strong>on</strong>ly<br />
after an adequate thermal processing, correct washing of fruit and vegetables<br />
• Measures of individual, envir<strong>on</strong>mental hygiene (active epidemiologic surveillance<br />
of risky professi<strong>on</strong>al domains)<br />
• Preventi<strong>on</strong> of infecti<strong>on</strong> transmissi<strong>on</strong> by blood transfusi<strong>on</strong> or organ transplant<br />
(exclusi<strong>on</strong> of seropositive d<strong>on</strong>ors)<br />
• Detecti<strong>on</strong> of acute infecti<strong>on</strong> and prompt treatment<br />
• Sanitary educati<strong>on</strong> of pregnant women<br />
• If the infecti<strong>on</strong> has occurred in the first trimester of pregnancy, therapeutic aborti<strong>on</strong><br />
is highly recommended, if the infecti<strong>on</strong> has occurred after the third m<strong>on</strong>th of<br />
pregnancy, the newborn is to be supervised and treated.<br />
478
The family doctors must be informed about the families where premature births,<br />
repeated aborti<strong>on</strong>s, malformed children or cases of sterility occurred, in order to<br />
perform an epidemiologic investigati<strong>on</strong> regarding Toxoplasma g<strong>on</strong>dii infecti<strong>on</strong>.<br />
Family planning offices as well as gynecologists must evaluate the possible<br />
toxoplasmic infecti<strong>on</strong> in future mothers.<br />
In Romania it is imperative to make the etiologic diagnosis by evidencing not <strong>on</strong>ly<br />
the specific IgM and IgG antibodies but also the antigenic structures which indicate the<br />
presence of living parasite inside the organism.<br />
REFERENCES<br />
1. J<strong>on</strong>es JL et all – C<strong>on</strong>genital toxoplasmosis a review, Obstet Gynecol Surv., 2001<br />
mai, 56(5): 296-305.<br />
2. Johns<strong>on</strong> LL. SCID mouse models <strong>on</strong> acute and relapsing chr<strong>on</strong>ic Toxoplasma<br />
g<strong>on</strong>dii infecti<strong>on</strong>s. Infect Immun. 1992;60:3719-3724.<br />
3. Kmiec K et all – C<strong>on</strong>genital toxoplasmosis in own studies, Wiad Parazytol,<br />
2001,47 Suppl 1:51-7<br />
4. Leport C, Remingt<strong>on</strong> JS. Toxoplasmose au cours du SIDA. Presse Med.<br />
1992;21:1165-1171.<br />
5. Mandell et all – Principles & Practice of Infectious Dieseases, 5 th editi<strong>on</strong> 2004,<br />
Toxoplasma G<strong>on</strong>dii, 2859-2882.<br />
6. Sinibaldi J, De Ramirez I. Incidence of c<strong>on</strong>genital toxoplasmosis in live<br />
Guatemalan newborns. Eur J Epidemiol. 1992;8:516-520.<br />
7. Subauste CS, de Waal Malefyt R, Fuh F. Role of CD80 (B7.1) and CD86 (B7.2) in<br />
the immune resp<strong>on</strong>se to an intracellular pathogen. J Immunol. 1998;160:1831-<br />
1840.<br />
8. Swartzberg JE, Remingt<strong>on</strong> JS. Transmissi<strong>on</strong> of Toxoplasma. Am J Dis Child.<br />
1995;129:777-779.<br />
9. Tirard V, Niel G, Rosenheim M, et al. Diagnosis of toxoplasmosis in patients with<br />
AIDS by isolati<strong>on</strong> of the parasite from the blood. N Engl J Med. 1991;324:632.<br />
479
BOTULISM: CLINICAL – EPIDEMIOLOGICAL STUDY<br />
Ildiko Lenard*, S<strong>on</strong>ia Draghici*, Viorica Coldea**, Mirela Indries*,<br />
Nicoleta Negrut*, Andrei Csep*, Timea Lenard***<br />
* University of Oradea, ** County Clinical Hospital of Oradea,<br />
*** County Clinical Hospital of Cluj-Napoca<br />
ABSTRACT<br />
Botulism is an alimentary toxiinfecti<strong>on</strong> caused by ex<strong>on</strong>eurotoxina, clinically<br />
characterized through bilateral and symmetric moti<strong>on</strong>al paralysis, secretor troubles,<br />
asthenia. OBJECTIVE: clinical – epidemiological assessment of patients with botulism<br />
hospitalized in the Clinical Hospital of Infectious Diseases, Oradea. PATIENTS AND<br />
METHODS: retrospective study <strong>on</strong> 29 patients with botulism hospitalized between<br />
January 1997 – December 2006, in whom there were analyzed epidemiological and<br />
clinical parameters. RESULTS: There was observed: an increased incidence of botulism<br />
in Bihor county, because of culinary customs, being predominate the cases in the<br />
m<strong>on</strong>ths of February and May (both at the end of winter and Easter period); there<br />
weren’t evidenced significant differences as c<strong>on</strong>cerns the sex and area of provenience<br />
distributi<strong>on</strong> of cases, with a morbidity level <strong>on</strong> the group of age of 15-24 years; most of<br />
the clinical forms were moderate , without being noticed particular forms of botulism;<br />
68,96% of cases were formally hospitalized in other medical services, because of the<br />
n<strong>on</strong> – recogniti<strong>on</strong> of the clinical symptomatology; the favorable evoluti<strong>on</strong> of the disease<br />
under treatment with antibotulinic serum.<br />
Key words: botulism, symmetrical paralysis, antibotulinic serum.<br />
1./ Introducti<strong>on</strong><br />
Botulism is a systemical disease, making the clinical table of alimentary toxiinfecti<strong>on</strong>,<br />
through the acti<strong>on</strong> of an ex<strong>on</strong>eurotoxina of protean nature, delivered by Clostridium<br />
botulinum (positive, anaerobe, sporuled and telluric gram bacillus which persists in the<br />
soil, marine bank, p<strong>on</strong>ds or rivers banks ). Clinically there is observed a bilateral and<br />
symmetrical moti<strong>on</strong>al paralysis of the ocular muscles, pharynx +/- respiratory muscles<br />
(by blocking the parasympathic colinergical synapses and the neuromuscular juncti<strong>on</strong>s<br />
by the botulinic toxin ) (Rummel et al, 2007), associated with secretor troubles and<br />
marked asthenia (Mandell et al, 2005; CDC 2005).<br />
Botulinic toxin is the most popular toxic biological substance, supposing that 200g<br />
of botulinic toxin could be enough to destroy the globe populati<strong>on</strong> (Villar et al, 2007).<br />
At present there are known 7 types of botulinic toxin (A, B, C, D, E, F, G), the<br />
frequency of human decays being caused by A, B and E types (Garcia-Rodriguez et al,<br />
2007). Botulism is a rare disease, with different spread around the world, its seriousness<br />
being in accordance with the type of botulinic toxin (A type from the vegetals and E<br />
type from the fish being the most virulent ) and the infecting doze. The transmissi<strong>on</strong> is<br />
made through c<strong>on</strong>taminated aliments: vegetables, fruits ( under juice form or tinned<br />
food ), meat and prepared meat (mostly pork), fish (Kirk, 2007). Depending <strong>on</strong> the<br />
transmissi<strong>on</strong> way there are described three particular forms of botulism, but with similar<br />
clinical tables Mandell et al, 2005; Mayo Clinic, 2006; Rebedea et al, 2000):<br />
1. botulism of ingesti<strong>on</strong>-the most frequent way, being resulted the toxiinfecti<strong>on</strong>;<br />
480
2. botulism of inoculati<strong>on</strong>-through the toxin’s penetrati<strong>on</strong> straight in the general flow<br />
from the level of a wound c<strong>on</strong>taminated with clostridies;<br />
3. infantile botulism-the botulinic toxin being produced in vivo, by col<strong>on</strong>izing the<br />
intestinal tract of the suckling, with Clostridium botulinum ( Arn<strong>on</strong> et al, 2006;<br />
Brett et al, 2005).<br />
2./ Objective:<br />
Assessment of clinical-epidemiological parameters <strong>on</strong> patients with botulism,<br />
hospitalized in the Clinical Hospital of infectious Diseases, Oradea between January<br />
1997 – December 2006.<br />
3./ Patients and methods<br />
The retrospective study in 29 patients with botulism hospitalized in the Clinical<br />
Hospital of Infectious Diseases, Oradea between January 1997 – December 2006, was<br />
made <strong>on</strong> the base of registered data in the clinical observati<strong>on</strong> papers of the patients.<br />
There were analyzed the following parameters:<br />
• demographic: age, sex, provenience areas;<br />
• epidemiological: way of transmissi<strong>on</strong>, source of c<strong>on</strong>taminati<strong>on</strong>;<br />
• clinical: incubati<strong>on</strong>, clinical forms of the disease, evoluti<strong>on</strong> under etiological<br />
treatment.<br />
The paraclinical c<strong>on</strong>firmati<strong>on</strong> of botulism was made through the test of specific<br />
ser<strong>on</strong>eutralizati<strong>on</strong> in the mouse.<br />
3./ Results<br />
In the period taken to be studied there were hospitalized in the Clinical Hospital of<br />
Infectious Diseases Oradea, a number of 912 patients with alimentary toxiinfecti<strong>on</strong>, of<br />
whom 29 patients were diagnosed with botulism (3.17% of the total cases of alimentary<br />
toxiinfecti<strong>on</strong>). The incidence of the cases of botulism was as follows:<br />
6<br />
11<br />
nr. Cases<br />
0 5<br />
Fig. nr.1. Incidence of cases<br />
481<br />
2<br />
3<br />
1<br />
1<br />
0<br />
1<br />
2<br />
3<br />
4<br />
5<br />
6<br />
7<br />
8<br />
9<br />
10
At the diagnosis moment, the patients had the average age of 32.1 years (limits 3-69<br />
years), with the median of 33 years. From this group, there were 15 men (52%), with<br />
the average age of 33.66 years and 14 women (48%) with the average age of 30.42<br />
years.<br />
48% male<br />
52%<br />
Fig. nr. 2. Repartiti<strong>on</strong> of the sex cases<br />
female<br />
The highest level of morbidity was described for the group of age of 15-24 years with a<br />
total of 9 cases (32%).<br />
10%<br />
14%<br />
14%<br />
3%<br />
0%3% 0%<br />
7%<br />
0-1year<br />
1-4 years<br />
17%<br />
32%<br />
Fig. nr. 3. Repartiti<strong>on</strong> <strong>on</strong> the age group<br />
482<br />
5-14years<br />
15-24year<br />
25-34year<br />
35-44year<br />
45-54year<br />
55-64year<br />
65-74year<br />
75-84year
The repartiti<strong>on</strong> of the provenience areas cases was dominated by the rural area with 16<br />
cases (55% ), versus the urban area with 13 cases ( 45% ), without being noticed any<br />
significant difference.<br />
45%<br />
55%<br />
Fig. nr. 4. Repartiti<strong>on</strong> of the provenience areas cases<br />
rural<br />
urban<br />
The m<strong>on</strong>ths with the most cases of botulism were February with 12 cases (41.4%) and<br />
may with 7 cases (24.1% ).<br />
The way of the transmissi<strong>on</strong> was digestive: in 22 cases (75.86% ) there were<br />
incriminated prepared meals from pork meat (smoked bac<strong>on</strong>, smoked sausages home<br />
prepared ), 4 cases (13.79 % ) were correlated with the c<strong>on</strong>sume of the vegetal tinned<br />
food and <strong>on</strong>ly in 3 cases (10.34% ) the epidemiological survey was negative. There<br />
were described 5 familial hotbeds (each of them with 2-5 pers<strong>on</strong>s ), the rest of cases<br />
evolving individually.<br />
The average period of time from the possible infecti<strong>on</strong> till the appearance of the<br />
first signs was of two days.<br />
All of the cases evolved with ocular paralyses (diplopia, troubles of adaptati<strong>on</strong>,<br />
palpebral ptosis, midriasis ), paralyses of the pharynx muscles (disphagia, disartry,<br />
disph<strong>on</strong>y, queasiness reflex diminuti<strong>on</strong> and the aboliti<strong>on</strong> of the velopalatin reflex) with<br />
secretor troubles (mucosa dryness) and marked asthenia. In 6 (26.8%) cases there were<br />
described paroxistical abdominal pains with meteorism, 7 (24.13%)cases developed<br />
urine acute retenti<strong>on</strong>, in <strong>on</strong>e case (3.44%) it was associated the paralysis of the<br />
respiratory muscles and in <strong>on</strong>e case (3.44%) it evolved with coma. 27 (93.10%) patients<br />
developed average forms of disease, in 2 (6.89%) cases the evoluti<strong>on</strong> being severe with<br />
respiratory paralysis, respectively coma.<br />
The evoluti<strong>on</strong> was favorable under the etiological treatment with polyvalent<br />
antibotulinic serum (in doze of 20-50ml/day <strong>on</strong> adult, and 10-25ml/day <strong>on</strong> child),<br />
antibiotherapy (in the 2 severe cases) symptomatic alimentati<strong>on</strong> through the gastric<br />
probe.<br />
483
4./ Discussi<strong>on</strong>s<br />
Botulism is a serious disease which not being treated leads to demise (Arn<strong>on</strong> et al, 2006,<br />
Mandell et al, 2005, Mayo Clinic, 2006). The historical study of the decays hotbeds<br />
prove the decrease in time of the number of the collective hotbeds, with the increase of<br />
the individual cases. In the present study 14 (48,27%) pers<strong>on</strong>s bel<strong>on</strong>g to the familial<br />
hotbeds, the rest of 15 (51,72%) ill people being individual cases of decay (Mandell et<br />
al, 2005; McLauchlin et al, 2006; WHO, 2006). It is also observed an increased<br />
incidence of botulism in Bihor county in the last 10 years, the maximum incidence<br />
being registered in 2004 with a total of 11 (37,93%) cases, fact that reveals the customs<br />
of preparing meat, fish and many other tinned food, frequently, incorrectly in precarious<br />
c<strong>on</strong>diti<strong>on</strong>s of hygiene.<br />
A feature of the present lot c<strong>on</strong>sists of the n<strong>on</strong>-recogniti<strong>on</strong> of the initial clinical<br />
table, 20 (68,96%) patients being hospitalized with different diagnoses in other medical<br />
services, being transferred in our service after a rigorous neurological exam and the<br />
exclusi<strong>on</strong> of the other neurological diseases; thus: 7 (24,13%) patients were formally<br />
hospitalized at Neurology and 1 (3,44%) case at Pediatry, digestive signs, marked<br />
asthenia and specific neurological signs, as well as the existence of <strong>on</strong>e or more<br />
members of the same family with characteristic symptomatology missing the initial<br />
examiners.<br />
The data from the literature point out a bigger frequency of botulism <strong>on</strong> men<br />
(60%) with the highest level of morbidity at 30-39 and 50-59 years (Mandell et a, 2005;<br />
Rebedea et al, 2000), unlike our lot where the report men/women tries to become equal<br />
with the highest level of morbidity at the group of age 15-24 years (from the category of<br />
pupils/students, with the frequency of the alimentary packs incorrectly preserved).There<br />
weren’t observed peculiar differences as c<strong>on</strong>cerns the provenience area of the patients<br />
or their level of educati<strong>on</strong>.<br />
Most of cases evolved as average clinical forms, the evoluti<strong>on</strong> being severe <strong>on</strong>ly in<br />
two cases, with the appearance of respiratory insufficiency because of respiratory<br />
muscular paralysis <strong>on</strong> a young woman patient of 20 years and of coma <strong>on</strong> a 3 year old<br />
girl, the clinical evoluti<strong>on</strong> being further favorable in both situati<strong>on</strong>s. The predominant of<br />
the average forms of the disease, as well as of the eradicators of pork meat involved in<br />
most of diseases, lead us to the associati<strong>on</strong> with the B type of exotoxin, frequent in<br />
Europe (Mandell et al, 2005; Rebedea et al, 2000), although the toxinopatia of the<br />
botulism cases couldn’t be made.<br />
A bigger frequency of the diseases in the m<strong>on</strong>ths of February and May is also<br />
correlated with the local customs ( the pig butchery in December, meat preservati<strong>on</strong> and<br />
that of other pork products being smoked during m<strong>on</strong>ths) and with the culinary customs<br />
for Easter.<br />
5./ C<strong>on</strong>clusi<strong>on</strong>s<br />
1. Botulism is a severe alimentary toxiinfecti<strong>on</strong> which, not being treated leads to<br />
demise; the preventi<strong>on</strong> of the decay c<strong>on</strong>sists of a harsh alimentary hygiene.<br />
2. The increased incidence of the decay (both under familial hotbeds and individual<br />
cases) in Bihor county because of the culinary customs specific to this geographic<br />
area.<br />
484
3. The patients hospitalizati<strong>on</strong>s most frequently in other medical services; through the<br />
n<strong>on</strong>-recogniti<strong>on</strong> of the clinical table.<br />
4. Almost equal distributi<strong>on</strong> <strong>on</strong> sexes and area of provenience of cases with the<br />
predominant affecti<strong>on</strong> of the group of age 15-24 years.<br />
5. Favorable evoluti<strong>on</strong> of cases under treatment with antibotulinic serum promptly<br />
introduced.<br />
REFERENCES<br />
1. .Arn<strong>on</strong> SS, Schechteck R, Maslanka SE, et al. Human botulism immune globulin<br />
for the treatment of infant botulism. N Engl J Med 2006 feb 2; 354(5): 462-71;<br />
2. Brett MM et al. 2005: A case of infant botulism with a possible link tom infeant<br />
formula milk powder: evidence for the presence of more than <strong>on</strong>e strain of<br />
Clostridium botulinum in clinical specimens and food.<br />
www.pathema.tigr.org/pathema/botulism.shtlm;<br />
3. Coordinating Center for Infectious Diseases/ Divisi<strong>on</strong> of Bacterial and Mycotic<br />
Diseases. 2005 oct 6. www.cdc.gov/ncdidod/dbmd/diseasesinfo/botulism<br />
4. Garcia-Rodriguez C, Leoy R, Arnat JW, et al. Molecular evoluti<strong>on</strong> of antibody<br />
croos-reactivity for two subtypes of type A botulinum neurotoxin. Nat Biotechnol<br />
2007 jan; 25(1): 107-16;<br />
5. Kirk M, Chan Tack MM. Botulism. 2007 mar 30. www.emedicine.com/med/topic<br />
238.htm;<br />
6. Mandell GL, Bennett JE, Dolin R: Clostridium botulinum. Principles and Practice<br />
of Infectious Diseases 5 th ed, 2005(2): 2822-26;<br />
7. Mayo Clinic Staff. Botulism – Comprehensive overview of causes, symptoms,<br />
treatments in adults and infants. 2006 mar 28.<br />
www.mayoclinic.com/botulism/article.htm;<br />
8. McLauchlin J, Grant KA. Food-borne botulism in the United Kingdom. J Public<br />
Health (Oxf). 2006 dec; 28(4):337-42;<br />
9. Peter PT, Kim J. Botulism. 2006 jun 6. www.emdicine.com/med/topic.htm;<br />
10. Rebedea I et al. Botulism. Boli Infectioase, 2000; 219-23;<br />
11. Rummel A, Eichner T, Weil T, et al. Identificati<strong>on</strong> of the protein receptor binding<br />
site of botulism neurotoxins B and G proves the double receptor c<strong>on</strong>cept. Proc Natl<br />
Acad Sci 2007 jan 2; 104(1):359-64;<br />
12. Villar RG, Elliot SP, Davenport KM. Botulism: the many faces botulinum toxin<br />
and its potential for bioterrorism. Infect Dis Clin N Am 2006; 20(1):313-27;<br />
13. World Health Organizati<strong>on</strong> (WHO): Outbreak news. Botulism, Thailand. Whly<br />
Epidemiol Rec 2006 mar 31; 81(13):118.<br />
485
STUDIES REGARDING THE INFLUENCE OF SALICYLIC ACID ON THE<br />
GERMINATION OF MAIZE CARYOPSIS (Zea Mays) AND ON THE<br />
PEROXIDASE ACTIVITY IN MAIZE PLANTLETS<br />
Cornelia Purcarea*, Dorina Cachita-Cosma **<br />
University of Oradea, Faculty for Envir<strong>on</strong>mental Sciences*<br />
Uiversity of Oradea, Faculty of Sciences**<br />
ABSTRACT<br />
In this paper we studied the influence of exogenous salicylic acid administrati<strong>on</strong> in<br />
different c<strong>on</strong>centrati<strong>on</strong>s (0.1mg/l, 1.0mg/l, 5.0mg/l), <strong>on</strong> maize caryopsis germinati<strong>on</strong> in<br />
comparis<strong>on</strong> with the germinati<strong>on</strong> of c<strong>on</strong>trol lots, wich was not treated with salicylic<br />
acid., and <strong>on</strong> the peroxidase activity in the maize plantlets resulted after 72, 96 and 120<br />
hours of germinati<strong>on</strong>. The results showed that the exogenous salicylic acid reduced the<br />
germinati<strong>on</strong> capacity of the maize caryopsis germinati<strong>on</strong> as compared to that of the<br />
reference lots, and intesified with 6.8 and 9.06% the peroxidase activity in the roots and<br />
with 5% in coleoptiles at 1.0 mg/l c<strong>on</strong>centrati<strong>on</strong> of the maize plantlets. The exogenous<br />
salicylic acid influences was dependent <strong>on</strong> the c<strong>on</strong>centrati<strong>on</strong> wich was used.<br />
Keywords: salicylic acid, maize, germinati<strong>on</strong>, peroxidase activity.<br />
1./ INTRODUCTION<br />
Salicylic acid (SA) that was named after Salix plant (willow), was first discovered as a<br />
major comp<strong>on</strong>ent in the extracts from willow tree bark that had been used as a natural<br />
anti-inflammatory drug from the ancient time to the 18th century. The name SA, from<br />
the Latin word Salix for willow tree, was given to this active ingredient of willow bark<br />
by Raffaele Piria in 1838 (Raskin, 1992)<br />
The salicylic acid or o-hydroxibenzoic acid, nowadays c<strong>on</strong>sidered to be an<br />
important signal molecule, involved in the development process of plants (Ryan and<br />
Farmer, 1992). Actually, SA is c<strong>on</strong>sidered a plant growth regulator, which influences<br />
many of the vegetal processes, such as: germinati<strong>on</strong> (Cutt and Klessig, 1992), stomatal<br />
behaviour (Larque-Saaveda, 1979), membrane permeability (Barkosky and Einhellig,<br />
1993), rate of photosynthesis (Khan and Prithiviraj, 2003), etc. In 1974, Cleland and<br />
Ajami, proved that the SA has an endogeneous regulator of flowering.<br />
In the opini<strong>on</strong> of Handro et al., (1997), the SA supports flowering and tuberizati<strong>on</strong><br />
of plants. They studied the in vitro effect of SA of flowering of Streptocarpus nobilis,<br />
but also of in vitro tuberizati<strong>on</strong> of Ullucus tuberosus. In the presence of 0,5-2,0 μM SA<br />
the number of Streptocarpus nobilis floral buds significantly increased and floral<br />
scapes were significantly l<strong>on</strong>ger as compared with the c<strong>on</strong>trols cultured without SA..<br />
Picomolar levels of SA, enhance cell growth and the embryogenesis in the cell<br />
suspensi<strong>on</strong> cultures of Coffea arabica,as compared with entreated c<strong>on</strong>trol cultures<br />
(Quiroz-Figueroa et al. 2001). They reached the c<strong>on</strong>clusi<strong>on</strong> that SA can activate as a<br />
morphoregulator signal, wich supports the hypothesis that SA is functi<strong>on</strong>ing as a growth<br />
regulator. The authors also suggested that it is possible that this phenolic compounds act<br />
as a signal to induce the differentiati<strong>on</strong> processes. An alternative explanati<strong>on</strong> is the<br />
486
possibility that, due to the chelating properties of these compounds, some inhibitors<br />
present in the embryogenic cultures are inactivated.<br />
The process of generating free radicals can be minimized by adding SA, which can<br />
chelating transiti<strong>on</strong>al metals, thus reducing the producti<strong>on</strong> of hydroxyl radicals (HO - ) or<br />
it can act directly as an antioxidant against these types of free radicals of oxygen<br />
(Antofie et al., 2003).<br />
Prachi et al. (2002) have found that additi<strong>on</strong> of 104 M SA, to callus cultures of<br />
Zingiber officinale produced the increased of peroxidase and β-1,3-glucanase activity,<br />
as well as the appearence of two new proteins in the calli.<br />
Some enzymes, such as guaiacol-peroxidase and glutati<strong>on</strong>-reductase, are activated<br />
by the treatments with SA soluti<strong>on</strong>s and others, such as catalase are inhibited as a result<br />
of SA treatments and changes in the patterns of its isoensimes can be noticed (Horvath<br />
et al., 2002).<br />
The aim of this work was to study the influence of the exogenous SA <strong>on</strong> maize<br />
caryopsiss germinati<strong>on</strong> (Zea Mays) and <strong>on</strong> the peroxidase activity in maize plantlets<br />
(roots and coleoptiles) derivates from its..<br />
2./ MATHERIALS AND METHODS<br />
Sample preparati<strong>on</strong>: SA treatments were applied to lots of 150 maize caryopsis<br />
sample, germinated into plastic recipients. The maize caryopsis used in this study have<br />
90% germinati<strong>on</strong> faculty.<br />
The germinati<strong>on</strong> was maded in plastic recipients, <strong>on</strong> a filter paper, moistened with<br />
20 ml:<br />
- tap water , for the c<strong>on</strong>trol sample,<br />
- salicylic acid soluti<strong>on</strong> 0.1mg/l<br />
- salicylic acid soluti<strong>on</strong> 1mg/l,<br />
- salicylic acid soluti<strong>on</strong> 5mg/l.<br />
The germinati<strong>on</strong> was maded at room temperature. Every day, the quantity of<br />
soluti<strong>on</strong> from the recipients was brought to the level of 20 ml. The germinati<strong>on</strong><br />
temperature was around 22 ± 2˚C.<br />
Germinati<strong>on</strong> energy, represents the speed with which the germinati<strong>on</strong> process begins.<br />
It is estimated by the number of germinated grains in 72 hours. The number of normally<br />
germinated grains over the three repetiti<strong>on</strong>s is divided by 100 for each plastic recipient<br />
to de<strong>term</strong>ine the germinati<strong>on</strong> energy (Andrei et al. 1981).<br />
Preparati<strong>on</strong> of enzyme extract: 1g for each samples separatelly (roots and coleoptiles)<br />
were collected from each variant, at 72, 96 and 120 hours of caryopsis put <strong>on</strong><br />
germinati<strong>on</strong>, and were blended with 4ml phosphate buffer soluti<strong>on</strong>, pH 7.0, diluted 1:9<br />
with distilled water, cooled at 4˚C.<br />
The samples were centrifuged at 10000 rpm, for 15 minutes at 4˚C, and supernatant<br />
were separated. The extract is kept in the refrigerator, for 2 hours for stabilizing and<br />
expressing enzyme activity.<br />
Enzyme assay: peroxidase activity (PA) was de<strong>term</strong>inated at 483nm wavelenght using<br />
phosphate buffer of 7.0 pH, hydrogen peroxide as substrate and p-phenilendiamine as<br />
chromogen (Hans-Luck, 1970 modified by Sipoş et al. 2003). The extensi<strong>on</strong> for each<br />
sample was noted after 30 sec<strong>on</strong>ds of reacti<strong>on</strong> period. For each extract were made four<br />
de<strong>term</strong>inati<strong>on</strong>s.<br />
487
The data obtained at the influence of the SA <strong>on</strong> PA were. The data were statistically<br />
evaluated using the standard deviati<strong>on</strong> (SD) and t-test methods.<br />
The values of the probabilities were de<strong>term</strong>ined from tables using the values of the<br />
“t” distributi<strong>on</strong> and the freedom degrees based <strong>on</strong> which the variance of the empiric<br />
series was calculated.<br />
3./ RESULTS AND DISCUSION<br />
Where germinati<strong>on</strong> energy is c<strong>on</strong>cerned, the data obtained after counting the germinated<br />
of 300 maize caryopsis, for each c<strong>on</strong>centrati<strong>on</strong> of SA, were mathematically processed<br />
obtaining the arithmetical mean of the three values. The obtained results were written<br />
in the diagram in figure. 1.<br />
Germinati<strong>on</strong> energy %<br />
100<br />
80<br />
60<br />
40<br />
20<br />
0<br />
90<br />
80.5<br />
C<strong>on</strong>trol SA 0.1mg/l SA 1.0mg/l SA 5.0mg/l<br />
SA soluti<strong>on</strong><br />
Figure.1– Percentage differences in the germinative energies of maize caryopsiss<br />
(Zea Mays) after 72 hours of germinati<strong>on</strong><br />
The germinati<strong>on</strong> energy (figure.1) of the maize caryopsis was lower when were used<br />
SA soluti<strong>on</strong>s of different c<strong>on</strong>centrati<strong>on</strong>s, as compared with the c<strong>on</strong>trol sample, a fact<br />
also emphasized in the reference literature by Cutt and Klessig (1992). When using<br />
1.0mg/l c<strong>on</strong>centrati<strong>on</strong> SA soluti<strong>on</strong>, the germinati<strong>on</strong> energy was 55% from germinati<strong>on</strong><br />
energy of c<strong>on</strong>trol lot. 5mg/l SA soluti<strong>on</strong> inhibit 100% the germinati<strong>on</strong> of maize<br />
caryopsis.<br />
Figure 2 shows an overview of the percentage differences in the extincti<strong>on</strong> which<br />
reflect the intensity of the peroxidase activity in roots and in coleoptiles of the maize<br />
plantlets originated from the maize caryopsis germinati<strong>on</strong> <strong>on</strong> SA soluti<strong>on</strong>s of 0.1mg/l,<br />
1.0mg/l and 5.0mg/l c<strong>on</strong>centrati<strong>on</strong>s, for 72, 96 and 120 hours in comparis<strong>on</strong> with the<br />
same parameter de<strong>term</strong>ined in the roots and coleoptiles of the c<strong>on</strong>trol sample, for which<br />
the extincti<strong>on</strong> was c<strong>on</strong>sidered to be 100%.<br />
488<br />
55<br />
0
difference of extincti<strong>on</strong> %<br />
120<br />
100<br />
80<br />
60<br />
40<br />
20<br />
0<br />
72 hour<br />
root<br />
96 hour<br />
root<br />
120<br />
hour<br />
root<br />
72 hour<br />
Cp<br />
time of germinati<strong>on</strong><br />
96 hour<br />
Cp<br />
120<br />
hour Cp<br />
c<strong>on</strong>trol<br />
SA 0.1mg/l<br />
SA 1.0mg/l<br />
SA 5.0mg/l<br />
Figure.2 – Percentage differences in extincti<strong>on</strong> which reflect the intensity of the<br />
peroxidase activity in the roots and coleoptiles of maize plantlets.<br />
Table 1. Results synthesis of peroxidase activity measurements in the vegetal extract<br />
obtained from the roots and coleoptiles of the plantlets after 72, 96 and 120 hours of<br />
germinati<strong>on</strong> <strong>on</strong> the substrated moisted with SA soluti<strong>on</strong>s of different c<strong>on</strong>centrati<strong>on</strong>s<br />
Parameter Hour of<br />
germinati<strong>on</strong><br />
Peroxidase<br />
extincti<strong>on</strong> in<br />
roots<br />
Peroxidase<br />
extincti<strong>on</strong> in<br />
coleoptiles<br />
72<br />
Salicylic acid mg/l c<strong>on</strong>centrati<strong>on</strong><br />
Average± SD<br />
C<strong>on</strong>trol SA 0.1mg/l SA 1.0mg/l<br />
0.844±0.029 0.889±0.007 0.924±0.011<br />
*<br />
***<br />
0.917±0.057<br />
n.s.<br />
n.s.<br />
0.741±0.014 0.758±0.006 0.791±0.005<br />
n.s.<br />
***<br />
0.931±0.039 0.992±0.0515 1.006±0.0105<br />
n.s.<br />
*<br />
0.908±0.044 0.929±0.034 0.977±0.015<br />
96 0.864±0.028 0.894±0.04<br />
120<br />
72<br />
96<br />
120<br />
n.s.<br />
0.761±0.021 0.788±0.0059<br />
n.s.<br />
*<br />
0.799±0.0139<br />
*<br />
Studying the peroxidase activity intensity separately in the roots and coleoptiles of the<br />
plantlets obtained from the germinati<strong>on</strong> of the maize caryopsis in SA soluti<strong>on</strong>s, after 72<br />
489
hours of germinati<strong>on</strong>, we observed an increase of the PA in the extract obtained from<br />
the roots as compared with the same parameter de<strong>term</strong>ined in the c<strong>on</strong>trol sample, the<br />
increase being very significant in the case of the germinati<strong>on</strong> in SA soluti<strong>on</strong> of 1.0mg/l,<br />
and significant when using a SA soluti<strong>on</strong> of 0.1mg/l c<strong>on</strong>centrati<strong>on</strong> (table 1).<br />
Studying the variati<strong>on</strong> of the same parameter after 96 hours, an insignificant<br />
increase of the peroxidase activity was observed as compared with the c<strong>on</strong>trol sample at<br />
c<strong>on</strong>centrati<strong>on</strong>s of 0.1 and 1.0 mg/l SA soluti<strong>on</strong> (table 1).<br />
After 120 hour of germinati<strong>on</strong>, we observed a very signifiant increase of the PA in<br />
the extract of the roots in the case of germinati<strong>on</strong> in 1.0mg/l SA soluti<strong>on</strong>, and an<br />
insignifiant increase for 0.1mg/l SA soluti<strong>on</strong>. (table1.)<br />
For the coleoptiles extracts, the intensity of the peroxidase activity after 72, 96 and<br />
120 hours of germinati<strong>on</strong> has insignificantly modified as compared with the same<br />
parameter of c<strong>on</strong>trol sample when we used 0.1mg/l SA soluti<strong>on</strong>, and significantly<br />
increase for 1mg/l SA soluti<strong>on</strong> for 72 and 120 hour of germinati<strong>on</strong> (table 1).<br />
4./ CONCLUSIONS<br />
It can be observed that the exogenous SA inhibits the germinati<strong>on</strong> energy of the maize<br />
caryopsis with 10 and 45% from c<strong>on</strong>trol lot, using SA soluti<strong>on</strong> with 0.1 and 1.0 mg/l<br />
c<strong>on</strong>centrati<strong>on</strong> and 100% in case of 5.0mg/l SA soluti<strong>on</strong>.<br />
The peroxidase activity of the vegetal extracts obtained from the roots of the<br />
plantlets obtained after germinati<strong>on</strong> <strong>on</strong> filter paper moistened with SA soluti<strong>on</strong> was<br />
very significantly increase after 72 and 120 hours from germinati<strong>on</strong> for 1.0 mg/l<br />
c<strong>on</strong>centrati<strong>on</strong> SA soluti<strong>on</strong> and significantly increase for 0.1mg/l SA soluti<strong>on</strong>, as<br />
opposed to the peroxidase activity of the extracts obtained from coleoptiles, where its<br />
activity changed insignificantly after 72 and 120 hours of germinati<strong>on</strong> for 0.1mg/l SA<br />
soluti<strong>on</strong>, and significantly for 1.0mg/l SA soluti<strong>on</strong>.<br />
In c<strong>on</strong>clusi<strong>on</strong> exogenous salicylic acid are influenced different physiological<br />
plant process.<br />
REFERENCES<br />
Andrei M., Anghel I., Popescu I., Stoica E., (1981): Lucrări practice de biologie<br />
vegetală, Editura didactică şi pedagogică Bucureşti.<br />
Antofie M., Carasan ME., Brezeanu A., (2003): Salicylic acid effects <strong>on</strong> two<br />
Gardenia Jasminoides cell lines. Acta Horti Bot. Bucureşti, 30:135-145.<br />
Barkosky RR., Einhellig FA., (1993): Effects of salicylic acid <strong>on</strong> plant water<br />
relati<strong>on</strong>ship. Journal of Chemical Ecolology, 19:237-247.<br />
Cleland FC., Ajami A., (1974) – Identificati<strong>on</strong> of the flower-inducing factor isolated<br />
from aphid h<strong>on</strong>ey dew as being salicylic acid. Plant Physiol. 54:904-906.<br />
Cutt JR. Klessig DF., (1992): Salicylic acid in plants. A changing perspective.<br />
Pharmacetical Technology, 16:25-34.<br />
Handro W., Mello CM., Manzano MA., Floh ES.,(1997): Promotive effects of<br />
Salicylic acid. R. Bras. Fisiol.Veg. 9(2): 139-142.<br />
Hans-Luck, (1970): Methdos of Enzymatic Analysis Verlag Chemie Gmb H.<br />
(Ed).Weinheim Bergstr. Academic Press, New york and L<strong>on</strong>d<strong>on</strong>.p.885-888.<br />
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Horváth E., Szalai G., Pál m., Páldi E, Janda T., (2002): Differences between the<br />
catalase isozymes of maize (Zea mays L.) in respect of inhibiti<strong>on</strong> by various<br />
phenolic compounds. Acta Biol Szeged 46 (3-4): 33-34.<br />
Khan W., Prithiviraj B., (2003): Photosynthetic resp<strong>on</strong>ses of corn and soybean to<br />
foliar applicati<strong>on</strong> of salicylates. Journal of Plant Physiology, p. 18<br />
Larque-Saaveda A., (1979): Stomatal closure in resp<strong>on</strong>se to salicylic acid treatment.<br />
Pflanzenphysiology, 93: 371-375.<br />
Prachi-Sharma TR., Singh BM., (2002): Salicylic acid induces Resistance to<br />
pathogen. European Journal of Plant Pathology 108 (1): 31-39.<br />
Quiroz-Figueroa F., Mendez-Zeel M., Larque-Saavedra., Loyola-Vargas VM.,<br />
(2001): Picomolar salicylate levels enhance cell growth and embryogenesis. Plant<br />
cell reports 20:679-684.<br />
Raskin I., (1992) – Salicylate a new plant horm<strong>on</strong>e. Plant Physiol. 99: 799-803.<br />
Ryan CA., Farmer EE, (1992): Oligosaccharide signals in plants: a current<br />
assessment. Ann Rev.Palnt Physiol. Molec.Biol., 42:651-674.<br />
Şipoş M.,Floriş C.,Chiriac C.,(2003): Activitatea peroxidazică în embri<strong>on</strong>ii şi<br />
plantulele de grâu (Triticum aestivum L.soiul Turda) rezultate prin germinaţia<br />
cariopselor submersate, în prealabil, în azot lichid -196˚C. Analele Univ. Oradea,<br />
Fasc.Biologie, Tom X., 315-320; 333-342.<br />
491
THE INFLUENCE OF SALICYLIC ACID ON CONTENT OF ASSIMILATORY<br />
PIGMENTS IN THE PRIMARY LEAVES OF WHEAT (Triticum aestivum)<br />
PLANTLETS<br />
Cornelia Purcarea * , Dorina Cachiţă-Cosma **<br />
*University of Oradea, Faculty for Envir<strong>on</strong>mental protevti<strong>on</strong><br />
**University of Oradea, Faculty of Sciences<br />
ABSTRACT<br />
Salicilyc acid (SA) is a phenolic compound recently recognized as plant growth<br />
regulator which is involved in many physiological processes including photosynthesis.<br />
In the present investigati<strong>on</strong> we were studied the influence of exogenous SA soluti<strong>on</strong> <strong>on</strong><br />
the c<strong>on</strong>tent of assimilatory pigments in the primary leaves of the wheat plantlets. The<br />
wheat grains were presoaked 6h with 0.01 mM, 0.1 mM, or 1.0 mM SA soluti<strong>on</strong>s and<br />
in water for the c<strong>on</strong>trol lots. After that, we planted the plantlets in sand and sprayed<br />
their coleoptiles and primary leaves, each day for an additi<strong>on</strong>al 7 days, with the same<br />
soluti<strong>on</strong>s. In the 14 th days of germinati<strong>on</strong> we de<strong>term</strong>ined the c<strong>on</strong>tent of assimilatory<br />
pigments extracted with N,N-dimethylformamide (DMF). The results showed that<br />
exogenous 0.01 mM, 0.1mM, or 1.0 mM SA soluti<strong>on</strong>s, enhanced clorophyll a,<br />
clorophyll b, and carotenoid pigments in the wheat plantlets primary leaves.<br />
Keywords: wheat, salicylic acid, primary leaves, clorophyll a, clorophyll b, carotenoid<br />
pigments.<br />
INTRODUCTION<br />
In aditi<strong>on</strong> to the classical plant horm<strong>on</strong>es, new natural growth substances with<br />
regulatory roles in tissue culture have been discovered in the last few years (Gross and<br />
Parthier, 1994). One of these substance is salicylic acid and its derivatives. Salicylic<br />
acid could be raised to the status of the above phytohorm<strong>on</strong>es because it has significant<br />
impact <strong>on</strong> the various aspects of the plant life (Hayat and Ahmad, 2007).<br />
Salicylic acid or ortho-hidroxibenzoic acid bel<strong>on</strong>gs to a diverse group of plant<br />
phenolics. These are compounds with an aromatic ring bearing a hydroxyl group or its<br />
functi<strong>on</strong>al derivative (Raskin, 1992). Salicylic acid is a natural signaling molecule<br />
involved in the regulati<strong>on</strong> of different physiological processes including photosynthesis.<br />
The metabolic aspect of plants, supplied with SA soluti<strong>on</strong> or its derivatives shifted<br />
to a varied degree depending <strong>on</strong> the plant type and the mode of applicati<strong>on</strong> of SA<br />
soluti<strong>on</strong>. The applicati<strong>on</strong> of SA soluti<strong>on</strong> (20 mg/ml) to the foliage of the plants of<br />
Brassica napus, improved the chlorophyll c<strong>on</strong>tents (Ghai et al., 2002). Similarly,<br />
soakink the grains of wheat in 10 -5 M of SA soluti<strong>on</strong> de<strong>term</strong>inated in the plants a higher<br />
pigment c<strong>on</strong>tents wich declived as the c<strong>on</strong>centrati<strong>on</strong> of SA was increased above that<br />
c<strong>on</strong>centrati<strong>on</strong> (Hayat et al., 2005). Moreover, 30 days old plants of Brassica juncea<br />
sprayed with 10 -5 M of SA soluti<strong>on</strong> possessed chlorophyll 20% higher than those<br />
sprayed by water <strong>on</strong>ly, however the maximum c<strong>on</strong>centrati<strong>on</strong> (10 -3 ) decreased the<br />
492
chlorophyll c<strong>on</strong>tents and the values were below that the water sprayed c<strong>on</strong>trol at 60<br />
days stage (Fariduddin et al., 2003).<br />
Soaking the seeds of Vigna mungo in aqueous soluti<strong>on</strong>s of SA (10-150μm) lead a<br />
decrease in the c<strong>on</strong>tent of chlorophyll and carotenoid in the leaves of subsequent plants,<br />
but supplementing SA throught irrigant did not prove as severe as seed treatment<br />
(Anandhi and Ramanujam, 1997).<br />
Salicylic acid activated the synthesis of carotenoids, xanthophylls and the rate of<br />
de-epoxidati<strong>on</strong> but decreased the level of clorophyll pigments, both in wheat and mo<strong>on</strong>g<br />
plants also the ratio of clorophyll a/b, in wheat plantlets (Moharekar et al., 2003).<br />
Jianping Xue et al., 2006, studied the effects of different c<strong>on</strong>centrati<strong>on</strong> of SA<br />
soluti<strong>on</strong> <strong>on</strong> the growth of Pinellia ternate. When the height of plant was about 10cm,<br />
sprayed them with different c<strong>on</strong>centrati<strong>on</strong> of SA soluti<strong>on</strong> and measured height, total<br />
chlorophyll c<strong>on</strong>tent, activity of SOD, MAD c<strong>on</strong>tent, photosynthesis speed, intercellular<br />
CO2 c<strong>on</strong>centrati<strong>on</strong>, the transpirati<strong>on</strong> speed and the leaf temperature. The results<br />
indicated that intercellular CO2 c<strong>on</strong>centrati<strong>on</strong> was increased, leaf temperature were<br />
decreased and photosynthesis speed was well in 0.5 mM SA soluti<strong>on</strong>. In c<strong>on</strong>clusi<strong>on</strong>, the<br />
c<strong>on</strong>centrati<strong>on</strong> of 0.5 mM SA soluti<strong>on</strong> was suitable for the growth of P. ternate.<br />
MATERIALS AND METHODS<br />
For study the acti<strong>on</strong> of SA treatments under laboratory c<strong>on</strong>diti<strong>on</strong>s, the wheat caryopsis<br />
were soaked for 6 hours in 0.01 mM, 0.1 mM, or 1.0 mM SA soluti<strong>on</strong>s, and in water<br />
for the c<strong>on</strong>trol lot. Then the grains were germinated for 7 days in plastic boxes. The<br />
germinati<strong>on</strong> was made <strong>on</strong> moistened filter paper with distilled water, at 22±2 °C.<br />
After 7 days of germinati<strong>on</strong> we planted the plantletss in sand, leaving them there<br />
for an additi<strong>on</strong>al 7 days, and sprayed theire coleoptiles and primary leaveas each day<br />
with 0.01 mM, 0.1 mM, or 1.0 mM SA soluti<strong>on</strong>s or with water for the c<strong>on</strong>trol lot.<br />
In the 14 th days we de<strong>term</strong>ined the c<strong>on</strong>tent of chlorophyllian and carotenoid<br />
pigments of the wheat plantlets primary leaves, using N,N-dimethylformamide, 99.9%,<br />
(Moran and Porath, 1980) for the extracti<strong>on</strong>. The extracti<strong>on</strong> of chlorophylls in higher<br />
plant tissue using N,N-dimethylformamide (DMF), expedits the process and enables the<br />
de<strong>term</strong>inati<strong>on</strong> of small samples with low pigment level (Moran, 1982). For extracti<strong>on</strong>,<br />
50 mg fresh weight of primary leaves, were collected separately from each samples, and<br />
were blended with 5ml DMF and then cooled at 4˚C for 72 hours. The supernatant was<br />
separated and the c<strong>on</strong>tent the pigment was de<strong>term</strong>ined using a Spekol 11 (Carl Zeiss<br />
Jena) spectrophotometer, at 664nm wave length for chlorophyll a, 647 nm for<br />
chlorophyll b, and 480 nm for carotenoids. For each sample we made 3 de<strong>term</strong>inati<strong>on</strong>s.<br />
The data obtained after the spectrophotometrycal de<strong>term</strong>inati<strong>on</strong>, were<br />
mathematically processed using formulae proposed by Moran and Porath (1982). For<br />
the de<strong>term</strong>inati<strong>on</strong> of the specific extincti<strong>on</strong> coefficients (SEC), they made pure<br />
chlorophyll a, chlorophyll b and carotenoid pigments soluti<strong>on</strong> similarly prepared from<br />
DMF extracts. The SEC were de<strong>term</strong>ined by the equati<strong>on</strong>:<br />
Aλ =ε cl<br />
Where Aλ is the absorbance at a given wavelength, ε is the SEC of the soluti<strong>on</strong> at<br />
wavwlength λ, c is the c<strong>on</strong>centrati<strong>on</strong> g/l, and l is the beam-path (1cm) in the measuring<br />
493
cuvette. Solving equati<strong>on</strong> for A664, A647 and A480 wavelength Moran obtained the<br />
formulae for de<strong>term</strong>inati<strong>on</strong> of chlorophyll a , chlorophyll b and carotenoids c<strong>on</strong>tents.<br />
Clorophyll a (mg/g SP) = (11.65 A664 – 2.69 A647) • v/SP<br />
Clorophyll b (mg/g SP) = (20.81 A647 – 4.53 A664) • v/SP<br />
Carotenoids (mg/g SP) = (1000 A480 – 0.89 clorophyll.a – 52.02 clorophyll.b)/ 245 •<br />
v/SP<br />
Where: A480 – the value read with a 480 nm filter<br />
A647 – the value read with a 647 nm filter<br />
A664 – the value read with a 664 nm filter<br />
v – ml of solvent used<br />
SP – mg of material used for <strong>on</strong>e extracti<strong>on</strong>/sample<br />
Chlorophyll a and b – quantity in mg calculated in the first two formulas<br />
The results obtained after the c<strong>on</strong>tent of assimilatory pigments de<strong>term</strong>inati<strong>on</strong> are<br />
averages of 3 de<strong>term</strong>inati<strong>on</strong> and were statistically processed using the “t- test”. The<br />
values of the probabilities were de<strong>term</strong>ined from tables using the values of the “t”<br />
distributi<strong>on</strong> and the freedom degrees based <strong>on</strong> which the variance of the empiric series<br />
was calculated.<br />
RESULTS AND DISCUSSION<br />
Studying the c<strong>on</strong>tent of chlorophyllian pigment (chlorophyll a and b) and carotenoids<br />
<strong>on</strong> the primary leaves of the wheat plantlets obtained from each experimental variants,<br />
we observed that the influence of the exogenous SA soluti<strong>on</strong>s treatment was dependent<br />
<strong>on</strong> the c<strong>on</strong>centrati<strong>on</strong> which was used and the type of pigment wich was analised. The<br />
results obtained were presented in table 1 and graphically represented in figure 1.<br />
The c<strong>on</strong>tent of chlorophyll a (table 1, figure 1.) increased significantly (with 17%<br />
from c<strong>on</strong>trol lot c<strong>on</strong>sidered 100%) after treatment with 0.01 mM SA soluti<strong>on</strong>. A very<br />
significant increased of chlorophyll a c<strong>on</strong>tents was observed, with 40% from the c<strong>on</strong>trol<br />
lot in the case of treatment with 0.1 mM SA soluti<strong>on</strong>, and with 23% from the c<strong>on</strong>trol lot<br />
in the case of treatment with 1.0 mM SA soluti<strong>on</strong>.<br />
In the case of the chlorophyll b c<strong>on</strong>tents (table 1., figure 1.) it could be observed a<br />
significant increased, with 20.9% from c<strong>on</strong>trol lot when using a 0.01 mM SA soluti<strong>on</strong>, a<br />
very significant increased, with 36% from the c<strong>on</strong>trol lot, in the case of treatment with<br />
0.1 mM SA soluti<strong>on</strong> and with 24.3% from the c<strong>on</strong>trol lot in the case of treatment with<br />
1.0 mM SA soluti<strong>on</strong>.<br />
Studying the carotenoids pigments c<strong>on</strong>tent (table 1., figure 1.), the results show<br />
that the accumulati<strong>on</strong> of these pigments in the leaves of wheat plantlets <strong>on</strong> the 14 th days<br />
of germinati<strong>on</strong>, increased significantly in comparis<strong>on</strong> with the same parameter<br />
de<strong>term</strong>ined from the c<strong>on</strong>trol lot, in the case of treatment with 0.01 mM SA soluti<strong>on</strong><br />
(with 38.8% from the c<strong>on</strong>trol lot), with 50% from the c<strong>on</strong>trol lot when the treatment<br />
was made with 0.1 mM SA soluti<strong>on</strong> and with 38.3% from the c<strong>on</strong>trol lot for treatment<br />
with 1.0 mM SA soluti<strong>on</strong>.<br />
494
Table 1.Estimative mean values for the c<strong>on</strong>tent of assimilatory pigments of the wheat<br />
plantlets (Triticum aestivum) leaves after treatment made with SA soluti<strong>on</strong>s of<br />
different c<strong>on</strong>centrati<strong>on</strong>s.<br />
Parameters<br />
Chlorophyll<br />
a mg/g<br />
Chlorophyll<br />
b mg/g<br />
Carotenoid<br />
pigm. mg/g<br />
C<strong>on</strong>trol<br />
lot<br />
0.65±0.03<br />
0.44±0.02<br />
0.18±0.01<br />
Salicylic acid soluti<strong>on</strong><br />
(mM c<strong>on</strong>centrati<strong>on</strong>)<br />
0.01mM 0.1mM 1.0 mM<br />
Average ± standard deviati<strong>on</strong><br />
0.76±0.01 0.91±0.22 0.801±0.012<br />
** *** ***<br />
0.532±0.02<br />
**<br />
0.25±0.02<br />
**<br />
0.60±0.014<br />
***<br />
0.27±0.021<br />
**<br />
0.547±0.016<br />
**<br />
0.249±0.01<br />
***<br />
p
and their coleoptiles and primary leaves were sprayed with the same c<strong>on</strong>centrati<strong>on</strong> SA<br />
soluti<strong>on</strong> or with water in the case of c<strong>on</strong>trol lot. The value for the c<strong>on</strong>trol lot was<br />
c<strong>on</strong>sidered 100%.<br />
CONCLUSION<br />
The exogenous 0.01 mM, 0.1 mM and 1.0 mM SA soluti<strong>on</strong>s treatments significantly<br />
increased the assimilatory pigments c<strong>on</strong>tents in leaves of wheat plantlets (Triticum<br />
aestivum) when the leaves and the coleoptiles was sprayed with this soluti<strong>on</strong>, in<br />
comparis<strong>on</strong> with the same parameters measured in leaves of wheat plantlets of the<br />
c<strong>on</strong>trol lot treated with water. The 0.1mM SA soluti<strong>on</strong> produced the highest increased<br />
(with 40% from the c<strong>on</strong>trol lot for chlorophyll a, with 36% from the c<strong>on</strong>trol lot for<br />
chlorophyll b and with 50% from the c<strong>on</strong>trol lot for carotenoid pigments).<br />
Salicylic acid, in the used c<strong>on</strong>centrati<strong>on</strong>, activated the synthesis of carotenoids,<br />
xanthophylls and the level of clorophyll pigments, in primary leaves of wheat plantlets.<br />
REFERENCES<br />
Anandhi, S., Ramanujam, M. P., 1997, Effect of salicylic acid <strong>on</strong> black gram (Vigna<br />
mungo) cultivars, Ind. J. Plant Physiol., 2, p138-141..<br />
Fariduddin, Q., Hayat , S., and Ahmad, A., 2003, Salicylic acid influences net<br />
photosynthetic rate, carboxylati<strong>on</strong> efficiency, nitrate reductase activity and seed<br />
yield in brasssica juncea, Photosynthetica, 41, p 281-284.<br />
Ghai, N.,Setia, R.C., 2002, Effect of paclobutrazol and salicylic acid <strong>on</strong> chlorophyll<br />
c<strong>on</strong>tent, hill activity and yield comp<strong>on</strong>ents in Brassica Napus L (cv. GSL-1),<br />
Phytomorphol., 52, p 83-87.<br />
Gross,D.,Parthier, B., 1994, Novell natural substances acting in plant growth<br />
regulati<strong>on</strong>, J.of Plant growth regulati<strong>on</strong>, 13, p 93-114.<br />
Hayat, S., Fariduddin, Q., Ali, B., and Ahmad, A., 2005, Effects of salicylic acid <strong>on</strong><br />
growth and enzymes activities of wheat caryopsislings, Acta.Agr<strong>on</strong>.Hung., 53, p<br />
433-437.<br />
Hayat S., Ahmad A., 2007, Salicylic Acid: A Plant Horm<strong>on</strong>e, SPRINGER (Ed)<br />
Dortrecht, The Netherlands.<br />
Khan, W., Prithiviraj, B., Smith, D.L, 2003, Photosynthetic resp<strong>on</strong>ses of corn and<br />
soybean to foliar applicati<strong>on</strong> of salicylates, Journal of Plant Physiology, 160, p<br />
485-492.<br />
Moharekar, S.T., Lokhande, S.D., Hara, T., Tanaka, R., Tanaka, A., and Chavan,<br />
P.D., 2003, Effect of salicylic acid <strong>on</strong> clorophyll and carotenoid c<strong>on</strong>tents of wheat<br />
and mo<strong>on</strong>g caryopsislings, Photosynthetica, 41, p 315-317.<br />
Moran, R., Porath, D., 1980, Clorophyll de<strong>term</strong>inati<strong>on</strong> in intact tissue using N,N-<br />
dimethylformamide, Plant Physiol, 65, p 478-479.<br />
Moran, R., 1982, Formulae for de<strong>term</strong>ianti<strong>on</strong> of chlorophyllous pigments extracted<br />
with N,N- dimethylformamide, Plant Physiol. 69 (6), p 1376-1381.<br />
Raskin I., 1992, Salicylate, a new plant horm<strong>on</strong>e, Plant Physiol., 99, p.799-803.<br />
Xue Jianping, Zhang Aimin, Fang Zh<strong>on</strong>gming, Sheng Wei, 2006, The effect of<br />
Salicylic acid <strong>on</strong> the growth of Pinellia Ternate, The 11 th <str<strong>on</strong>g>Internati<strong>on</strong>al</str<strong>on</strong>g> C<strong>on</strong>gress of<br />
Plant Tissue culture & Biotechnologie, China, p 96.<br />
496
TREE GROWTH EQUATIONS IN DIFFERENT SITES FOR MIXED STANDS<br />
OF SESSILE OAK WITH BEECH FROM THE MIDDLE BASIN OF THE<br />
CRISUL REPEDE RIVER<br />
Sorin Dorog<br />
Faculty of Envir<strong>on</strong>mental Protecti<strong>on</strong>, University of Oradea<br />
ABSTRACT<br />
The de<strong>term</strong>ining of some growth equati<strong>on</strong>s, which comprise simultaneously increase<br />
factors and diameters in relative values, is of interest because these can be used<br />
afterwards for stands which are similar in structure and vegetate in the same<br />
envir<strong>on</strong>mental c<strong>on</strong>diti<strong>on</strong>s.<br />
The paper presents three equati<strong>on</strong>s for three types of sites mostly encountered in<br />
mixed stands of sessile oak with beech from the middle basin of the Crisul Repede<br />
river. These equati<strong>on</strong>s are representative for the types of sites presented, their applying<br />
in other areas of the country and eventually, in other types of sites, presents verrifyings<br />
and ulterior adjustments of the mathematical pattern. The paper also presents the<br />
intervals of the diameter categories for which these equati<strong>on</strong>s are viable..<br />
Key words: equati<strong>on</strong>s, tree growth, mixed stands, beech, sessile oak.<br />
1. INTRODUCTION<br />
The dendrometrical characteristics of the even-aged stands present a larger or smaller<br />
variability in relati<strong>on</strong> with the species and the envir<strong>on</strong>mental c<strong>on</strong>diti<strong>on</strong>s. The diameters<br />
have according to the experts in this field varioti<strong>on</strong> factors comprised between 20 – 30<br />
%, this variability being caused by radial increases (Giurgiu V., 1967).<br />
The stands formed by light demanding species are characterised by high<br />
correlati<strong>on</strong> coefficients compared to shade and half – shade demanding species (Leahu<br />
I., 1994). Relatively even – aged stands have a decrease of the radial growth rythm for<br />
the superior diametre categories (Giurgiu V., 1979). In such circumstances there is a<br />
difference between radial growth average <strong>on</strong> each stand and the radial growth of the<br />
medium diameter category.<br />
The liniarity after which the variati<strong>on</strong> of the radial growth can be ajusted in relati<strong>on</strong><br />
with the diameters at even – aged stands, was dem<strong>on</strong>strated an emphasized by the F test.<br />
The de<strong>term</strong>ining of some growth equati<strong>on</strong>s, which can comprise at the same time<br />
growth factors and diameters in relative values, shows interest because of the easy way<br />
with which these can be used afterwards at stands, which are not <strong>on</strong>ly structurally<br />
similar, but also having similar envir<strong>on</strong>mental c<strong>on</strong>diti<strong>on</strong>s (Cook E., 1985).<br />
Once these equati<strong>on</strong>s have been established, they provide the users accurate results<br />
c<strong>on</strong>cerning the growth and samples are taken with the boring bit from the medium<br />
category of diameters (Cook E., 1990).<br />
This paper was based up<strong>on</strong> the analysis of mixed even – aged and relatively even –<br />
aged stands of sessile oak and beech, in three types of sites, representative with regards<br />
to the offered c<strong>on</strong>diti<strong>on</strong>s and percentage surface. In the studied stands trial squares of<br />
2500 m 2 were placed and in these places all the trees were inventoried, starting with the<br />
8 cm diameter. For the medium diameter tree category, samples were taken with the<br />
Pressler’ s borer (Dorog S., 2006).<br />
497
2. WORKING METHODS<br />
Radial increases have different characteristics, which have had an impact up<strong>on</strong> what<br />
working method was used. Thus, in order to establish the diameter categories from<br />
which samples were taken with the help of the Pressler’s borer, the following issues<br />
were taken into account:<br />
• For the same category of diameters, the variati<strong>on</strong> coefficients of growth have<br />
reduced values, comprised between 20-30%;<br />
• In the same diameter category, the species has little influence up<strong>on</strong> the variati<strong>on</strong><br />
factors;<br />
• The variati<strong>on</strong> coefficients of the radial growths decrease from the inferior diameter<br />
categories to the superior <strong>on</strong>es, as a size order, these are situated between 20-25%;<br />
• Within the same category of diameters, the variati<strong>on</strong> amplitude of the growths is<br />
comprised somewhere in between 0,1 and 1,9;<br />
• The variati<strong>on</strong> coefficients of growth for the whole stand are larger, with regards to<br />
values, than the variati<strong>on</strong> factors established for the diameter categories, superior<br />
to the medium diameter of the stand (Giurgiu V., 1972).<br />
The selecti<strong>on</strong> of the trees, for the taking of the growth sample, was made as<br />
follows(Dorog S., 2006):<br />
• all the trees from the trial square were inventoried, established from the 8 cm<br />
diameter upwards;<br />
- the medium diameter of the trees from the test area was established;<br />
- the trees, from which samples were to be taken, were selected; these trees<br />
were chosen so that they would be a part of diameter categories, with values around the<br />
arithmetic average.<br />
The samples obtained with the Pressler’s borer have necessitated preliminary<br />
procedures, before their actual measurement. The preliminary procedures were the<br />
following:<br />
• the c<strong>on</strong>versi<strong>on</strong> through grinding with abrasive paper (granulati<strong>on</strong> 180) of the<br />
samples;<br />
• establishing by visual analysis which parts from the sample come from the inside<br />
and which from the outside of the radial secti<strong>on</strong>.<br />
The c<strong>on</strong>versi<strong>on</strong> and measurement of the samples included the following steps (Dorog<br />
S., 2006):<br />
• the scaning of the samples with a specialised program, sec<strong>on</strong>dary to the scaner and<br />
saving he pictures in a *bmp format.<br />
• the annual rings were established by means of a specialised programme of picture<br />
analysis;<br />
498
Image no.1 ImageTool programme was used to get the width of the annual rings<br />
• the width of the annual rings was obtained with the Distance opti<strong>on</strong> from the<br />
ImageTool programme;<br />
• saving the results and transforming the size of the annual rings from dpi into<br />
milimeters.<br />
For this process of transformati<strong>on</strong> of the pixels into milimeters, the following pattern<br />
was used: Ring width (mm) = 0,042333*Ring width (dpi)<br />
The equati<strong>on</strong> coefficients (Leahu I., 1990) were established by the following steps:<br />
• The obtaining of the medium growth for each particular tree;<br />
• Establishing the growth average for all the samples taken from a type of<br />
envir<strong>on</strong>ment;<br />
• The establishing of the medium diameter for the diameter categories and within<br />
these, there were trees from which samples were taken with Pressler’s borer;<br />
• The calculating of the relative values for the entire string of diameter values and of<br />
the relative values of the medium growths, respectively.<br />
3. RESULTS AND CONCLUSIONS<br />
On the basis of the algorythm presented previously, after the calculati<strong>on</strong>s, the following<br />
values of the regressi<strong>on</strong> equati<strong>on</strong> coefficients were obtained. These coefficients are<br />
presented in the table below.<br />
Table nr. 1Regressi<strong>on</strong> equati<strong>on</strong> coefficients and the string of values which they are<br />
suitable for<br />
The linear regressi<strong>on</strong> equati<strong>on</strong> coefficients<br />
(y = ax+b)<br />
Type of site<br />
Coefficients Limit diameters<br />
a b Minimum maximum<br />
5153 0,0021 0,8486 16 40<br />
5242 0,0462 0,7431 34 66<br />
5243 0,0059 0,8274 24 64<br />
499
In the linear equati<strong>on</strong> y represents the radial relative increases and x represents the<br />
relative values of the diameters. The relative values for both characteristics were<br />
established in relati<strong>on</strong> with their medium values.<br />
The images corresp<strong>on</strong>ding to each type of envir<strong>on</strong>ment were made simultaneously with<br />
the values of the a and b coefficients.<br />
Picture nr.2. The variati<strong>on</strong> of the relative increases in relati<strong>on</strong> with the relative<br />
values of the diameters<br />
relative increases /<br />
relative diameters<br />
Valorile cresterilor<br />
relative<br />
The variati<strong>on</strong> of the relative increases and of<br />
the relative diameter patterns<br />
3<br />
2<br />
1<br />
0<br />
1 8 15 22 29 36 43 50 57 64<br />
number of observati<strong>on</strong>s<br />
Variatia cresterilor relative radiale in raport cu<br />
Valorile diametrelor relative<br />
2,5<br />
2<br />
1,5<br />
1<br />
0,5<br />
0<br />
1<br />
12<br />
23<br />
34<br />
45<br />
56<br />
67<br />
78<br />
89<br />
100<br />
111<br />
Valorile diametrelor relative<br />
Adatsor1<br />
Adatsor2<br />
Picture nr. 3. The variati<strong>on</strong> of the relative increases and of the relative diameter<br />
patterns at stands from the 5243 type of site<br />
500<br />
122<br />
133<br />
144
Following the results obtained, these c<strong>on</strong>clusi<strong>on</strong>s occur:<br />
• The present equati<strong>on</strong>s go with the pattern of variati<strong>on</strong> of the previously presented<br />
diameters<br />
• The most significant variati<strong>on</strong>s of the radial growths occur in mixed stands<br />
• The equati<strong>on</strong>s can be used in the previously established c<strong>on</strong>diti<strong>on</strong>s to establish<br />
relative radial growths at other stands, which vegetate in similar envir<strong>on</strong>mental<br />
c<strong>on</strong>diti<strong>on</strong>s in the middle basin of the Crisul Repede river<br />
• The equati<strong>on</strong>s are viable <strong>on</strong>ly locally, in other areas, they need to be verified by<br />
further research<br />
• These equati<strong>on</strong>s were elaborated for even-aged stands, their applying to relatively<br />
even-aged stands must be verified.<br />
REFERENCES<br />
1. Cook E., 1990, A c<strong>on</strong>ceptual linear aggregate model for tree rings, In Cook, E.R.<br />
Kairiukstis L.A. (eds). Methods of dendrochr<strong>on</strong>ology. Applicati<strong>on</strong>s in the<br />
envir<strong>on</strong>mental sciences, Kluwer Academic Publishers, Dordrecht 98-104<br />
2. Cook E., 1985, A time series analysis aproach to tree ring standardizati<strong>on</strong>, School<br />
of renewable natural resources, The university of Ariz<strong>on</strong>a<br />
3. Dorog S., 2006, C<strong>on</strong>siderati<strong>on</strong> up<strong>on</strong> the increment of sessile oak and commun<br />
beech forests in the middle watershed of Crişul Repede river, Natural resources and<br />
sustainable development pag. 857-864 ISBN (10) 973-759-158-5, ISBN (13) 978-<br />
973-759-158-6, HU ISBN-10: 963-9274-99-2, HU ISBN-13: 978-963-9274-99-0<br />
4. Dorog S., 2006, De<strong>term</strong>inarea unor ecuaţii pentru creşterea arboretelor amestecate<br />
de gorun cu fag din bazinul mijlociu al Crişului Repede, Analele Universităţii din<br />
Oradea, Fascicula Silvicultură vol XI, pag. 129-134 ISSN. 1453-9489<br />
5. Dorog S., 2006, Compararea creşterii goruneto-făgetelor din bazinul mijlociu al<br />
Crişului Repede cu tabelele de producţie româneşti, Analele Universităţii din<br />
Oradea, Fascicula Silvicultură vol XI, pag. 135-144 ISSN. 1453-9489<br />
6. Giurgiu V., 1979, Dendrometrie şi auxologie forestieră, Editura Ceres, Bucureşti<br />
7. Giurgiu V., 1972, Metode ale statisticii matematice aplicate în silvicultură, Editura<br />
Ceres, Bucureşti<br />
8. Giurgiu V., 1967, Studiul creşterilor la arborete, Editura Agro-silvică, Bucureşti<br />
9. Leahu I.,1990, O modalitate de exprimare a relaţiilor interspecifice în raport cu<br />
particularităţile auxologice ale arboretelor, în: Metode şi tehnologii moderne în<br />
cultura şi exploatarea pădurilor, Universitatea din Braşov<br />
10. Leahu I., 1994, Dendrometrie, Editura Didactică şi Pedagogică, Bucureşti<br />
11. Leahu, I.,1994, Metode şi modele biometrice aplicate în dendrometrie,<br />
Universitatea “Transilvania” din Braşov<br />
501
THE SIMULATION AND EVALUATION OF A BREAK IN MIXED<br />
STANDS OF BEECH WITH SESSILE OAK<br />
Sorin Dorog<br />
Faculty of Envir<strong>on</strong>mental Protecti<strong>on</strong>, University of Oradea<br />
ABSTRACT<br />
The work discusses the simulati<strong>on</strong> and evaluati<strong>on</strong> of a break for mixed stands of sessile<br />
oak(Quercus petraea) with beech(Fagus sylvatica). Through this break, the structure of<br />
the stand is modified and, to this extent, the work wants to establish the optimal<br />
structure through successive simulati<strong>on</strong>s.<br />
The structure of the stands offers complex informati<strong>on</strong> with regards to the<br />
programme and the future development of the stand, thus, knowing the structure and the<br />
way the break influences it, becomes very important. This paper analysed the<br />
experimental distributi<strong>on</strong>s, obtained <strong>on</strong> the basis of the field data , in comparis<strong>on</strong> with<br />
those recommended in other works of speciality (normal distributi<strong>on</strong>, Charlie’s<br />
distributi<strong>on</strong> and Beta distributi<strong>on</strong>).<br />
Key words: structure, distributi<strong>on</strong>, mixed stands, beech, sessile oak<br />
1. INTRODUCTION<br />
The structure of the stands is the <strong>on</strong>e that gives the most complex informati<strong>on</strong> about the<br />
programme of the stand and about its future development, if essential<br />
changes(unexpected) do not occur, changes that can modify entirely the initial<br />
programme. In this way, knowing the structure of the stands becomes extremely<br />
important, also from the perspective of the future breaks. In order to have a point of<br />
refference, this paper analysed experimental distributi<strong>on</strong>s, obtained from the field data,<br />
in relati<strong>on</strong> to the distributi<strong>on</strong>s recommended in other works.<br />
Applying different silvotechnical measurement systems influences not <strong>on</strong>ly the<br />
size of the growth, but also the quality of the wood producti<strong>on</strong>, that is the relati<strong>on</strong>ship<br />
between the volume of thick or thin trees, bel<strong>on</strong>ging to different species – which are<br />
harvested and of various quality and also the efficiency of the social and ecological<br />
functi<strong>on</strong>s, that the stands have to fulfill (Leahu I.,2001). C<strong>on</strong>sidering as representative<br />
for this area the even-aged mixed stands or relatively even-aged stands of beech with<br />
sessile oak, we will try to define the stands in c<strong>on</strong>necti<strong>on</strong> with the proporti<strong>on</strong> of the<br />
species and also with the structure of the stands <strong>on</strong> diameter categories.<br />
The simulati<strong>on</strong> of the break offers informati<strong>on</strong> about the directi<strong>on</strong> up<strong>on</strong> which the<br />
stand has to be taken in order to improve its compositi<strong>on</strong>, structure, the optimal<br />
functi<strong>on</strong>ing of its given attributes and its guidance towards the domestic aims,<br />
established through a management plan.<br />
2. WORKING METHOD<br />
To evaluate the structure of mixed stands of sessile oak with beech, some stands were<br />
selected, stands which vegetate in proper c<strong>on</strong>diti<strong>on</strong>s with regards to envir<strong>on</strong>mental<br />
c<strong>on</strong>diti<strong>on</strong>s for the two species menti<strong>on</strong>ed above.<br />
The stands were chosen so that situati<strong>on</strong>s, in which they were partially or totally<br />
derrived, would be avoided. The field was devised in rectangular test areas of 2.500 sq.<br />
502
ms. The placement of the areas was selected in such a way so as to render the structure<br />
of the stand as accurately as possible.<br />
Within each area the diameters of all the trees from different species were<br />
measured and also the heights of the trees at the medium central diameter, for each<br />
particular species. After collecting the data from the field, the structure of the stand, for<br />
every species and diameter categories were discussed. With the help of the data, the<br />
structure of the stands was evaluated according to the diameter categories, using<br />
distributi<strong>on</strong>s recommended by specialists in this field: Gauss distributi<strong>on</strong>, A type<br />
Charlier distributi<strong>on</strong> and Beta distributi<strong>on</strong> (Giurgiu V., 1979, Giurgiu V., 1972).<br />
Hi-square criterium was applied to assess the quality of the adjustments <strong>on</strong> the<br />
experimental distributi<strong>on</strong>, after the theoretical <strong>on</strong>es. After the structure of the stands was<br />
revealed, there was a simulati<strong>on</strong> of some breaks(thinnings), through which it was<br />
intended to promote the most valuable pieces of stand, in what species and quality were<br />
c<strong>on</strong>cerned, to improve the quantitative producti<strong>on</strong> and especially the qualitative<br />
<strong>on</strong>e(Florescu I., Nicolescu N., 1998).<br />
The simulati<strong>on</strong> of a break has been accomplished relying <strong>on</strong> the data from the<br />
field and also by using data, obtained <strong>on</strong> the basis of the adjustment of experimental<br />
distributi<strong>on</strong>s with those recommended by the specialists. The quality of the simulated<br />
break was verified by calculating the dispersi<strong>on</strong> indicators, the indicators of the form of<br />
the distributi<strong>on</strong>, the intensity of the breaks c<strong>on</strong>nected to the number of trees, the basic<br />
area and the volume, Hart-Becking index (spacing factor) before and after the break.<br />
The volume was calculated with the help of the relative volume method, a method<br />
which is recommended by the specialised literature.<br />
The structural differentiati<strong>on</strong> in a stand is a c<strong>on</strong>sequence of the integrality state and<br />
leads, eventually, to the self-thinning of the trees, following the self-improvement trait<br />
of the biological systems.<br />
3. RESULTS AND DISCUSSIONS<br />
To describe the simulati<strong>on</strong> and the evaluati<strong>on</strong> of a break in a stand, some synthetical<br />
data is presented in the following lines, data which has been calculated before and after<br />
the simulated break in a.u. 78C U.P.III, F.D. Dobreşti.<br />
Table no. 1 The calculating of the medium diameter, dispersi<strong>on</strong> indicators and of the<br />
distributi<strong>on</strong> form case 1<br />
Calculated characteristics Before the break After the break<br />
Medium diameter(cm) 22,04188482 26<br />
Variance 67,44612783 65,92<br />
Standard – Deviati<strong>on</strong> 8,212559152 8,1191133<br />
Variati<strong>on</strong> factor 0,372588788 0,3122736<br />
Asimetry -0,142394868 -0,9295828<br />
Excess 1,89798843 2,5502924<br />
From the above data <strong>on</strong>e can notice that the simulated break c<strong>on</strong>sisted of the extracti<strong>on</strong><br />
of the trees from the inferior diameter categories, mostly white beech trees, but also<br />
503
sessile oak and dying oak trees, which have remained in the inferior c<strong>on</strong>tent. The tree<br />
number intensity is close to the value of 32%, fact which indicates a moderate to str<strong>on</strong>g<br />
thinning , while the volume intensity is somewhere around 11% , which points to a low<br />
intensity thinning. Thus it can be c<strong>on</strong>cluded that a low thinning was simulated in the<br />
stand. The spacing factor modifies itself also, meaning that the experimental value<br />
increases after the break by 3%.<br />
Table no. 2 Species proporti<strong>on</strong> case 1<br />
Basic area propoti<strong>on</strong> Volume proporti<strong>on</strong> Tree number<br />
Species<br />
Before After Before After<br />
proporti<strong>on</strong><br />
Before After<br />
break break break break break break<br />
FA 33,60 34,98 35,32 39,91 27,96 41,27<br />
GO 42,29 43,69 43,74 49,06 26,88 38,89<br />
CA 20,51 7,23 17,37 7,01 43,01 12,70<br />
ST 3,60 3,75 3,56 4,02 2,15 3,17<br />
CE 0,00 10,36 0,00 0,00 0,00 3,97<br />
CI 9,95 0,00 8,30 0,00 2,69 0,00<br />
Table no. 3 The calculati<strong>on</strong> of the break intensity and of Hart-Becking index case 1<br />
Calculated characteristics Values<br />
Tree number intensity 32,25806452<br />
Basic area intensity 3,924714353<br />
Volume intensity 11,48726754<br />
Spacing factor ( Hart-Becking index)<br />
Before break<br />
14,273994<br />
After break<br />
17,3427<br />
Numar de arbori (buc)<br />
Distributia experimentala, distributia dupa<br />
simularea interventiei si ajustarea acestora dupa<br />
distributia normala, Charlier tip A,Beta<br />
20<br />
18<br />
16<br />
14<br />
12<br />
10<br />
8<br />
6<br />
4<br />
2<br />
0<br />
d8<br />
d12<br />
d16<br />
d20<br />
d24<br />
d28<br />
d32<br />
d36<br />
d40<br />
d44<br />
Categorii de diametre (cm)<br />
Distrib exp. Distrib. Sim interv..(ni) ni distrib. Norm.<br />
ni Charlier tip A ni Beta<br />
Image no 1 – Experimental distributi<strong>on</strong>, the distributi<strong>on</strong> after the simulati<strong>on</strong> of the<br />
break and adjusting after the normal law, Charlier type A and Beta, case 1<br />
504<br />
d48<br />
d52<br />
d56
Another possible simulated break in the stand from the a.u. 78C is the <strong>on</strong>e in which<br />
acti<strong>on</strong> is taken up<strong>on</strong> the trees from the inferior diameter category, as well as up<strong>on</strong> the<br />
trees from the medium and superior diameter categories. Thus, the obtained data is<br />
shown as follows:<br />
Table no. 4 The calculati<strong>on</strong> of the medium diameter, dispersi<strong>on</strong> and distributi<strong>on</strong><br />
form indicators, case 2<br />
Calculated characteristics Before break After break<br />
Medium diameter(cm) 22,04188482 21,338583<br />
Variance 67,44612783 50,105542<br />
Standard – Deviati<strong>on</strong> 8,212559152 7,0785268<br />
Variati<strong>on</strong> Factor 0,372588788 0,3317243<br />
Asimetry -0,142394868 0,1765171<br />
Excess 1,89798843 2,2001468<br />
Table no. 5 Species proporti<strong>on</strong> case 2<br />
Basic area proporti<strong>on</strong> Volume proporti<strong>on</strong> Tree number<br />
Species<br />
Before After Before After<br />
proporti<strong>on</strong><br />
Before After<br />
break break break break break break<br />
FA 33,60 33,26 35,88 35,15 27,96 35,43<br />
GO 42,29 43,40 44,43 45,70 26,88 29,92<br />
CA 20,51 20,46 16,08 16,23 43,01 33,07<br />
ST 3,60 2,88 3,62 2,92 2,15 1,57<br />
CE 0,00 0,00 0,00 0,00 0,00 0,00<br />
CI 9,95 0,00 8,43 0,00 2,69 0,00<br />
Table no. 6 The calculati<strong>on</strong> of the break intensity and of Hart-Becking index case 2<br />
Calculated characteristics Values<br />
Tree number intensity 31,72043011<br />
Basic area intensity 36,4963218<br />
Volume intensity 36,99021667<br />
Spacing factor ( Hart-Becking index)<br />
Before break<br />
14,273994<br />
After break<br />
17,274286<br />
In this simulated break it can be noticed that the medium diameter does not grow<br />
artificially, as seen in the first case, which emphasizes that by breaking in all diameter<br />
categories, the medium diameter of the stand remains aproximately c<strong>on</strong>sistent. Further<br />
more, in this case, the diffferences related to tree number, basic area and volume<br />
intensities are not too big.<br />
505
Table no. 7 The calculati<strong>on</strong> of the medium diameter, dispersi<strong>on</strong> and distributi<strong>on</strong> form<br />
indicators , case 3<br />
Calculated characteristics Before break After break<br />
Medium Diameter(cm) 22,04188482 23,173913<br />
Variance 67,44612783 65,020477<br />
Standard deviati<strong>on</strong> 8,212559152 8,0635276<br />
Variati<strong>on</strong> factor 0,372588788 0,3479571<br />
Asimetry -0,142394868 -0,2028422<br />
Excess 1,89798843 2,0031626<br />
Table no. 8 Species proporti<strong>on</strong> case 3<br />
Basic area proporti<strong>on</strong> Volume proporti<strong>on</strong> Tree number<br />
Species<br />
Before After Before After<br />
proporti<strong>on</strong><br />
Before After<br />
break break break break break break<br />
FA 33,60 34,62 35,32 35,73 27,96 36,23<br />
GO 42,29 47,54 43,74 48,55 26,88 34,78<br />
CA 20,51 13,51 17,37 11,49 43,01 26,09<br />
ST 3,60 4,33 3,56 4,23 2,15 2,90<br />
CE 0,00 0,00 0,00 0,00 0,00 0,00<br />
CI 9,95 0,00 8,30 0,00 2,69 0,00<br />
Table no. 9 The calculati<strong>on</strong> of the break intensity and of Hart-Becking index case 3<br />
Calculated characteristics Values<br />
Tree number intensity 25,80645161<br />
Basic area intensity 16,79057756<br />
Volume intensity 15,80365993<br />
Spacing factor ( Hart-Becking index)<br />
Before break<br />
14,273994<br />
After break<br />
16,571523<br />
The last case describes a str<strong>on</strong>g thinning(5), which is recommendable in the actual<br />
situati<strong>on</strong>, because of the fact that an intensity, that is too big, could destablize the<br />
internal structure of the stand.<br />
This pattern for the simulati<strong>on</strong> and evaluati<strong>on</strong> of the break puts at the user’s disposal<br />
also the comparis<strong>on</strong> of experimental distributi<strong>on</strong>s with those recommended as normal<br />
by the speciality works(1,2). In c<strong>on</strong>tinuati<strong>on</strong>, the distributi<strong>on</strong> formulas of the tree<br />
number with regards to quality grades, for the above examples, are presented:<br />
- Experimental distributi<strong>on</strong> after Beta<br />
, 3<br />
0446 , 1 2<br />
0, 01592 × 10 x − 7 57 − x<br />
y = ( ) ( ) 7517<br />
- distributi<strong>on</strong> after break case 1<br />
0153 , 2 2<br />
436, 064 × 10 x − 7 57 − x<br />
y = ( ) ( ) 9197 , 3<br />
506
- distributi<strong>on</strong> after break case 2<br />
y = ( ) ( ) 1281 , 2<br />
0706 , 1 2<br />
10, 3337 × 10 x − 7 43 − x<br />
- distributi<strong>on</strong> after break case 3<br />
y = ( ) ( ) 3354 , 2<br />
0882 , 1 2<br />
2, 9207 × 10 x − 7 49 − x<br />
in which x represents the diameter categories and y the number of trees corresp<strong>on</strong>ding<br />
to each diameter category.<br />
Numar de arbori (buc)<br />
Distributia experimentala, distributia dupa<br />
simularea interventiei si ajustarea acestora dupa<br />
distributia normala, Charlier tip A,Beta<br />
20<br />
18<br />
16<br />
14<br />
12<br />
10<br />
8<br />
6<br />
4<br />
2<br />
0<br />
d8<br />
d12<br />
d16<br />
d20<br />
d24<br />
d28<br />
d32<br />
Distrib exp.<br />
Categorii de diametre (cm)<br />
Distrib. Sim interv..(ni) ni distrib. Norm.<br />
ni Charlier tip A ni Beta<br />
Image 2 – Experimental distributi<strong>on</strong>, the distributi<strong>on</strong> after the simulati<strong>on</strong> of the<br />
break and the adjusting after the normal law, Charlier type A and Beta, case 2<br />
4. CONCLUSIONS<br />
From the c<strong>on</strong>verted data in the mixed stands of beech with sessile oak there are<br />
interesting aspects in relati<strong>on</strong> to both the interspecific competiti<strong>on</strong> between the two<br />
species and between these and the comm<strong>on</strong> hornbeam, the lime-tree and other species<br />
of mixed hardwood. In order to develop the stand according to the management, aims<br />
established through a working-plan, the silvotechnical breaks will follow the change in<br />
the compositi<strong>on</strong> and structure of the stands in the directi<strong>on</strong> given by the established<br />
social-ec<strong>on</strong>omical and ecological objectives.<br />
The pattern for the evaluati<strong>on</strong> and the simulati<strong>on</strong> of the break in a stand has the<br />
advantages that, before the break is performed, informati<strong>on</strong> related to the tree number<br />
structure, the basic area, volume, species proporti<strong>on</strong>, Hart-Becking coefficient values is<br />
available, which allows for the possibility of analysing and improving of the break<br />
according to the objectives established by the management plan.<br />
507<br />
d36<br />
d40<br />
d44<br />
d48<br />
d52<br />
d56
The value of the spacing factor increases with each thinning, its value can modify with<br />
up to 5% at each break (Florescu I., 1998). The data presented as examples in the<br />
previous pages support our point.<br />
The simulati<strong>on</strong> of a low thinning is reckognised by the fact that tree number<br />
intensities of over 25-30% occur, while in what the basic area and the volume are<br />
c<strong>on</strong>cerned, they are situated between 5-10%, which is normal, if we take into account<br />
that trees from the inferior categories are removed, trees left behind with regards to their<br />
growth and placed in the lower c<strong>on</strong>tent. The remaining sessile oak or Turkey oak trees<br />
in the lower c<strong>on</strong>tent are declined at the tree-foot and they are the first to be<br />
recommended for extracti<strong>on</strong> through tending processes. The combined thinning implies<br />
the removal of the trees from all diameter categories, this can be noticed by the fact that<br />
number of trees, basic area and volume intensities, respectively, are aproximately equal,<br />
or differing by no more than 4 or 5%.<br />
REFERENCES<br />
1. Chiţea Gh., 1997, Biostatistică, Editura Universităţii Transilvania, Braşov<br />
2. Chiţea Gh., 2001, Biostatistică, Editura Universităţii Transilvania, Braşov<br />
3. Cristea Maria, 2004, Riscurile climatice din Bazinul hidrografic al Crişurilor,<br />
Editura Abaddaba Oradea<br />
4. Florescu I., Nicolescu N., 1998, Silvicultura, vol. I – Silvobiologia, Editura<br />
Universităţii Transilvania, Braşov<br />
5. Florescu I., Nicolescu N., 1998, Silvicultura, vol. II – Silvotehnica, Editura<br />
Universităţii Transilvania, Braşov<br />
6. Giurgiu V., 1979, Dendrometrie şi auxologie forestieră, Editura Ceres, Bucureşti<br />
7. Giurgiu V., 1972, Metode ale statisticii matematice aplicate în silvicultură, Editura<br />
Ceres, Bucureşti<br />
8. Leahu I., 1994, Dendrometrie, Editura Didactică şi Pedagogică, Bucureşti<br />
9. Leahu I., 2001, Amenajarea pădurilor, Editura Didactică şi Pedagogică, Bucureşti<br />
508
THE FREQUENCY OF DAYS WITH DIFFERENT TEMPERATURES IN THE<br />
CRISUL REPEDE HYDROGRAPHICAL BASIN<br />
Ana Cornelia Moza<br />
University of Oradea, Faculty for Envir<strong>on</strong>mental Protecti<strong>on</strong><br />
ABSTRACT<br />
The particularities of the air temperature’s regime are indicated by a series of elements,<br />
calculated or extracted graphically from the four stati<strong>on</strong>s histograms. Some of these<br />
elements are: frequency of frosty nights, frequency of frosty days, winter days, summer<br />
days and tropical days. These elements were analyzed during 1970-2005 at all 4<br />
stati<strong>on</strong>s.<br />
INTRODUCTION<br />
The air temperature presents a series of particularities: depending <strong>on</strong> the altitude,<br />
described by the decrease of the air temperature with the rise of altitude, and depending<br />
<strong>on</strong> the orientati<strong>on</strong> and the gradient of the slopes, described by their different warming.<br />
While describing the climate of a regi<strong>on</strong>, the focus is <strong>on</strong> m<strong>on</strong>thly and annual<br />
frequency of days and nights over or under certain characteristic limits, represented in<br />
number of days.<br />
Over the year, the air temperature drops below or rises above certain limits,<br />
according to which there were assigned frosty nights, frosty days, winter days, summer<br />
days and tropical days.<br />
MATERIAL AND METHOD<br />
In order to point out the characteristics of the temperature, the data during 1970-2005<br />
were used, obtained from the observati<strong>on</strong>s made at the four meteorological stati<strong>on</strong>s<br />
(Oradea, Săcueni, Borod, Stâna de Vale), situated both in plain areas and mountain<br />
areas, so that they cover the entire researched area.<br />
The number of days with certain temperatures is extracted from the meteorological<br />
tables for every m<strong>on</strong>th. Adding the respective values and dividing the result to the<br />
number of m<strong>on</strong>ths in the investigated years, we obtain the average number of days in a<br />
m<strong>on</strong>th with characteristic temperatures, which is graphically represented by bars.<br />
RESULTS AND DISCUSSIONS<br />
1. The frequency of frosty nights<br />
In the cold seas<strong>on</strong>, the nocturnal cooling reaches the highest values, so that the<br />
minimum air temperatures may drop below -10ºC, situati<strong>on</strong> in which the nights are<br />
c<strong>on</strong>sidered frosty.<br />
In the Crisul Repede hydrographical basin, the frequency of frosty nights rises<br />
directly proporti<strong>on</strong>al to the altitude from 11-12 cases in Săcueni, Oradea, up to 52 cases<br />
in Stâna de Vale ( because of the flow of cold air from surrounding peaks and its<br />
stagnati<strong>on</strong> <strong>on</strong> the bottom of the depressi<strong>on</strong> for several days in a row) (fig.1).<br />
Through out a year, frosty nights are recorded at Stâna de Vale meteorological<br />
stati<strong>on</strong> from October until April, while at the other stati<strong>on</strong>s (Borod, Oradea, Săcueni)<br />
509
the frosty nights begin in November and last until March. The maximum number of<br />
frosty nights is recorded in January: 4.7 and 4.9 days in Săcueni and Oradea, 6.4 days in<br />
Borod and 14.9 days in Stâna de Vale (fig.1).<br />
55<br />
50<br />
45<br />
40<br />
35<br />
30<br />
25<br />
20<br />
15<br />
10<br />
5<br />
0<br />
Figure no.1: Variati<strong>on</strong> of the number of days with<br />
minimum temperature ≤10ºC (frosty nights)<br />
I II III IV V VI VII VIII IX X XI XII An<br />
Borod<br />
Oradea<br />
Sacueni<br />
Stâna de Vale<br />
2. The average frequency of freezing days (minimum temperature≤0ºC)<br />
The freezing days have a higher share from October up to April, but low values (0.1<br />
days) are recorded too from September to May in plain areas and depressi<strong>on</strong>, with<br />
freezing days all year l<strong>on</strong>g in mountain areas.<br />
The frequency of freezing days increases with the rise of altitude, reaching the<br />
maximum value at Stâna de Vale (1108m) with 182 freezing days per year, while<br />
lower, at Borod (333m) the value is 107.8 days per year, dropping to 91.4 days of<br />
freezing days per year at Săcueni (fig.2).<br />
In the studied area, over a 36 years period, under the influence of the<br />
atmosfere’s general circulati<strong>on</strong> and the local c<strong>on</strong>diti<strong>on</strong>s, during the year the highest<br />
number of freezing days appear in January 23-24 days in the plain areas, 25.4 days at<br />
Borod, while at Stâna de Vale the maximum value of 29.6 days of freezing are<br />
recorded. At Stâna de Vale stati<strong>on</strong>, freezing days are found all year l<strong>on</strong>g, but with a<br />
lower frequency of 0.3 days in August and <strong>on</strong>ly 0.1 days in July.<br />
510
200<br />
190<br />
180<br />
170<br />
160<br />
150<br />
140<br />
130<br />
120<br />
110<br />
100<br />
90<br />
80<br />
70<br />
60<br />
50<br />
40<br />
30<br />
20<br />
10<br />
0<br />
Figure no.2: Variati<strong>on</strong> of the number of the frosty days<br />
(minimum temperature ≤ 0ºC) in the Crisul Repede<br />
hydrographical basin during the period 1970-2005<br />
I II III IV V VI VII VIII IX X XI XII An<br />
Borod<br />
Oradea<br />
Sacueni<br />
Stâna de Vale<br />
3. Average frequency of winter days (maximum temperature≤0ºC)<br />
Winter days are recorded from November to March, in the cold seas<strong>on</strong>, their frequency<br />
rises al<strong>on</strong>g with the increasing activities of the Scandinavian, Greenland and especially<br />
the Siberian anticycl<strong>on</strong>e, which c<strong>on</strong>diti<strong>on</strong>s the arctic air masses movement to the South.<br />
511
50<br />
45<br />
40<br />
35<br />
30<br />
25<br />
20<br />
15<br />
10<br />
5<br />
0<br />
Figure no.3: Variati<strong>on</strong> of the medium number of the<br />
winter days (maximum temperature ≤0ºC) in the<br />
Crisul Repede hydrographical basin during the<br />
period 1970-2005<br />
I II III IV V VI VII VIII IX X XI XII An<br />
Borod<br />
Oradea<br />
Sacueni<br />
Stâna de Vale<br />
Throughout the year, the winter days can be produced from November until March at<br />
low altitudes and from October to April at high altitudes. The number of winter days<br />
from the Crisul Repede hydrographical basin reaches the highest value of 48.8 days at<br />
Stâna de Vale meteorological stati<strong>on</strong> and the lowest value of 22.9 days in Oradea.<br />
Throughout the year, the highest frequency of these days is recorded in January 13.9<br />
days at Stâna de Vale, when the entire year’s highest value is reached, while the lowest<br />
number of winter days is recorded in March at Oradea 0.8 days. Winter days also take<br />
place in large numbers in February and December, while starting with March, al<strong>on</strong>g<br />
with the general warming of the atmosphere, to drop significantly at plain stati<strong>on</strong>s and<br />
starting with April to decrease almost totally at Stâna de Vale (1.1 days) (fig.3).<br />
4. Average frequency of summer days (maximum temperature.≥25ºC)<br />
High temperatures are usually recorded in case of anticycl<strong>on</strong>e regime, described by the<br />
invasi<strong>on</strong> of dry c<strong>on</strong>tinental air from medium latitudes or tropical air from Central Asia<br />
or Northern Africa. In these c<strong>on</strong>diti<strong>on</strong>s a very hot weather sets in, accompanied by dry<br />
winds. Usually the summer days settle in during the April-October period.<br />
An average number of 80-90 days per year are recorded in plain meteorological<br />
stati<strong>on</strong>s, 61.1 days in depressi<strong>on</strong>s, while as the altitude increases, the average number of<br />
days drops, reaching at 9.3 days at Stâna de Vale (fig.4).<br />
Over the year, the maximum number of summer days is reached in July (17.8 days<br />
at Borod, 22.8 days at Oradea, 21.7 days at Săcueni) at low altitudes and in August, at<br />
high altitudes, (4 days at Stâna de Vale) (fig. 4). The favourable period for summer days<br />
decreases with the rise of altitude, so that at low altitudes it lasts from March until<br />
512
October, and at higher altitudes (0.3 days at Stana de Vale) until September (0.4 days at<br />
Stâna de Vale).<br />
90<br />
85<br />
80<br />
75<br />
70<br />
65<br />
60<br />
55<br />
50<br />
45<br />
40<br />
35<br />
30<br />
25<br />
20<br />
15<br />
10<br />
5<br />
0<br />
Figure no. 4: Variati<strong>on</strong> of the medium number of<br />
summer days (maximum temperature ≥25ºC) in the<br />
Crişul Repede hydrographical basin during the period<br />
1970-2005<br />
I II III IV V VI VII VIII IX X XI XII An<br />
Borod<br />
Oradea<br />
Sacueni<br />
Stâna de Vale<br />
5. Average frequency of tropical days (maximum temperature.≥30ºC)<br />
High temperatures (≥30ºC) are c<strong>on</strong>diti<strong>on</strong>ed by the movement of dry tropical air masses<br />
within the country’s territory. The tropical days are days in which the maximum air<br />
temperature is greater or equal with 30ºC.<br />
The maximum number of tropical days in the Crisul Repede hydrographical basin,<br />
as an average, is recorded at Oradea, 24.3 days, and at Săcueni, 20.3 days, then they<br />
begin to drop as the altitude rises, 10.3 days at Borod, while at Stâna de Vale they<br />
disappear (fig.5).<br />
The favourable annual interval for producing tropical days takes place between<br />
May and September, with a maximum number of days in August at Borod 4.2 days, in<br />
July at Oradea 9.1 days and 7.6 days at Săcueni (fig.5).<br />
513
25<br />
24<br />
23<br />
22<br />
21<br />
20<br />
19<br />
18<br />
17<br />
16<br />
15<br />
14<br />
13<br />
12<br />
11<br />
10<br />
9<br />
8<br />
7<br />
6<br />
5<br />
4<br />
3<br />
2<br />
1<br />
0<br />
Figure no.5: Variati<strong>on</strong> of the medium number of tropical<br />
days (maximum temperature ≥30ºC in Crişul Repede<br />
hydrogaphical basin during the period 1970-2005<br />
I II III IV V VI VII VIII IX X XI XII An<br />
Borod<br />
Oradea<br />
Sacueni<br />
Stâna de Vale<br />
CONCLUSIONS:<br />
1. The frequency of frosty nights rises directly proporti<strong>on</strong>al with the height in 11-12<br />
cases/year in the plains and up to 52 cases/year in the mountain area. Frosty nights<br />
appear from November until March in the plains and from October to April in the<br />
mountains, with a maximum value in January, when the highest value of 14.9 days<br />
is recorded at Stâna de Vale, and the lowest value is 4.7 days at Săcueni.<br />
2. The frequency of freezing days raises proporti<strong>on</strong>al with the height in 91-96 cases in<br />
the plains, in about 108 cases in the depressi<strong>on</strong> area of Borod and up to 182 cases in<br />
the mountains. Throughout the year the highest number of frosty days is produced<br />
in January, 23-24 days in the plains, 25 days in the depressi<strong>on</strong> area of Borod and<br />
almost 30 days at Stâna de Vale.<br />
3. The frequency of winter days increases as the altitude rises, starting with 23<br />
days/year at Oradea, up to approximately 50 days/year at Stâna de Vale. Over the<br />
year, winter days are found from November to March at Borod, Oradea, Săcueni<br />
and from October to April in the mountains. The highest frequency of winter days<br />
is in January, when the records show about 14 days in the mountains, about 9 days<br />
in the depressi<strong>on</strong> area of Borod and 10-12 days in the plains.<br />
4. The frequency of summer days is between 80-86 days/year in the plains, about 61<br />
days/year at Borod, while in the mountain area the frequency is <strong>on</strong>ly about 9 days/<br />
year. Throughout the year the highest number of summer days is recorded in July<br />
(between 22-23 days) in the plains, about 18 days in the depressi<strong>on</strong> area of Borod,<br />
while in the mountains the maximum number of <strong>on</strong>ly 4 days is recorded in August.<br />
514
5. The frequency of tropical days ranges between 20-24 days in the plains, is about 10<br />
days in the depressi<strong>on</strong> area of Borod, while in the mountains they lack. Over the<br />
year the favourable interval for tropical days is set between May and September.<br />
The highest frequency is recorded in July in the plains and in August, about 4 days,<br />
in the depressi<strong>on</strong> area of Borod.<br />
REFERENCES<br />
1 Berindei O., Pop Gr., Măhăra Gh., Posea Aurora, (1977), Câmpia Crişurilor, Crişul<br />
Repede, Ţara Beiuşului, Cercetări în geografia României, Editura Ştiinţifică şi<br />
Enciclopedică, Bucureşti.<br />
2 Cristea Maria (2003), Temperatura aerului în bazinul hidrografic al Crişurilor,<br />
Analele Universităţii din Oradea, Seria Geografie, Tom.XIII, pag.77-80.<br />
3 Cristea Maria (2004), Riscurile climatice din bazinul hidrografic al Crişurilor, Edit.<br />
Abaddaba, Oradea, ISBN 973-8102-19-7<br />
4 Gaceu O.(2002), Elemete de climatologie practică, Editura Universităţii din<br />
Oradea, ISBN 973-613-216-1.<br />
5 Gaceu O.(2005), Clima şi riscurile climatice din Munţii Bihor şi Vlădeasa, Editura<br />
Universităţii din Oradea, ISBN 973-759-008-2.<br />
6 Măhăra Gh. (1974), Variaţiile neperiodice ale temperaturii aerului în spaţiul<br />
Câmpiei Crişurilor, Lucr.Ştiinţifice Seria A, Matematică-Fizică-Geografie, Oradea,<br />
pag.37-45.<br />
515
THE RISK OF INUNDATIONS INDUCED BY THE FLOODS ON THE<br />
RIVERS FROM ALMAS-AGRIJ DEPRESSION<br />
AND CLUJ - DEJ HILLS<br />
Alina-Daciana Dumitra<br />
Faculty of Geography, Babes-Bolyai University – Cluj-Napoca<br />
ABSTRACT<br />
The aim of this study was to analyse several aspects of the floods <strong>on</strong> the rivers from<br />
Almas-Agrij Depressi<strong>on</strong> and Cluj and Dej Hills, because they often generated<br />
catastrophic inundati<strong>on</strong>s.<br />
In this work it were used series of data for 28 years within the interval 1976-2004<br />
of nine hydrometrical stati<strong>on</strong>s: Almaş, Hida, Românaşi, in the Almaş-Agrij Depressi<strong>on</strong>,<br />
and: Căpuşu Mare, Mera ( Rădaia), Aghireş, Maia, Borşa and Luna de Jos, in the Cluj<br />
and Dej Hills, <strong>on</strong> the: Almaş, Agrij, Căpuş, Nadăş, Olpret, Borşa and L<strong>on</strong>ea rivers. The<br />
main purpose of the study was to distinguish some aspects of floods that would<br />
c<strong>on</strong>stitute a very important help for the predicti<strong>on</strong>, the preventi<strong>on</strong> and the diminishment<br />
of the hydrological risks. Thus, the work presents the floods formati<strong>on</strong>, analyses their<br />
frequency of occurrence and describes some characteristics of flash floods (debits,<br />
durati<strong>on</strong>) from the area. The study represents a preliminary stage for further analysis<br />
regarding the hydrological risks in the Almaş-Agrij Depressi<strong>on</strong> and the Cluj and Dej<br />
Hills.<br />
Key words: floods, risk of inundati<strong>on</strong>s<br />
INTRODUCTION<br />
From the beginnings the human societies were attracted by the riversides and took the<br />
risk of living there (Pandi, 2002).<br />
The inundati<strong>on</strong>s represent the most often spread hasard <strong>on</strong> Earth, with numerous<br />
losses of human lives and material damages of huge proporti<strong>on</strong>s. Inundati<strong>on</strong> means the<br />
temporary cover of an area with stagnating or moving waters, as a c<strong>on</strong>sequence of the<br />
raise in levels of a river, lake or other mass of water (Sorocovschi, 2002).<br />
Catastrophic inundati<strong>on</strong>s are often generated by river floods. Floods represent the<br />
significant growth and decrease of water discharge in the river bed (Mustăţea, 2005),<br />
which, al<strong>on</strong>g with large waters, are phases of the flood flow that could take place in any<br />
period of the year. They represent natural phenomena that are part of the chain of<br />
normal events c<strong>on</strong>cerning water discharge.<br />
The spatial allocati<strong>on</strong> and amplitude of the floods are de<strong>term</strong>ined by natural and<br />
also by antropic factors, depending <strong>on</strong> which their effects are different.<br />
The climate has the main role in the formati<strong>on</strong> of floods. The general circulati<strong>on</strong><br />
of the atmosphere is the genetic factor which de<strong>term</strong>ines the n<strong>on</strong>-periodical variati<strong>on</strong> of<br />
hydrological regime. Rainfall play the decisive part in producing the highest discharges.<br />
Heavy rains usually affect small areas and de<strong>term</strong>ine floods in small catchement areas.<br />
Morphology exerts its influence indirectly up<strong>on</strong> the discharge by: de<strong>term</strong>ining the<br />
vertical z<strong>on</strong>ality, the c<strong>on</strong>figurati<strong>on</strong>, fragmentati<strong>on</strong> and expositi<strong>on</strong> of slopes.<br />
516
At the same time, the genesis of floods is indirectly linked to the geological factor: the<br />
lythology and the structure both influence the movement of water and the development<br />
of the type of drainage.<br />
Because of its hydrical and physical properties, the soil is an important factor<br />
regarding water flow <strong>on</strong> its surface. The vegetati<strong>on</strong> acts directly up<strong>on</strong> discharge by<br />
influencing the regime of soil humidity, and indirectly by protecting against erosi<strong>on</strong> .<br />
Antropic activity acts against the genetic factors of flow (by clearance, plantati<strong>on</strong>,<br />
pasture etc.) <strong>on</strong> the <strong>on</strong>e hand, and against the flow from the riverbed <strong>on</strong> the other hand<br />
(by the mitigati<strong>on</strong> of debits, embankments, water c<strong>on</strong>sume usages).<br />
MATERIALS AND METHODS<br />
To emphasize the territorial and temporal differences between the floods of the Almaş-<br />
Agrij Depressi<strong>on</strong> and the floods of the Cluj and Dej Hills, there were analysed and<br />
processed data that originate from nine hydrometrical stati<strong>on</strong>: Almaş, Hida, Românaşi<br />
(Almaş-Agrij Depressi<strong>on</strong>), Căpuşu Mare, Mera (Rădaia), Aghireş, Maia, Borşa and<br />
Luna de Jos (Cluj and Dej Hills) <strong>on</strong> the: Almaş, Agrij, Căpuş, Nadăş, Olpret, Borşa and<br />
L<strong>on</strong>ea rivers. The data are resulted from the observati<strong>on</strong>s and measurements performed<br />
over a period of 28 years (between 1976 and 2004), and they are archived in the Annual<br />
Hydrological Studies of the Somes-Tisa Water Department of Cluj-Napoca.<br />
The time frequency of floods within the studied time interval <strong>on</strong> the rivers of<br />
Almas- Agrij Depressi<strong>on</strong> and Cluj-Dej Hills has been analysed based <strong>on</strong> these data. The<br />
seas<strong>on</strong>al frequency of floods has been graphically represented, calculating the total<br />
number of floods in each seas<strong>on</strong> at each hydrological stati<strong>on</strong>. The m<strong>on</strong>thly frequency of<br />
floods has been represented the same way using the same method, the calculati<strong>on</strong> being<br />
made for each m<strong>on</strong>th.<br />
Then, also based <strong>on</strong> these data, it were described two types of parametres of flood<br />
(the maximal debit and the durati<strong>on</strong>). The main characteristic parameters of a flood are:<br />
debits (maximum debit, base debit), durati<strong>on</strong> (total durati<strong>on</strong> or total time, increasing<br />
time, decreasing time), water volumes, the form coefficient of flood, the medium flowed<br />
layer, etc. Maximum debit ( Qmax m 3 /s) represents the parameter of the maximum<br />
interest, because this is the moment of the biggest danger, the maximum overflow. The<br />
durati<strong>on</strong> of floods is an important element of which depends the size of the effects<br />
which could be generated (Sorocovschi, 2005). The whole durati<strong>on</strong> or total time is<br />
composed by increasing time and decreasing time<br />
The maximal debits ( Qmax ) of floods <strong>on</strong> the rivers of Almas-Agrij Depressi<strong>on</strong> and<br />
Cluj-Dej Hills have been graphically represented for the period between 1976 and<br />
2004. The years with highest and lowest risk of inundati<strong>on</strong>s have been clearly<br />
underlined.<br />
The total durati<strong>on</strong> (Tt) of the flood has been obtained by calculating the number of<br />
hours, from the time when the debit overpassed the basic debit, until the time it came<br />
back to the value of the basic debit (of course, after reaching the maximal debit - Qmax).<br />
The increasing time of flood has been obtained by adding the number of hours<br />
from the time the debits started to grow until the time the maximal debit (Qmax) has been<br />
recorded.<br />
517
RESULTS AND DISCUSSIONS<br />
In Almaş-Agrij Depressi<strong>on</strong> and in Cluj and Dej Hills, <strong>on</strong> the majority of rivers, the<br />
registered floods were originated in pluvial (50 – 55% of the analyzed floods), followed<br />
by a much smaller number of floods originated in pluvial-nival.<br />
floods<br />
winter spring summer autum<br />
seas<strong>on</strong>s<br />
Almasu<br />
Hida<br />
Romanasi<br />
Figure 1. The seas<strong>on</strong>al flood frequency in Depresiunea Almaş-Agrij (1976-2004)<br />
floods<br />
35<br />
30<br />
25<br />
20<br />
15<br />
10<br />
5<br />
0<br />
winter spring summer autum<br />
seas<strong>on</strong><br />
Capusu Mare<br />
Aghires<br />
Mera(Radaia)<br />
Figure 2. The seas<strong>on</strong>al flood frequency in Cluj - Dej Hills (1976-2004)<br />
Figure 1,2 and 3 shows that, regarding the seas<strong>on</strong>s, the maximum frequency is observed<br />
in spring (35-55% of the entire number of floods), followed by summer (17 – 33%). In<br />
518
winter the floods frequency has smaller values (included within the 14 – 25% of total<br />
floods). The most reduced frequency of the floods (with values between 14 and 25 % of<br />
the total floods) is recorded in the Almaş-Agrij Depressi<strong>on</strong> but also in the Cluj - Dej<br />
Hills in autumn.<br />
floods<br />
35<br />
30<br />
25<br />
20<br />
15<br />
10<br />
5<br />
0<br />
winter spring summer autum<br />
seas<strong>on</strong><br />
Borsa<br />
Luna de Jos<br />
Maia<br />
Figure 3. The seas<strong>on</strong>al flood frequency in Cluj and Dej Hills (1976-2004)<br />
It is noticed, regarding the floods frequency during the seas<strong>on</strong>s, a spatial differentiati<strong>on</strong><br />
<strong>on</strong> the Cluj and Dej Hills, thus: <strong>on</strong> the rivers from the east of the unit (Olpret, Borşa,<br />
L<strong>on</strong>ea) the number of floods during the summer has a very close value to that number<br />
which represents the floods during the winter (34 floods during the summer, and 32<br />
floods in winter); <strong>on</strong> the Nadăş and Căpuş Streams (Cluj Hills) the difference is<br />
remarkable, the number of summer floods being much higher than the number of winter<br />
floods (20 floods, 46 floods respectively). More over, this c<strong>on</strong>siderable difference is<br />
found again in Almaş-Agrij Depressi<strong>on</strong> (29 floods, 45 floods respectively).<br />
The m<strong>on</strong>thly frequency of floods is maximum in March <strong>on</strong> rivers of the Cluj and<br />
Dej Hills (Olpret, L<strong>on</strong>ea, Borşa, Nadăş, Căpuş), representing 17–23 % of all selected<br />
floods. High values of the m<strong>on</strong>thly frequency of floods formati<strong>on</strong> have been recorded in<br />
April and May. The lowest frequencies of floods took place <strong>on</strong> Olpret, Borşa and L<strong>on</strong>ea<br />
rivers in October and November. There were no floods <strong>on</strong> Căpuş and Nadăş rivers in<br />
November, within the studied interval, and in December there were recorded minimal<br />
values of the floods frequency (1.9% of total) (Fig.4, Fig. 5)<br />
519
%<br />
floods<br />
14<br />
12<br />
10<br />
8<br />
6<br />
4<br />
2<br />
0<br />
14<br />
12<br />
10<br />
I II III IV V VI VII VIII IX X XI XII<br />
m<strong>on</strong>ths<br />
Capusu Mare<br />
Aghires<br />
Mera(Radaia)<br />
Figure 4. M<strong>on</strong>thly floods frequency in Cluj- Dej Hills (1976-2004)<br />
8<br />
6<br />
4<br />
2<br />
0<br />
I II III IV V VI VII VIII IX X XI XII<br />
m<strong>on</strong>ths<br />
Borsa<br />
Luna de Jos<br />
Maia<br />
Figure 5. M<strong>on</strong>thly floods frequency in Cluj- Dej Hills (1976-2004)<br />
The m<strong>on</strong>thly frequency of floods formati<strong>on</strong> presents a maximum point in May, <strong>on</strong> the<br />
Almaş and Agrij rivers, in the Almaş-Agrij Depressi<strong>on</strong> (15 – 23% of the entire number<br />
of sorted floods), followed by March and June (with the same number of floods). The<br />
minimal frequency of floods or even their absence was recorded in November (Fig. 6)<br />
520
%<br />
14<br />
12<br />
10<br />
8<br />
6<br />
4<br />
2<br />
0<br />
I II III IV V VI VII VIII IX X XI XII<br />
m<strong>on</strong>ths<br />
Almasu<br />
Hida<br />
Romanasi<br />
Figure 6. M<strong>on</strong>thly floods frequency in Almas- Agrij Depressi<strong>on</strong> (1976-2004)<br />
Figure 7 shows the maximum debits <strong>on</strong> rivers in Almaş-Agrij Depressi<strong>on</strong>. Here the<br />
maximum debits had values between 150m 3 /s <strong>on</strong> Almaş river, in Hida, and 220 m 3 /s, <strong>on</strong><br />
Agrij river, in Românaşi. The highest risk of inundati<strong>on</strong>s took place in the year 1989<br />
and ten years after, in 1999, when the maximal debits of floods have been recorded. In<br />
1982 and 2001 the risk was serious, too. However the maximal debits of floods had<br />
minimal values in some years (such as in 1984).<br />
250<br />
200<br />
150<br />
100<br />
50<br />
0<br />
1981<br />
1982<br />
1983<br />
1984<br />
1985<br />
1986<br />
1987<br />
1988<br />
1989<br />
1990<br />
1991<br />
1992<br />
1993<br />
1994<br />
1995<br />
1996<br />
1997<br />
1998<br />
1999<br />
2000<br />
2001<br />
2002<br />
2003<br />
2004<br />
Almas<br />
Hida<br />
Romanasi<br />
Figure 7. Maximum debits of floods <strong>on</strong> rivers in Almas-Agrij Depressi<strong>on</strong> (1981-<br />
2004)<br />
The maximum debits de<strong>term</strong>ined in Cluj and Dej Hills within years 1976 and 2004 had<br />
values between 44.1 m 3 /s <strong>on</strong> the Nadăş river, in Aghireş, and 146 m 3 /s <strong>on</strong> L<strong>on</strong>ea river,<br />
in Luna de Jos.<br />
In the Almaş-Agrij Depressi<strong>on</strong> and in Cluj and Dej Hills the multiannual maximum<br />
debits have varied between very large limits, depending <strong>on</strong> numerous factors<br />
(climacteric c<strong>on</strong>diti<strong>on</strong>s, receiving basin surface etc.) The maximum debits were<br />
521
ecorded with a high level frequency <strong>on</strong> the streams in the Cluj and Dej Hills and in the<br />
Almaş-Agrij Depressi<strong>on</strong> in May and June.<br />
On most rivers in Almaş-Agrij Depressi<strong>on</strong> and Cluj and Dej Hills the most<br />
c<strong>on</strong>siderable frequency represented those floods that maintained their total durati<strong>on</strong><br />
between 5 and 15 days (30 – 45 % of all cases). The streams that had more frequent<br />
floods with durati<strong>on</strong> over 20 days (Borşa river) were excepted. The floods with a less<br />
durati<strong>on</strong> than 5 days had the most c<strong>on</strong>siderable frequency in Agrij, Almaş and Nadăş<br />
basins, being de<strong>term</strong>ined by the torrential nature of the rainfalls, the reduced surface of<br />
the basin (Nadăş to Aghires, 39.1 km 2 ) and by the relative large slope.<br />
The increasing time of the floods is different, depending <strong>on</strong> the nature of rainfalls,<br />
varying, in the analysed rivers, between 4 and 130 hours.<br />
The maximum debit, increasing time, total time and the volume of floods are the<br />
main elements of the flood hydrograph. They c<strong>on</strong>stitute the bench mark for flood.<br />
CONCLUSIONS<br />
The results presented above let us come to the following c<strong>on</strong>clusi<strong>on</strong>: the knowledge<br />
regarding floods would help solving several issues like: the understanding of the<br />
nature, the development of some protecti<strong>on</strong> or adaptati<strong>on</strong> programmes etc. Establishing<br />
the frequency and the amplitude of floods c<strong>on</strong>stitutes very important aspects<br />
c<strong>on</strong>cerning the assessment of risk of inundati<strong>on</strong>s.<br />
After the floods, it is well known that the set back to normal c<strong>on</strong>diti<strong>on</strong>s and the<br />
rehabilitati<strong>on</strong> of the damaged envir<strong>on</strong>mental factors involves material effort and<br />
important collective acti<strong>on</strong>s, sometimes difficult to perform (Stanescu, Drobot, 2002).<br />
REFERENCES<br />
1. Mustăţea, A., (2005), Viituri Excepţi<strong>on</strong>ale pe teritoriul României, Bucureşti;<br />
2. Pandi, G., (2002), Riscul în activitatea de apărare împotriva inundaţiilor, Riscuri şi<br />
Catastrofe, vol. I, Editura Casa Cărţii de Ştiinţă, Cluj-Napoca, p. 131-142;<br />
3. Sorocovschi, V., (2002), Hidrologia uscatului, vol II, Editura Casa Cărţii de Ştiinţă,<br />
Cluj-Napoca;<br />
4. Sorocovschi, V., (2005), Câmpia Transilvaniei – Studiu hidrogeografic, Editura Casa<br />
Cărţii de Ştiinţă Cluj-Napoca;<br />
5. Stănescu, V. Al., Drobot, R., (2002), Măsuri nestructurale de gestiune a inundaţiilor,<br />
Editura HGA Bucureşti;<br />
6. Studii Hidrologice Anuale, 1976-2004, Arhiva Direcţiei Apelor Someş-Tisa, Cluj-<br />
Napoca.<br />
522
RESEARCH REGARDING MAIN CHEMICAL REACTIONS IN<br />
PREDEFECATION OF DIFFUSION JUICE<br />
Gheorghe Sarca<br />
University of Oradea, Faculty for Envir<strong>on</strong>mental Protecti<strong>on</strong><br />
ABSTRACT<br />
The quality of sugar is the decisive factor in ensuring the sugar commercializing<br />
through the c<strong>on</strong>servati<strong>on</strong> of old markets and expansi<strong>on</strong> towards new markets. The sugar<br />
quality is also the key element which de<strong>term</strong>ines the price maintenance in according<br />
with the expenditures performed or even the diminuti<strong>on</strong> of the price as a c<strong>on</strong>sequence of<br />
the quality lessening.<br />
INTRODUCTION<br />
The main chemical reacti<strong>on</strong>s occurred in the predefecati<strong>on</strong> of diffusi<strong>on</strong> juice are the<br />
following:<br />
• Neutralisati<strong>on</strong> of free acids, especially the neutralisati<strong>on</strong> of the organic acids of<br />
sugar beet and of the mineral acids c<strong>on</strong>tained in the extracti<strong>on</strong> water:<br />
2 R COOH + Ca(OH)2→ (R COO) 2Ca + H2O<br />
• I<strong>on</strong>s exchange between magnesium and calcium which can be mainly described by<br />
the in<strong>term</strong>ediate of a chemical reacti<strong>on</strong> such as:<br />
2 R COOMe + Ca(OH)2+ Ca(OH)2→ (R COO) 2Ca+2MeOH<br />
At pre-defecated juice defecati<strong>on</strong> the following chemical reacti<strong>on</strong>s take place:<br />
R CONH2 + H2O → R COONH4<br />
H2O + R CONH2 → R COOH + NH2<br />
H2O + CO2 → H2CO3<br />
H2O + CaO → Ca(OH)2<br />
Reducti<strong>on</strong> substances + Ca(OH)2 → Colouring Substances + Organic Acids<br />
During defecated juice carb<strong>on</strong>izing the following reacti<strong>on</strong>s take place:<br />
CO2 + H2O → H2CO3<br />
CO2 + OH → HCO?<br />
H2Co3 + Ca(OH)2 → CaCO3 + H2O<br />
An important role in providing the adequate c<strong>on</strong>diti<strong>on</strong>s for the development of these<br />
reacti<strong>on</strong>s is taken by the calcium oxide solubility in water and in the sugar-in-water<br />
impure soluti<strong>on</strong>s. As higher level of calcium oxide solubility is as better the purificati<strong>on</strong><br />
effect would be. In the chart 1 the solubility of the oxide calcium solved in water and in<br />
sucrose soluti<strong>on</strong>s is presented.<br />
523
Table 1.The solubility of the calcium oxide in water<br />
and in the in sucrose soluti<strong>on</strong>s in water<br />
Sucrose The solubility of the calcium oxide in the in sucrose soluti<strong>on</strong>s in<br />
water, soluti<strong>on</strong> expressed in g CaO/100 mg, at temperature: [ºC]<br />
g/100 ml<br />
water<br />
0 20 40 60 80<br />
0,0 0,135 0,126 0,109 0,088 0,071<br />
1,5 0,252 0,175 0,132 0,118 -<br />
3,0 0,476 0,280 0,200 0,145 -<br />
6,0 1,172 0,660 0,310 0,212 0,130<br />
12,0 2,082 1,969 0,936 0,422 0,204<br />
18,0 4,140 3,553 1,942 1,168 0,356<br />
MATERIALS AND METHODS<br />
In order to obtain the largest amount of high-quality crystal sugar from the diffusi<strong>on</strong><br />
juice its purificati<strong>on</strong> and removing the n<strong>on</strong>-sugar are required. Actually, the current<br />
systems designed for diffusi<strong>on</strong> juice purificati<strong>on</strong> ensure a n<strong>on</strong>-sugar parts removal<br />
reaching at max. 45% of the n<strong>on</strong> sugar of diffusi<strong>on</strong> juice.<br />
With the purpose of diffusi<strong>on</strong> juice purificati<strong>on</strong> <strong>on</strong>e must make use of cheap<br />
methods and of those substances which would not alter the sugar and are not to cause<br />
sugar loses while n<strong>on</strong> sugar precipitated with these substances are to be easily separated<br />
out of the juice mass and subsequently eliminated. Nowadays, for diffusi<strong>on</strong> juice<br />
purificati<strong>on</strong> <strong>on</strong> utilise calcium oxide and carb<strong>on</strong> dioxide. That is the reas<strong>on</strong> for which<br />
the current technological process of purificati<strong>on</strong> is called “calco-carb<strong>on</strong>ic purificati<strong>on</strong>”.<br />
During the technological process of sugar beet processing, the aim of calco-carb<strong>on</strong>ic<br />
purificati<strong>on</strong> of the diffusi<strong>on</strong> juice is well settled and established. The amount of n<strong>on</strong><br />
sugar eliminated during the purificati<strong>on</strong> process favorably influences the subsequent<br />
development of the technological process in particular those operati<strong>on</strong>s accomplished at<br />
high-level temperatures e.g. the c<strong>on</strong>centrati<strong>on</strong> through vaporizati<strong>on</strong>, boiling and<br />
crystallizati<strong>on</strong> but also the outturn and the quality of the sugar obtained.<br />
The diffusi<strong>on</strong> juice purificati<strong>on</strong> may be accomplished through other methods e.g.<br />
i<strong>on</strong> exchange, ultra-filtrati<strong>on</strong>, i<strong>on</strong>-exclusi<strong>on</strong>, electro-dialysis and inverse-osmosis.<br />
Nevertheless, due to some reas<strong>on</strong>s of ec<strong>on</strong>omic effectiveness, the diffusi<strong>on</strong> juice<br />
purificati<strong>on</strong> obtained by the help of these methods is not used at industrial scale yet.<br />
By definiti<strong>on</strong>, diffusi<strong>on</strong> juice purificati<strong>on</strong> is the operati<strong>on</strong> which aims the optimal<br />
development of the subsequent phases of the technological process. The phases and the<br />
operati<strong>on</strong>s which c<strong>on</strong>stitute the calco-carb<strong>on</strong>ic purificati<strong>on</strong> process and the<br />
technological parameters at the level of which these operati<strong>on</strong>s are executed are all<br />
established functi<strong>on</strong> of the technological quality of the sugar beet and of the diffusi<strong>on</strong><br />
juice processed. The purificati<strong>on</strong> of the diffusi<strong>on</strong> juice must always have a positive<br />
influence up<strong>on</strong> the entire producti<strong>on</strong> process’ effectiveness, an influence materialized as<br />
following:<br />
524
• Ensuring the juice’ thermal stability at c<strong>on</strong>centrati<strong>on</strong> through vaporizati<strong>on</strong>, a<br />
stability materialized by the avoidance of Ph dropping, by preventing juice intense<br />
coloring and by the sucrose hydro-dialysis avoidance;<br />
• Avoiding the intense frothing which may lead to great losses of sucrose al<strong>on</strong>g with<br />
the froth which is produced;<br />
• Minimal losses of sucrose as a c<strong>on</strong>sequence of its instability and its thermal<br />
decompositi<strong>on</strong> during its c<strong>on</strong>centrati<strong>on</strong> through vaporizati<strong>on</strong>;<br />
• Minimal values for the sugar remained in molasses;<br />
• Ensuring the quality sugar producti<strong>on</strong> in order to maintain its stability during<br />
depositing;<br />
• Lending itself for subsequent use in the industries employing crystal sugar.<br />
In the chart 2 the influence of the calco-carb<strong>on</strong>ic purificati<strong>on</strong> up<strong>on</strong> the diffusi<strong>on</strong> juice<br />
n<strong>on</strong> sugar is sketchily presented.<br />
N<strong>on</strong> sugar of the diffusi<strong>on</strong> juice<br />
N<strong>on</strong> sugar N<strong>on</strong> sugar N<strong>on</strong> sugar N<strong>on</strong> sugar<br />
n<strong>on</strong> influenced precipitated decomposed absorbed<br />
by by by and adsorbed<br />
Calco-carb<strong>on</strong>ic calcium oxide Ca(OH)2 and by limest<strong>on</strong>e<br />
Purificati<strong>on</strong> temperature<br />
Sugar<br />
remained<br />
Sugar n<strong>on</strong>-absorbed and n<strong>on</strong>-adsorbed by Sugar reabsorbed<br />
in thin juice the calcium carb<strong>on</strong>ate (limest<strong>on</strong>e) out of the<br />
limest<strong>on</strong>e<br />
Chart 2 The acti<strong>on</strong> of the calco-carb<strong>on</strong>ic purificati<strong>on</strong> up<strong>on</strong> the diffusi<strong>on</strong> juice’<br />
n<strong>on</strong>-sugar and the n<strong>on</strong>-sugar circulating during the diffusi<strong>on</strong> juice purificati<strong>on</strong><br />
The amount of sucrose remained in molasses depends up<strong>on</strong> the compositi<strong>on</strong> and<br />
c<strong>on</strong>centrati<strong>on</strong> of the thin juice’ n<strong>on</strong>-sugar. That is why, the eliminati<strong>on</strong> of the n<strong>on</strong>-sugar<br />
from the diffusi<strong>on</strong> juice is a c<strong>on</strong>diti<strong>on</strong> sine qua n<strong>on</strong> for obtaining some crystal sugar’<br />
good outturns.<br />
525
The classical calco-carb<strong>on</strong>ic purificati<strong>on</strong> of the diffusi<strong>on</strong> juice c<strong>on</strong>sists in the juice’<br />
treatment with lime milk and the subsequent precipitati<strong>on</strong> of the excess (surfeit) of<br />
Ca(OH)2. The resulted mud’ separati<strong>on</strong> raises some special problems. That is the reas<strong>on</strong><br />
for which, in practice, there have been introduced some purificati<strong>on</strong> methods to which<br />
some compromises are being made between the purificati<strong>on</strong> effect and the separati<strong>on</strong><br />
capacity, through decantati<strong>on</strong> or through mud filtering.<br />
RESULTS AND DISCUSSIONS<br />
The technological role of the diffusi<strong>on</strong> juice’ calco-carb<strong>on</strong>ic purificati<strong>on</strong> in the process<br />
crystal sugar obtaining c<strong>on</strong>sists in the eliminati<strong>on</strong> of a part of the diffusi<strong>on</strong> juice’ n<strong>on</strong>sugar<br />
substance employing some operati<strong>on</strong>s specific to each n<strong>on</strong>-sugar groups and<br />
which would provide ,at the given momentum, some optimal c<strong>on</strong>diti<strong>on</strong>s for the<br />
destructi<strong>on</strong> and eliminati<strong>on</strong> of a certain sugar type. Punctually, the role of the calcocarb<strong>on</strong>ic<br />
purificati<strong>on</strong> c<strong>on</strong>sists in:<br />
• Neutralizing the acidity of the diffusi<strong>on</strong> juice and precipitating the calcium salts;<br />
• Precipitating and separating diffusi<strong>on</strong> juice’ colloids;<br />
• Decompounding the reducing substances (agents) and the amides and obtaining the<br />
heat-proof (thermo stable) juices;<br />
• N<strong>on</strong>-sugar absorbing and adsorbing <strong>on</strong> the calcium carb<strong>on</strong>ate (limest<strong>on</strong>e)<br />
• Mud decanting (separating) out of the purified juice;<br />
• Sweetening the mud;<br />
• Thin juice decalcifying and preventing the c<strong>on</strong>centrating stati<strong>on</strong>’ peeling through<br />
vaporizati<strong>on</strong>.<br />
In order to obtain the diffusi<strong>on</strong> juice purificati<strong>on</strong> <strong>on</strong>e employ calcium oxide under the<br />
lime milk shape and carb<strong>on</strong>adoing gas c<strong>on</strong>taining carb<strong>on</strong> dioxide.<br />
In the chart 3 there are represented the characteristics of the lime milk used for<br />
purificati<strong>on</strong> in sugar factories.<br />
Table 3. The characteristics of the lime milk used for<br />
calco-carb<strong>on</strong>ic purificati<strong>on</strong> in sugar factories<br />
C<strong>on</strong>centrati<strong>on</strong> in dry Specific CaO c<strong>on</strong>tent Ca(OH)2 c<strong>on</strong>tent<br />
substance mass<br />
ºBe ºBx g/cm 3 g/l g/100g g/l g/100g<br />
1,49 2,69 1,0084 9 0,98 13,1 1,30<br />
4,69 8,39 1,0314 39 3,87 52,7 5,12<br />
7,59 13,69 1,0534 69 6,64 92,4 8,78<br />
10,34 18,69 1,0749 99 9,29 132,0 12,28<br />
15,54 28,19 1,1184 159 14,29 211,3 18,89<br />
20,34 37,09 1,1614 219 18,93 290,6 25,02<br />
25,49 46,79 1,2124 289 23,91 383,1 31,60<br />
For an accurate evaluati<strong>on</strong> of the functi<strong>on</strong> of the diffusi<strong>on</strong> juice purificati<strong>on</strong>’ stati<strong>on</strong><br />
from a sugar factory it is necessary to be aware of the exact amount of carb<strong>on</strong>adoing<br />
526
mud which is evacuated out of the process. This quantity depends up<strong>on</strong> the calcium<br />
dioxide dose actively employed for purificati<strong>on</strong> and it has a direct influence up<strong>on</strong> the<br />
following several indicators:<br />
1. The sugar losses in the mud;<br />
2. The calco-carb<strong>on</strong>ic purificati<strong>on</strong> effect of which the efficiency of the purificati<strong>on</strong><br />
depends;<br />
3. Providing the adequate c<strong>on</strong>diti<strong>on</strong>s for separati<strong>on</strong> thorough decantati<strong>on</strong> and mud<br />
filtering out of the turbid juice from 1 st (I) saturati<strong>on</strong>.<br />
REFERENCES<br />
1. Banu C., 1992 – „Progrese tehnice, tehnologice şi ştiinţifice în industria<br />
alimentară”, Ed. Tehnică, Bucureşti.<br />
2. Beceanu Dumitru, Chira Adrian, 2003 – „Tehnologia produselor agricole”, Ed.<br />
Ec<strong>on</strong>omică, Bucureşti.<br />
3. Sarca Gheorghe, 2004 – „Materii prime vegetale”, Ed. Universităţii din Oradea.<br />
4. <str<strong>on</strong>g>Internati<strong>on</strong>al</str<strong>on</strong>g> Sugar Journal, 2006 december.<br />
5. Popescu V., Stroia A., 1998 – „Tehnologii moderne de industrializarea sfeclei de<br />
zahăr”, Ed. Ceres, Bucureşti.<br />
527
DETERMINATION OF NITROGEN-CONTAINING POLYNUCLEAR AROMATIC<br />
HYDROCARBONS IN SMOKED FOODS BY LIQUID CHROMATOGRAPHY<br />
Lucian Bara<br />
University of Oradea, Faculty of Envir<strong>on</strong>mental Protecti<strong>on</strong><br />
26, General Magheru Street, Oradea<br />
ABSTRACT<br />
Several smoked foods were analyzed for basic nitrogen-c<strong>on</strong>taining polynuclear aromatic<br />
hydrocarb<strong>on</strong> (NPAH) c<strong>on</strong>tent by a relatively rapid liquid chromatographic (LC) technique.<br />
The analyzed products included both domestic and imported market basket commodities.<br />
Nanogram quantities of NPAH standards were detected by UV and fluorescence detectors<br />
c<strong>on</strong>nected in series. The NPAHs were extracted from basic aqueous ethanolic soluti<strong>on</strong> into<br />
cyclohexane, extracted from cyclohexane into 6N HCl, and extracted back into cyclohexane<br />
after neutralizati<strong>on</strong> of the acid.<br />
The NPAHs were then purified by filtering the extract through deactivated basic<br />
alumina. The eluate from this step was c<strong>on</strong>centrated to dryness, and the residue was<br />
dissolved in 95% ethanol and analyzed by LC, using a Vydac C-18 column and acet<strong>on</strong>itrilewater<br />
(9 + 1) as the mobile phase. Recoveries of 3 NPAHs, 5,7-dimethylbenz acridine,<br />
dibenz acridine, and dibenz acridine, each added to salm<strong>on</strong> and sausage at the 5 ppb level,<br />
ranged from 62 to 101% by fluorescence measurement and from 64 to 106% by UV<br />
measurement. N<strong>on</strong>e of the NPAHs used, as standards were found by either fluorescence or<br />
UV detecti<strong>on</strong> at levels greater than or equal to 5 ppb in any of the foods analyzed.<br />
INTRODUCTION<br />
Interest in the producti<strong>on</strong> of petroleum substitutes from shale oil and coal has increased. The<br />
increasing use of coal and synthetic fuels may significantly increase the levels of<br />
heterocyclic nitrogen compounds in the envir<strong>on</strong>ment because of the charasterically higher<br />
level of organic nitrogen in coal and its by-products, compared to other fossil fuels. Many<br />
nitrogen bases, thought to be products of incomplete combusti<strong>on</strong> of organic matter that<br />
c<strong>on</strong>tains nitrogen or to be derived from hydrolytic processes similar to those producing<br />
polynuclear aromatic hydrocarb<strong>on</strong>s, have been shown by various types of skin tests to be<br />
toxic, carcinogenic, and mutagenic to terrestrial and aquatic organisms.<br />
Many azaarenes have been identified in sources such as tobacco, automobile exhaust,<br />
high boiling petroleum distillates, crude oil, shale oil, and lake sediments.<br />
The purpose of our investigati<strong>on</strong> was to develop an analytical method for the<br />
extracti<strong>on</strong>, cleanup, and LC analysis of the basic NPAH fracti<strong>on</strong>s obtained from some<br />
smoked foods. The foods analyzed included both domestic and imported products.<br />
528
MATERIALS AND METHOD<br />
Apparatus<br />
Clean all glassware with chromatic acid/sulfuric acid soluti<strong>on</strong>s, rinse thoroughly with<br />
distilled water, oven-dry at 120 o C, and solvent-rinse before use to minimize interferences.<br />
a) Meat chopper – Launders, Frary and Clark Co. or equivalent.<br />
b) Digesti<strong>on</strong> apparatus – 1) Heating mantle – 500 mL; 2) Friedrich c<strong>on</strong>denser; 3)<br />
Boiling flask<br />
c) Separatory funnels<br />
d) Flasks<br />
e) Flash evaporator<br />
f) Filter funnel<br />
g) Sample clarificati<strong>on</strong> kit<br />
h) Liquid chromatograph<br />
i) Recording data processors<br />
Reagents<br />
a) Solvents – Ethanol, 190 proofs, distill before use; cyclohexane, distilled-in-glass;<br />
acet<strong>on</strong>itrile, LC grade; double-distilled water.<br />
b) NPAH standards – 5,7-Dimethylbenze acridine, dibenz acridine. Each standard<br />
was purified by reverse phase LC, and the identity of each was verified by GC/MS<br />
before use in this study.<br />
c) Standard soluti<strong>on</strong>s – 1) Stock soluti<strong>on</strong>s – 0.1 mg/mL. Dissolve 1 mg of each<br />
NPAH standard in 10mL ethanol. 2) Working standard soluti<strong>on</strong> dilute to volume with<br />
ethanol.<br />
d) Alumina – aluminum oxide 90, active basic, activity stage 1<br />
e) Deactivated alumina – Prepared from alumina above<br />
f) Soluti<strong>on</strong>s – 6N HCI, 10N KOH, ethanol-methylene chloride.<br />
Extracti<strong>on</strong><br />
Place 25 g homogenized smoked food into 500 mL boiling flask. Add 100 mL ethanol, 4 g<br />
KOH, and boiling chips. Insert Friedrich c<strong>on</strong>denser and reflux at rapid rate for 2 h. let digest<br />
cool for 20 min, remove c<strong>on</strong>denser, and filter c<strong>on</strong>tents of flask through ethanol-rinsed glass<br />
wool pad into 1 L separatory funnel. Shake funnel vigorously for 2 min. let layers separate;<br />
then draw lower aqueous layer into sec<strong>on</strong>d 1L separatory funnel. Repeat extracti<strong>on</strong> with 50<br />
mL cyclohexane.<br />
Add 20 mL 6N HCI and shake funnel for 2 min. let layers separate and draw off<br />
bottom acidic layer into sec<strong>on</strong>d 1L funnel. Repeat extracti<strong>on</strong> in first funnel with 20 mL<br />
porti<strong>on</strong>s of 6N HCI, and combine each extract with first extract in sec<strong>on</strong>d funnel. Discard<br />
cyclohexane layer remaining in first funnel.<br />
Carefully add 50 mL 10 N KOH to acidic extracts while cooling funnel under cold tap<br />
water. Add 50 mL cyclohexane and shake funnel for 2 min. Draw off lower aqueous layer<br />
into sec<strong>on</strong>d funnel and repeat extracti<strong>on</strong> with 50 mL cyclohexane.<br />
529
Liquid Chromatography<br />
Inject 20 μL c<strong>on</strong>centrate or standard soluti<strong>on</strong> <strong>on</strong>to Vydac 201TP54 column and run mobile<br />
phase c<strong>on</strong>sisting of acet<strong>on</strong>itrile-water at flow rate of 1 mL/min. Between injecti<strong>on</strong>s, flush<br />
injecti<strong>on</strong> loop and port passages with 3 mL acet<strong>on</strong>itrile-water to prevent crossc<strong>on</strong>taminat<strong>on</strong>.<br />
For tentative identificati<strong>on</strong>, compare retenti<strong>on</strong> times of any peaks observed<br />
with those of known NPAH standards chromatographed under same c<strong>on</strong>diti<strong>on</strong>s.<br />
RESULTS AND DISCUSSIONS<br />
All of the domestic smoked foods analyzed in this study were previously analyzed for their<br />
polynuclear aromatic hydrocarb<strong>on</strong> c<strong>on</strong>tent; ham, bac<strong>on</strong>, sausage, whiting, salm<strong>on</strong>, and trout<br />
represent this group. Some important smoked foods were also analyzed; oysters, clams,<br />
sardines, red snapper, and brislings represent this group. The 3 NPAHs used as standards for<br />
this study were chosen primarily because they were readily available and easily purified.<br />
These compounds represent some of the carcinogenic basic NPAHs, which might be found<br />
in c<strong>on</strong>taminated smoked foods.<br />
CONCLUSIONS<br />
A relatively short, straightforward LC method has been developed for the analysis of several<br />
smoked foods for NPAHs. Although no NPAHs were isolated from any of the smoked foods<br />
analyzed, the recovery data indicate that the method is reliable. A study of the distributi<strong>on</strong><br />
ratios of DMBaAc, DbajAc, and DbahAc between cyclohexane and the aqueous ethanolic<br />
sap<strong>on</strong>ificati<strong>on</strong> digest indicates that the best recovery of the azaarenes occurred when the<br />
water-ethanol ratio was 1:1. Future studies will include oil-c<strong>on</strong>taminated foods.<br />
REFERENCES<br />
1. Buchanan, M. V., Ho, C., Guerin (1981) in Chemical Analysis and Biological Fate:<br />
Polynuclear Aromatic Hydrocarb<strong>on</strong>s, Battelle Press, Columbus, OH, p. 133-144.<br />
2. Hertz, H. S., et al. (1980) Anal. Chem. 52, 1650-1657.<br />
3. Colin, H., Schmitter, J., & Guioch<strong>on</strong>, G. (1981) Anal. Chem. 53, 625-631.<br />
4. Joe, F. L., Jr., Salemme, J., & Fazio, T. (1984) J. Assoc. Off. Anal. Chem. 67, 1076-<br />
1082.<br />
530
THE IMPORTANCE OF VOLATILE N-NITROSAMINES IN FOOD-<br />
PRODUCTS<br />
Lucian Bara, Camelia Bara, Vasile Bara, Dana Marele<br />
University of Oradea, Faculty of Envir<strong>on</strong>mental Protecti<strong>on</strong><br />
26, General Magheru Street, Oradea<br />
ABSTRACT<br />
A variety of food products c<strong>on</strong>taining nitrite were analyzed for 14 volatile N-nitrosamines<br />
by using a method dem<strong>on</strong>strated to be sensitive to 10 ppb. A total of 121 food samples were<br />
screened for volatile N-nitrosamine c<strong>on</strong>tent. N-Nitrosopyrrolidine was c<strong>on</strong>firmed in fried<br />
bac<strong>on</strong> at levels up to 139 ppb. N-Dimethylnitrosamine, N-nitrosopyrrolidine, and Nnitrosopiperidine<br />
were also c<strong>on</strong>firmed in spice-cure mixtures at levels ranging from 50 to<br />
2000 ppb.<br />
INTRODUCTION<br />
The many publicati<strong>on</strong>s in the literature during the last years pertaining to the study of<br />
volatile Af-nitrosamines in the envir<strong>on</strong>ment and in food supplies attest to the c<strong>on</strong>cern of the<br />
scientific community with these potent carcinogens. It has been shown that nitrosamines are<br />
formed by the acti<strong>on</strong> of nitrite <strong>on</strong> sec<strong>on</strong>dary and tertiary amines. The widespread use of<br />
nitrite as a preservative and color fixative, coupled with the natural occurrence of amines in<br />
foods, prompted the surveillance of products so preserved to de<strong>term</strong>ine the extent of these<br />
compounds in our food supply. The food products investigated to date include alcoholic<br />
beverages, bac<strong>on</strong>, fish, frankfurters, ham, spinach, bologna, prepared meat products, and<br />
meat spice-cure mixtures. This report c<strong>on</strong>tinues the study of volatile N-nitrosamines in our<br />
food supply. The results of the analysis of 121 samples are given.<br />
MATERIALS AND METHOD<br />
Apparatus and Reagents: The silica gel and the solvents, methylene chloride, pentane, and<br />
ethyl ether, were purified and tested prior to use to ensure the absence of interfering peaks.<br />
Procedure: Fourteen compounds can be de<strong>term</strong>ined: N-nitroso-dimethylamine, -<br />
methylethyl-amine, -diethylamine, -methylpropylamine, -ethyl-propylamine, -<br />
dipropylamine, -ethylbutylamine, -propylbutylamine, -methylamylamine, -dibutylamine, -<br />
piperidine, -pyrrolidine, -morpholine, and -diamylamine.<br />
Briefly, the comminuted food sample is digested in methanolic potassium hydroxide<br />
and the nitrosamines are extracted with methylene chloride in a liquid-liquid extractor,<br />
distilled from alkaline soluti<strong>on</strong>, further isolated by extracti<strong>on</strong> and column chromatographic<br />
techniques, and de<strong>term</strong>ined by gas-liquid chromatography (GLC), using a modified<br />
thermi<strong>on</strong>ic detector.<br />
531
When a peak was observed at the retenti<strong>on</strong> time of <strong>on</strong>e of the 14 nitrosamines, the identity<br />
of this compound was c<strong>on</strong>firmed by mass spectrometry (MS). To ensure adequate cleanup<br />
for this analysis, the c<strong>on</strong>centrated methylene chloride eluate from the silica gel column was<br />
passed through an acid-Celite column.<br />
The identities of the nitrosamines isolated were c<strong>on</strong>firmed with a combined GLC-MS<br />
system. A Varian Model 5600X, 2-stage membrane separator was interfaced to an Atlas<br />
CH-4 mass spectrometer. The separator is an integral part of the Varian Model 1700 gas<br />
chromatograph. This unit has the facility of venting the solvent to the atmosphere and<br />
maintaining the MS vacuum. Accordingly, large quantities of solvent can be injected <strong>on</strong>to<br />
the GLC column, vented, and prevented from entering the mass spectrometer. In this study,<br />
30 /A of the final methylene chloride extract (equivalent to 100 ng nitrosamine) was injected<br />
<strong>on</strong>to the GLC column.<br />
RESULTS AND DISCUSSION<br />
Before the recovery studies, the individual food products were analyzed for the 14 volatile<br />
N-nitrosamines. In instances where a nitrosamine was suspected, recovery values were ad<br />
justed appropriately. As shown in Table 1, recoveries of the 14 volatile N-nitrosamines<br />
added at 10 ppb (/µg/kg) to 25 g of the food products ranged from 67 to 100%, with an<br />
overall average of 84%.<br />
A total of 121 food samples were analyzed during this survey, including bac<strong>on</strong>, lard, baby<br />
food, total diet samples, miscellaneous pork products, spice-cure mixtures, imported<br />
cheeses, and Icelandic nati<strong>on</strong>al foods.<br />
Table 1. Recoveries of nitrosamines a added at levels of 10 ppb to 25 g food product<br />
Rec., %<br />
Sample Product Range Av.<br />
1 Bac<strong>on</strong>, hams, other pork<br />
products (liver) 77-95 82<br />
2 Baby foods 76-95 83<br />
3 Cheeses (imported) 71-94 85<br />
4 Fats and oils 67-100 83<br />
5 Total diet products 75-100 86<br />
6 Spice-cure mixtures 74-87 81<br />
7 Icelandic foods 84-100 91<br />
a 14 volatile A/-nitrosamines as described in text.<br />
All bac<strong>on</strong> samples were purchased at local retail markets. Bac<strong>on</strong> was analyzed raw and also<br />
after frying at 340 °F in an electric frypan for 3 min <strong>on</strong> each side. Excess fat was removed<br />
from the bac<strong>on</strong> by blotting with paper toweling. As shown in Table 2, N-nitrosopyrrolidine<br />
was found in fried bac<strong>on</strong> at levels ranging from 7 to 139 ppb.<br />
532
The fat cooked out of some of the bac<strong>on</strong> samples was analyzed and was found to c<strong>on</strong>tain<br />
from 19 to 92 ppb N-nitrosopyrrolidine. All values in Table 2 were c<strong>on</strong>firmed by combined<br />
GLC-MS, except Sample 16, which had a low c<strong>on</strong>centrati<strong>on</strong>.<br />
Table 2. N-Nitrosopyrrolidine (ppb) found in commercial bac<strong>on</strong><br />
Sample Raw Fried Fat cooked out<br />
1 0 67 92<br />
2 0 91<br />
3 0 74, a 64 a 111," 129*<br />
4 0 30<br />
5 0 119, 139<br />
6 0 129, c 104 c<br />
7 0 33, 33, 33 47<br />
8 0 34 48<br />
9 0 10<br />
10 0 16, 13 24<br />
11 0 10<br />
12 0 28, 29<br />
13 0 42, 51<br />
14 0 28,30 55,44<br />
15 0 22, 20 19<br />
16 0 7, d 7 d<br />
17 0 23, b 19 c<br />
18 0 13, 12, 13<br />
19 0 8<br />
20 0 42,34 49,44<br />
21 0 36, c 29 c<br />
22 0 65, 59, 72, 64<br />
a<br />
Held at 72°F for 8 hr.<br />
b<br />
Held at 72°F for a total of 34 hr.<br />
c<br />
Held at 72°F for 36 c<strong>on</strong>secutive hr.<br />
d<br />
Not c<strong>on</strong>firmed.<br />
No nitrosamines were found in the raw bac<strong>on</strong>. One sample of dry-cured bac<strong>on</strong> (not shown<br />
in the table) was analyzed; no nitrosamines were found in either the fried or the raw product.<br />
It is evident that pan-frying does induce the formati<strong>on</strong> of N-nitrosopyrrolidine in bac<strong>on</strong>, but<br />
the actual mode of formati<strong>on</strong> during the cooking process has not been definitely established.<br />
It may be through the formati<strong>on</strong> of nitrosoproline from proline and nitrite and subsequent<br />
decarboxylati<strong>on</strong> to nitrosopyrrolidine, or by direct interacti<strong>on</strong> of pyrrolidine, which could<br />
arise from proline or putrescine and nitrite.<br />
Four bac<strong>on</strong> samples were also analyzed after having been kept at room temperature<br />
(72°F) for 34-36 hr, before frying. It has been suggested that bacterial acti<strong>on</strong> <strong>on</strong> meat tissue<br />
could result in higher c<strong>on</strong>centrati<strong>on</strong>s of polyamines. In the presence of nitrite these could, in<br />
533
turn, react to produce larger c<strong>on</strong>centrati<strong>on</strong>s of N-nitrosamines up<strong>on</strong> frying. Samples 3, 6, 17,<br />
and 21 in Table 2 were held at 72°F 34 hr above<br />
that of Samples 2 and 16, while Samples 6 and 21 showed little or no increase. A reas<strong>on</strong> for<br />
these apparent inc<strong>on</strong>sistencies could be the amount of residual nitrite present in the bac<strong>on</strong>.<br />
Producers use different c<strong>on</strong>centrati<strong>on</strong>s of nitrite in their cures. Bac<strong>on</strong> with a low residual<br />
nitrite does not show an increase in nitrosamine formati<strong>on</strong> after storage at room temperature,<br />
since there is an insufficient quantity of nitrite available to react with the additi<strong>on</strong>al amines<br />
formed. On the other hand, bac<strong>on</strong> with a high residual nitrite, which can c<strong>on</strong>tinue to react<br />
with the amines as they form at room temperature, may show a greater formati<strong>on</strong> of<br />
nitrosamines. After frying, the bac<strong>on</strong> was found to c<strong>on</strong>tain increased levels of Nnitrosopyrrolidine.<br />
Table 3 shows the results of N-nitrosamine analyses <strong>on</strong> a variety of food products. Two<br />
types of baby foods that c<strong>on</strong>tained meat cured with nitrite were analyzed, but no<br />
nitrosamines were found (Samples 27 and 28). Of the 5 variety meat samples analyzed<br />
(Samples 29-34), <strong>on</strong>ly the fried canned bac<strong>on</strong> (Sample 32) showed N-nitrosopyrrolidine to<br />
be present. A peak at the retenti<strong>on</strong> time of N-nitrosodimethylamine (DMNA) was also<br />
observed but the c<strong>on</strong>centrati<strong>on</strong> was too low to c<strong>on</strong>firm its identity.<br />
Six total diet samples were analyzed (Samples 35-40) comprising (1) grain and cereal<br />
products; (2) meat, fish, and poultry; (3) leafy vegetables; (4) root vegetables; (5) oils, fats,<br />
and shortenings; and (6) dairy products. (These composites represent the typical diet of an<br />
American male teenager between 15 and 19 years of age.) Because of the physical nature of<br />
the oils, fats, and shortenings sample (Sample 40) and the dairy products sample (Sample<br />
35), they were analyzed by the procedure for fat cooked out of bac<strong>on</strong>. N<strong>on</strong>e of the 6 total<br />
diet samples analyzed was found to c<strong>on</strong>tain N-nitrosamines.<br />
Since lard is produced from rendered pork fat (in some instances a cured product) and<br />
high levels of N-nitrosopyrrolidine have been found in fried bac<strong>on</strong> and fat cooked out of<br />
bac<strong>on</strong>, it was decided to analyze a sample of lard (Samples 41 and 42). No N-nitrosamines<br />
were found, even after heating at 340°F for 10 min.<br />
Several workers have reported finding DMNA and N-nitrosodiethylamine in cheese,<br />
which had been processed with nitrite. However, n<strong>on</strong>e of the data had been c<strong>on</strong>firmed. In<br />
order to verify these findings we surveyed 10 imported cheeses (Samples 43-52). N<strong>on</strong>e of<br />
the 14 volatile N-nitrosamines was found in any of these products.<br />
Lamb is prepared by soaking it in a similar salt soluti<strong>on</strong> and further treated by smoking<br />
for 6-8 hr over sheep manure. The high c<strong>on</strong>centrati<strong>on</strong> of amines normally found in fish<br />
coupled with the added nitrite and unusual mode of food processing should have provided<br />
an ideal envir<strong>on</strong>ment for nitrosamine formati<strong>on</strong>. As shown in Table 3, 18 different samples<br />
were analyzed (Samples 53-70). Despite the apparently ideal envir<strong>on</strong>ment, no nitrosamines<br />
were found in any of the samples.<br />
534
A problem c<strong>on</strong>cerning the formati<strong>on</strong> of nitrosamines in meat spice-cure mixtures has<br />
recently been investigated. Salts, including nitrite, are added to a variety of spices in the<br />
formulati<strong>on</strong> of meat spice-cure mixtures. Amines present in spices could react with the<br />
added nitrite and form nitrosamines. It was c<strong>on</strong>firmed DMNA, N-nitrosopyrrolidine, and Nnitrosopiperidine<br />
in meat curing mixtures at levels up to 25 ppm.<br />
Until recently spice-cure mixtures have been marketed in several forms and formulati<strong>on</strong>s,<br />
such as single packaging of spices and curing salts, "piggy-back" (separate packaging of<br />
spices and curing salts), with and without sodium carb<strong>on</strong>ate buffers, and with and without<br />
protective coating of the nitrite crystals (zein corn protein).<br />
A survey of 32 spice-cure mixtures was undertaken to de<strong>term</strong>ine the effectiveness of<br />
the 3 marketing processes and also to de<strong>term</strong>ine the effect of storage time and temperature<br />
<strong>on</strong> nitros-amine formati<strong>on</strong>. The 9 samples found to c<strong>on</strong>tain nitrosamines are shown in Table<br />
4. All data in the table were c<strong>on</strong>firmed by MS.<br />
Three samples were part of a time-temperature stability study. Each was buffered with<br />
sodium carb<strong>on</strong>ate. The spice-cure mixtures were analyzed at room temperature and 100°F at<br />
0, 30, and 60 days storage. Samples 1 and 2 in Table 4 were nitrosamine-free when first analyzed,<br />
but after heat and storage, both c<strong>on</strong>tained N-nitrosopiperidine from 150 to 420 ppb<br />
and N-nitrosopyrrolidine from 50 to 95 ppb. The third sample (a c<strong>on</strong>trol not shown in Table<br />
4) did not c<strong>on</strong>tain nitrite and, as expected, no nitrosamines were found.<br />
Table 3. Food products analyzed: no nitrosamines detected<br />
Sample No. of analyses Product<br />
Pork Products<br />
23<br />
24<br />
25<br />
26<br />
27<br />
28<br />
29<br />
30<br />
31<br />
32a<br />
33<br />
34<br />
2<br />
1<br />
1<br />
1<br />
5<br />
6<br />
2<br />
2<br />
1<br />
1<br />
1<br />
1<br />
pork liver (raw)<br />
pork liver (fried)<br />
Polish style sausage (raw)<br />
Polish style sausage (boiled)<br />
Baby Foods<br />
beans, potatoes, and ham<br />
casserole creamed potatoes and ham<br />
Variety Meats<br />
Crumbled bac<strong>on</strong><br />
crumbled ham<br />
bac<strong>on</strong> (imported canned; raw)<br />
bac<strong>on</strong> (imported canned; fried)<br />
beer sticks<br />
smoked pepper<strong>on</strong>i<br />
Total Diet Samples<br />
35 1 dairy products<br />
535
36<br />
37<br />
38<br />
39<br />
40<br />
41<br />
42<br />
1<br />
1<br />
1<br />
1<br />
1<br />
1<br />
1<br />
meat, fish, and poultry<br />
grain and cereal<br />
leafy vegetables<br />
root vegetables<br />
fats, oils, and shortenings<br />
Fats and Oils<br />
lard (uncooked)<br />
lard (heated) b<br />
Cheese<br />
43 2 Edam<br />
44 2 Gouda<br />
45 2 Samsoe<br />
46 2 Danbo<br />
47 2 Gouda (baby)<br />
48 2 Blue<br />
49 2 Tilsit Sq. Havarti<br />
50 1 Port Salut<br />
51 1 Tybo<br />
52 1 Tilsit Havarti<br />
53<br />
54<br />
55<br />
56<br />
57<br />
58<br />
59<br />
60<br />
61<br />
62<br />
63<br />
64<br />
65<br />
66<br />
69<br />
70<br />
1<br />
1<br />
2<br />
2<br />
1<br />
1<br />
1<br />
3<br />
1<br />
2<br />
2<br />
1<br />
1<br />
1<br />
1<br />
1<br />
Icelandic Nati<strong>on</strong>al Foods<br />
smoked lamb's leg<br />
salted lamb<br />
smoked mutt<strong>on</strong> sausage<br />
singed lamb's head<br />
smoked salm<strong>on</strong><br />
smoked lamb's leg<br />
singed lamb's leg<br />
smoked mutt<strong>on</strong> sausage<br />
smoked trout<br />
putrescent skate<br />
singed lamb's head (oil flame)<br />
smoked lamb (birchwood)<br />
smoked lamb (beechwood)<br />
smoked salm<strong>on</strong><br />
salted lamb<br />
smoked mutt<strong>on</strong> sausage<br />
a C<strong>on</strong>tained 6 ppb A/-nitrosopyrrolidine, c<strong>on</strong>firmed by GLC-MS. b Heated at 340°F 10 min.<br />
536
Sample 3, found to c<strong>on</strong>tain 343 ppb DMNA, 730 ppb N-nitrosopyrrolidine, and 2000 ppb<br />
N-nitrosopiperidine, was a spice-cure mix being used at the time in a cured meat product.<br />
From the data shown in the table it is evident that buffering with sodium carb<strong>on</strong>ate does not<br />
block nitrosamine formati<strong>on</strong>. Samples 4, 7, and 9 were buffered and still DMNA, Nnitrosopiperidine,<br />
and N-nitrosopyrrolidine were found. Ascorbate was also found to be<br />
ineffective in blocking nitrosamine formati<strong>on</strong> as shown by Samples 7 and 8, which<br />
c<strong>on</strong>tained this compound and yet had levels of N-nitrosopiperidine and N-nitrosopyrrolidine<br />
up to 232 ppb. Similar results with ascorbate were obtained. Two samples of spice-cure<br />
mixtures utilizing the piggyback method of packaging were analyzed. No nitrosamines were<br />
found. A limited study of 4 samples of spice-cure mixtures c<strong>on</strong>taining nitrite crystals coated<br />
with zein were analyzed and all 4 were found to be free of nitrosamines. Our studies have<br />
indicated that piggy-back packaging is the most effective method of preventing nitrosamine<br />
formati<strong>on</strong>, since the react ants do not come in c<strong>on</strong>tact until they are mixed with the<br />
comminuted meat. Federal laws in the United States and Canada now prohibit the marketing<br />
of meat spice-cure mixtures in any way other than the piggy-back method.<br />
Table 4. Analysis of spice-cure mixtures for nitrosamines (ppb)'<br />
Buffer 0 days 30 days 60 days<br />
Sample 72°F 72°F 100°F 72°F 100°F<br />
1 Na2C03 0 0 0 0 50 NPYR<br />
2 Na,C03 0 0 150 NPIP 0 420 NPIP 95<br />
3 Na2C03 343<br />
730<br />
2000<br />
DMNA<br />
NPYR<br />
NPIP<br />
4 Na4P207 72 DMNA<br />
5 n<strong>on</strong>e 106<br />
314<br />
28<br />
6 n<strong>on</strong>e 768<br />
144<br />
DMNA<br />
NPIP<br />
NPYR<br />
NPIP<br />
NPYR<br />
7 b Na2C03 232 NPIP<br />
d b n<strong>on</strong>e 160 24 NPIP<br />
9 Na2C03 29 91<br />
49<br />
NPYR<br />
DMNA<br />
NPIP<br />
NPYR<br />
a DMNA, N-nitrosodimethylamine; NPYR, N-nitrosopyrrolidine; NPIP, A/nitrosopiperidine.<br />
b With ascorbate added.<br />
537<br />
NPYR
REFERENCES<br />
1. Castegnaro, M., Pignatelli, B., & Walker, E. A. (1974) Analyst 99, 156-162<br />
2. Collins-Thomps<strong>on</strong>, D. L., Chang, P. C, Davids<strong>on</strong>, C. M., Larm<strong>on</strong>d, E., & Pivnick, H.<br />
(1974) /. Food Sci. 39, 607-609<br />
3. Kushnir, I., Feinberg, J. I., Pensabene, J. W., Piotrowski, E. G., Fiddler, W., &<br />
Wasserman,<br />
4. Sen, N. P., Iyengar, J. R., D<strong>on</strong>alds<strong>on</strong>, B. A., & Panalaks, T. (1974) J. Agric. Food<br />
Chem. 22, 540-541<br />
5. Sen, N. P., Schwinghamer, L., D<strong>on</strong>alds<strong>on</strong>, B. A, & Miles, W. F. (1972) J. Agric.<br />
Food Chem. 20, 1280-1281<br />
6. Fiddler, W., Doerr, R. C, Ertel, J. R., & Wasserman, A. E. (1971) JAOAC 54, 1160-<br />
1163<br />
7. Heisler, E., Siciliano, J., Krulick, S., Fein-berg, J., & Schwartz, J. H. (1974) J. Agric.<br />
Food Chem. 22, 1029-1032<br />
8. Palumbo, S. A., Smith, J. L., Gentilcore, K. M., & Fiddler, W. (1974) /. Food Sci. 39,<br />
1257-1258<br />
9. Sen, N. P, Miles, W. F., D<strong>on</strong>alds<strong>on</strong>, B. A, Panalaks, T., & Iyengar, J. R. (1973) Nature<br />
L<strong>on</strong>d<strong>on</strong> 245, 104-105<br />
538
LUNG CANCER CAUSED BY SMOKING AND CADMIUM AND CHROMIUM IN<br />
HUMAN LUNG TISSUE<br />
Camelia Bara<br />
University of Oradea, Faculty of Envir<strong>on</strong>mental Protecti<strong>on</strong><br />
26, General Magheru Street, Oradea<br />
ABSTRACT<br />
The pulm<strong>on</strong>ary Cd and Cr c<strong>on</strong>tent were de<strong>term</strong>ined from 53 lung cancer patients operated<br />
for cancer and from 39 patients who died of n<strong>on</strong>-malignant diseases. The results were<br />
correlated with smoking habits, pulm<strong>on</strong>ary emphysema and occupati<strong>on</strong>al history. Both the<br />
pulm<strong>on</strong>ary Cd and Cr increased with the amount of smoking. In ex-smokers the Cr c<strong>on</strong>tent<br />
in lung tissue did not diminish with the time since stopping smoking, but the Cd did follow<br />
the half-life of about 9 years. The pulm<strong>on</strong>ary Cd, compared with smoking habits, behaved<br />
similarly in both the lung cancer and c<strong>on</strong>trol patients, whereas Cr in the lung cancer patients<br />
could not be explained solely by smoking, but some of the cancer patients may have been<br />
occupati<strong>on</strong>ally exposed to Cr. Speciati<strong>on</strong> was not studied.<br />
Keywords: Cadmium, chromium, smoking, occupati<strong>on</strong>al hygiene, lung cancer, pulm<strong>on</strong>ary<br />
emphysema, lobe samples, sample preparati<strong>on</strong>, direct current plasma atomic emissi<strong>on</strong><br />
spectrometry (DCP-AES).<br />
INTRODUCTION<br />
Lung cancer patients have usually been exposed to tobacco smoke for several decades. In<br />
additi<strong>on</strong> to organic compounds, tobacco c<strong>on</strong>tains carcinogenic metals such as As, Cd, Cr<br />
and Ni. These metals may functi<strong>on</strong> as an indicati<strong>on</strong> of tobacco smoking in lung tissue, and<br />
in this respect especially Cd 2 and Cr 3 have proved promising. The main source of Cd in<br />
tobacco is c<strong>on</strong>sidered to be phosphate fertilizers. Cr is a comm<strong>on</strong> envir<strong>on</strong>mental pollutant<br />
originating from the burning of coal and from metallurgical chromium and cement<br />
producing plants, and c<strong>on</strong>centrating in tobacco leaves from the c<strong>on</strong>taminated soil. In this<br />
work the pulm<strong>on</strong>ary Cd and Cr c<strong>on</strong>tent was compared with the smoking habits and<br />
occupati<strong>on</strong>al history of patients with and without lung cancer.<br />
Table 1. Age and smoking habits of the lung cancer and c<strong>on</strong>trol patients<br />
Cancer C<strong>on</strong>trol<br />
Number 53 39<br />
Age 61 ±10 67±9<br />
Smoking time (years) 39+11 37± 12<br />
Pack years 41 + 15 36+17<br />
Time since stopping smoking (years) 4 + 7 8±11<br />
539
MATERIALS AND METHODS<br />
The material c<strong>on</strong>sisted of 53 male lung cancer patients operated for the cancer, and of 39<br />
c<strong>on</strong>trol patients who died of n<strong>on</strong>-malignant diseases and were autopsied. Because<br />
pulm<strong>on</strong>ary emphysema is a good indicator for smoking, the c<strong>on</strong>trol material was collected<br />
to include cases with various degrees of emphysema. The smoking habits and occupati<strong>on</strong>al<br />
history were required from the patients or next of kin and from the hospital records (Table<br />
1). The severity of emphysema was graded in the radiographs of excised air-inflated lungs<br />
or lobes of lungs.<br />
Fresh lung tissue for analyses was taken from the (apico) posterior and anterior<br />
segment of the upper lobe and from the superior (apical) segment and basal part of the lower<br />
lobe. The samples, weighing 0.5-2 g, c<strong>on</strong>tained no pleural surface or cancer tissue. Ninety<br />
percent of the Cd analyses were made from the anterior segment and 80% of the Cr analyses<br />
from the (apico) posterior segment of the upper lobe. The samples were prepared under a<br />
laminar flow hood avoiding c<strong>on</strong>taminati<strong>on</strong>. The fresh, vacuum-dried samples were ashed in<br />
glass cups with a blank and NBS Bovine Liver (SRM 1577) c<strong>on</strong>trol for 48-72 h in a low<br />
temperature oxygen plasma asher (BIO-RAD Plasma Asher E 2000, 100-120 °C). The<br />
tissue residue was dissolved in a total of 3 ml of 50 % ethanol and transferred into a Tefl<strong>on</strong><br />
cup and dried at 80 °C for 24 h. The residue was digested in a mixture of 3 ml of<br />
c<strong>on</strong>centrated nitric acid and 0.2 ml of percloric acid. The soluti<strong>on</strong> was allowed to stand<br />
overnight and dry-heated at moderate temperature (100-150 °C) in a sand-bath. Finally the<br />
ash was diluted in 3-6 ml of high quality water. The Cd and Cr c<strong>on</strong>tents were de<strong>term</strong>ined <strong>on</strong><br />
a direct current plasma atomic emissi<strong>on</strong> spectrometer (DCP-AES) (SpectraSpan IIIB<br />
coupled with a Hewlett-Packard 85 data processor) not c<strong>on</strong>sidering speciati<strong>on</strong>. A standard<br />
additi<strong>on</strong> method was employed for Cr to eliminate matrix interference. The analytical<br />
characteristics are shown in Table 2.<br />
Table 2. Analytical characteristics<br />
Cd Cr______<br />
Accuracy<br />
NBS 1577 Bovine Liver 0.36 0.25<br />
(0.274 ±0.086) a (0.125±0.057) a<br />
Lan<strong>on</strong>orm-Metalle 2 0.028<br />
(0.021-0.030) b<br />
RSD (0.02 ppm) % 3.9 2.4<br />
Detecti<strong>on</strong> limit (ppm) 0.010 0.017<br />
_________________________________________________________<br />
RESULTS AND DISCUSSION<br />
Both the total pulm<strong>on</strong>ary Cd and Cr c<strong>on</strong>tent were higher in the lung cancer (Cd 3.0 ± 2.4<br />
and Cr 6.1 ± 4.2 µg/g dry weight) than in the c<strong>on</strong>trol patients (Cd 2.1 ± 1.9 and Cr 4.1 ± 4.0<br />
540
µg/g dry weight). The pulm<strong>on</strong>ary Cd increased with the amount of smoking measured in<br />
pack years (r = 0.32, p = 0.005) in both the lung cancer and c<strong>on</strong>trol patients. Cr correlated<br />
positively with the smoking time in the c<strong>on</strong>trol (r = 0.41, p
FORMATION OF AFLATOXIN M- TRIFLUOROACETIC ACID DERIVATIVE IN<br />
OPTIMUM CONDITIONS<br />
Lucian Bara<br />
University of Oradea, Faculty of Envir<strong>on</strong>mental Protecti<strong>on</strong><br />
26, General Magheru Street, Oradea<br />
ABSTRACT<br />
Because thin-layer chromatographic (TLC) c<strong>on</strong>firmati<strong>on</strong> of identity and reverse-phase<br />
liquid chromatographic (LC) de<strong>term</strong>inati<strong>on</strong> with fluorescence detecti<strong>on</strong> of aflatoxin M1,<br />
both require the derivative formed in the reacti<strong>on</strong>s c<strong>on</strong>diti<strong>on</strong>s were studied to obtain<br />
complete derivative formati<strong>on</strong>. Of the various organic solvents tested, the reacti<strong>on</strong> between<br />
M1 and TFA preceded best in the n<strong>on</strong>polar solvents hexane and isooctane. Other parameters<br />
investigated were reacti<strong>on</strong> temperature and time, aflatoxin M1 c<strong>on</strong>centrati<strong>on</strong>, and solvent<br />
volume. The following procedure is c<strong>on</strong>sidered optimum: 200 µL each of hexane and<br />
trifluoroacetic acid are mixed with M1 standard in a silylated glass vial or with milk residue<br />
in a regular glass vial with a Tefl<strong>on</strong>-lined screw cap and heated 10 min at 40ºC. The mixture<br />
is evaporated to dryness under N2, and the derivative is saved for TLC or LC. No unreacted<br />
aflatoxin M1 was detected by reverse-phase LC after this procedure was incorporated for<br />
analysis of milk samples.<br />
INTRODUCTION<br />
The hemiacetal derivatives of aflatoxins B1 and G1 (B2a and G2a) have been used for thinlayer<br />
chromatographic (TLC) c<strong>on</strong>firmati<strong>on</strong> of identity since 1971. The derivatives are<br />
highly fluorescent and used extensively for the quantitati<strong>on</strong> of these aflatoxins by reversephase<br />
liquid chromatography (LC), because B1 and G1 are <strong>on</strong>ly weakly fluorescent in<br />
aqueous mobile phases. Derivatives B2a and G2a are readily prepared by the trifluoroacetic<br />
acid (TFA)-catalyzed hydrati<strong>on</strong> of aflatoxins B1 and G1 are at room temperature. This is not<br />
true with aflatoxin M1 because the reacti<strong>on</strong> rarely goes to completi<strong>on</strong>. The first reversephase<br />
LC method for de<strong>term</strong>ining aflatoxin M1 in milk and milk products, which measured<br />
the fluorescence of the M1-TFA derivative (designated M2a), was reported by Beebe and<br />
Takahashi in 1980. No formal evidence has been published that definitely identifies this<br />
derivative as the hemiacetal; however, for c<strong>on</strong>sistency with the authors, it is reffered to here<br />
as M2a. Beebe and Takahashi were successful in the reproducible derivatizati<strong>on</strong> of aflatoxin<br />
M1 to M2; however, other scientists have encountered incomplete derivatizati<strong>on</strong> and have<br />
incorporated different reacti<strong>on</strong> c<strong>on</strong>diti<strong>on</strong>s. C<strong>on</strong>sequently, some of these workers have<br />
chosen to quantitate aflatoxin M1 directly, even though M1 is less fluorescent than M2a.<br />
In this report, the reacti<strong>on</strong> parameters for derivatizati<strong>on</strong> of aflatoxin M1 or M2a are<br />
examined, and the procedure that produces complete c<strong>on</strong>versi<strong>on</strong> of M1 to the M2a derivative<br />
is given.<br />
542
MATERIALS AND METHOD<br />
Apparatus and Reagents<br />
(a) Solvents. – All are reagent grade.<br />
(b) Trifluoroacetic acid (TFA). – Purity 99 + %.<br />
(c) Dichlorodimethysilane (DDS). – Prepare 5% (v/v) soluti<strong>on</strong> in toluene.<br />
(d) Aflatoxin standard soluti<strong>on</strong>s. – Aflatoxin M1 standards were 0.5, 7.04, and 8.37<br />
μg/mL in acet<strong>on</strong>itrile-benzene (1 + 9). Aflatoxin B1 and G1 standard soluti<strong>on</strong> was 8.0<br />
μg each/mL in acet<strong>on</strong>itrile-benzene (2+98).<br />
(e) Mobile phase. – Mix water-isopropyl alcohol-acet<strong>on</strong>itrile (80+12+8). Degas with<br />
ultras<strong>on</strong>ic probe.<br />
(f) Heating block. – Reacti-therm heating module or equivalent.<br />
(g) Silylated vials. – Add 5% DDS soluti<strong>on</strong> to 1-1.5 dram vials and heat ca 40 min at<br />
45-55°C. Discard soluti<strong>on</strong> and rinse vials 3 times with toluene and then 3 times with<br />
methanol. Heat vials at 75°C for 20-30 min to evaporate methanol. Cap vials (with<br />
Tefl<strong>on</strong> liners) and store.<br />
(h) LC system. – Spectra-Physiscs Model 8700, equipped with injector (Rheodyne<br />
Model 7125) with 2.0 mL loop, recorder/integrator (Spectra-Physics SP4270),<br />
fluorescence detector (Kratos FS970), set at 365 nm for excitati<strong>on</strong> and 418 nm for<br />
emissi<strong>on</strong>, and 4.3 mm id X 25 cm Zorbax ODSLC column. Mobile phase, waterisopropyl<br />
alcohol-acet<strong>on</strong>itrile (80 + 12 + 8), 1.0 mL/min.<br />
Initial Derivative Formati<strong>on</strong><br />
The following were mixed in a Tefl<strong>on</strong>-lined screw-cap vial: 200 µL solvent, 50 µL TFA,<br />
and 50 µL aflatoxin M1 standard. The mixture was allowed to react at room temperature for<br />
15 min and then was evaporated under nitrogen. The residue was dissolved in 2000 µL<br />
water-acet<strong>on</strong>itrile (75 + 25) for LC. The injecti<strong>on</strong> volume was 50 µL.<br />
RESULTS AND DISCUSSION<br />
This study was initiated by duplicating the reacti<strong>on</strong> c<strong>on</strong>diti<strong>on</strong>s of Beebe and Takahashi.<br />
They used n-hexane as a base solvent to dissolve the dairy extract residue c<strong>on</strong>taining<br />
aflatoxin M1. Although hexane is an excellent solvent for this purpose, it is a poor solvent<br />
for the polar aflatoxin M1, when present in pure form (e.g., aflatoxin M1 standard).<br />
Therefore, other solvents were substituted for hexane to de<strong>term</strong>ine if any improvement<br />
colud be obtained. Solvents were chosen that scanned the elutropic series, with values of -<br />
0.25 to 0.95. The data indicate that the solvents that produced n-hexane, isooctane, and<br />
1,1,2-trichlorotrifluoroethane. Generally, as the solvent polarity increased, the M1,<br />
c<strong>on</strong>versi<strong>on</strong> decreased. Unreacted aflatoxin M1 was detected with all solvents. When water<br />
was added to the reacti<strong>on</strong> mixture, poor yields of M2a were obtained. Even though aflatoxin<br />
M1 is less soluble in n<strong>on</strong>polar solvents, the reacti<strong>on</strong> was more complete with them. In this<br />
study, aflatoxin M1 standard was added to the solvent-TFA mixture in 50 µL aliquots. This<br />
technique prevented adsorpti<strong>on</strong> of M1 to the glass vial – a problem encountered with<br />
aflatoxin standard soluti<strong>on</strong>s and discussed in more detail later.<br />
543
Next, the optimum reacti<strong>on</strong> temperature was de<strong>term</strong>ined for hexane and isooctane as base<br />
solvents. 1,1,2-Trichlorotrifluoroethane was not tested because it is not a comm<strong>on</strong><br />
laboratory chemical. The reacti<strong>on</strong> c<strong>on</strong>diti<strong>on</strong>s were identical to those given in the text and for<br />
the data in Table 1, except the temperature was varied from 10 to 50°C. As the temperature<br />
increased, aflatoxin M2a was produced in greater yield until a decrease was detected at 50°C.<br />
The optimum reacti<strong>on</strong> temperature was 40°C. No unreacted M1 could be detected at<br />
temperatures above ambient; however, some must have been present at 30 and 50°C<br />
because less M2a was present. Aflatoxin M1 is not as fluorescent as M2a in polar mobile<br />
phases, so small, undected amounts of M1 could be present.<br />
The reacti<strong>on</strong> temperature was held at 40°C, and hexane was selected as the residue<br />
solvent because most aflatoxin laboratories use it routinely in their assays. No unreacted M1<br />
was found after 15 min, and maximum aflatoxin M2a peak areas occurred at 30-45 min. This<br />
was a l<strong>on</strong>ger reacti<strong>on</strong> time than desired, so the TFA volume was increased from 50 to 200<br />
µL, and the experiment was repeated. With equal volumes (200 µL) of hexane and TFA, the<br />
reacti<strong>on</strong> was complete at 10 min, and <strong>on</strong>ly M2a, was detected. On the basis of these data, it<br />
appears that the best c<strong>on</strong>diti<strong>on</strong>s for forming M2a, from M1 are mixing equal 200 µL porti<strong>on</strong>s<br />
of hexane and TFA with the dairy extract residue and letting the mixture react at 40ºC for 10<br />
min.<br />
These reacti<strong>on</strong> parameters were tested with increasing quantities of aflatoxin M1 t<br />
de<strong>term</strong>ine the maximum amount of toxin that would react. Only aflatoxin M2a was visible<br />
when quantities of less than or equal to 150 ng or 300 200 µL M1 standard were treated with<br />
TFA. The data suggest that this volume (300 µL) of standard soluti<strong>on</strong> diluted the reacti<strong>on</strong><br />
mixture excessively. To de<strong>term</strong>ine if this was true, a c<strong>on</strong>centrated M1 soluti<strong>on</strong> (1675 ng in<br />
200 µL) was treated, and a very small unreacted M1 peak-63 area counts – was detected.<br />
Therefore, the quantity of aflatoxin M1 is not a critical factor in the derivatizati<strong>on</strong>, but<br />
diluti<strong>on</strong> of the reactants is to be avoided. With most of the current methods, the dairy<br />
product extract is evaporated before it is treated with hexane and TFA; therefore, the<br />
proposed technique will successfully form the M2a derivative.<br />
If aflatoxin M1 standards are evaporated in a glass vial, M1 is irreversibly adsorbed to<br />
the glass. Glass vials should be silylated to avoid adsorpti<strong>on</strong> of M1 during evaporati<strong>on</strong>.<br />
Standard soluti<strong>on</strong>s (benzene-acet<strong>on</strong>itrile) of less than 200 µL can be added to the hexane-<br />
TFA mixture directly and the reacti<strong>on</strong> will go to completi<strong>on</strong>.<br />
The derivatizati<strong>on</strong> procedure was tried also with standard soluti<strong>on</strong>s c<strong>on</strong>taining<br />
aflatoxins B1 and G1 and it c<strong>on</strong>verted them completely to B2a and G2a. In a recent<br />
internati<strong>on</strong>al collaborative study, four collaborators obtained incomplete reacti<strong>on</strong> of<br />
aflatoxin M1 with TFA, probably because the specified procedure did not use heat. Since<br />
then, the 4 collaborators have successfully used the procedure given in this paper (private<br />
communicati<strong>on</strong>s).<br />
In summary, the optimum c<strong>on</strong>diti<strong>on</strong>s for c<strong>on</strong>verting aflatoxin M1 to aflatoxin M2a are to<br />
add equal 200 µL volumes of hexane and TFA to dry dairy extract in a screw-cap vial, mix<br />
well, heat the vial at 40ºC for 10 min, evaporate the mixture under N2, and save the residue<br />
544
for either TLC or reverse-phase LC de<strong>term</strong>inati<strong>on</strong>s. This procedure also will successfully<br />
form aflatoxins soluti<strong>on</strong>s should be transferred to silylated vials, prior to forming the<br />
derivatives, to prevent irreversible adsorpti<strong>on</strong> of the aflatoxins to the glass; however, sample<br />
extracts do not need silylated vials.<br />
REFERENCES<br />
1. Przybylski, W. (1975) J. Assoc. Off. Anal. Chem. 58, 163-164<br />
2. Beebe, R.M. &Takahashi, D.M. (1980) J.Agric, Food Chem.28, 481-482<br />
3. Tuinstra, L.G.M., Th., &Haasnoot, W. (1982) Fresenius Z. Anal.Chem.312, 622-623.<br />
545
DETERMINATION OF “CARCINOGENESIS” BY ALTERNATIVE BIOASSAYS<br />
Camelia Bara, Lucian Bara<br />
University of Oradea, Faculty of Envir<strong>on</strong>mental Protecti<strong>on</strong><br />
26, General Magheru Street, Oradea<br />
ABSTRACT<br />
Chhabra et al. (1988) describe a “carcinogenesis” bioassay that appeared to show the<br />
inhibiti<strong>on</strong> of sp<strong>on</strong>taneous tumors by 4-hexylresorcinol in rats and mice. This is not an<br />
unusual finding. It is even possible to find c<strong>on</strong>sistent patterns am<strong>on</strong>g these<br />
“antitumorigens”. All four of the antidiabetic drugs tested in the program and all but <strong>on</strong>e of<br />
the antioxidants bel<strong>on</strong>g to the group with statistically significant dose-related reducti<strong>on</strong>s in<br />
sp<strong>on</strong>taneous tumors <strong>on</strong>ly; however, the group also includes a dioxin.<br />
Chhabra et al. suggest that their test compound might be investigated as an<br />
antineoplastic agent. Since there are more than 40 other compounds with the same property,<br />
should they also be investigated as antineoplastic agents? If we think of the<br />
“carcinogenesis” bioassay in a symmetric fashi<strong>on</strong> (as capable of showing both tumorigenic<br />
and antitumorigenic activity), a finding that a compound is a potential “carcinogen” have the<br />
same validity as the finding that a compound is a potential “antineoplastic” agent.<br />
In the following secti<strong>on</strong>s, an argument is made that, with the current design (maximum<br />
tolerated dose and 18-24 m<strong>on</strong>ths exposure), it can be expected that every compound tested<br />
will produce an effect <strong>on</strong> the incidence of tumors, regardless of whether the compound is a<br />
potential “carcinogen”, “antineoplastic” agent, or neither. Next, a statistical procedure is<br />
outlined that can be used to test this hypothesis, and the procedure is applied to six<br />
bioassays that were run <strong>on</strong> similar compounds in the same laboratory during the same<br />
period. Finally, the implicati<strong>on</strong>s of this hypothesis for interpretati<strong>on</strong> of these bioassays are<br />
discussed.<br />
INTRODUCTION<br />
The philosophical foundati<strong>on</strong> of toxicology as a science rests <strong>on</strong> a thought experiment<br />
attributed to the Renaissance alchemist Paracelsus (1493-1541). In this thought experiment,<br />
a potential victim is given ever smaller doses of a “pois<strong>on</strong>” until a dose is found that has no<br />
apparent effect. Then, the potential victim is given ever-larger doses of some “n<strong>on</strong>pois<strong>on</strong>”<br />
until serious biological damage results. The c<strong>on</strong>clusi<strong>on</strong> is the aphorism, “Solely the dose<br />
de<strong>term</strong>ines that a thing is not a pois<strong>on</strong>”.<br />
There is a modern versi<strong>on</strong> of this thought experiment. Let as c<strong>on</strong>sider giving a rat or a<br />
mouse a dose of a biologically active drug, such as a B-adrenergic receptor-blocking agent.<br />
Let the dose be large enough to produce almost 100 % blockade. The catecholamines that<br />
the body produces in the normal course of life will fail to have the effects expected, and<br />
homeostasis will be affected, perhaps calling forth greater output of catecholamines. These<br />
agents will then bind to sec<strong>on</strong>dary receptor sites, producing resp<strong>on</strong>ses, which, in turn, will<br />
546
trigger still other resp<strong>on</strong>ses. In this way, the animal may eventually reach some state of<br />
equilibrium but in the presence of a changed biochemical milieu.<br />
If, now, we c<strong>on</strong>tinue treating this animal with the 100 % blocking dose for most of its<br />
natural life span, we can expect this changed biochemical milieu to be reflected in the<br />
patterns of lesi<strong>on</strong>s that are associated with the aging process. Thus, the treated animals will<br />
tend to have a different syndrome of chr<strong>on</strong>ic lesi<strong>on</strong>s than untreated c<strong>on</strong>trols. If the strains of<br />
rat or mouse chosen are pr<strong>on</strong>e to tumor, untreated, then we can expect that this shift in<br />
senile lesi<strong>on</strong>s will involve a shift in neoplastic lesi<strong>on</strong>s.<br />
Suppose we used some other drug, like e calcium channel blocker, or a m<strong>on</strong>oamine<br />
oxidase (MAO) inhibitor, or any other drug at a dose sufficiently high to produce a<br />
sustained biological effect. The same argument holds, that the resulting change in<br />
biochemical milieu will produce a shift in senile lesi<strong>on</strong>s (including tumor patterns).<br />
Suppose, instead of a drug, we use an insecticide, or a dye precursor, or any other agent, but<br />
at a dose sufficiently high ti produce a discernible biological effect (like a 10 % reducti<strong>on</strong> in<br />
weight). The same argument holds.<br />
Thus, we can expect, just from the design of these studies, that the “carcinogenesis”<br />
bioassay will show a treatment-related shift in neoplastic lesi<strong>on</strong>s for every compound tested.<br />
MATERIALS AND METHOD<br />
A. Using Data from the “Carcinogenesis”Bioassay<br />
The standard statistical analyses of the “carcinogenesis” bioassay require that we compare<br />
counts of animals with specific tumor types between treatment groups. As a result, the<br />
unsual NTP study involves formal hypothesis tests comparing tumor incidences for 10-15<br />
tumor types, each comparis<strong>on</strong> involving a test of c<strong>on</strong>trols versus low dose; c<strong>on</strong>trols versus<br />
high dose; two dose-resp<strong>on</strong>se tests, adjusted for early deaths; and n<strong>on</strong>linearity hypothesis<br />
tests per sex per species.<br />
The problem of multiple testing and inflati<strong>on</strong> of the false-positive error rate has had<br />
c<strong>on</strong>siderable discussi<strong>on</strong> in the literature. However, there is another side of the problem, the<br />
false-negative error rate. There are <strong>on</strong>ly 50 animals per treatment, sex, species, and a<br />
statistical test that compares the number of animals with the a given lesi<strong>on</strong> cannot detect a<br />
slight shift in the probability of lesi<strong>on</strong>. For instance, if the background incidence of a<br />
particular tumor is 20 % and if the treatment reduces that to 7%, half of the studies would<br />
show a “significant” difference and half would not. Thus, the failure to show “significant”<br />
differences between treatments and c<strong>on</strong>trols for some of the compounds that have been<br />
tested is no proof that there was no effect. In fact, a rather dramatic increase or reducti<strong>on</strong> in<br />
a particular tumor type could have occurred without producing a “significant” result.<br />
There are ways of strengthening the power of statistical hypothesis tests. One way is to<br />
allow each animal to provide more informati<strong>on</strong>. When we compare the counts of animals<br />
with a specific tumor type, each animal provides <strong>on</strong>ly 0 or a 1, the least amount of<br />
informati<strong>on</strong> available from a single experimental unit. The fact that we look at several<br />
547
different tumor types does not help. In fact, because of multiple testing we have to adjust the<br />
nominal p value and make the test even less powerful. Since the individual animal is the unit<br />
of experimentati<strong>on</strong>, the greatest power can be gained by using the entire pattern of lesi<strong>on</strong>s<br />
seen in a given animal and by using <strong>on</strong>ly <strong>on</strong>e or two hypothesis tests for the entire sex X<br />
species experiment.<br />
The appendix describes a method of multivariate analysis that does it. General<br />
multivariate methods foe analyzing chr<strong>on</strong>ic toxicity studies can be found in Salsburg. The<br />
specific method described in the Appendix was applied to six AZO dyes, whose bioassays<br />
took place at the same site during the same period of time, and these analyses are discussed<br />
in the next secti<strong>on</strong>.<br />
If the Paracelsean thought experiment is correct, and with it the hypothesis that any<br />
treatment has an effect, then a powerful statistical test should detect a significant treatment<br />
effect in almost all studies – “almost all” because no statistical test has 100 % power.<br />
B. C<strong>on</strong>structing Additi<strong>on</strong>al Studies to Test the Hypothesis<br />
Statistical tests that show a shift in tumor patterns for all or almost all compounds, sexes,<br />
and species in these studies provide str<strong>on</strong>g evidence in favor of the hypothesis. However,<br />
scientific hypotheses are not necessarily proven by the reanalysis of old data. We usually try<br />
to c<strong>on</strong>struct new experiments that provide an opportunity to refute the hypothesis.<br />
We can c<strong>on</strong>struct such studies by taking compounds that appear to show a complicated<br />
shift in tumors and trying to find the biochemical or tissue changes that occur as a result of<br />
treatment. We have to be careful to recognize that this is a very general hypothesis, and we<br />
should not restrict the search for changes in milieu to specific horm<strong>on</strong>es or to specific<br />
histopathological events. We also have to be careful that such studies are used primarily to<br />
examine the validity of the Paracelsean hypothesis. If, for instance, we find a set of changes<br />
that seem to be associated with <strong>on</strong>e class of chemical compounds, and if that class tends to<br />
be mutagenic or ‘carcinogenic” in other models, we cannot assume that we have found<br />
marker or proximate events unique to that class, and far more work will be needed to<br />
establish such findings as indicative of that class. I propose a set of experiments of<br />
following type: We pick 10-20 of the compounds that have shown paradoxical shifts in<br />
tumors in the NCI/NTP program (increase in tumor incidence for <strong>on</strong>e type of tissue<br />
associated with decreases in tumor incidence of another type of tissue, where the<br />
paradoxical events may be in different sexes or species). Using the same doses and the same<br />
strains of animal, we redo the feeding study for a small number of animals over a relatively<br />
short period (1-3 m<strong>on</strong>ths). At the end of this time, we sacrifice the animals and examine the<br />
target tissues (tissues in which paradoxical tumor increases or decreases have occurred).<br />
The hypothesis predicts that we will find differences in “milieu”. There are rough tools<br />
for comparing general patterns of proteins or fragments of active agents between two sets of<br />
tissue, which have been used in the primary screening of undifferentiated biological<br />
products where we are looking for something “new”. It would not be necessary to identify<br />
these differences. It should be sufficient to show that there are discernible differences. If<br />
548
these general patterns appear to be identical between treatment groups, then that would be<br />
evidence against this hypothesis.<br />
SIX AZO DYES<br />
As part of its <strong>on</strong>going program applying a “carcinogenesis” bioassay to chemical<br />
compounds now in use, the NTP has reported <strong>on</strong> studies involving six AZO dyes. These<br />
assays were c<strong>on</strong>ducted at a single site. Each dye was subjected to the same protocol, in<br />
which male and female rats and mice were divided into three groups each: c<strong>on</strong>trols, low<br />
dose, and high dose. There was an excepti<strong>on</strong> to the standard protocol. For three of the dyes,<br />
single c<strong>on</strong>trol groups of 90 animals each were used in the male and female rat porti<strong>on</strong>s of<br />
the assay. Otherwise, there were separate c<strong>on</strong>trols for each sex/species/compound of 50<br />
animals each).<br />
This provides us with a repeated experiment against which to test the hypothesis<br />
proposed above, using the method described in subsecti<strong>on</strong> A in the preceding secti<strong>on</strong>.<br />
Three of the six dyes were declared “carcinogenic” for at least <strong>on</strong>e speciesor sex. The<br />
marginal counts of animal with tumors in specific sites (the usual NTP analysis) suggested<br />
dose-related changes for all but 3 of the 48 sex X species comparis<strong>on</strong>s. As indicated above,<br />
however, multiple testing for tumors in specific sites raises the questi<strong>on</strong> of how to adjust the<br />
nominal p values to keep the false-positive rate from going too high. Other suggesti<strong>on</strong>s have<br />
been made based <strong>on</strong> the use of B<strong>on</strong>fer<strong>on</strong>ni bounds and the statistical characteristics of the<br />
tests used. Most of this amount to requiring a lower level of significance (usually around<br />
0.01) for frequently occurring tumors. When this is d<strong>on</strong>e, all six dyes have significant doserelated<br />
shifts in tumors for at least <strong>on</strong>e sex or species and 36 of the 48 sex X species<br />
combinati<strong>on</strong>s still show significant dose-related changes.<br />
Analyses like these have been criticized because they are based up<strong>on</strong> multiple testing<br />
and because the p-value adjustments tend to be of an ad hoc nature. To get around such a<br />
criticism, I ran overall tests of shifts in tumor patterns as described in the preceding secti<strong>on</strong><br />
for all six studies.<br />
Using methods described in the Appendix, data for each animal were c<strong>on</strong>verted to a<br />
vector of 25 numbers, representing the maximum degree of dysplasia discovered in specific<br />
organ systems, al<strong>on</strong>g with the number of weeks the animal was <strong>on</strong> study. Organized this<br />
way, the 150 animals in a given sex X species X compound study can be thought of as a<br />
cloud of points in multidimensi<strong>on</strong>al space. Animals with similar patterns of neoplastic<br />
lesi<strong>on</strong>s will tend to provide points close together.<br />
If there were no effect of treatment <strong>on</strong> neoplastic lesi<strong>on</strong>s, then the cloud of points<br />
would be such that animals in a given treatment group would be randomly scattered thought<br />
the space. The points closest to a given animal’s point would bel<strong>on</strong>g to animals associated<br />
with any of the three treatment groups with equal probability. However, if treatment had an<br />
effect <strong>on</strong> neoplastic lesi<strong>on</strong>s, we would expect animals in a given treatment group to be<br />
clustered together more so than might be expected at random.<br />
549
The appendix describes two tests for homogeneous scatter, <strong>on</strong>e al<strong>on</strong>g the first principal<br />
comp<strong>on</strong>ent of the data, the other al<strong>on</strong>g the sec<strong>on</strong>d principal comp<strong>on</strong>ent. The principal<br />
comp<strong>on</strong>ents are vectors that identify the directi<strong>on</strong>s of greatest spread am<strong>on</strong>g the points. The<br />
linear sum of elements that defines the projecti<strong>on</strong> of a point al<strong>on</strong>g a principal comp<strong>on</strong>ent is<br />
uncorrelated with the sum that defines the projecti<strong>on</strong> al<strong>on</strong>g another principal comp<strong>on</strong>ent.<br />
Because of this, the nominal significance test associated with two such tests can be adjusted<br />
to ensure a false-positive rate of 5%. In this situati<strong>on</strong>, a nominal p value of 0,0253 or less for<br />
either of the tests implies an overall significance level of 5 %.<br />
For each of the six tested compounds, there are significant treatment-related clusters in<br />
at least <strong>on</strong>e sex or species.<br />
The results of this analysis support the c<strong>on</strong>tenti<strong>on</strong> that all tested compounds will have<br />
an effect, in the current design of these studies.<br />
DISCUSSION<br />
If the hypothesis that every compound tested will cause some sort of a shift in neoplastic<br />
lesi<strong>on</strong>s is true, then it will be necessary to rec<strong>on</strong>sider the interpretati<strong>on</strong> of these studies. A<br />
treatment-related increase in tumors of <strong>on</strong>e type is no more indicative, by itself, of<br />
“carcinogenesis” than a decrease in tumors of another type is indicative of an<br />
“antineoplastic” effect. The cause of these shifts is a complex biological interacti<strong>on</strong> of a<br />
living animal seeking homeostasis in the face of a biochemical derangement of its milieu.<br />
The several hundred chr<strong>on</strong>ic toxicity tests that have been run in the past 10-15 years<br />
provide the science of toxicology with an unprecedented opportunity to investigate the<br />
nature of biological homeostasis and the resp<strong>on</strong>se of a living animal to toxic pressure.<br />
Regardless, of the eventual use of these studies to identify “carcinogens”, there is a wealth<br />
of useful informati<strong>on</strong> in them, waiting to be uncovered.<br />
In the meantime, can these studies be used to identify “carcinogens” (or even<br />
“antineoplastic” agents), if the hypothesis of universal effect is true?<br />
The answer, clearly, is no. In fact, the hypothesis of universal effect brings the problem<br />
of finding “carcinogens” back am<strong>on</strong>g the traditi<strong>on</strong>al problems of toxicology. In traditi<strong>on</strong>al<br />
toxicology, we recognized that chemical compounds cannot be divided into two classes,<br />
pois<strong>on</strong>s, (or toxic chemicals) and n<strong>on</strong>pois<strong>on</strong>s (or n<strong>on</strong>toxic chemical). All chemical will<br />
cause biological damage at sufficiently high doses, and the dual task of toxicology is to<br />
identify the nature of that damage as a functi<strong>on</strong> of dose and de<strong>term</strong>ine dose levels that have<br />
a good chance of being “safe”. So, if all chemicals will cause a derangement of senile<br />
neoplastic lesi<strong>on</strong>s, then it mekes no sense to divide the universe of chemicals into<br />
“carcinogens” and “n<strong>on</strong>carcinogens”. This derangement of neoplastic lesi<strong>on</strong>s is more<br />
complicated than the usual type of biological damage seen in subchr<strong>on</strong>ic and acute studies.<br />
There are problems due to latency of expressi<strong>on</strong> and irreversibility of some of the lesi<strong>on</strong>s.<br />
But, the problems of “carcinogenic” effects still bel<strong>on</strong>g in the general class of toxicological<br />
problems. We need to find parallels to the two tasks described in the previous paragraph.<br />
550
The questi<strong>on</strong> of de<strong>term</strong>ining a “safe” dose has been badly distorted in the statistical<br />
literature. Traditi<strong>on</strong>ally, toxicologists did not de<strong>term</strong>ine a “safe” dose by c<strong>on</strong>sidering doses<br />
at which no animals had a particular lesi<strong>on</strong>. It was not a case of naively assuming that<br />
seeing nothing in a small number of animals meant that nothing would occur. Instead, the<br />
toxicologist examined the high-dose animals to de<strong>term</strong>ine a pattern of lesi<strong>on</strong>s that could be<br />
associated with the compound at test. Then, he or she projected a set of putative precursors<br />
of this syndrome of lesi<strong>on</strong>s. The traditi<strong>on</strong>al “maximum safe dose” was the dose at which<br />
n<strong>on</strong>e of the lesi<strong>on</strong>s nor any of their putative precursors were seen in any animal. The<br />
statisticians assumed that each animal was given a 0 (no lesi<strong>on</strong>) or 1 (lesi<strong>on</strong>). In fact, each<br />
animal is scored in a complicated but biologically meaningful way. A “safe” dose was <strong>on</strong>e<br />
in which all the animals had scores less than the least score that could be associated with a<br />
treatment effect.<br />
The first toxicological problem, describe the “toxic” effect as a functi<strong>on</strong> of dose, is a<br />
much more difficult <strong>on</strong>e. We have addressed this problem elsewhere but <strong>on</strong>ly by suggesting<br />
multivariate statistical methods that might be useful. If we do not use maximum tolerated<br />
doses and reduce the length of studies to 12 m<strong>on</strong>ths, then we may be able to describe a dose<br />
resp<strong>on</strong>se for frank carcinogens like the nitrosamines. But, when we use very high doses and<br />
follow the animals for most of their natural life span, so that the “effect” is to shift lesi<strong>on</strong><br />
patterns, increasing the incidence of <strong>on</strong>e and decreasing the incidence of another, there is no<br />
clear “directi<strong>on</strong>” to the graph of resp<strong>on</strong>se versus dose. To “solve” this problem, we will<br />
need to think of dose resp<strong>on</strong>se in some different fashi<strong>on</strong> or we will have to aband<strong>on</strong> the<br />
current design of these studies because they produce uninterpretable results.<br />
APPENDIX: STATISTICAL METHODOLOGY<br />
The individual animal tabulati<strong>on</strong>s in many of the NTP studies define organ and organsystems<br />
that were routinely examined. In additi<strong>on</strong>, there is a category of any organ not<br />
routinely examined but for which histopathology was run because of a grossly observable<br />
lump or bump. In the tabulati<strong>on</strong>, individual neoplastic lesi<strong>on</strong>s are described in <strong>term</strong>s of<br />
tissues type, degree of malignancy, and (possible) source. Tissue type designati<strong>on</strong>s tend to<br />
be pathologist specific, so these were ingnored in this analysis. Instead, for each organ or<br />
organ system a given animal was scored as follows:<br />
0= no neoplastic lesi<strong>on</strong><br />
1= worst neoplastic lesi<strong>on</strong> was benign<br />
2= worst neoplastic lesi<strong>on</strong> was malignant but in situ<br />
3= worst neoplastic lesi<strong>on</strong> was invasive or metastatic<br />
This produced a vector of 20-25 numbers for each animal (depending <strong>on</strong> sex and<br />
species). An additi<strong>on</strong>al comp<strong>on</strong>ent was added by including the number of weeks <strong>on</strong> trial for<br />
that individual animal.<br />
All the animals in a given sex X species X AZO dye group, including the c<strong>on</strong>trols, the lowdose<br />
animals, and the high-dose animals, were combined, and the first and sec<strong>on</strong>d principal<br />
comp<strong>on</strong>ents were computed for the 150 points.<br />
551
The vector for a given animal was c<strong>on</strong>verted to the linear combinati<strong>on</strong> that represented the<br />
projecti<strong>on</strong> of that vector <strong>on</strong>to the first principal comp<strong>on</strong>ent. These projecti<strong>on</strong>s were then<br />
ordered; a Kruskal-Wallis test was run the ordered projecti<strong>on</strong>s. The same thing was then<br />
d<strong>on</strong>e for the sec<strong>on</strong>d principal comp<strong>on</strong>ent.<br />
REFERENCES<br />
1. Chhabra, R.S., Haseman, J., Hall, A., and Baskin, C. (1988), Inhibiti<strong>on</strong> of some<br />
sp<strong>on</strong>taneous tumors by 4-hexylresorcinol in F344 rats and B6C3F mice, Fundam.<br />
Appl.Toxicol.11, 685-690<br />
2. Haseman, J.K. (1983), A Reexaminati<strong>on</strong> of false positive rates for carcinogenesis<br />
studies, Fundam. Appl. Toxicol.3, 334-339.<br />
3. Salsburg, D. S. (1983), The lifetime feeding study in mice and rats – An examinati<strong>on</strong> of<br />
its validity as a bioassay for human carcinogens, Fundam. Appl. Toxicol. 3, 63-67.<br />
552
CLINICAL AND EPIDEMIOLOGICAL ASPECTS OF<br />
HEPATIC HYDATID CYST<br />
Mirela Indrieş, Viorica Coldea, Cristian Brisc, Ildiko Lenard, S<strong>on</strong>ia Drăghici,<br />
Andrei Csep, Daciana Sabău, Nicoleta Negruţ<br />
University of Oradea, Faculty of Medicineand Pharmacy Oradea<br />
ABSTRACT<br />
Human echinococcosis is a zo<strong>on</strong>otic infecti<strong>on</strong> caused by the tapeworm of the genus<br />
Echinococcus. Human disease is a „dead end” which interrupts the life cycle of the<br />
parasite. Hepatic localizati<strong>on</strong> is the most frequent (63%), because the infestati<strong>on</strong> occurs by<br />
digestive way, and liver is the first filter <strong>on</strong> this way. Hydatid disease may have any<br />
localizati<strong>on</strong>. Cystic echinococcosis has public health importance not <strong>on</strong>ly in areas of<br />
endemicity but also in countries or regi<strong>on</strong>s without endemicity due to the migrati<strong>on</strong> of<br />
infected people and livestock exchanges.<br />
Keywords: Echinococcus granulosus, hepatic hydatic cyst (HHC).<br />
INTRODUCTION<br />
Human echinococcosis is a zo<strong>on</strong>otic infecti<strong>on</strong> caused by the tapeworm of the genus<br />
Echinococcus. Of the 4 known species of Echinococcus, 3 are of medical importance in<br />
humans. These are Echinococcus granulosus, causing cystic echinococcosis; Echinococcus<br />
multilocularis, causing alveolar echinococcosis; and Echinococcus vogeli. Echinococcus<br />
granulosus is the most comm<strong>on</strong> of the three. Echinococcus multilocularis is rare, but is the<br />
most virulent and Echinococcus vogeli is the most rare (Table 1) (Brunetti E, 2006).<br />
Table 1: Echinococcus Species<br />
Echinococcus Species Echinococcosis Diseases<br />
1. Echinococcus granulosus Cystic Echinococcosis (cystic hydatid disease)<br />
2. Echinococcus multilocularis Alveolar Echinococcosis<br />
3. Echinococcus vogeli Polycystic Echinococcosis<br />
AS 4. Echinococcus oligarthus Polycystic Echinococcosis<br />
AS=mal Species<br />
Incidence<br />
Echinococcosis is a very rare disease in northern Europe. The disease is endemic in the<br />
Balkan Area and in Eastern Europe. The endemic areas are the Mediterranean countries and<br />
the Middle East. The intensive endemic areas are the southern part of South America,<br />
Iceland, Australia, New Zealand and southern parts of Africa. The incidence of Cystic<br />
553
Echinococcosis in endemic areas ranges from 1-220 cases per 100,000 inhabitants. In USA,<br />
echinococcosis remains a very rare disease (
Figure 3.-Life Cycle of Echinococcus granulosus.<br />
Structure<br />
Adult Echinococcus granulosus tapeworms are relatively very small, c<strong>on</strong>sisting of 3 to 5<br />
segments, and usually are less than 1 cm l<strong>on</strong>g. Dogs and wild canids are the <strong>on</strong>ly final hosts<br />
in which the adults are found, often adhering in great numbers to the small intestinal<br />
mucosa. The scolex has four suckers and is crowned with a circle of hooks as in Taenia<br />
solium. It is followed by a germinative neck regi<strong>on</strong>, <strong>on</strong>e developing segment, and usually<br />
<strong>on</strong>e gravid segment c<strong>on</strong>taining several hundred eggs (Figure 2 A, B).<br />
Figure 2. A- Adult Echinococcus granulosus worm, B- scolex showing hooks, C-<br />
hydatid brood capsule with protoscolices<br />
The hydatid larva is found in sheep, in many other herbivores and in humans. In humans<br />
the cyst is slow-growing, but in a period of years may reach a diameter of 30 cm with a 1<br />
mm thick, laminated sheath surrounded by fibrous reactive host tissue. The cyst is usually<br />
fluid-filled and, if viable, has a germinative inner lining from which many thousands of<br />
555
scoleces are budded off into the lumen or remain attached to the germinative wall (Figure 2<br />
C). The floating scoleces often enlarge, become vesicular, and develop into daughter<br />
floating col<strong>on</strong>ies within the parent cyst. These in turn may bud off a third generati<strong>on</strong> of<br />
cysts within themselves. The result is an enclosed sac c<strong>on</strong>taining myriads of future adult<br />
worms ready to infect a dog or other susceptible carnivore that feeds <strong>on</strong> the hydatid-infected<br />
animal and the scolex-filled cyst fluid (Adams<strong>on</strong> ML, 1994).<br />
Clinical Manifestati<strong>on</strong>s<br />
Echinococcosis (hydatid disease) results from the presence of <strong>on</strong>e or more massive cysts, or<br />
hydatids, which can develop in any tissue site, including the liver, lungs (25%), heart, brain<br />
(1%), kidneys (2%), spleen (1%), tyroid and l<strong>on</strong>g b<strong>on</strong>es (3%). Hepatic localizati<strong>on</strong> is the<br />
most frequent (63%), because the infestati<strong>on</strong> occurs by digestive way, and liver is the first<br />
filter <strong>on</strong> this way. Hydatid disease may have any localizati<strong>on</strong>. The clinical manifestati<strong>on</strong>s of<br />
this infecti<strong>on</strong> therefore vary greatly, depending <strong>on</strong> the site and size of the cyst, but resemble<br />
those of a slow-growing tumor that causes gradually increasing pressure. Infecti<strong>on</strong>s in the<br />
liver, lungs, or subcutaneous tissue sites may be asymptomatic for many years, but pressure<br />
effects eventually develop. In sensitive or vital areas, hydatids produce a panoply of<br />
symptoms, chiefly owing to mechanical compressi<strong>on</strong> or blocking effects but also include<br />
collapse of infected l<strong>on</strong>g b<strong>on</strong>es, blindness, and epileptiform seizures. The rupture of a<br />
hydatid cyst may induce sudden anaphylactic shock. Sec<strong>on</strong>dary complicati<strong>on</strong>s may occur as<br />
a result of infecti<strong>on</strong> of the cyst or leakage of the cyst. Unilocular cysts are frequently single<br />
(75%). However, they can also have a multifocal or multiorgan localizati<strong>on</strong>. The frequency<br />
of multiple localizati<strong>on</strong> ranges between 11% and 30% from 2 % to 14%. Lung involvement<br />
is solitary in 70% of the cases, and multiple in 30% Unilocular cysts can remain<br />
asymptomatic for 5-20 years. The diagnosis is established, with few excepti<strong>on</strong>s, by<br />
paraclinical investigati<strong>on</strong>s, especially by imaging techniques (ultras<strong>on</strong>ography, c<strong>on</strong>venti<strong>on</strong>al<br />
radiology, computed tomography) as well as by immunological studies (ELISA). Although<br />
drug treatment has been followed by many failed attempts, today we can witness its revival<br />
due to the new drugs (mebendazol, albendazole) which completely destroy the univesicular<br />
hydatid cyst in 50-80% of the cases after 3 m<strong>on</strong>ths of treatment. Some authors reported a<br />
sterilizati<strong>on</strong> rate of 95%. Nevertheless, the treatment of choice remains surgical.<br />
Patients and Methods: We studied a lot of 18 cases with age between 0-69 years,<br />
hospitalized at the Clinical Hospital Infectious Diseases of Oradea, between years 1998-<br />
2006, with a hepatic hydatid cyst, established by clinical and paraclinical data. The study<br />
was performed <strong>on</strong>: the patients observati<strong>on</strong> files; the biochemical methods (a complete<br />
blood count), the serologic methods (detecti<strong>on</strong> of the anti-Echinococcus antibodies with<br />
ELISA method) and imaging techniques (ultras<strong>on</strong>ography, c<strong>on</strong>venti<strong>on</strong>al radiology).<br />
The Epidemiological Particularities:<br />
The Number of cases with Hepatic Hydatid Cyst<br />
From analyzing the annual incidence of the Hepatic Hydatid Cyst (HHC) between years<br />
1998-2006, hospitalized the Clinical Hospital Infectious Diseases of Oradea, we observed<br />
556
that the values are 1 case at 600.000 inhabitans of Bihor county in 1998 and it grows at 4<br />
cases at 600.000 inhabitans in even years 2002, 2004, 2006 (Figure 4).<br />
Nr. de cazuri<br />
25<br />
20<br />
15<br />
10<br />
5<br />
Incidenţa MNI între 1987-2003<br />
0<br />
1986 1988 1990 1992 1994 1996 1998 2000 2002 2004<br />
Ani<br />
Incidenţa MNI între 1987-2003<br />
Figure 4 -The annual distributi<strong>on</strong> of HHC cases in Bihor county, Romania.<br />
It is specified that the real annual incidence of HHC could not be proper for the following<br />
reas<strong>on</strong>s: the most part of infecti<strong>on</strong>s evolves as subclinic or inaparent infecti<strong>on</strong>s, in this way<br />
remaining undiagnosed and unreported.<br />
The Distributi<strong>on</strong> of Cases <strong>on</strong> Age Groups.<br />
The highest incidence of HHC infecti<strong>on</strong> is to be found the age group of 61-70 years old<br />
patients (33,33%), followed by the age group of 51-60 years old patients (22,22%), 49-50<br />
years old patients (22,22%) and 31-40 years old patients (16,67%).<br />
33,33%<br />
22,22%<br />
0,00%<br />
Age Groups<br />
5,56%<br />
16,67%<br />
22,22%<br />
Figure 5 -The distributi<strong>on</strong> of cases <strong>on</strong> age groups.<br />
0-10 years<br />
11-20 years<br />
21-30 years<br />
31-40 years<br />
41-50 years<br />
51-60 years<br />
61-70 years<br />
The Distributi<strong>on</strong> of the Cases According to Sex<br />
From the total of 18 patients with HHC, <strong>on</strong>ly 22,22% are men patients and 77,78% are<br />
women patients (that is 14 of them).<br />
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Men<br />
22,22%<br />
Women<br />
77,78%<br />
Rural<br />
77,78<br />
%<br />
Urba<br />
n<br />
22,22<br />
%<br />
Figure 6 -The distributi<strong>on</strong> of casesaAccording to sex and to geographic origin.<br />
The Distributi<strong>on</strong> of Cases According to Geographic Origin<br />
The majority of HHC cases come from the rural area 77,78% (that is 14 patients), most<br />
probably due to a precarious accessibility to diagnosis and treatment units as well as to the<br />
educati<strong>on</strong> of the patients.<br />
Clinical-biological particularities of the Hepatic Hydatid Cyst hospitalized at Clinical<br />
Hospital Infectious Diseases of Oradea between 1998-2006<br />
On the first places have situated asthenia, hephatalgia and unappetite and <strong>on</strong>ly in 2 cases<br />
have presented vomiting and fever (Table 2).<br />
94,44<br />
77,77<br />
66,66<br />
Asthenia Hepathalgy Headache Unappetite Vomiting Fever<br />
Figure 7 -Clinical symptoms of HHC<br />
In the majority of the cases the patients presented hepathomegaly and in 3 cases appeared<br />
jaundice and hyperchrome urinary emissi<strong>on</strong>s.<br />
558<br />
50<br />
33,33<br />
11,11<br />
%
Paraclinical data<br />
From the laboratory analyses we detected the followings: leucocitosis appears in 2 cases,<br />
eosinophilia (≥ 4 %) in 13 cases, anemia (Hg
3. The HHC is more frequently at women (77,78 %) than men, is more frequently in rural<br />
area than in urban area and is more frequently at domestic animal holder/farmer.<br />
4. Hepatic localizati<strong>on</strong> is the most frequent (60%), because the infestati<strong>on</strong> occurs by<br />
digestive way, and liver is the first filter <strong>on</strong> this way.<br />
5. In the majority of the cases the patients presented asthenia, hephatalgia, unappetite,<br />
hepathomegaly and <strong>on</strong>ly in 3 cases appeared jaundice and hyperchrome urinary<br />
emissi<strong>on</strong>s.<br />
6. In the majority of the cases HHC were single and <strong>on</strong>ly in 2 cases HHC have evoluated<br />
with multiple intrahepatic localisati<strong>on</strong>s.<br />
7. The lethal risk results in case of HHC breake followed by anaphylactic shock.<br />
8. The true prophilaxy of domestic animal illness should prevent the existence of the<br />
Echinoccocosis disease at human and should decrease the costs of treatment .<br />
9. With a universal outspread, the HHC remains permanently an open problem.<br />
REFERENCES:<br />
1. Eckert J., Deplazed P.: Biological, epidemiological and clinical aspects of echinooccosis,<br />
a zo<strong>on</strong>osis of increasing c<strong>on</strong>cern, Clin Microbiol Rev. 2004;17: 107-125.<br />
2. G. Mandell, J. Bennett, & R. Dolin (Eds.) Principles and practices of infectious diseases<br />
(5th ed.); 3205-3207.<br />
3. Brunetti E, Filice C, Mer<strong>on</strong>i V: Comment <strong>on</strong> percutaneous treatment of liver hydatid<br />
cysts. AJR Am J Roentgenol 2006 Apr; 186(4): 1198-9.<br />
4. Rogan MT, Hai WY, Richards<strong>on</strong> R: Hydatid cysts: does every picture tell a story?<br />
Trends Parasitol 2006 Sep; 22(9): 431-8.<br />
5. T. Vaida: Parazitologie medicală, Universitatea Oradea, 1993: 76-80.<br />
6. M J Kumar, K Toe and R D Banerjee: Hydatid cyst of liver, Postgraduate Medical<br />
Journal 2003;79:113-114.<br />
7. Sréter T, Széll Z, Egyed Z, Varga I. Echinococcus multilocularis: an emerging<br />
pathogen in Hungary and central eastern Europe. Emerg Infect Dis [serial <strong>on</strong>line] 2003<br />
Mar . Available from: URL: http://www.cdc.gov/ncidod/EID/vol9no3/02-0320.htm<br />
8. Ammann RW, Eckert J.: Cestodes. Echinococcus.Gastroenterol Clin North Am. 1996<br />
Sep; 25(3):655-89.<br />
9. Radulescu S. Parazitologie medicala, Ed. All Educati<strong>on</strong>al, Bucuresti, 2000: 282-95. 10.<br />
Zanc V. Parazitologie clinica, Ed. Sincr<strong>on</strong>, 2001, Cluj-Napoca: 108-155.<br />
10. WHO - Grupul neoficial de lucru OMS in problema hidatidozei. Directive privind<br />
tratamentul hidatidozei chistice si alveolar la om. Buletin OMS, 1996.<br />
11. Codreanu R., Sim<strong>on</strong>a R.: Orientari actuale in tratamentul bolii hidatice, Revista romana<br />
de parazitologie, 2002(XII): 30-38.<br />
12. Ş. Georgescu1, L. Dubei and al: Minimally Invasive Treatment of Hepatic Hydatid<br />
Cysts, Romanian Journal of Gastroenterology, September 2005 (14 ); 249-252.<br />
13. Adams<strong>on</strong> ML, Caira JN. Evoluti<strong>on</strong>ary factors influencing the nature of parasite<br />
specificity. (Review) Parasitology, 1994 (109);85-95,<br />
560
THIN LAYER CHROMATOGRAPHY OF STERIGMATOCYSTIN IN CHEESE<br />
Bara Lucian<br />
University of Oradea, Faculty of Envir<strong>on</strong>mental Protecti<strong>on</strong><br />
ABSTRACT<br />
A <strong>on</strong>e-dimensi<strong>on</strong>al thin layer chromatographic method has been developed for de<strong>term</strong>ining<br />
sterigmatocystin in cheese. Cheese is extracted with acet<strong>on</strong>itrile-4% KCl (85 + 15). A<br />
simplified liquid-liquid partiti<strong>on</strong> cleanup is used, and the sample extract is passed through a<br />
cupric carb<strong>on</strong>ate column for final purificati<strong>on</strong>. Sterigmatocystin is visualized by spraying<br />
the plate with aluminium chloride. The fluorescence of the spot is enhanced 10-fold by<br />
additi<strong>on</strong>al plate spraying with silic<strong>on</strong>e-either mixture, enabling sterigmatocystin detecti<strong>on</strong><br />
and quantitati<strong>on</strong> at 2 and 5 μ/kg, respectively. Average recoveries were 88.3 and 86.4% at<br />
the 10 and 25 μ/kg levels, respectively.<br />
Keywords: cromatography, cheese<br />
INTRODUCTION<br />
Sterigmatocystin is a mycotoxin that has potent toxic and carcinogenic effects. This<br />
metabolite is produced by several species of Aspergillus and at least <strong>on</strong>e species of<br />
Bipolaris. Sterigmatocystin occurs naturally in grains, green coffee, fruit juices and fruitbased<br />
infant foods, and cheese. Since sterigmatocystin-producing fungi are so ubiquitously<br />
distributed, this metabolite must be c<strong>on</strong>sidered as an important c<strong>on</strong>taminant in these<br />
commodities as well as in other foods and feeds. Thus, interest was aroused in analyzing<br />
cheese for sterigmatocystin to de<strong>term</strong>ine levels and incidences of c<strong>on</strong>taminati<strong>on</strong>.<br />
Methods have been published for the de<strong>term</strong>inati<strong>on</strong> of sterigmatocystin in grains and cereals<br />
and have been used for other commodities; however, these methods were found to be<br />
unsatisfactory for this project. The method of van Egm<strong>on</strong>d et al., which is essentially their<br />
earlier method with modificati<strong>on</strong>s, has been developed for the de<strong>term</strong>inati<strong>on</strong> of the<br />
sterigmatocystin in cheese, reportedly with a 5 μ/kg limit of detecti<strong>on</strong>. I tested this method<br />
and c<strong>on</strong>cluded that it was complex and time c<strong>on</strong>suming, and had a relatively high limit of<br />
detecti<strong>on</strong> al<strong>on</strong>g with poor spiked sample recoveries. Experimental data collected during the<br />
method evaluati<strong>on</strong> indicated the potential for the development of a more satisfactory<br />
procedure. The thin layer chromatographic (TLC) method described here resulted.<br />
MATERIALS AND METHODS<br />
Apparatus<br />
a) Chromatographic tube – Plain, 22 x 250 mm<br />
b) TLC plates – precoated, Sil G-25 HR No. 66-14-600-6. Do not activate plates.<br />
c) TLC apparatus – Developing tank with cover for use with 20 x 20 cm glass<br />
plates;spotting template; 10 μL syringe; l<strong>on</strong>gwave 15 watt UV lamp (use with<br />
absorbing eyeglasses) or Chromato-Vue cabinet equipped with <strong>on</strong>e or two 15 watt<br />
561
lamps; if available, a thin layer plate densitometer with fluorescent capability,<br />
excitati<strong>on</strong> wavelength 365 nm and emissi<strong>on</strong> wavelength > 400 nm.<br />
d) TLC plate sprayer – K<strong>on</strong>tes, K-422540<br />
e) Blender – waring, equipped with 1 L jar and lied<br />
f) Filter paper – Whatman 2V folded circles, 24 cm<br />
g) Vials – 5 mL, c<strong>on</strong>ical, tapered inside, with Tefl<strong>on</strong>-lined screw cap<br />
Reagents<br />
a) Solvents – reagent grade acet<strong>on</strong>itrile, hexane, methylene chloride, benzene, methanol,<br />
acetic acid, and anhydrous ethyl ether. (Cauti<strong>on</strong>: Benzene is a suspected carcinogen.<br />
Use necessary safety measures when handling this chemical)<br />
b) Silic<strong>on</strong>e DC-200 – 12 500 centistokes<br />
c) Silic<strong>on</strong>e-ether spray – silic<strong>on</strong>e-anhydrous ether (18 + 82). If weighing silic<strong>on</strong>e is more<br />
c<strong>on</strong>venient, 1g = 1mL<br />
d) Potassium chloride soluti<strong>on</strong> – dissolve 4g KCl in 100 mL water<br />
e) Aluminium chloride soluti<strong>on</strong> – Dissolve 15 g AlCl, in ethanol and dilute to 100mL<br />
f) Calcium chloride – anhydrous, granular (8mesh). Prepare in advance CaCl2 –water (1 +<br />
2, w/v). Hasten soluti<strong>on</strong> by placing in ultras<strong>on</strong>ic bath. Remove slight cloudiness by<br />
filtering through Whatman 2V paper<br />
g) Curpic carb<strong>on</strong>ate, Cu (II) – Reagent grade powder<br />
h) Diatomaceous earth – Hyflo Super- Cel<br />
i) Sterigmatocystin standards – Stock standard – Prepare according to sec. 26.133.<br />
Working standard – Dilute stock standard to ca ng/ μL benzene<br />
j) Boiling chips – Si C. Float off fines and extraneous particles with water, wash with<br />
acet<strong>on</strong>e, and dry.<br />
Extracti<strong>on</strong><br />
Weigh 36 G composited sample into 1L Waring blender jar. Add 170 mL acet<strong>on</strong>itrile and<br />
30 mL KCl soluti<strong>on</strong>; secure lid and blend 3 min at high speed. Decant extract through<br />
Whatman 2V paper. Transfer 50 mL filtrate to 250 mL separatory funnel, add 50 Ml CaCl2<br />
soluti<strong>on</strong>, vigorously shake ca 30 s, and let stand until layers separate; separati<strong>on</strong> usually<br />
occurs within 5 min.<br />
Discard bottom layer. Remove excess water from inside the stem of the separatory<br />
funnel by drying with laboratory wiping tissue, such as Kimwipes. Place a small wad of<br />
glass wool into the stem of the separatory funnel to serve as a filter and drain the remaining<br />
phase into a sec<strong>on</strong>d separatory funnel. Add 50 Ml water to remaining phase and mix. Add<br />
50 mL hexane, vigourously shake about 1 min, and let layers separate. Transfer bottom<br />
layer to another 250 mL separatory funnel. Extract, using 50 and 25 mL porti<strong>on</strong>s of<br />
methylene chloride, by shaking first extracti<strong>on</strong> 1 min and sec<strong>on</strong>d extracti<strong>on</strong> 10 s. Again,<br />
complete separati<strong>on</strong> usually occurs within 5 min. Collect methylene chloride extracts<br />
separately and save for column chromatography.<br />
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Column Chromatography<br />
Loosely place glass wool plug in bottom of chromatographic tube and add ca 15 g<br />
anhydrous sodium sulfate. Add 4 g cupric carb<strong>on</strong>ate-diatomaceous earth (1 + 2); lightly<br />
pack column by holding in upright positi<strong>on</strong> and by allowing it to drop gently <strong>on</strong> its tip <strong>on</strong>to<br />
book or writing pad. Add 20 g sodium sulfate to top of column.<br />
Sequentially pass methylene chloride extracts through column, beginning with 50 mL<br />
porti<strong>on</strong>. Collect that eluate and each successive eluate in 250 mL Philips beaker. Add 25 mL<br />
porti<strong>on</strong> to column when first porti<strong>on</strong> has drained to below top layer of sodium sulfate. Rinse<br />
column with 10 mL methylene chloride.<br />
Add 4-5 SiC boiling chips to Philips beaker and evaporate solvent nearly to dryness <strong>on</strong><br />
steam bath under gentle stream of nitrogen. Cool to room temperature and quantitatively<br />
transfer with small porti<strong>on</strong>s of methylene chloride to 5 mL vial. Evaporate solvent to<br />
dryness under gentle stream of nitrogen. Save sample residue for TLC.<br />
This Layer Chromatography<br />
Add 100 μL benzene (screen < 10 ppb) or 200 μL benzene (screen1<br />
h before using.<br />
Spot plate as previously described and develop using benzene-methanol (95 + 5).<br />
Solvent fr<strong>on</strong>t travels 13-15 cm in 45 min.<br />
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Table 1. Recovery of sterigmatocystin added to Gouda cheese<br />
Det. No. Found, μg/kg<br />
10μg/kg Level<br />
Rec., %<br />
1<br />
0, c<strong>on</strong>trol<br />
2<br />
10.0<br />
100<br />
3<br />
9.9<br />
99<br />
4<br />
8.4<br />
84<br />
5<br />
8.6<br />
86<br />
6<br />
7.9<br />
79<br />
7<br />
8.2<br />
82<br />
Mean<br />
8.8<br />
88.3<br />
Std. Dev.<br />
0.89<br />
Coeff. Of var., %<br />
10.2<br />
25 μg/kg Level<br />
1<br />
0, c<strong>on</strong>trol<br />
2<br />
27.6<br />
110<br />
3<br />
21.4<br />
86<br />
4<br />
19.7<br />
79<br />
5<br />
20.1<br />
81<br />
6<br />
20.0<br />
80<br />
7<br />
20.8<br />
83<br />
Mean<br />
21.6<br />
86.4<br />
Std. Dev.<br />
2.99<br />
Coeff. Of var., %<br />
13.9<br />
Calculati<strong>on</strong><br />
Calculate c<strong>on</strong>centrati<strong>on</strong> of sterigmatocytin:<br />
μg/kg = ng/g = (S x Y x V) / (X x W)<br />
where S = μL standard equal to unknown, Y = c<strong>on</strong>centrati<strong>on</strong> of standard (ng/ μL), V = final<br />
volume of sample extract (μL), X = μL sample extract spotted giving fluorescent intensity<br />
equal to S, and W = g sample in final extract (9g).<br />
RESULTS AND DISCUSSIONS<br />
Table 1 shows the percentage recovery of sterigmatocystin from spiked Gouda cheese after<br />
extracti<strong>on</strong> and cleanup, as measured by <strong>on</strong>e-dimensi<strong>on</strong>al TLC. The mean recoveries of<br />
sterigmatocystin added at levels of 10 and 25 μg/kg were 88.3 and 86.4%, respectively. The<br />
coefficients of variati<strong>on</strong> were 10.2 and 13.9%.<br />
While trying to resolve the problem of sterigmatocystin spots fading <strong>on</strong> TLC, I noticed<br />
that spots, particularly those of 30 ng and less, were very visible, especially when the plate<br />
was still warm; however, the spots faded rapidly up<strong>on</strong> cooling of the plate. I also observed<br />
564
that spot intensity could be regenerated by reheating the plate. This fading was unlike fading<br />
shown by aflatoxins, in that aflatoxin fading is minimized when plates are protected from<br />
light and, in some cases, from atmospheric c<strong>on</strong>taminants such as oz<strong>on</strong>e. With<br />
sterigmatocystin, I surmised that the activity of the silica gel was causing the observed<br />
behaviour. I c<strong>on</strong>cluded that the problem would be solved if the silica was held in an<br />
activated state, which could be accomplished by sealing the plate with paraffin or silic<strong>on</strong>e.<br />
Various mixtures of silic<strong>on</strong>e-anhydrous ether were applied by spraying before arriving at the<br />
mixture described here.<br />
In additi<strong>on</strong> to sealing the plate, the silic<strong>on</strong>e also enhanced the fluorescence of the<br />
sterigmatocystin spots approximately 10-fold. This type of fluorescent spot enhancement<br />
had been noted earlier by other researchers (17), who used viscous organic solvents such as<br />
mixtures of liquid paraffin and n-hexane as a spray.<br />
No significant loss in fluorescent intensity of sterigmatocystin was observed in plates<br />
that were stored unprotected from light and moisture for 2 weeks.<br />
A sterigmatocystin standard curve gave a linear resp<strong>on</strong>se in the range from 3 to 106 ng, with<br />
a correlati<strong>on</strong> coefficient (r) of 0.9994. One ng sterigmatocyctin standard is easily detected,<br />
while 3 ng represents the lowest amount quantitated at a signal-to-noise ratio of 3:1.<br />
The proposed method is simple, practical, rapid, ec<strong>on</strong>omical, and precise. Incorporati<strong>on</strong><br />
of calcium chloride soluti<strong>on</strong> into the sample extract before hexane defatting and methylene<br />
chloride partiti<strong>on</strong>ing solved emulsi<strong>on</strong> problems previously encountered when using other<br />
methods. The eliminati<strong>on</strong> of emulsi<strong>on</strong> formati<strong>on</strong> during separatory funnel cleaup and<br />
partiti<strong>on</strong>ing has possibly provided the breakthrough necessary to improve analytical<br />
methods used for the de<strong>term</strong>inati<strong>on</strong> of other mycotoxins. The c<strong>on</strong>centrati<strong>on</strong> of calcium<br />
chloride used may vary am<strong>on</strong>g commodities. The minimum c<strong>on</strong>centrati<strong>on</strong> required to be<br />
effective in this method has not been thoroughly investigated. Remaining TLC interferences<br />
are eliminated by passing the methylene chloride extract through the cupric carb<strong>on</strong>ate<br />
column. Further work is in progress <strong>on</strong> the use of the method to de<strong>term</strong>ine other<br />
mycotoxins.<br />
REFERENCES<br />
1. 1.Purchase, I., F., H., & van der Watt, J. J. (1970), Food Cosmet, Toxicol. 8, 289-296.<br />
2. 2.van der Watt, J.J. (1974) Mycotoxins, Elsevier Scientific Publishing Co., Amsterdam,<br />
The Netherlands, pp. 369-382<br />
3. 3.Bartos, J. & Matyas, Z. (1983) Vet. Med. (Prague) 28, 189-192<br />
4. 4.Nowotny, P., Baltes, W., Kroenert, W., & Weber, R. (1983) Lebensmittelchem.<br />
Gerichtl. Chem 37, 71-72<br />
5. 5.Official Methods of Analysis (1984) 14 th Ed., AOAC, Arlingt<strong>on</strong>, VA<br />
565
TISSUE LEVELS AND BIOLOGICAL EFFECTS OF N-<br />
NITROSODIMETHYLAMINE IN MICE DURING CHRONIC LOW OR HIGH<br />
DOSE EXPOSURE WITH OR WITHOUT ETHANOL<br />
Bara Camelia<br />
1 University of Oradea, Faculty of Envir<strong>on</strong>mental Protecti<strong>on</strong><br />
ABSTRACT<br />
In a study of the metabolism, dispositi<strong>on</strong>, and hepatotoxicity of the envir<strong>on</strong>mental<br />
carcinogen N-nitrosodimethylamine (NDMA), as a functi<strong>on</strong> of dose in the drinking water<br />
and of c<strong>on</strong>comitant administrati<strong>on</strong> of ethanol, outbred Swiss mice were given NDMA for 1-<br />
4 weeks at levels of 50-0.5 ppm, with or without 10, 20, or 30% ethanol. NDMA, assayed in<br />
blood, liver, kidney, lung, and brain by thermal energy analysis after methylene chloride<br />
extracti<strong>on</strong>, was detectable (.0.5 ppb) in tissue levels of NDMA (1-4 ppb) near the detecti<strong>on</strong><br />
limit of 0.5 ppb; this was also found to be the minimal c<strong>on</strong>centrati<strong>on</strong> causing significant<br />
numbers of lung tumors in strain A mice after treatment for 16-18 weeks. Co-administrati<strong>on</strong><br />
of ethanol caused an increase in blood and tissue levels of NDMA at all levels of both<br />
chemicals, often by a factor of 10 or more. Ethanol also partially alleviated the<br />
morphological hepatotoxic effects of NDMA at 50 ppm (centrilobular hemorrhage and<br />
necrosis). These results are c<strong>on</strong>sistent with competitive inhibiti<strong>on</strong> of metabolic activati<strong>on</strong> of<br />
NDMA by ethanol. Ten per cent ethanol did not induce liver NDMA demethylase activity<br />
significantly and did not prevent loss of this activity from the livers of mice receiving 5-50<br />
ppm NDMA. Thus, inhibiti<strong>on</strong>, rather than inducti<strong>on</strong>, of NDMA metabolism was the<br />
predominant effect of ethanol, with increased levels of NDMA in blood and other tissues as<br />
a c<strong>on</strong>sequence.<br />
Keywords: N-nitrosodimethylamine, dose exposure<br />
INTRODUCTION<br />
The potent carcinogen NDMA is present in a variety of human c<strong>on</strong>tact sources.<br />
Furthermore, low levels of NDMA, usually in the 0.1-1-ppb range, have been reported in<br />
human blood and urine. Since the meaning of these human blood NDMA levels for cancer<br />
risk is of interest, we have measured amounts of NDMA in blood and other tissues of mice<br />
exposed chr<strong>on</strong>ically to various c<strong>on</strong>centrati<strong>on</strong>s of NDMA in drinking water or liquid diet,<br />
and correlated these values with incindence of tumors in the lung.<br />
A sec<strong>on</strong>d objective of the study was examinati<strong>on</strong> of the effect of chr<strong>on</strong>ic coadministrati<strong>on</strong><br />
of ethanol <strong>on</strong> tissue levels and hepatotoxicity of chr<strong>on</strong>ic NDMA, of c<strong>on</strong>cern<br />
because of the occurrence of NDMA in some alcoholic beverages, frequent c<strong>on</strong>comitant<br />
exposure of humans to nitrosamine-c<strong>on</strong>taining cigarette smoke al<strong>on</strong>g with ethanol and<br />
NDMA, as related to carcinogenicity and toxicity, have revealed that this and other alcohols<br />
have complex effects <strong>on</strong> the metabolism, pharmacokinetics, and biological effectiveness of<br />
NDMA and other nitrosamines. When present simultaneously with NDMA, ethanol inhibits<br />
566
its metabolism by oxidative pathways, in vivo, in perfused liver, in intact liver cells in vitro,<br />
in liver slices, and in cell-free preparati<strong>on</strong>s of microsomes. The inhibiti<strong>on</strong> is competitive in<br />
nature. C<strong>on</strong>versely, chr<strong>on</strong>ic treatment of animals with ethanol results in inducti<strong>on</strong> of the low<br />
Km form of NDMA demethylase. With regard to the biological c<strong>on</strong>sequences of these<br />
effects of ethanol, liver cells or microsomes from rats pretreated with ethanol were more<br />
effective than c<strong>on</strong>trols in catalyzing mutagenesis or DNA damage by nitrosamines in vitro,<br />
but similar effects were not seen in vivo. Pretreatment of rats with isopropanol 24 hours<br />
before NDMA led to potentiati<strong>on</strong> of hepatotoxicity, whereas rats given ethanol in a liquid<br />
diet for 3 weeks before NDMA experienced a reducti<strong>on</strong> in hepatotoxicity compared to<br />
c<strong>on</strong>trols. Effects <strong>on</strong> neoplasia are similarly complex. Ethanol given simultaneously with Nnitrosodiethylamine<br />
or N-nitrosomorpholine caused an increase in number and size of<br />
preneoplastic liver foci, but ethanol given simultaneously with various nitrosamines reduced<br />
the number of liver tumors, sometimes with a c<strong>on</strong>comitant increase in tumors of distal<br />
targets such as esophagus or nose. Ethanol administered with N-nitros<strong>on</strong>ornicotine to rats<br />
resulted in fewer esophageal but more nasal cavity tumors than in rats given the carcinogen<br />
al<strong>on</strong>e.<br />
No studies have been reported <strong>on</strong> the short-<strong>term</strong> toxic effects of ethanol given<br />
c<strong>on</strong>tinuously and simultaneously with NDMA, even though this is presumably an exposure<br />
mode comm<strong>on</strong>ly encountered by the human. Under these c<strong>on</strong>diti<strong>on</strong>s, both inducti<strong>on</strong> and<br />
competitive inhibiti<strong>on</strong> of NDMA demethylase would be possible, and either acti<strong>on</strong> could<br />
influence the activati<strong>on</strong> and / or detoxificati<strong>on</strong> of the chemical. The nature of the biological<br />
phenomena resoving from this complexity of possibilities required empirical de<strong>term</strong>inati<strong>on</strong>.<br />
In the experiments reported here, we inquired as to whether co-treatment with ethanol<br />
during exposure of mice to a low but tumorigenic dose of NDMA in the drinking water, or<br />
to a high, hepatotoxic dose of this agent, would influence expressi<strong>on</strong> of hepatotoxicity or<br />
levels of NDMA demethylase in the liver and, c<strong>on</strong>comitantly, circulating levels of NDMA<br />
in the blood and in organs distal to the liver.<br />
MATERIALS AND METHODS<br />
Lung Tumorigenesis Assay. Male strain A/J mice were housed in plastic cages with<br />
hardwood shavings as bedding at 24+2 C, 40-60% humidity, and with a 12-hour fluorescent<br />
light / dark cycle. Soluti<strong>on</strong>s of 500 ppb NDMA were diluted daily in distilled water from a<br />
stock soluti<strong>on</strong> of 10 ppm NDMA; the later was kept at 4C in the dark and<br />
prepared m<strong>on</strong>thly. The drinking water soluti<strong>on</strong>s of NDMA were c<strong>on</strong>tained in amber bottles.<br />
All bottles and sipper tubes were rinsed with distilled water before use. C<strong>on</strong>trol animals<br />
received distilled water.<br />
567
Expt. Diet Treatm<br />
. time<br />
(weeks)<br />
A/J<br />
mice<br />
A/JCr<br />
mice<br />
Purina<br />
5002<br />
Purina<br />
5002<br />
Purina<br />
5015<br />
Table 1. Lung tumorigenesis in strain A mice by 500 ppb NDMA<br />
12<br />
14<br />
16<br />
18<br />
18<br />
Treatmen<br />
t<br />
NDMA<br />
C<strong>on</strong>trol<br />
NDMA<br />
C<strong>on</strong>trol<br />
NDMA<br />
C<strong>on</strong>trol<br />
NDMA<br />
C<strong>on</strong>trol<br />
NDMA<br />
C<strong>on</strong>trol<br />
Tumor<br />
Beares /Total<br />
8/49<br />
8/50<br />
14/50<br />
8/49<br />
23/50 a<br />
10/50 a<br />
27/99 a<br />
18/100 a<br />
33/100 a<br />
17/100 a<br />
Average No. of<br />
Tumors (+ SD)<br />
0.16+0.37<br />
0.16+0.37<br />
0.28+0.45<br />
0.18+0.44<br />
0.48+0.54 b<br />
0.22+0.46 b<br />
0.33+0.61<br />
0.20+0.45<br />
0.43+0.70 b<br />
0.18+0.41 b<br />
a 2<br />
Significant differences between NDMA-treated and c<strong>on</strong>trol mice, p
Exposure<br />
time<br />
weeks<br />
1<br />
2<br />
3<br />
Table 2. Effects of 50 ppm NDMA in the drinking water: water c<strong>on</strong>sumpti<strong>on</strong>, body weights,<br />
NDMA demethylase, and pathological change<br />
Treat-<br />
ment<br />
N<strong>on</strong>e<br />
10% EtOH<br />
50ppmNDMA<br />
NDMA+EtOH<br />
N<strong>on</strong>e<br />
10% EtOH<br />
50ppmNDMA<br />
NDMA+EtOH<br />
N<strong>on</strong>e<br />
10% EtOH<br />
50ppmNDMA<br />
NDMA+EtOH<br />
Average<br />
Water<br />
C<strong>on</strong>sumpti<strong>on</strong><br />
(ml)<br />
3.6+0.6 (4) a<br />
4.5+0.4 (4)<br />
2.4+0.1 (4)<br />
2.6+0.3 (4)<br />
4.0+0.3 (4)<br />
4.0+0.8 (4)<br />
2.3+0.6 (4)<br />
1.9+0.4 (4)<br />
4.9+1.7 (4)<br />
2.9+0.3 (4)<br />
2.5+0.6 (4)<br />
1.8+0.3 (4)<br />
Average Body<br />
Weight<br />
(g)<br />
26.2+1.7 (10) a<br />
26.8+1.9 (10)<br />
23.4+1.9 (20)<br />
22.4+2.4 (20)<br />
26.8+1.5 (10)<br />
25.9+1.9 (10)<br />
24.6+2.3 (20) d<br />
20.1+2.3 (20) d<br />
27.9+1.7 (10)<br />
28.6+1.7 (10)<br />
24.6+2.3 (10) d<br />
22.4+2.0 (10) d<br />
NDMA<br />
Demethylase<br />
(Nmol<br />
CH2O/mg<br />
Pathological Change<br />
protein/20 min)<br />
Gross Histological<br />
ND b<br />
ND<br />
1.0+0.7 (6) c<br />
2.0+1.0 (6)<br />
ND<br />
ND<br />
1.8+0.7 (6)<br />
1.6+0.8 (6)<br />
ND<br />
ND<br />
0.9+0.6 (6) c<br />
0.5+0.4 (6) c<br />
a<br />
All values are + standard deviati<strong>on</strong>. Numbers of parantheses are numbers of de<strong>term</strong>inati<strong>on</strong>s<br />
b<br />
ND, not d<strong>on</strong>e<br />
c<br />
significantly different from value in untreated mice, 2.4+1.0 (N=7), p
The c<strong>on</strong>centrati<strong>on</strong> of the chemical was c<strong>on</strong>firmed by chemical analysis (extracti<strong>on</strong> with<br />
methylene chloride followed by measurement by gas chromatography).<br />
In experiment 1, the mice were fed Purina Certified Rodent Chow. Treatment was<br />
started at 4 weeks of age and c<strong>on</strong>tinued for 12 , 14 or 16 weeks. For experiment 2, two diets<br />
were formulated to be of defined compositi<strong>on</strong> with regard to essential nutrients, so that these<br />
could be reproducibly manufactured if desired. One of these was similar to Chow 5002, as<br />
used in experiment 1. The other was similar to Purina Mouse Chow 5015, a high fat, low<br />
fiber diet. Treatment was started at 4 weeks of age and c<strong>on</strong>tinued for 18 weeks. Lungs and<br />
stomachs were fixed in Bouin’s soluti<strong>on</strong> and were examined under a dissecting microscope<br />
for tumors. Lungs were examined grossly and after hand-secti<strong>on</strong>ing into 1-mm slices. Any<br />
questi<strong>on</strong>able lesi<strong>on</strong>s were subjected to histological verificati<strong>on</strong> (7- μm secti<strong>on</strong>s, hematoxylin<br />
and eosin staining). Statistical analyses included the x 2 test for differences in percentages of<br />
tumor-bearing mice and the two-tailed Student’s t test for differences in average number of<br />
tumors per mouse.<br />
Subchr<strong>on</strong>ic Exposure of Mice. The mice were housed in polycarb<strong>on</strong>ate cages with wire<br />
inserts and fed NIH autoclavable diet 31. Envir<strong>on</strong>mental c<strong>on</strong>diti<strong>on</strong>s included a temperature<br />
of 25+2 C, humidity of 50+10% and 12 changes of room air / hour. Fluorescent lights were<br />
automatically timed at 12 hours <strong>on</strong> / 12 hours off. Treatment was started when the mice<br />
were 5 weeks of age. Drinking water was shielded from room lighting with aluminium foil<br />
and was changed and measured twice weekly. NDMA and ethanol were diluted with<br />
dei<strong>on</strong>ized water.<br />
At kill, blood was collected after decapitati<strong>on</strong> into cryotubes c<strong>on</strong>taining a few crystals<br />
of ascorbic acid and 50 units of amm<strong>on</strong>ium heparin. The carcass was placed <strong>on</strong> ice and the<br />
livers, kidneys, lungs, and brains were quickly removed to cryotubes. The tissues were<br />
frozen in liquid nitrogen and stored at –20 C until analyzed. In some experiments secti<strong>on</strong>s of<br />
liver were fixed in 10% formalin for histopathology. Porti<strong>on</strong>s of liver to be analyzed for<br />
c<strong>on</strong>tent of NDMA demethylase activity were stored at –80 C prior to analysis.<br />
Gross pathology was estimated semiquantitatively at kill by scoring <strong>on</strong> the basis of 0-3<br />
(0 = normal, 1 = focal change in liver appearance and / or slight perit<strong>on</strong>eal fluid; 2 = general<br />
liver change (pits, yellowing, or darkening) and / or moderate perit<strong>on</strong>eal fluid; 3 =<br />
pr<strong>on</strong>ounced change in liver morphology and extensive perit<strong>on</strong>eal fluid). Similarly,<br />
histopathological change was estimated semiquantitatively <strong>on</strong> a basis of 0-4 (0 = normal or<br />
minimal focal change; 1 = minimal centrilobular hemorrhage; 2 = mild centrilobular<br />
hemorrhage; 3 = moderate centrilobular hemorrhage with focal necrosis; 4 = moderate<br />
centrilobular hemorhhage with massive necrosis).<br />
In order to assess the effects of ethanol <strong>on</strong> tissue levels of NDMA under c<strong>on</strong>diti<strong>on</strong>s of<br />
c<strong>on</strong>trolled nutriti<strong>on</strong>, Lieber-DeCarli Liquidiets were employed. Weanling female mice were<br />
allowed to become accustomed to the diet for 5 weeks, until their weights stabilized. The<br />
mice were then divided into two groups and those of <strong>on</strong>e group were offered diet with<br />
gradually increasing amounts of ethanol, according to the instructi<strong>on</strong>s of the supplier. The<br />
mice accepted the ethanol-c<strong>on</strong>taining diet <strong>on</strong>ly poorly. After an additi<strong>on</strong>al 6 weeks of<br />
570
attempted accommodati<strong>on</strong> to the diet, it was decided to carry out the experiment with twothirds<br />
of the recommended ethanol level (4.3 % of the diet, 24% of total calories). NDMA<br />
was included in both ethanol and c<strong>on</strong>trol diets at 10 ppm, and the mice were pair-fed for 2<br />
weeks so that mice were given NDMA-c<strong>on</strong>trol diet in amounts equal to that c<strong>on</strong>sumed by<br />
the mice given NDMA –ethanol diet.<br />
Estimati<strong>on</strong> of Tissue C<strong>on</strong>tent of NDMA. The amount of NDMA in samples of blood and<br />
other organs (pooled from 5-10 mice) was estimated by methylene chloride extracti<strong>on</strong>thermal<br />
energy analysis as described previously. The limit of detecti<strong>on</strong> of NDMA by this<br />
procedure was 0.5-1 ppb, depending <strong>on</strong> the amount of starting material. Water blanks and<br />
tissues from c<strong>on</strong>trol mice were repeatedly assayed and did not c<strong>on</strong>tain measurable levels of<br />
NDMA (
RESULTS<br />
Tumorigenesis by 500 ppb NDMA. These experiments were designed to test whether a<br />
significant increase in lung tumors could be reproducibly obtained after exposure of male<br />
strain. A mice to 500 ppb NDMA in the drinking water for 16-18 weeks as previously<br />
reported. The results presented in table 1 answer this questi<strong>on</strong> affirmatively: in three<br />
separate trials involving strain A mice from different sources and fed widely differing diets,<br />
exposure to 500 ppb NDMA resulted in significant increases in incidences of lung tumors<br />
compared with c<strong>on</strong>trols. In experiment 1 it was found that 16 weeks of exposure was the<br />
minimum time required for a significant difference to be manifested. The strain A mice in<br />
experiment 2 exhibited somewhat fewer tumors after NDMA treatment than did those in<br />
experiment 1. In spite of the large differences in diet compositi<strong>on</strong> for the two groups in<br />
experiment 2, resulting in those given the 5002 diet weighing <strong>on</strong> average 19% less and<br />
drinking 34% more than those given the 5015 diet (both differences significant; data not<br />
shown), the diet compositi<strong>on</strong> did not significantly affect the incidences of lung tumors in the<br />
c<strong>on</strong>trol or NDMA-treated mice.<br />
Forestomach papillomas were countered in experiment 1 as a possibly useful indicator of<br />
tumorigenesis. The incidences of these tumors were not treatment related (table 1)<br />
Effects of Subchr<strong>on</strong>ic Administrati<strong>on</strong> of 50 ppm NDMA with and without Ethanol. In<br />
order to test whether exposure to a high, hepatotoxic dose of NDMA would result in<br />
measurable tissue levels of NDMA, and whether this level might be influenced by the<br />
simultaneous presence of ehanol, mice were given 50 ppm NDMA with or without 10%<br />
ethanol for 1, 2, or 4 weeks. The NDMA was clearly hepatotoxic, as expected, giving clear<br />
gross and histopathological signs of centrilobular liver damage and leading to a decline in<br />
liver NDMA N-demethylase activity between 1 and 4 weeks of exposure (table 2). NDMA<br />
was measurable in blood and all organ samples (table 3). Inclusi<strong>on</strong> of 10% ethanol al<strong>on</strong>e in<br />
the drinking water had no effect <strong>on</strong> water c<strong>on</strong>sumpti<strong>on</strong>, body weight, or liver histopathology<br />
(table 2). However, the ethanol significantly affected the interacti<strong>on</strong>s of NDMA with liver.<br />
The mice who received ethanol al<strong>on</strong>g with the NDMA experienced noticeably less liver<br />
damage, as indicated by both gross appearance and histopathology, especially after 1 or 4<br />
weeks. Furthermore, the amounts of NDMA recovered from blood and organs were<br />
c<strong>on</strong>sistently greater, by factors of 2-40, in the mice given ethanol as well, at all three time<br />
points (table 3).<br />
Subchr<strong>on</strong>ic Administrati<strong>on</strong> of Low Doses of NDMA with and without Ethanol. In male<br />
and female mice given 5 ppm NDMA in the drinking water for 2 weeks, low levels (1-4<br />
ppb) of NDMA were detected in the blood samples and in <strong>on</strong>e sample each of liver and<br />
brain (table 4). Co-treatment with 30% ethanol resulted in a dramatic increase in amounts of<br />
the NDMA in blood and organs, to about 20 ppb in most cases; livers, especially of the<br />
treated males, were the <strong>on</strong>ly excepti<strong>on</strong> to this pattern. Similar results were obtained in<br />
experiment 2 (table 4) when 10% ethanol was used. In this case the amounts of NDMA<br />
appearing in the blood and tissues of the mice co-treated with ethanol were less than after<br />
30% ethanol, indicating dose dependency of the effect of ethanol. In experiment 3 of this<br />
572
series, a dose of 0.5 ppm NDMA was given with or without ethanol. In the absence of<br />
ethanol, NDMA was detected at low levels in half of the samples analyzed. When ethanol<br />
was present, all samples c<strong>on</strong>tained NDMA at a rather uniform c<strong>on</strong>centrati<strong>on</strong> of about 2-4<br />
ppb.<br />
Effects of Ethanol <strong>on</strong> NDMA Tissue Levels in Mice Pair-Fed a Liquid Diet. In order to<br />
ascertain whether the effects of ethanol in the drinking water might be due to nutriti<strong>on</strong>al<br />
differences between these mice and those not receiving ethanol, two groups with or without<br />
4 % ethanol. Inlcusi<strong>on</strong> of ethanol resulted in the appearance of significant levels of NDMA<br />
in all tissues sampled, whereas, in the absence of ethanol, NDMA was detected at a low<br />
level <strong>on</strong>ly in kidney.<br />
NDMA Demethylase Activity in Livers of Mice Given Various Doses of NDMA with or<br />
without Ethanol. The results presented in table 2 suggested that 10% ethanol in the<br />
drinking water had little inductive effect <strong>on</strong> NDMA demethylase activity in the presence of<br />
50 ppm NDMA, and that chr<strong>on</strong>ic treatment with this high dose of NDMA resulted in a loss<br />
of enzyme activity. Treatment with 10% ethanol al<strong>on</strong>e for 2 weeks resulted in <strong>on</strong>ly a small,<br />
insignificant increase in NDMA demethylase activity, compared with c<strong>on</strong>trols. Levels of<br />
enzyme activity decreased in the mice given 5 or 50 ppm NDMA, significantly so in those<br />
receiving ethanol also. Lack of a similar decrease in those given 20 ppm NDMA may have<br />
been related in part to reducti<strong>on</strong> in water c<strong>on</strong>sumpti<strong>on</strong> at the higher doses of chemical (an<br />
average of 4.3 ml/day compared with 5.7 ml/day for the mice given 5 ppm NDMA).<br />
DISCUSSIONS<br />
In a previous investigati<strong>on</strong> it was found that exposure of male strain A mice to 500 ppb<br />
NDMA for 16 weeks resulted in lung tumors in 44%, compared with 8% of c<strong>on</strong>trols;<br />
increases in lung tumor incidence with lower doses of NDMA were not of statistical<br />
significance. The findings of the present study are in good agreement: 500 ppb NDMA for<br />
16 weeks yielded lung tumors in 48% of the mice. The results further showed that a<br />
significant effect of 500 ppb NDMA can be dem<strong>on</strong>strated with a different substrain of A<br />
mice and with different diets. Thus, 500 ppb NDMA may be the lowest level that gives an<br />
easily measurable tumorgenic effect in adult strain. A mouse lung, but it does so<br />
reproducibly. It is of c<strong>on</strong>siderable interest that this was also a minimally effective<br />
c<strong>on</strong>centrati<strong>on</strong>, in our subchr<strong>on</strong>ic trials, in giving organs. In mice given 500 ppb NDMA for<br />
2 weeks, about half of the tissue sample assayed had measurable NDMA, in the low ppb<br />
range. Although great cauti<strong>on</strong> must be employed in extrapolating from mice to man, thses<br />
results suggest c<strong>on</strong>siderati<strong>on</strong> of the possibility that humans with blood levels of 0.1-1 ppb<br />
NDMA may be at risk of tumor initiati<strong>on</strong> in genetically sensitive peripheral target organs.<br />
573
Table 4. Tissue levels of NDMA in mice given low doses of NDMA in drinking water with or without ethanol.<br />
Expt. NDMA<br />
Treatmen<br />
t<br />
1a (males)<br />
1b<br />
(females)<br />
2<br />
(females)<br />
3<br />
(females)<br />
5<br />
5<br />
5<br />
5<br />
5<br />
5<br />
0.5<br />
0.5<br />
Ethanol Average<br />
Water<br />
C<strong>on</strong>sumpti<br />
<strong>on</strong> a<br />
3.8+0.7<br />
30% 4.7+1.1.<br />
30%<br />
10%<br />
20%<br />
4.2+0.7<br />
4.7+0.4<br />
3.9+0.7<br />
4.5+1.1<br />
4.2+1.5<br />
5.4+1.2<br />
Average<br />
Body<br />
Weight a<br />
25.4+1.9<br />
20.1+2.7<br />
19.2+1.5<br />
17.6+1.7<br />
20.5+2.1<br />
20.5+1.9<br />
20.7+4.3<br />
22.5+2.0<br />
574<br />
NDMA C<strong>on</strong>tent<br />
Blood Liver Kidney Lun<br />
g<br />
1<br />
22<br />
4<br />
26<br />
0.6,2 b<br />
4,12 b<br />
At 500 ppb and at all higher doses, inclusi<strong>on</strong> of ethanol in the drinking water led to<br />
c<strong>on</strong>sistent increases in the amounts of NDMA in the blood and other organs. Although<br />
there was quantitative variati<strong>on</strong>, as might be expected in an experiment where the<br />
chemical dose was received in the drinking water, the differences in NDMA levels with<br />
and without ethanol were in many cases of c<strong>on</strong>siderable magnitude. The obvious<br />
explanati<strong>on</strong> for this effect of ethanol is competitive inhibiti<strong>on</strong> of the metabolism of<br />
NDMA, as reported by other workers.This inhibiti<strong>on</strong>, causing a reducti<strong>on</strong> in the rate of<br />
formati<strong>on</strong> of cytotoxic metabolites, probably also accounts for the alleviati<strong>on</strong> of<br />
morphologically apparent hepatotoxic effects of NDMA as well. The ethanol did not,<br />
however, prevent the loss of NDMA demethylase activity caused by the higher doses of<br />
NDMA but, in fact, seemed to c<strong>on</strong>tribute to this loss. Although ethanol is an inducer of<br />
NDMA demethylase activity in rats after administrati<strong>on</strong> in drinking water, at most, <strong>on</strong>ly<br />
a small inductive effect was seen in our mice. Inhibiti<strong>on</strong> of NDMA metabolism was<br />
clearly the dominant acti<strong>on</strong>.<br />
In studies employing ethanol in the drinking water, the argument may be raised<br />
that the observed effects are due to nutriti<strong>on</strong>al imbalance sec<strong>on</strong>dary to ethanol<br />
c<strong>on</strong>sumpti<strong>on</strong>, rather than the chemical itself; pair-feeding of liquid diets is thus<br />
recommended. The experiment c<strong>on</strong>firmed that the observed effects of ethanol <strong>on</strong> blood<br />
and organ c<strong>on</strong>tent of NDMA were due directly to the ethanol and not to nutriti<strong>on</strong>al<br />
differences, since both groups c<strong>on</strong>sumed the same amount of nutriti<strong>on</strong>ally equivalent<br />
diet. Other workers have similarly found that administrati<strong>on</strong> of ethanol in the drinking<br />
water is a sound experimental approach.<br />
In sum, chr<strong>on</strong>ic co-treatment of mice with ethanol, al<strong>on</strong>g with NDMA, resulted in<br />
a loweing of <strong>on</strong>e hepatotoxic effect of a high dose of NDMA, centrilobular necrosis, but<br />
enhancement of another effect, destructi<strong>on</strong> of NDMA demethylase activity. At all doses<br />
the ethanol resulted in marked increases in circulating levels of NDMA. The<br />
c<strong>on</strong>tributi<strong>on</strong> of this effect to tumorgenesis by low doses of NDMA in peripheral target<br />
organs will be an interesting subject for future experiments.<br />
REFERENCES<br />
1. D.H. Fine, “Nistrosamine in the general envir<strong>on</strong>ment and food”. In “ Nistrosamine<br />
and Human Cancer” (Banbury Report 12) (P.N. Magee, ed.), pp. 199-210. Cold<br />
Spring Harbor Laboratory, New York, 1982<br />
2. L. Lakritz, R.A. Gates, A.M. Gugger and A.E. Wasserman, “Nitrosamine levels in<br />
human blood, urine and gastric aspirate following ingesti<strong>on</strong> of foods c<strong>on</strong>taining<br />
potential nistrosamine precursors of preformed nistrosamines”, Food Chem.<br />
Toxicol. 20, 455-459 (1982)<br />
3. T.A. Gough, K.S.Webb and P.F. Swann, “An examinati<strong>on</strong> of human blood for the<br />
pressure of volatile nistrosamines”, Food Chem. Toxicol. 21, 151-156.<br />
575
DETERMINATION OF METALLOTHIONEIN, CADMIUM, COPPER AND ZINC<br />
LEVELS IN HUMAN AND RAT TISSUES<br />
Camelia Bara<br />
University of Oradea, Faculty of Envir<strong>on</strong>mental Protecti<strong>on</strong>, Oradea<br />
26, General Magheru Street, Oradea<br />
ABSTRACT<br />
Metallothi<strong>on</strong>ein (MT), zinc (Zn), copper (Cu) and cadmium (Cd) were de<strong>term</strong>ined in 10<br />
tissues (brain, heart, kidney cortex, liver, lung, muscle, pancreas, small intestine, spleen and<br />
stomach) from human autopsies (10 male individuals, mean age 43 ± 9 years, all smokers)<br />
and Wistar rats. The mean tissue c<strong>on</strong>centrati<strong>on</strong>s of MT in the human samples varied<br />
between 3.8 and 495 µg/g wet weight in spleen and kidney cortex, respectively. In most<br />
tissues human MT levels were high as compared to rats; particularly in liver and kidney<br />
cortex human MT levels exceeded those of rats about 25- and 10-fold, respectively. Positive<br />
linear relati<strong>on</strong>ships were observed between Zn or Cu and MT in human liver and between<br />
Cd and MT in human kidney cortex.<br />
INTRODUCTION<br />
Metallothi<strong>on</strong>ein (MT), a low molecular weight protein with high cysteine and metal c<strong>on</strong>tent,<br />
binds both essential (Zn, Cu) and n<strong>on</strong>-essential metals (Cd, Hg). In view of the ability of<br />
MT to be induced by metals, the protein plays a crucial role in the homeostasis and toxicity<br />
of metals. The biological activity of metals thus is de<strong>term</strong>ined by both the c<strong>on</strong>centrati<strong>on</strong> in<br />
the tissue and the fracti<strong>on</strong> bound to MT. However, there are few reports as to the MT<br />
c<strong>on</strong>centrati<strong>on</strong>s in human tissues. In a previous study we found high MT levels in human<br />
kidney cortex, which were even higher in smokers. In these kidney cortices the amount of<br />
cadmium bound to MT directly correlated with the c<strong>on</strong>tent of renal cadmium but not with<br />
zinc or copper. In a study, Onosaka et al. reported similarly high MT levels in human liver<br />
and a str<strong>on</strong>g positive relati<strong>on</strong>ship between hepatic Zn and MT c<strong>on</strong>centrati<strong>on</strong>s.<br />
To further elucidate the relati<strong>on</strong>ship between tissue levels of MT and metals, we<br />
investigated in this study the levels of MT, zinc, copper and cadmium in 10 different organs<br />
from human autopsies and rats. Some of the rat data have been previously reported.<br />
MATERIALS AND METHODS<br />
Samples of human tissues were obtained from 10 male individuals (20-50 years old, mean<br />
age 43 ± 9, all smokers) necropsied within 2 days after sudden death. N<strong>on</strong>e of the<br />
individuals received medicati<strong>on</strong> before death. Samples were stored at -80°C until being<br />
processed further. Tissues of humans and rats (Wistar strain, males, 180-200 g) were<br />
homogenized with a Potter-Elvehjem or Ultra-Turrax in 4 or 9 vols, of 30 mM Tris-HCl<br />
buffer pH 7.4, and subsequently centrifuged at 18 000 x g. MT was de<strong>term</strong>ined in the<br />
576
supernatants by a modified cadmium saturati<strong>on</strong> method as described previously. Zn, Cu and<br />
Cd were detected in tissue homogenates by atomic emissi<strong>on</strong> spectroscopy with inductively<br />
coupled plasma excitati<strong>on</strong> or atomic absorpti<strong>on</strong> spectroscopy after wet ashing of the<br />
samples at 140°C with nitric acid.<br />
RESULTS<br />
MT was detected in all human tissues investigated. The highest MT tissue levels were found<br />
in kidney cortex. In all tissues except brain and small intestine, human samples showed<br />
higher MT levels than those of rats. Particularly in liver and kidney, human MT levels<br />
exceeded those of rats about 25- and 10-fold, respectively.<br />
Human tissues showed moderate differences in their Zn or Cu levels whereas regarding Cd<br />
levels pr<strong>on</strong>ounced variati<strong>on</strong>s were observed. The highest Cd c<strong>on</strong>centrati<strong>on</strong>s (29.7 ±13.6<br />
µg/gwet weight) were observed in the kidney cortex.<br />
In most human and rat tissues the levels of Zn and Cu were approximately similar<br />
within a factor of 2. In comparis<strong>on</strong> with the very low levels of Cd in rat tissues, the Cd<br />
levels of all human tissues were remarkably higher. The Cd levels of all rat tissues were<br />
lower than the detecti<strong>on</strong> limit of 20 ng Cd/g tissue wet weight. In human liver, positive<br />
relati<strong>on</strong>ships between the c<strong>on</strong>centrati<strong>on</strong>s of both Zn and Cu and the amount of MT were<br />
observed. The regressi<strong>on</strong> coefficients were r = 0.871 (MT (µg) = 9.8 Zn (µg) - 257) and r =<br />
0.955 (MT (µg) = 100 Cu (µg) — 359), respectively.<br />
In human kidney cortex, a positive relati<strong>on</strong>ship was observed between Cd and MT with<br />
a regressi<strong>on</strong> coefficient of r = 0.779 (MT (µg) = 13.4 Cd (µg) + 98.5). Metallothi<strong>on</strong>ein<br />
levels in human kidney and liver were not significantly correlated.<br />
DISCUSSION<br />
MT was detected in all rat and human tissues investigated. The rat data c<strong>on</strong>firm earlier<br />
findings of others, and us although lower levels of brain MT were obtained with a<br />
radioimmunoassay. In most human tissues higher levels of MT were observed than in rats.<br />
Moreover, human MT levels, particularly in liver and kidney showed greater interindividual<br />
variati<strong>on</strong>s, e.g., the c<strong>on</strong>centrati<strong>on</strong>s of human hepatic MT varied between 11 and 1000 µg/g.<br />
The extraordinarily high levels of MT in human kidney and liver may reflect a different<br />
transcripti<strong>on</strong>al c<strong>on</strong>trol of the protein, the occurrence of multiple subtypes of human MT<br />
isoforms, or the high exposure of man to envir<strong>on</strong>mental metals. In accordance with the<br />
latter, levels of zinc, copper and particularly cadmium in human liver and levels of zinc and<br />
cadmium in human kidney clearly exceeded those of the rat. With a ratio of 7 mol Cd per<br />
mol of MT, MT of human liver and kidney cortex has the capacity to bind total tissue<br />
cadmium. This supports the assumed protective role of MT in the detoxificati<strong>on</strong> of Gd.<br />
However, in the renal cortex of both smokers and n<strong>on</strong>-smokers, MT was shown to bind<br />
about 50% of tissue Cd <strong>on</strong>ly. The remaining n<strong>on</strong>-MT Cd may represent a significant toxic<br />
potential. Zinc, copper and cadmium are potent inducers of MT in animal tissues. Although<br />
there is no clear evidence for the inducti<strong>on</strong> of MT by these metals in human tissues, the<br />
577
present findings <strong>on</strong> positive correlati<strong>on</strong>s between renal MT and cadmium, and hepatic MT<br />
and zinc support this view.<br />
Similarly, a close relati<strong>on</strong>ship between renal MT and tissue cadmium of smokers and<br />
n<strong>on</strong>-smokers and a positive correlati<strong>on</strong> between MT and zinc in human liver have been<br />
observed. These data indicate that MT in human kidney is mainly induced by cadmium<br />
whereas in liver it may be preferentially induced by zinc and possibly copper. However,<br />
since inducti<strong>on</strong> of MT has also been reported after various stresses, factors like the<br />
horm<strong>on</strong>al and nutriti<strong>on</strong>al status and even the type of death might c<strong>on</strong>tribute as well.<br />
There seems to be no correlati<strong>on</strong> between hepatic and renal MT levels which we have<br />
also observed in a recent investigati<strong>on</strong> with 145 postmortem samples of liver and kidneys.<br />
Except in 2 individuals, MT levels in human kidney cortex exceeded those of the liver.<br />
However, in these cases hepatic Zn and renal Cd levels were more than twice and less than<br />
50% of the average values, respectively. This supports the relevance for MT inducti<strong>on</strong> of the<br />
high Cd c<strong>on</strong>tent of the kidney as compared to the liver. In c<strong>on</strong>trast, MT levels of rat liver<br />
and kidney positively correlated with each other either after acute or subchr<strong>on</strong>ic exposure to<br />
cadmium and zinc or the acute administrati<strong>on</strong> to iodoacetate. Therefore the chr<strong>on</strong>ic<br />
exposure of man to low levels of metals, in particular cadmium, and the preferential<br />
distributi<strong>on</strong> of Cd-MT into the kidney may explain the observed lack of correlati<strong>on</strong>.<br />
In c<strong>on</strong>clusi<strong>on</strong>, these results dem<strong>on</strong>strate that c<strong>on</strong>siderable amounts of MT are present<br />
in various human tissues. The particularly high levels of MT in human liver and kidney also<br />
in comparis<strong>on</strong> to rats and the high levels of metals in these tissues suggest that the<br />
expressi<strong>on</strong> of MT in human tissues may resp<strong>on</strong>d to envir<strong>on</strong>mental factors, especially<br />
chr<strong>on</strong>ic low-level exposure to metals.<br />
REFERENCES<br />
1. E.C Foulkes (Ed.), Biological Roles of Metallothi<strong>on</strong>ein, Elsevier/North-Holland, New<br />
York, 1982.<br />
2. R.J. Cousins, Metallothi<strong>on</strong>ein - aspects related to copper and zinc metabolism, J.<br />
Inherit. Metab. Dis., 6 (Suppl. 1) (1983) 15-21.<br />
3. K.H. Summer, G.A. Drasch and H.E. Heilmaier, Metallothi<strong>on</strong>ein and cadmium in<br />
human kidney cortex: influence of smoking, Hum. Toxicol., 5 (1986) 27-33.<br />
4. S. Onosaka, K.S. Min, C Fukuhara, K. Tanaka, S.I. Tashiro, I. Shimizu, M. Furuta, T.<br />
Yasutomi, K. Kobashi and K.I. Yamamoto, C<strong>on</strong>centrati<strong>on</strong>s of metallothi<strong>on</strong>ein and<br />
metals in malignant and n<strong>on</strong>-malignant tissues in human liver, Toxicology, 38 (1986)<br />
261-268.<br />
5. H.E. Heilmaier and K.H. Summer, Metallothi<strong>on</strong>ein c<strong>on</strong>tent and zinc status in various<br />
tissues of rats treated with iodoacetic acid and zinc, Arch. Toxicol., 56 (1985) 247-251.<br />
6. S. Onosaka and M.G. Cherian, The induced synthesis of metallothi<strong>on</strong>ein in various<br />
tissues of rat in resp<strong>on</strong>se to metals. I. Effect of repeated injecti<strong>on</strong> of cadmium salts,<br />
Toxicology, 22 (1981) 91-101.<br />
7. C.V. Nolan and Z.A. Shaikh, De<strong>term</strong>inati<strong>on</strong> of metallothi<strong>on</strong>ein in tissues by<br />
578
adioimmunoassay and by cadmium saturati<strong>on</strong> method, Anal. Biochem., 154 (1986)<br />
213-223.<br />
8. C.J. Schmidt and D.H. Hamer, Cell specificity and an effect of ras <strong>on</strong> human<br />
metallothi<strong>on</strong>ein gene expressi<strong>on</strong>, Proc. Natl. Acad. Sci. U.S.A., 83 (1986) 3346-3350.<br />
9. J. Chung, N.O. Nartey and M.G. Cherian, Metallothi<strong>on</strong>ein levels in liver and kidney of<br />
Canadians - A potential indicator of envir<strong>on</strong>mental exposure to cadmium, Arch.<br />
Envir<strong>on</strong>. Health, 41 (1986) 319-323.<br />
579
THE ROLE OF WATER QUALITY IN INFANTILE ALIMENTATION<br />
Andrei Csép, Draghici S<strong>on</strong>ia, Lenard Ildiko, Mirela Indries, Nicoleta Negrut<br />
University of Oradea, Faculty of Medicine and Farmacy<br />
ABSTRACT<br />
The study makes c<strong>on</strong>crete reference to the risks linked to the water c<strong>on</strong>sume, which doesn’t<br />
corresp<strong>on</strong>d from a chemical point of view, through the presence and the c<strong>on</strong>centrati<strong>on</strong> over<br />
the limits admitted by nitrates and nitrites in the water used at milk powder preparati<strong>on</strong>,<br />
distributed through the nati<strong>on</strong>al programme and used in the alimentati<strong>on</strong> of babies.<br />
Keywords: water, nitrate c<strong>on</strong>tent, intoxicati<strong>on</strong>, Faecal coliforms, Faecal streptococci<br />
INTRODUCTION<br />
Nitrate intoxicati<strong>on</strong> was firstly described and named well water intoxicati<strong>on</strong> in 1945<br />
(quoted from Biro O.). It is also known under the name of infantile intoxicati<strong>on</strong>, infantile<br />
cyanotic methemoglobinemia, or the newborn’s blue disease, because the dominant<br />
symptom is cyanosis, symptom that can lead to the rapid diagnosticati<strong>on</strong> of the disease.<br />
(Chiotan, 1999)<br />
The nitrates enter the organism altogether with the foodstuff of vegetal and animal origin,<br />
but mainly through drinking water. They can have a double origin in water:<br />
a) They can come from soils rich in nitrogen salts, when their origin is c<strong>on</strong>sidered natural;<br />
b) As a c<strong>on</strong>sequence of water polluti<strong>on</strong>, that comes either directly from nitrates (e.g.<br />
industrial and agricultural polluti<strong>on</strong> with nitrogen-based fertilizers), or from organic<br />
substances, which through decompositi<strong>on</strong> release nitrates.<br />
Nitrates are not toxic by themselves. To achieve this quality, they have to undergo a<br />
reducti<strong>on</strong> process and be transformed in nitrites.<br />
Transformati<strong>on</strong> may take place:<br />
a) Exogenously (in water), but <strong>on</strong>ly in the warm periods of the year, and in the presence<br />
of a reductive flora;<br />
b) Endogenously (in organism), under the acti<strong>on</strong> of the same reductive flora;<br />
This reductive flora either gets up in the distal digestive tract in the proximal regi<strong>on</strong> where<br />
the absorpti<strong>on</strong> takes place in case of digestive perturbati<strong>on</strong>s (coliform germs), or gets down<br />
from nasal-pharynges in the case of some diseases, such as: rhinitis, t<strong>on</strong>silitis, otitis etc,<br />
especially if they are caused by streptostaphilococci.<br />
The nitrites that entered the body combine with the haemoglobin (Hb) forming<br />
methemoglobin (MHb), leading to a deficit of oxygen. The seriousness of the disease<br />
(clinical forms) is given by the quantity of bloked haemoglobin (that is the percent of MHb)<br />
(Biro, 1993):<br />
• The superficial form appears between 10 – 25% and is characterized by cyanosis of<br />
face (lips, nose), extremities (nails, finger tips) and mucous membrane<br />
580
• The in<strong>term</strong>ediary form appears between 25 – 45%, and dyspnoea, tachycardia, cardiac<br />
breath, headaches are added to the symptoms characteristic to the superficial form<br />
• The severe form appears over 50%, c<strong>on</strong>vulsi<strong>on</strong>s, arhytmia, coma and even death (blue<br />
death due to asphyxia) being added to the previously menti<strong>on</strong>ed symptoms<br />
characteristic for superficial and in<strong>term</strong>ediary forms of the disease<br />
The disease is present almost exclusively in little children, from the first year of their life,<br />
who are fed artificially (Ciofu E., Ciofu C, 1999, Apllet<strong>on</strong> & Lange, 1993), due to the<br />
following special physiological characteristics of the sucklings:<br />
• Gastric acidity is lower in the first m<strong>on</strong>ths of life;<br />
• 80% of the suckling’s Hb is of fetal type, being more unstable and easier to be oxidized<br />
in comparis<strong>on</strong> with the adult’s Hb.;<br />
• Water quantity (hence of nitrates) ingested by the baby, with respect to the blood<br />
volume, is about 12 times higher than in the case of an adult;<br />
• Some enzymatic deficiencies (deficit of glucoso-6-phosphate-dehydrogenase) favour<br />
the appearance of the disease;<br />
• A high frequency disease rate in athrepsic suckling, with gastro-intestinal perturbati<strong>on</strong>s<br />
and infecti<strong>on</strong>s.<br />
MATERIALS AND METHODS<br />
The study was made <strong>on</strong> 17 sucklings from some rural places of Bihor county, hospitalized<br />
in 2006. The methodology that was used during the experiment was imposed by Ministry of<br />
Family and Health, by studying t€he medical records sent by the health units with beds to<br />
the Public Health Directorate of Bihor County.<br />
The data collected from these medical records are presented in the following tables:<br />
Table 1. Data collected from the patients’ medical records<br />
Case No. Sex Age till the<br />
debut<br />
(m<strong>on</strong>ths)<br />
Alimentati<strong>on</strong> Associati<strong>on</strong><br />
with BDA<br />
Clinical<br />
form<br />
1 male 1-2 natural no Sever<br />
2 female 0-1 mixed no Medium<br />
3 female 0-1 Natural no Sever<br />
4 male 0-1 Artificial no Sever<br />
5 female 7-8 Artificial no Medium<br />
6 female 2-3 Natural no Sever<br />
7 female 0-1 Mixed yes Sever<br />
8 male 1-2 Artificial yes Sever<br />
9 female 0-1 natural no Medium<br />
10 male 2-3 artificial no Medium<br />
581
11 female 0-1 artificial yes Sever<br />
12 female 14-15 artificial no Medium<br />
13 male 0-1 mixed no Medium<br />
14 male 2-3 natural no Mediumj<br />
15 male 0-1 mixed no Sever<br />
16 female 0-1 natural no medium<br />
17 male 0-1 artificial no medium<br />
Case no. Sanitary<br />
protecti<strong>on</strong> of<br />
well<br />
Table 2. Data regarding well water quality<br />
Chemical polluti<strong>on</strong> Microbiological polluti<strong>on</strong><br />
% Nitrates<br />
in water<br />
(mg/l)<br />
% Nitrites<br />
in water<br />
(mg/l)<br />
Faecal<br />
coliforms<br />
Faecal<br />
streptococci<br />
1 no 45 0-0,3 ≥10 ≥10<br />
2 no 45 0-0,3 ≥10 ≥10<br />
3 no 46-100 0-0,3 ≥10 ≥10<br />
4 no 101-400 0-0,3 ≥10 ≥10<br />
5 no 101-400 ≥0,3 ≥10 ≥10<br />
6 yes 45 0-0,3 ≥10 ≥10<br />
7 yes 46-100 0-0,3 ≥10 ≥10<br />
8 no 46-100 0-0,3 ≥10 ≥10<br />
9 no 46-100 0-0,3 ≥10 ≥10<br />
10 no 45 0-0,3 ≥10 ≥10<br />
11 no 101-400 0-0,3 ≥10 ≥10<br />
12. no 45 0-0,3 ≥10 ≥10<br />
13 no 46-100 0-0,3 ≥10 ≥10<br />
14 no 45 0-0,3 ≤2 ≤2<br />
15 no 46-100 0-0,3 ≥2 ≤2<br />
16 no 46-100 0-3 ≥10 ≥10<br />
17 no 45 ≥3 ≤2 ≤2<br />
RESULTS AND DISCUSSIONS<br />
1. Out of 192 water probes sampled from public and individual wells for lab analyses in<br />
2006, 57 probes (29.68%) were inadequate as regards the nitrate c<strong>on</strong>tent, and the rest<br />
of 107 probes (55.72%) were inadequate as regards the nitrite c<strong>on</strong>tent.<br />
2. Out of the 17 cases, 9 cases (52.94%) involved female sucklings and 8 cases (47.05%)<br />
involved malesucklings, fact that proves the disease doesn’t appear <strong>on</strong>ly at a certain<br />
582
sex. In fact, there are no data in the specilized literature regarding a variable sensibility<br />
for <strong>on</strong>e sex or another.<br />
47%<br />
53%<br />
Figure 1. Suckling distributi<strong>on</strong> <strong>on</strong> sexes.<br />
male<br />
female<br />
3. By having a look at the age of sucklings till the moment of disease appearance, it is<br />
noticed that, out of 17 cases, 15 cases (88.23%) are recorded for sucklings of 0 - 3<br />
m<strong>on</strong>ths old, <strong>on</strong>e case (5.88%) for sucklings of 6 -12 m<strong>on</strong>ths old, <strong>on</strong>e case (5.88%) for<br />
sucklings of 12 - 36 m<strong>on</strong>ths old; out of the 15 cases involving sucklings of 0-3 m<strong>on</strong>ths<br />
old, 10 cases (58.82%) are with sucklings of 0-1 m<strong>on</strong>th old, two cases (11.76%) with<br />
sucklings of 1-2 m<strong>on</strong>ths old and three cases (17.64%) with sucklings of 2-3 m<strong>on</strong>ths<br />
old.<br />
4. Out of 17 cases, six sucklings (35.29%) were feeeded naturally, seven sucklings<br />
(41.17%) were feeded artificially (cow milk) and four sucklings were feeded in a<br />
mixed manner (23.52%).<br />
5. In most cases, that is 88.23% (15 cases), the serious form of disease appeared after the<br />
c<strong>on</strong>sumpti<strong>on</strong> of well water without sanitary protecti<strong>on</strong>, even if the distance<br />
recommended by norms, with respect to possible sources of c<strong>on</strong>taminati<strong>on</strong>, was<br />
respected (latrines, manure and fertilizer deposits);<br />
6. Out of 17 probes, seven probes (41,17%) have the nitrate c<strong>on</strong>centrati<strong>on</strong> within the<br />
limits imposed by the sanitary norms (under 45mg/l), the rest of them (58.82%) exceed<br />
the maximum admitted c<strong>on</strong>centrati<strong>on</strong> (MAC), as it follows: seven probes (41.17%)<br />
have the nitrate c<strong>on</strong>centrati<strong>on</strong> between 46-100mg/l and three probes (17.64%) have the<br />
c<strong>on</strong>centrati<strong>on</strong> between 101-400mg/l. It is noticed that there were superficial and severe<br />
clinical forms regardless the nitrate c<strong>on</strong>centrati<strong>on</strong> in the water;<br />
7. Most of the probes, that is, 15 probes (88.23%) had the nitrite c<strong>on</strong>centrati<strong>on</strong> within the<br />
limits imposed by the sanitary norms (0-0.3mg/l), <strong>on</strong>ly two probes (11.76%) had a<br />
583
c<strong>on</strong>centrati<strong>on</strong> higher than 0.3 mg/l. There were also superficial and severe clinical<br />
forms regardless the nitrite c<strong>on</strong>centrati<strong>on</strong> in the water.<br />
8. In 14 cases (82.35%), there is recorded an outrunning of the sanitary norms as regards<br />
the microbiological quality of water (faecal coliforms and streptococci, indicators of<br />
faecal polluti<strong>on</strong> over the admitted limit), fact that highlights double water polluti<strong>on</strong>:<br />
chemical and biological, which favor the increase of nitrite-nitrate c<strong>on</strong>centrati<strong>on</strong> in<br />
water.<br />
CONCLUSIONS<br />
In Bihor County, the water from individual wells is polluted with nitrites – nitrates in<br />
different rural localities (spread all over the county), without recording predisposed<br />
regi<strong>on</strong>s).<br />
An increased morbidity is noticed due to acute intoxicati<strong>on</strong> with nitrates at the age<br />
group 0-1 year, which can be avoided by the following measures that can be addressed to all<br />
the factors involved:<br />
• Intensificati<strong>on</strong> of the water quality c<strong>on</strong>trol to maintain MAC of nitrates under toxic<br />
values;<br />
• Mother training regarding the advantages of natural alimentati<strong>on</strong>, at least during the<br />
first 3-4 m<strong>on</strong>ths, and the preventi<strong>on</strong> of digestive and respiratory infecti<strong>on</strong>s of the<br />
suckling, as well as the informati<strong>on</strong> of the families regarding the above-menti<strong>on</strong>ed<br />
risks;<br />
• Counselling of pregnant women from the rural areas to take a water sample from the<br />
well for c<strong>on</strong>trol;<br />
• Training the medical-sanitary pers<strong>on</strong>nel from the rural health unit to know the situati<strong>on</strong><br />
of the water supplies from the area, and to recognize the disease, which is often<br />
c<strong>on</strong>fused with other diseases associated with cyanosis (heart and pulm<strong>on</strong>ary diseases);<br />
• C<strong>on</strong>trolled usage and storage of the natural and chemical fertilizers, m<strong>on</strong>itoring the<br />
removal of solid and liquid wastes in order to protect the water supplies.<br />
REFERENCES<br />
1. Apllet<strong>on</strong> & Lange, 1993 – Current Medical Diagnosis and Treatment, Publishing<br />
Divisi<strong>on</strong> of Prentice Hall, C<strong>on</strong>necticut, USA<br />
2. Biro G., 1993 – Elelmiszer-higienia, Agroinform Kiado, Budapest<br />
3. Chiotan M., 1999 – Boli infectioase, Editura Nati<strong>on</strong>al, Bucuresti<br />
4. Ciofu E., Ciofu C., 1999 – Esentialul in Pediatrie, Editura Medicala Amaltea, Bucuresti<br />
584
SOME ASPECTS OF SHELTER REGENERATION OF QUERCUS SPP.<br />
FOREST STANDS FROM PRODUCTION UNIT II VALEA LUNGĂ,<br />
FORESTRY DISTRICT, LUGOJ (TIMIŞ COUNTY)<br />
Crsitian C. Crainic<br />
University of Oradea, Romania<br />
ABSTRACTS<br />
The paper presents some characteristics of shelter regenerati<strong>on</strong> of Quercus spp. stands<br />
from Producti<strong>on</strong> Unit II Valea Lungă, Forestry District Lugoj (Timiş County) as a<br />
c<strong>on</strong>sequence of the treatment with repeated logging. The obtained results permit some<br />
theoretical c<strong>on</strong>siderati<strong>on</strong>s <strong>on</strong> shelter wood treatment of Quercus spp.<br />
Stands vegetating in particular site c<strong>on</strong>diti<strong>on</strong>s.<br />
Key words: shelter regenerati<strong>on</strong>, stand, treatment, oaks, ecosystem<br />
INTRODUCTION<br />
Choosing and promoting the treatments represent a fundamental problem for the forest<br />
district. The extensive treatments, which are characterised by a possibility of an integral<br />
mechanizati<strong>on</strong> and a relatively reduced technicality, offer fewer assortments and<br />
str<strong>on</strong>gly affect the dynamic balance of the ecosystem. The intensive treatments need a<br />
high degree of technicality, a proper equipping with transport machines, and the quality<br />
of the products obtained after their applicati<strong>on</strong> is superior. Also, when choosing the<br />
treatments we have to take into account the stati<strong>on</strong>’s particularities, the physiological<br />
and ecological particularities of the species and not in the end the local experience.<br />
The natural envir<strong>on</strong>ment, the social-ec<strong>on</strong>omic requests and the traditi<strong>on</strong> of the<br />
Romanian forestry, plead for preserving the forests’ authenticity within the Carpathian-<br />
Danube-P<strong>on</strong>tic frame, in all its compositi<strong>on</strong>al diversity, for managing this very complex<br />
but efficient bio-cybernetic system optimised by nature, <strong>on</strong> some criteria compatible<br />
with the social-ec<strong>on</strong>omic exigency of the future (Giurgiu V, 2005)<br />
MATERIALS AND METHODS<br />
In order to point out the applicati<strong>on</strong> manner of the regenerating cuttings in the<br />
producti<strong>on</strong> unit (U.P.) II Valea Lungă, Forestry District Lugoj, it was analysed the way<br />
of c<strong>on</strong>ducting the cuttings, the intensity of the interventi<strong>on</strong>s, the periodicity of the<br />
interventi<strong>on</strong>s and the dynamic of the natural regenerati<strong>on</strong>, from the valuating documents<br />
and from the records at the forest district.<br />
The distributi<strong>on</strong>s of the numbers of trees <strong>on</strong> diameter category were carried out for<br />
the forest stands implied in the regenerati<strong>on</strong> process <strong>on</strong> the basis of the centralizing<br />
records from the valuating documents. The stati<strong>on</strong>al c<strong>on</strong>diti<strong>on</strong>s and the vegetati<strong>on</strong> from<br />
U.P. II Valea Lungă, Forestry District Lugoj were analysed. In order to be able to<br />
present the objective reality from the site, digital photographic recordings were carried<br />
out, <strong>on</strong> magnetic support using a digital camera brand H.P., with a 3,2 MP resoluti<strong>on</strong>.<br />
In the present case study, from objective reas<strong>on</strong>s, it is analysed the dynamic of the<br />
regenerati<strong>on</strong> cuttings at three representative forest stands, in the forest planning units<br />
(u.a.) 3A, 4A şi 11C.<br />
585
RESULTS<br />
Following the results of the data from the site, a series of results were reached at, results<br />
presented in the following lines. If the stati<strong>on</strong>al c<strong>on</strong>diti<strong>on</strong>s are analysed – especially the<br />
ecologic factors (D<strong>on</strong>iţă N., et. al., 1977) represented by precipitati<strong>on</strong>s and respectively<br />
temperature, it can be noticed that they are premises for establishing the seedlings for<br />
usage in favourable c<strong>on</strong>diti<strong>on</strong>s – table 1.<br />
Table 1: M<strong>on</strong>thly average temperatures and precipitati<strong>on</strong>s at Lugoj meteorological<br />
stati<strong>on</strong> (*)<br />
M<strong>on</strong>th I II III IV V VI VII VIII IX X XI XII<br />
0 1 2 3 4 5 6 7 8 9 10 11 12<br />
T[°C] -1 1 5 12 17 20 22 22 18 13 6 2<br />
Pp [mm] 40 40 50 60 80 80 60 60 50 60 50 50<br />
Tma = 11[°C]<br />
Pp = 600 [mm], where:<br />
Tma – annual average temperature;<br />
Pp – annual average precipitati<strong>on</strong>s;<br />
Pp<br />
I ar = , where: (1)<br />
Tma<br />
+ 10<br />
Iar – de Mart<strong>on</strong>ne dryness index<br />
600<br />
I ar = = 28,<br />
5 ≅ 29<br />
11+<br />
10<br />
The value of de Mart<strong>on</strong>ne dryness index calculated with formula 1 suggests the fact that<br />
the forest vegetati<strong>on</strong> can be developed in favourable c<strong>on</strong>diti<strong>on</strong>s. In this c<strong>on</strong>text it can be<br />
noticed the growth dynamic of samples from cerris species (Q. cerris) developed from<br />
seedlings in the forest stand from u.a. 11C, analysing the picture in figure 1.<br />
In the oaks forest stand from U.P. II Valea Lungă n<strong>on</strong>-uniform regenerati<strong>on</strong> cuttings are<br />
applied – progressive cuttings (in gaps), and as a result, the regenerati<strong>on</strong> benefits from<br />
the best bio ecological c<strong>on</strong>diti<strong>on</strong>s if the treatment technique is strictly applied and it<br />
adapts correctly to the state and structure of the working forest (Florescu I.I., Nicolescu<br />
N. V., 1998).<br />
During 1995 - 2004, in u.a. 3A, 4A and 11C from U.P. II Valea Lungă interventi<strong>on</strong>s in<br />
n<strong>on</strong>-uniform regenerati<strong>on</strong> cuttings were applied – progressive cuttings (in gaps), and<br />
were correlated to the fructificati<strong>on</strong> years at the main cvernicee species (Q. robur, Q.<br />
cerris, Q. frainetto, Q. petraea, Stănescu V., et. al.,1997) .<br />
586
In the following lines it is presented the dynamic of the interventi<strong>on</strong>s in u.a. 3A, 4A and<br />
11C in U.P. II Valea Lungă, analysing the distributi<strong>on</strong> of the number of trees <strong>on</strong><br />
diameter categories and respectively <strong>on</strong> the intensity of interventi<strong>on</strong>s <strong>on</strong> volume (fig. 2,<br />
3, 4, 5, 6, 7).<br />
Fig. 1 – Growth vigour at cerrus regenerati<strong>on</strong> developed from seedling<br />
Analysing figures 2 and 4, it can be observed a surplus of trees of small diameters, trees<br />
that were not totally extracted at the cultural operati<strong>on</strong>s (especially <strong>on</strong> the occasi<strong>on</strong> of<br />
the first thinning), at the preparatory cutting, but the extracti<strong>on</strong> was d<strong>on</strong>e step by step, at<br />
the applicati<strong>on</strong> of favourable workings, the seedlings establishing, <strong>on</strong> ecologic grounds,<br />
in order to prevent the establishing of the grassy vegetati<strong>on</strong>, the soil sodding and the<br />
occurrence of the forest vegetati<strong>on</strong> successi<strong>on</strong>, <strong>on</strong> those areas.<br />
The presence of trees with small diameters in an increased number, that c<strong>on</strong>stitutes<br />
an element of a specific forest stand, can be explained by the fact that in the gaps<br />
accidentally opened, at the extracti<strong>on</strong> of the accidental and hygiene products, there were<br />
established seedlings that developed at the shelter of the existent forest stand.<br />
Due to the fact that the interventi<strong>on</strong>s have a complex character (character of<br />
preparing, of gaps opening, of gaps enlargement and c<strong>on</strong>necti<strong>on</strong> – <strong>on</strong> some areas), they<br />
will be called P1 respectively progressive 1, P2 – progressive 2, etc.<br />
587
Număr de arbori<br />
700<br />
600<br />
500<br />
400<br />
300<br />
200<br />
100<br />
0<br />
0<br />
-100<br />
20 40 60 80 100<br />
Diametru [cm]<br />
97 98 2001 2004 Arboret<br />
Fig. 2 – Dynamic of the trees number distributi<strong>on</strong> <strong>on</strong> diameter categories when<br />
applying the regenerati<strong>on</strong> cuttings in u.a. 3 A<br />
R<br />
18%<br />
P3<br />
54%<br />
P1<br />
15%<br />
P2<br />
13%<br />
P1<br />
P2<br />
P3<br />
R<br />
Fig. 3 – Intensity of interventi<strong>on</strong>s <strong>on</strong> volume in u.a. 3 A,<br />
P1 the first interventi<strong>on</strong> (with a preparing character), P2 – the sec<strong>on</strong>d interventi<strong>on</strong>,<br />
P3 – the third interventi<strong>on</strong>, R – the last interventi<strong>on</strong> of c<strong>on</strong>necting<br />
From the analysis of the data presented in the diagram in figure 3, we can notice that the<br />
intensity of interventi<strong>on</strong>s at the regenerati<strong>on</strong> cuttings in the forest stand from u.a. 3A is<br />
almost equal in P1, P2 and R, in P3 extracting almost half of the initial forest stand,<br />
aspect proved by the presence of a very abundant fructificati<strong>on</strong> at the oak species in the<br />
years 1999-2000 (Maxim I. C., 2003).<br />
588
Număr de arbori<br />
160<br />
140<br />
120<br />
100<br />
80<br />
60<br />
40<br />
20<br />
0<br />
0<br />
-20<br />
20 40 60 80 100<br />
Diametru [cm]<br />
1998 2000 2001 Arboret<br />
Fig. 4 - Dynamic of the trees number distributi<strong>on</strong> <strong>on</strong> diameter categories when<br />
applying the regenerati<strong>on</strong> cuttings in u.a. 4 A<br />
From the analysis of the data presented in the diagram in figure 5, we can notice that the<br />
intensity of interventi<strong>on</strong>s at the regenerati<strong>on</strong> cuttings is relatively equal in the three<br />
interventi<strong>on</strong>s, the forest stand being grown in a period of about 4 – 5 years, due to the<br />
fact that it occurred two very abundant fructificati<strong>on</strong>s at the oak species in the years<br />
1993-1994, and respectively 1999-2000, because of the favourable stati<strong>on</strong>al c<strong>on</strong>diti<strong>on</strong>s<br />
(Florescu I.I., Nicolescu N. V., 1996).<br />
30%<br />
32%<br />
38%<br />
Fig. 5 – The intensity of the interventi<strong>on</strong>s <strong>on</strong> volume in u.a. 4 A<br />
589<br />
P1<br />
P2<br />
R
From the analysis of the experimental distributi<strong>on</strong>s presented in figure 6, it can be<br />
noticed the fact that in u.a. 11 C the number of small trees is relatively reduced<br />
compared to u.a. 3A, but there is a surplus, their extracti<strong>on</strong> being carried out step by<br />
step during the interventi<strong>on</strong>s, <strong>on</strong> the reas<strong>on</strong>s menti<strong>on</strong>ed before.<br />
Număr de arbori<br />
100<br />
90<br />
80<br />
70<br />
60<br />
50<br />
40<br />
30<br />
20<br />
10<br />
0<br />
-10 0 20 40 60 80 100<br />
Diametru [cm]<br />
1995 1998 2000 2001 Arboret<br />
Fig. 6 - Dynamic of the trees number distributi<strong>on</strong> <strong>on</strong> diameter categories when<br />
applying the regenerati<strong>on</strong> cuttings in u.a. 11 C<br />
17%<br />
24%<br />
25%<br />
34%<br />
Fig. 7 - The intensity of the interventi<strong>on</strong>s <strong>on</strong> volume in u.a. 11 C<br />
590<br />
P1<br />
P2<br />
P3<br />
R
Analysing the data presented in the diagram from figure 7, it can be noticed that the first<br />
interventi<strong>on</strong> – respectively P1 has intensity <strong>on</strong> volume of 34%, P2 an intensity of 25%,<br />
P3 has an intensity of 17% and at the c<strong>on</strong>necting cutting it will raise at 24% from the<br />
forest stand. This dynamic of the intensity of interventi<strong>on</strong>s carried out at the<br />
regenerati<strong>on</strong> cuttings is explained by the fact that the fructificati<strong>on</strong> at oak species was<br />
abundant and the regenerati<strong>on</strong> is d<strong>on</strong>e in the best c<strong>on</strong>diti<strong>on</strong>s.<br />
DISCUSSION<br />
Some ec<strong>on</strong>omical imperatives have imposed the carrying out of interventi<strong>on</strong>s at<br />
relatively short periods and with relatively increased interventi<strong>on</strong> intensities. For<br />
reaching a massif state in a relatively short period, it was carried out works for<br />
completing the natural regenerati<strong>on</strong> using rapidly growing species (red oak).<br />
In the regenerati<strong>on</strong> compositi<strong>on</strong>s there were promoted some species that regenerated<br />
relatively easy from seeds – field maple, Banat field maple, etc., that favour the<br />
reaching at a massif state in best c<strong>on</strong>diti<strong>on</strong>s and ensure the stability of the ecosystems<br />
where they are situated (Enescu V., et. al., 1997); The need of silvotechnical<br />
interventi<strong>on</strong>s at the new forest stand is imperative because in some situati<strong>on</strong>s the<br />
shrubby vegetati<strong>on</strong> becomes overwhelming, being able to spoil the compositi<strong>on</strong> of the<br />
new forest stand.<br />
The need of helping works for a natural regenerati<strong>on</strong> (favouring works of establishing<br />
the natural regenerati<strong>on</strong>) is evident, for a successful valuing of fructificati<strong>on</strong> at basic<br />
species figure 8 a and b.<br />
a) b)<br />
Fig. 8 – Favouring works for establishing a natural regenerati<strong>on</strong> in the forest<br />
stand in u.a. 4A<br />
591
The wild boars had a special c<strong>on</strong>tributi<strong>on</strong> for the acorn incorporati<strong>on</strong> in the soil, which<br />
in the same time influenced the forest soil’s loosening process in the cvernicee forest<br />
stand that fructificated abundantly.<br />
The local experience must be highly valued, thus, having some benefits from the<br />
abundant fructificati<strong>on</strong> of the main oak species, the carrying <strong>on</strong> of some interventi<strong>on</strong>s of<br />
high intensity (punctiform interventi<strong>on</strong>s) correlated with a proper system of collecting<br />
establishments allowed a reducti<strong>on</strong> of the regenerati<strong>on</strong> period up to 5-8 years.<br />
REFERRENCES<br />
D<strong>on</strong>iţă N., Purcelean Şt., Ceianu Ig., Beldie A., 1977, Ecologie forestieră, Editura<br />
Ceres, Bucureşti;<br />
Enescu V., Cherecheş D., Bândiu C., 1997, C<strong>on</strong>servarea biodiversităţii şi a resurselor<br />
genetice forestiere, Editura Agris - Redacţia revistelor agricole, Bucureşti;<br />
Florescu I.I., Nicolescu N. V., 1996, Silvicultura vol. I Studiul pădurii, Editura Lux-<br />
Libris, Braşov;<br />
Florescu I.I., Nicolescu N. V., 1998, Silvicultura vol. II Silvotehnica, Editura<br />
universităţii Transilvania Braşov;<br />
Giurgiu V., 2005, Principii şi criterii pentru alegerea speciilor, proiectarea şi realizarea<br />
de compoziţii optime ale arboretelor, Editura Ceres, Bucureşti;<br />
Maxim I. C., 2003, Lucrare de diplomă, Universitatea din Oradea, Facultatea de<br />
Protecţia Mediului;<br />
Stănescu V., Şofletea N., Popescu O., 1997, Flora forestieră lemnoasă a României,<br />
Editura Ceres, Bucureşti;<br />
*Amenajamentul Unităţii de Producţie II Valea Lungă, Ocolul Silvic Lugoj, Direcţia<br />
Silvică Timişoara;<br />
**Norme tehnice privind alegerea şi aplicarea tratamentelor, 2000, Ministerul Apelor,<br />
Pădurilor şi Protecţiei Mediului, Bucureşti;<br />
592
WASTEWATER TREATMENT TEHNOLOGIES OF BEER INDUSTRY OF<br />
ROMANIA<br />
Pantea Emilia Valentina*, Mirel I**., Romocea Tamara*, Mitrasca Mihaela*<br />
*University of Oradea, ** University of Timisoara<br />
ABSTRACT<br />
For beer industry, the implementati<strong>on</strong> of the (EMS) ISO 14001 protecti<strong>on</strong> managerial<br />
system is primordial, de<strong>term</strong>inant for the study of wastewater treatment optimizati<strong>on</strong>.<br />
Beer factories are using a lot of water, so it is very important that they choose<br />
efficient methods for treating the wastewater in different stages of the producti<strong>on</strong> with a<br />
minimum impact <strong>on</strong> the envir<strong>on</strong>ment.<br />
At first, in our country, an anaerobe treatment method was chosen, either for an<br />
anaerobe treatment of this water, but comparing the efficiency of these two processes as<br />
well as the process combined of anaerobe pre-treatment followed by passing the<br />
anaerobe effluent through an anaerobe treatment stage, it was noticed that this choice is<br />
much more advantageous for used water from beer industry.<br />
This paper is trying to exemplify this technology in the case of a beer factory from<br />
Romania.<br />
Keywords: aerobic, anaerobic, brewery effluent, wastewater treatment<br />
INTRODUCTION<br />
Al<strong>on</strong>g time were recorded the effects of waste water coming from breweries <strong>on</strong> the<br />
envir<strong>on</strong>ment and especially <strong>on</strong> the effluents in which they are discharged. Protecting the<br />
envir<strong>on</strong>ment, especially the water sources, has become <strong>on</strong>e of the primordial objectives<br />
of the government, in this way the adopti<strong>on</strong> of a more efficient water management was<br />
attempted.<br />
Beer breweries from Romania produce different sorts of beer, their number has<br />
been increasing after 1990. The quantity and quality of waste water coming from these<br />
breweries vary between 3-5 and 40-50 hectolitres, being dependant <strong>on</strong> a series of factors<br />
specific to producti<strong>on</strong> process.(Pantea et al., 2005)<br />
The treatment of industrial waste water is c<strong>on</strong>sidered as a n<strong>on</strong>-productive activity,<br />
so that many industries have tried to minimize the investments in water treatment plants.<br />
But, in the last years, together with our EU adhering the legal framework has become<br />
more restrictive, so it has de<strong>term</strong>ined the installati<strong>on</strong> and performing of water treatment<br />
plants of industrial wastewater which should be prerequisite for each industry.<br />
WASTEWATER TREATMENT<br />
Breweries and malt factories are part of the fermentative industries category which use<br />
raw materials like: barley, corn, wheat, hop and which use big quantities of water for the<br />
technological process.<br />
Due to the high c<strong>on</strong>tent of substances easily fermentable, there are some acid<br />
fermentati<strong>on</strong> phenomena, which give off unpleasant smell and develop fungi and the<br />
c<strong>on</strong>tent in matters in suspensi<strong>on</strong> favour the make up of sediments which affect the<br />
piscicultural envir<strong>on</strong>ment by using the oxygen. (Negulescu C, 1987)<br />
593
The managerial plan of breweries c<strong>on</strong>sist in introducing some measures and methods<br />
meant to assure a minimum c<strong>on</strong>sumpti<strong>on</strong> and a maximum recirculati<strong>on</strong>, achieved<br />
through integrated systems such as: wastewater treatment by recovering and reusing<br />
both the water and the sub-products resulted after the treatment process.<br />
The main wastewater treatment methods from breweries are based <strong>on</strong>: mechanical<br />
purificati<strong>on</strong>, chemical treatment and biological purificati<strong>on</strong> assuring in this way the<br />
removal of CBO5 to maximum 30%, 75% and respectively 95%.<br />
Mechanical treatment is based <strong>on</strong> physical proprieties aiming at the separati<strong>on</strong> of<br />
suspensi<strong>on</strong>s and other materials; it uses decanters.<br />
Chemical purificati<strong>on</strong> aims at separating or transforming the settling substances or<br />
in suspensi<strong>on</strong> by using chemical reactives, assuring in this way the neutralizati<strong>on</strong>, the<br />
separati<strong>on</strong> of colloidal and dissolved substances, etc.<br />
Biological purificati<strong>on</strong> is based <strong>on</strong> the use of dissolved and undissolved organic<br />
substances by the micro organisms for multiplicati<strong>on</strong> and c<strong>on</strong>servati<strong>on</strong>. According to<br />
the presence or lack of oxygen, the transformati<strong>on</strong> of organic substances by bacteria can<br />
be achieved in aerobe and/or anaerobe processes.<br />
The residues discharged from breweries represent around 39 kg/hl of produced<br />
beer and are mainly made up of 55% carb<strong>on</strong> hydrates, 27,8% proteins, 6,7% cellulose<br />
and 6,4% grease. The evacuated wastewater charge is reduced to 70-90% as a result of<br />
fermentati<strong>on</strong> process, the resulted fermented sludge is a very good fertilizer.<br />
Am<strong>on</strong>g biological treatment systems <strong>on</strong>e can distinguish between anaerobic and<br />
aerobic processes. Anaerobic treatment is characterized by biological c<strong>on</strong>versi<strong>on</strong> of<br />
organic compounds (COD) into biogas(CH4 70-85% and CO2 15-30% with traces of<br />
hydrogen sulphide). During aerobic treatment oxygen is supplied to oxidize the COD<br />
into CO2 and H2O. Both biological processes produce new biological biomass.<br />
The overall basic reacti<strong>on</strong>s are:<br />
Anaerobic: COD → CH4 + CO2 + anaerobic biomass<br />
Aerobic: COD + O2 → CO2 + H2O + aerobic biomass<br />
Table 1 presents a general comparis<strong>on</strong> between anaerobic and c<strong>on</strong>venti<strong>on</strong>al aerobic<br />
biological treatment systems.(Driessen W. et al., 2003)<br />
Table 1: Anaerobic treatment as compared to aerobic treatment<br />
AEROBIC SYSTEMS ANAEROBIC<br />
SYSTEMS<br />
COD removal 90 - 98% 70 – 85%<br />
Nutrients (N/P) removal high low<br />
Biosolides producti<strong>on</strong> high low<br />
Space requirement high low<br />
Energy c<strong>on</strong>sumpti<strong>on</strong> high low<br />
Energy producti<strong>on</strong> no yes<br />
Anaerobic pre-treatment followed by aerobic post – treatment will result in a positive<br />
energy balance, reduced sludge producti<strong>on</strong> and space saving.<br />
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As a result of the comparis<strong>on</strong>s between the aerobe and anaerobe processes the<br />
advantages of introducing the anaerobe processes for the treatment of wastewater with<br />
medium and high charges were discovered.( Minovsky J.,2002).<br />
CASE STUDY<br />
Brewery’s purificati<strong>on</strong> plant was initially designed for the purificati<strong>on</strong> of wastewater<br />
using the aerobe biological purificati<strong>on</strong> process, but because the results were not<br />
satisfactory, they decided to upgrade it and to introduce an anaerobe pre-purificati<strong>on</strong><br />
phase achieved in a reactor UASB type and then the aerobe treatment of the effluent<br />
corresp<strong>on</strong>ding to this phase.<br />
THE DESCRIPTION OF THE TECHNOLOGICAL PROCESS<br />
In table 2 the main characteristics of the process are introduced:<br />
Table 2: Parameters specific to purificati<strong>on</strong> process<br />
Parameters Unit of measurement Values<br />
COD mg/l 10500<br />
DEBIT mg/l 4200<br />
COD mg/l 2181<br />
TSS mg/l 750<br />
Temperature<br />
o<br />
C 30<br />
pH - 5-8<br />
The wastewater from the brewery with an 86 m 3 /h output will be pumped to a buffer<br />
tank with a 780 m 3 volume, <strong>on</strong>ly after a separati<strong>on</strong> of the solid substances of higher<br />
volume in a static filter, the suspended solids being collected in a c<strong>on</strong>tainer for wastes.<br />
The water is passed from the buffer tank where it is c<strong>on</strong>stantly mixed with the help of<br />
two submersible mixers. The period of water retenti<strong>on</strong> in the buffer tank is 13 h.<br />
After the evaluati<strong>on</strong> of the costs and for their reducti<strong>on</strong>, it was c<strong>on</strong>cluded that the<br />
integrati<strong>on</strong> of a neutralizati<strong>on</strong> tank inside the buffer tank was a good soluti<strong>on</strong>.<br />
The chemical dosage and the recirculati<strong>on</strong> of the anaerobe effluent will be d<strong>on</strong>e in<br />
this neutralizati<strong>on</strong> tank. The measuring of the pH in this tank is c<strong>on</strong>tinuous. From the<br />
correcti<strong>on</strong> tank the water is pumped for the anaerobe treatment with the help of two<br />
pumps. The influent flow is c<strong>on</strong>stantly maintained with the help of an electromagnetic<br />
flow measuring device. The output of the influent is 111 m 3 /h.<br />
The reactors active volume is estimated at about 370 m 3 /reactor, so there is a total<br />
active volume of 740 m 3 . This volume releases a volumetric COD charging of 6,1<br />
kg/m 3 . In the anaerobe reactors a 80 –85% purificati<strong>on</strong> productivity should be achieved,<br />
the methane producti<strong>on</strong> is 0,3 m 3 CH4/COD removed for a c<strong>on</strong>centrati<strong>on</strong> of 80% CH4<br />
out of the biogas producti<strong>on</strong>.<br />
The characteristics of the anaerobe effluent are:<br />
- COD: 436 mg/l<br />
- BOD: 260 mg/l<br />
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The effluent from the anaerobe reactors flows due to gravity towards the tank where the<br />
aerobe purificati<strong>on</strong> of the anaerobe pre-treated water takes place. A smaller flow will be<br />
recirculated towards the correcti<strong>on</strong> tank through a recirculati<strong>on</strong> pipe.<br />
The percolating filters existent before the plant was extended will be used as<br />
aerati<strong>on</strong> tanks. When the biofilter is filled up to a height of 5,1 m (the tank’s volume is<br />
770 m 3 ), o decrease of BOD is produced for an anaerobe productivity of 80% of: 0,71<br />
kg BOD / m 3 . The four aerati<strong>on</strong> tanks are used in a series c<strong>on</strong>figurati<strong>on</strong>.<br />
The anaerobe effluent will flow together with the recirculati<strong>on</strong> course of the sludge<br />
decanters in the first aerati<strong>on</strong> compartment, passing then through all the four<br />
compartments. The dissolved oxygen will be measured with the help of a measuring<br />
device placed in the third and forth aerati<strong>on</strong> compartment. The aerati<strong>on</strong> is achieved with<br />
the help of a bubble aerati<strong>on</strong> system, supplied by two bellows, the retenti<strong>on</strong> type being<br />
2,9 hours. The tank is aerated with a capacity of 334 Nm 3 /h.<br />
At the forth compartment outlet of aerobe treatment, a sludge/effluent separati<strong>on</strong> is<br />
achieved with the help of sludge decanters, with a total area of 226 m 2 .<br />
The sludge resulted after the purificati<strong>on</strong> should be processed by thickening, as a<br />
result the thickened sludge is separated and the excess water from the sludge thickener<br />
will flow in the pH correcting tank.<br />
The decanted effluent has the following parameters: Flow: 86 m 3 /h<br />
BOD: 82 mg/l<br />
TSS: 158 mg/l<br />
pH : 6,5 – 8,5<br />
temperature: < 30 o C<br />
Figure 1. The scheme of the technological process<br />
596
CONCLUSIONS<br />
Anaerobe-aerobe treatment is the most efficient alternative of wastewater treatment for<br />
breweries, the effluents’ qualities which leave the water treatment stati<strong>on</strong> are greatly<br />
improved.<br />
Combined anaerobic/aerobic treatment of brewery effluent has important<br />
advantages over complete aerobic treatment especially regardind: reduced sludge<br />
producti<strong>on</strong>, significant low space requirements and a pozitive energy balance.<br />
REFERENCES<br />
1. Dima M., Meglei V., Dima B., Badea C., - 2002- Bazele epurării biologice ale<br />
apelor uzate, ETP Tehnopress, Iaşi<br />
2. Driessen, W., Habets, L. Verejiken, T. – 1997- Novel anaerobic – aerobic process<br />
to meet strict effluent plant design requirements, Ferment. Vol.10. No.4, U.K.,<br />
pp.243 – 250<br />
3. Driessen W., Vereijken T.,- 1972 - Recent developments in biological treatment of<br />
brewery effluent – The Institute and Guild of Brewering C<strong>on</strong>venti<strong>on</strong>, Livingst<strong>on</strong>e,<br />
Zambia, March 2-7, 2003, Nashville, Tennessee<br />
4. Ianculescu, O., I<strong>on</strong>escu GH., Racoviţeanu Raluca,- 2001 -Epurarea apelor uzate,<br />
Editura Matrix Rom, Bucureşti<br />
5. Minovsky J., Svatopluk C.,- 2002 - Compararea tratării anaerobe şi aerobe a apei<br />
uzate provenită de la industria alimentară, Sinaia<br />
6. Negulescu M. Ant<strong>on</strong>iu R., Negulescu M.,- 1987 - Epurarea apelor uzate<br />
industriale, Editura Tehnică, Bucureşti<br />
7. Pantea E., Mirel I., Romocea T., - 2005 - The anaeroba-aerobe purge system, an<br />
efficient alternative for the purge of warn-out waters from the beer industry,<br />
C<strong>on</strong>ferinţa internaţi<strong>on</strong>ală „Tehnologii moderne pentru mileniul III”, Oradea,<br />
pp.115-120<br />
597
Figure 2. Wastewater treatment in a brewery<br />
598
REGIONAL DEVELOPMENT USING CONSTRUCTED WETLANDS FOR<br />
WASTEWATER TREATMENT IN VALEA IERULUI (ÉRMELLÉK) REGION<br />
Bot<strong>on</strong>d L. Pete<br />
City Hall of Oradea<br />
P-ţa Unirii nr. 1, Oradea, Romania<br />
ABSTRACT<br />
Valea Ierului (Érmellék) lays <strong>on</strong>e the Romania - Hungary border regi<strong>on</strong>, and joins the<br />
settlements of the two country sides. As the level of development is not too high in the<br />
regi<strong>on</strong>, new soluti<strong>on</strong>s are needed to be found, especially in the domain of the<br />
wastewater treatment. The most suitable soluti<strong>on</strong> for communal wastewater treatment<br />
should be the epurati<strong>on</strong> with c<strong>on</strong>structed wetlands, because this technology is less<br />
expensive then the classic methods, and it makes possible to save funds by making<br />
smaller investments. Using this technology, the saved funds can be returned and used in<br />
other projects in the c<strong>on</strong>cerning of regi<strong>on</strong>al development. The most suitable wetland<br />
plants that can be used in the regi<strong>on</strong> are the Phragmites australis (communis),<br />
Schoenoplectus lacustris, Typha latifolia.<br />
Keywords: regi<strong>on</strong>al development, Valea Ierului, wastewater treatment, c<strong>on</strong>structed<br />
wetlands<br />
INTRODUCTION<br />
One of the major problems of Europe became the issue of wastewater treatment in the<br />
local communities. The irreversible growth of the cities results in a higher water<br />
c<strong>on</strong>sumpti<strong>on</strong> and the increase of the wastewater quantity. Due to the nati<strong>on</strong>al increase of<br />
the Romanian life standards more wastewater treatment systems will be needed and this<br />
fact will raise more problems for the local authorities. Starting with the year of 2007<br />
every village with more than 2000 citizens, must have an adequate wastewater treatment<br />
system. In the same time, the increasing use of chemical substances, in special those<br />
from the domestic use, make necessary the modernizati<strong>on</strong> of the purificati<strong>on</strong> plants. The<br />
building of a modern infrastructure is very expensive, and the major part of the small<br />
communities does not have the necessary financial resources.<br />
A short presentati<strong>on</strong> of Valea Ierului (Érmellék) Regi<strong>on</strong><br />
Valea Ierului (Érmellék) area can be defined as a regi<strong>on</strong> between the foothills of the<br />
Occidental Carpathians, the Bükk, Meszes and the Réz-hegység Mountains, and the<br />
Tisza’s lowland. The bigger part of the regi<strong>on</strong> is represented by the Sălaj (Szilágy) and<br />
Bihor (Bihar) counties from Romania and just a smaller part bel<strong>on</strong>gs to Hungary. On<br />
the north side it is bordered by the Crasna’s basin, <strong>on</strong> the east side by the hills of Sălaj<br />
county, <strong>on</strong> south by the valley of Barcău (Berettyó) river. The town of Săcuieni<br />
(Székelyhíd) can be c<strong>on</strong>sidered the geographical center of the regi<strong>on</strong>.<br />
Valea Ierului (Érmellék) as a regi<strong>on</strong> was described in more research studies, but<br />
the geographical limits were not established exactly until now. Some of the specialists<br />
c<strong>on</strong>sider that Valea Ierului (Érmellék) is such a regi<strong>on</strong> that includes Tasnad (Tasnad)<br />
and the others claim that Carei (Nagykaroly) is also included. According to this theory<br />
599
Valea Ierului (Érmellék) has 150.000 residents. In other specialists’ opini<strong>on</strong> the regi<strong>on</strong><br />
of Valea Ierului (Érmellék) does not include Carei, Tasnad and other localities, as the<br />
number of the residents did not exceed 77.000 inhabitants. As there are more c<strong>on</strong>cepts<br />
related, we have to choose <strong>on</strong>e of them. Therefore, we c<strong>on</strong>sider that Valea Ierului<br />
(Érmellék) is a regi<strong>on</strong> that has 80 localities and 77.000 inhabitants. The largest localities<br />
are the following: Carei (with 45.000 inhabitants), Marghita (Margita – 29.000<br />
inhabitants), Valea lui Mihai (Ermihalyfalva – 21.000 inhabitants), Tăsnad (Tasnad –<br />
20.000 inhabitants), Sacuieni (Szekelyhid – 18.000 inhabitants), Diosig (Bihardioszeg –<br />
9.000 inhabitants), and <strong>on</strong> the Hungarian regi<strong>on</strong> border, Letavertes (12.000 inhabitants).<br />
Figure 1: The map of Valea Ierului (Érmellék)<br />
(Veliky J, 2005)<br />
The natural structure of Valea Ierului (Érmellék)<br />
The regi<strong>on</strong> is characterized - as it is situated in the middle of the Carpathian basin – by a<br />
c<strong>on</strong>tinental climate and by some fluctuati<strong>on</strong>s of the temperature, though the average<br />
value does not pass 10°C. According to the measurements made at the Sacuieni<br />
Meteorological Stati<strong>on</strong> in 1996 the average precipitati<strong>on</strong> wasn’t more than 800mm.<br />
The value of the atmospheric pressure is not more than 1000 milibar, the number<br />
of sunny hours are approximately 2000, and this fact indicates the equality of the<br />
number of sunny and cloudy hours.<br />
The surface water system of Valea Ierului (Érmellék) is composed by three rivers.<br />
The most important river is the Ier (Ér), which crosses the area from the north-east side<br />
towards south-west. The other rivers are Crasna (Kraszna) in the north and Barcău<br />
(Berettyó) in the south-west.<br />
The Valea Ierului (Érmellék) of our days is not the same with the <strong>on</strong>e from the<br />
40’s. In those times the regi<strong>on</strong> was a marshy, swampy area with a rich biodiversity,<br />
many floral and fish species. Some specialists c<strong>on</strong>sidered that in that time the regi<strong>on</strong><br />
could compete with the biodiversity from the Danube’s Delta. The flora was<br />
600
characterized by marshy plants like the comm<strong>on</strong> reed, sedge and the rush. These plants<br />
are also present today, but unfortunately in a small number.<br />
Related to the characteristics of the soil we have to menti<strong>on</strong> that in this area we can<br />
find chemozem, but the oxygen poor soils with an important c<strong>on</strong>tent of clay are<br />
dominating the area (general characteristics of the marshes). The sandy soil can also be<br />
found in some places, but that is not characteristic.<br />
Wastewater treatment with c<strong>on</strong>structed wetlands<br />
The wastewater purificati<strong>on</strong> with c<strong>on</strong>structed wetlands is not a very new water cleaning<br />
technology. It has already been used since the 60’, when some German scientists<br />
discovered the purificati<strong>on</strong> capacity of the natural wetlands, and after that the idea of<br />
wastewater cleaning with plants was spread all of the word. Nowadays, in Hungary<br />
many systems like that are working and their efficiency is really high. The reducti<strong>on</strong> of<br />
P, NOx, NH3, K, metals, phenols and other elements can compete in many cases with<br />
the classic wastewater treatment systems.<br />
The wetlands are very successful wastewater purificati<strong>on</strong> systems all over the<br />
world, and they are very various. They can be characterized by many criteria, as their<br />
genesis, locati<strong>on</strong>, functi<strong>on</strong>ing.<br />
By their genesis wetlands can be:<br />
• Marshes<br />
• Bugs<br />
• Flood area<br />
• Marginalized wetlands<br />
• Periodic wetlands<br />
From another point of view they can be natural or artificial wetlands, lakes or marshes,<br />
and some of them are complex piped systems, with a surface or a subsurface flow. In<br />
these complex systems the input wastewater generally enters in the pipe lines flowing<br />
through under the root z<strong>on</strong>e of the plants (generally comm<strong>on</strong> reed), and after that is<br />
evacuated in the nature. At this time the water has to be cleaned and purified.<br />
The technology that is going to be used depends <strong>on</strong> the situati<strong>on</strong> of the villages, the<br />
number of residents, the compositi<strong>on</strong> and quantity of the wastewater, external factors as<br />
climate, type of the soil and many other factors. The origin of the wastewater is another<br />
criteri<strong>on</strong> that has to be taken into c<strong>on</strong>siderati<strong>on</strong> in order to select the suitable methods,<br />
because there is a significant difference between the wastewaters coming from<br />
communal, industrial or agricultural use.<br />
The system of c<strong>on</strong>structed wetlands is based <strong>on</strong> the absorpti<strong>on</strong> process of the<br />
pollutants by the emergent, submersed and floating plants, using the biofilters created<br />
under the water surface, and in that way the toxic elements are going to be eliminated<br />
from the water.<br />
There exist more physical, chemical and biological processes that make the<br />
treatment efficient. These processes generally are the following: sedimentati<strong>on</strong>,<br />
filtrati<strong>on</strong>, adsorpti<strong>on</strong> and absorpti<strong>on</strong>, biological decomposing, and there are many other<br />
processes that assure together a complex purificati<strong>on</strong>.<br />
601
The advantages of this technology are numerous:<br />
• These are ecological and natural systems, and they can represent a habitat for<br />
other species that do not take part in the purificati<strong>on</strong> process (plants, birds and<br />
other animals);<br />
• The system doesn’t need too much maintenance work, big stuff (numerous<br />
employees), and compared to the classic technology it has low operating costs;<br />
• It is not as expensive to be built as the classic systems;<br />
• C<strong>on</strong>structed wetlands can reach the same high efficiency than the classic<br />
systems;<br />
• They can be used as a z<strong>on</strong>e of recreati<strong>on</strong>, of entertainment, sport and other<br />
events for people;<br />
• Corrects the panorama of the regi<strong>on</strong>.<br />
General aspects regarding the plants that can be used in the purificati<strong>on</strong> process<br />
The world’s flora is divided in more floral z<strong>on</strong>es that are organized in systems and<br />
subsystems, <strong>on</strong> each c<strong>on</strong>tinent. Valea Ierului (Érmellék)’s aquatic flora bel<strong>on</strong>gs to a<br />
temperate climatic z<strong>on</strong>e and it is characterized by a vegetati<strong>on</strong> period in winter time. In<br />
this period, when the water loses its flowing capacity because it freezes, the life<br />
processes of the plants are going to slow down. After that some enzymatic processes<br />
start to take place and the result is the development of the vegetative organs.<br />
There are more types of c<strong>on</strong>structed wetlands taking into c<strong>on</strong>siderati<strong>on</strong> the plants<br />
that are going to be used: wetlands with emergent, submersed and floating plants; they<br />
can also use the both types of plants. The natural growth is influenced by many factors<br />
as: the water’s depth, the extensi<strong>on</strong> of the water surface, debit and change of the water<br />
level. As some sec<strong>on</strong>dary factors we can menti<strong>on</strong> the atmospheric factors (temperature,<br />
precipitati<strong>on</strong>, wind speed, evaporati<strong>on</strong>, number of hours with sunlight, etc.), pedological<br />
factors (type of the soil, clay quantity, etc.), biological factors (the presence of certain<br />
floral and fauna species) and characters of the bulked wastewater (qualitative and<br />
quantitative compositi<strong>on</strong>, ph, temperature, turbidity, c<strong>on</strong>tent of the organic material).<br />
These factors are going to be researched in the next period of our scientific activity.<br />
The emergent plants are the most frequent plants in the aquatic systems. They can<br />
be found in the marshes and bogs, where the plants are fixed in the sediments with their<br />
roots, having their stems and leaves above the water surface. In the water they create<br />
some biological associati<strong>on</strong>s, named biofilters, and in that way they c<strong>on</strong>tribute to the<br />
cleaning process. Such plants are the comm<strong>on</strong> reed (Phragmatis australis, Phragmatis<br />
comunis), the rush (Schoenoplectus lacustris ) and the bulrushes (Typha latifolia, T.<br />
augustipfolia, T. minima).<br />
Other types of the macropytes are the floating plants, which – due to their special<br />
cellular c<strong>on</strong>stituti<strong>on</strong> and their air filled leaves – are drifting <strong>on</strong> the water surface. They<br />
have two subcategories, some of them being unattached and others attached floating<br />
plants.<br />
The floating unattached plants are drifting <strong>on</strong> the water surface without touching<br />
the bottom with their 20-30 cm l<strong>on</strong>g roots. The Pistia Stratiotes or the Eichornia<br />
Crassipes bel<strong>on</strong>g to this category. The floating attached plants are also floating <strong>on</strong> the<br />
602
watersurface, but their roots are touching the bottom sediments of the water. Such plants<br />
are: the Nymphaea alba, the Nuphar lutea, the Lemna ssp. and the Wolffia arrhiza.<br />
The submersed plants are another category of the aquatic flora, which are fixed in<br />
the underwater ground (in some of the cases the roots are missing), and their leaves and<br />
stems are entirely under the water surface. They play several roles in an aquatic habitat<br />
like: producti<strong>on</strong> of the organic material, oxygen liberati<strong>on</strong>, transfer of material<br />
providing of life place for the flora and fauna of the underwater ground. Many plants<br />
bel<strong>on</strong>g to this category, but we will menti<strong>on</strong> just some of them like: Potamoget<strong>on</strong><br />
pectinátu, Elodea canadensis, Chara ssp., Lemna trisulca and Ceratophyllum<br />
demersum.<br />
Final c<strong>on</strong>clusi<strong>on</strong>, remarks<br />
The wastewater treatment with c<strong>on</strong>structed wetlands is the same, or is similarly efficient<br />
as the classical purificati<strong>on</strong> systems, so that they could be used in the localities from the<br />
Valea Ierului (Érmellék) regi<strong>on</strong>. Taking into c<strong>on</strong>siderati<strong>on</strong> that in this area the most<br />
localities d<strong>on</strong>’t have the infrastructure to perform a wastewater treatment, we think, that<br />
this kind of technology could assure a good soluti<strong>on</strong>, or at least an alternative for the<br />
problems. The building of the system is not as expensive as the building of the classic<br />
<strong>on</strong>e and thus the capital that has not been used can be reinvested and used in other area<br />
of regi<strong>on</strong>al development.<br />
In c<strong>on</strong>clusi<strong>on</strong> this technology can solve in a direct way the wastewater<br />
treatment issue in the regi<strong>on</strong>, and in an indirect way, <strong>on</strong> l<strong>on</strong>g <strong>term</strong>, can str<strong>on</strong>gly<br />
c<strong>on</strong>tribute to the local and regi<strong>on</strong>al development.<br />
REFERENCES<br />
• Veliky J. (2005): Érmellék Kalauz. Hatvani István Szakkolégium, Debrecen. 21-27,<br />
79-10 p.<br />
603
STUDY OF VERTICAL MOVEMENT OF SOME MICROELEMENTS<br />
IN THE SOIL<br />
László Szegedi (1) – Lajos Szabó (2.) – Mária Takács Hájos (2.)<br />
(1.) Károly Róbert College Department of Envir<strong>on</strong>mental Management and Agr<strong>on</strong>omy,<br />
(2.) Tessedik Sámuel College, Institute of Envir<strong>on</strong>mental Sciences<br />
ABSTRACT<br />
A l<strong>on</strong>g <strong>term</strong> field experiment was established with different doses of some heavy metals<br />
(As, Cr, Cd, Cu, Hg, Pb, Zn) to study their behaviour in the soil-plant system. The soil<br />
of the experimental site was characterized by pH (H2O) 6.2, pH (KCI) 5.4, soil organic<br />
matter c<strong>on</strong>tent 3%, and clay mineral c<strong>on</strong>tent 30-35%. To check the vertical movement<br />
of the studied elements, two years after applicati<strong>on</strong> soil samples were taken from the 0-<br />
30, 30-60 and 60-90 cm layers of plots treated with the highest heavy metal load (270<br />
kg element/ha). Both total (cc.HNO3+H2O2 extractable) and available (NH4 – acetate +<br />
EDTA extractable) element c<strong>on</strong>tent was de<strong>term</strong>ined using the ICP-MS technique.<br />
Vertical movement of Cr (VI) was detected int he full soil profile. Therefore Cr<br />
c<strong>on</strong>taminati<strong>on</strong> of the soil profile suggest a significant risk for the quality of groundwater.<br />
As and Pb remained mostly int he 0-30 cm layer. There was <strong>on</strong>ly a slight<br />
c<strong>on</strong>taminati<strong>on</strong> int he 30-60 cm layer. Cd, Hg, Cu and Zn were fixed in the plough layer.<br />
Both total c<strong>on</strong>tent and available fracti<strong>on</strong> of these elements were detected in the upper 0-<br />
30 cm layer. These c<strong>on</strong>taminants seem to be resistant to leaching.<br />
INTRODUCTION<br />
Envir<strong>on</strong>mental polluti<strong>on</strong> is placing an ever-increasing load <strong>on</strong> the various resources of<br />
our envir<strong>on</strong>ment, including soil. Tests and experiments involving heavy metals and<br />
other toxic elements play a very important role in our envir<strong>on</strong>mental research programs.<br />
Soils polluted with heavy metals can be found near industrial areas, metropolises, al<strong>on</strong>g<br />
major transportati<strong>on</strong> routes, roads, and areas treated with waste-water sludge, but soils<br />
can also be ”polluted” geologically (1). Soils are able to accumulate heavy metals for<br />
many years without the obvious signs of their acute toxic effect. However, the<br />
filtering/buffering capacity of soils is finite and above a certain level soils are no l<strong>on</strong>ger<br />
able to absorb these elements and become sources of polluti<strong>on</strong> themselves. Toxic<br />
elements are released into water, absorbed by cultivated crops and plants; they are<br />
assimilated into vegetative and generative organs, and enter the food-chain where they<br />
cause l<strong>on</strong>g-<strong>term</strong> harm (2,3).<br />
The system of relati<strong>on</strong>s between heavy metals-soils-plants can be accurately and<br />
reliably studied in targeted field experiments. The Ministry of Envir<strong>on</strong>ment and the Soil<br />
Research Institute of the Hungarian Academy of Science launched a research program<br />
in 1991, titled ”Heavy metal loading of the envir<strong>on</strong>ment”. The goal of the research<br />
program is to study the effect of certain heavy metals and other potentially toxic<br />
elements in the soil –plant system and in the food chain. The study is carried out in l<strong>on</strong>g<br />
<strong>term</strong> field experiments in major soil types (carb<strong>on</strong>ate-loam, carb<strong>on</strong>ate-sand, acidicloam,<br />
acidic-sand) (4). The aim of the present scientific paper is to examine the<br />
following:<br />
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• Which of the heavy metals that are dispresed over the soil accumulate in the upper<br />
(tilled) strata/horiz<strong>on</strong>s?<br />
• Are the heavy metals that are dispresed over the soil, leached into deeper horiz<strong>on</strong>s<br />
and as a result, do these elements endanger ground water?<br />
MATERIAL AND METHODS<br />
A field experiment was set up at the K.R. College Farm in 1994, to study the effect of 8<br />
heavy metals (Al, As, Cd, Cu, Hg, Pb, Zn) at three dosage levels (0/30, 90 and 270<br />
kg/ha of each element). These were repeated three times <strong>on</strong> 35 m 2 plots (Table 1).<br />
Element<br />
Al<br />
As<br />
Cd<br />
Cr<br />
Cu<br />
Hg<br />
Pb<br />
Zn<br />
Table 1. Field experiments with heavy metal load, Gyöngyös, 1994<br />
1<br />
Load levels kg of element/ha<br />
2 3<br />
0<br />
90<br />
270<br />
30<br />
90<br />
270<br />
30<br />
90<br />
270<br />
30<br />
90<br />
270<br />
30<br />
90<br />
270<br />
30<br />
90<br />
270<br />
30<br />
90<br />
270<br />
30<br />
90<br />
270<br />
Form of salts<br />
applied<br />
Al(NO3)3 · 9H2O<br />
NaAsO2<br />
3CdSO4 · 8H2O<br />
K2CrO4<br />
CuSO4 · 5H2O<br />
HgCl2<br />
Pb(NO3)2<br />
ZnSO4 · 7H2O<br />
The soil type used for the experiment is mildly acidic chernozem brown forest soil<br />
formed <strong>on</strong> alkaline sediment (andesite, andesite tuff). Key soil parameters are the<br />
following: pH(H2O)=6.2; pH(KCl)=5.4;y1=9.5; CaCO3%=0; humus%=3; KA=45;<br />
L%=70; hy=4.8. Particle size of the soil is dominated by clay and silt fracti<strong>on</strong>. The<br />
textural type is clay-loam with a bulk density of 1.20 g/cm 3 . Soil surface is slightly<br />
sloping, elevati<strong>on</strong> is 150 m above sea level. Water table depth is 10 m, therefore the<br />
probability of polluti<strong>on</strong> through surface leaching is minimal. The soil has good water<br />
absorpti<strong>on</strong>, c<strong>on</strong>ducti<strong>on</strong> and storage capability. In the event of heavy rainfall, rill erosi<strong>on</strong><br />
occurs <strong>on</strong>t illed surfaces.<br />
The soil in the plots was examined and tested each year. Samples were taken from<br />
the following strata of the plots treated with the maximum dosage (270 kg/ha): 0-30, 30-<br />
60, 60-90 cm. Samples were taken from bored holes in cartridges. In each plot, 5 bores<br />
were made at each depth range to obtain at an average sample. The samples were dried,<br />
ground and homogenised before de<strong>term</strong>ining the ”total” element c<strong>on</strong>tent estimated with<br />
the cc.HNO3+cc.H2O2 extracti<strong>on</strong> and the available element c<strong>on</strong>tent with the Lakanen-<br />
Erviö’s method (6). The analysis of samples for elements was carried out by the ICP<br />
laboratory at the Soil Research Institute (RISSAC) of the Hungarian Academy of<br />
Science, for 25 elements, using the ICP-AES method.<br />
RESULTS AND DISCUSSION<br />
As was dominantly present in the upper stratum (0-30 cm) int he 2nd year of the<br />
experiment ( Figure 1). The 0-60 cm stratum was also polluted as shown in the figure by<br />
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the ”total” and available element c<strong>on</strong>tent. Enrichment/accumulati<strong>on</strong> in deeper soil<br />
horiz<strong>on</strong>s/strata is not significant, with respect to the tolerance of sampling, it is<br />
practically negligible. Leaching, and the extent and dynamics of leaching cannot be<br />
de<strong>term</strong>ined from this data.<br />
Figure 1. The effect of 270 kg/ha arsenic load <strong>on</strong>t he arsenic c<strong>on</strong>tent of the soil<br />
profile (Chernozem brown forest soil, Gyöngyös, 1996)<br />
Figure 2. The effect of 270 kg/ha cadmium load <strong>on</strong> the cadmium c<strong>on</strong>tent of the soil<br />
profile (Chernozem brown forest soil, Gyöngyös, 1996)<br />
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Figure 2 shows that Cd also is c<strong>on</strong>centrated int he upper, tilled stratum of soil. Both<br />
”total” and available c<strong>on</strong>tent is present int he place of input, deeper horiz<strong>on</strong>s show no<br />
sign of accumulati<strong>on</strong>. It seems that this extremely toxic element is highly resistant to<br />
leaching in loamy soils int he initial years. According to literature, Cd is highly mobile<br />
in the soil-plant system (5, 7).<br />
The mobility of Cr(VI) is unambiguously evident (Figure 3). In comparis<strong>on</strong> with<br />
unpolluted c<strong>on</strong>trol soil, accumulati<strong>on</strong> in the deeper strata between 30-60 and 60-90 cms<br />
is obvious, both in <strong>term</strong>s of ”total” and available c<strong>on</strong>tent. Heavy rainfall in 1995 and<br />
1996 may also have c<strong>on</strong>tributed to leaching. It is also true that the available Cr c<strong>on</strong>tent<br />
of the deeper strata is <strong>on</strong>e magnitude less than the values measured at the input point.<br />
The fact that Cr polluti<strong>on</strong> was, although more moderately, apparent between 60-90 cms<br />
after the sec<strong>on</strong>d year draws attenti<strong>on</strong> to the importance of examining the mobility of<br />
Cr(VI) in the horiz<strong>on</strong>s bey<strong>on</strong>d 1-meter depth. We will need to m<strong>on</strong>itor the limits of Cr<br />
leaching in the future with deeper test bores.<br />
Figure 3. The effect of 270 kg/ha chromium load <strong>on</strong>t he chromium c<strong>on</strong>tent of the soil<br />
profile (Chernozem brown forest soil, Gyöngyös, 1996)<br />
The majority of Pb remained at the point of input, however it is not realistic to assume<br />
that this element will remain completely immobilised in acidic forest soils. Hence Pb<br />
may pose a moderate danger to soil water and the food chain (8). According to the<br />
results of soil examinati<strong>on</strong>s, Hg is not leached. Literature shows that HgCl2 is rapidly<br />
b<strong>on</strong>ded by organic matter and clay fracti<strong>on</strong>s or precipitated in the form of insoluble<br />
salts. Solubility may increase under acidic c<strong>on</strong>diti<strong>on</strong>s, thereby Hg in soils may become<br />
mobilised. Cu and Zn are b<strong>on</strong>ded in the tilled horiz<strong>on</strong>, mobility was not experienced.<br />
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CONCLUSION<br />
1. The rate of transformati<strong>on</strong> and immobilisati<strong>on</strong> of toxic pollutants in soils varied<br />
according to elements. A clear distincti<strong>on</strong> can be made between elements that are<br />
mobile pollutants in soils (Cd, Zn, Pb, Cu) and those that transform rapidly into<br />
insoluble forms and become b<strong>on</strong>ded (As, Hg, Cr).<br />
2. Heavy metals and other pollutants that are spread over soils accumulate in the tilled<br />
soil horiz<strong>on</strong>s where root mass is highest. Cd, Cu, Pb, Hg and Zn are b<strong>on</strong>ded in the<br />
tilled horiz<strong>on</strong>s, while As moderately polluted the deeper horiz<strong>on</strong>s between 30-60<br />
cms.<br />
3. The leaching of Cr input into soils in the form of chromate is a rapid process in<br />
mildly acidic brown forest soils, the element was detected at depths of 60-90 cms<br />
after 2 years. Subterranean water sources are endangered by this element as a result<br />
of rapid leaching.<br />
REFERENCES<br />
1. Csathó, P. 1994. Heavy metal polluti<strong>on</strong> and agricultural producti<strong>on</strong>. Thematic<br />
literature review. Akaprint, Budapest<br />
2. Fodor, L. 1977. Mobility of some toxic elements in soil-plant system <strong>on</strong> brown<br />
forest soil. Proceedings of <str<strong>on</strong>g>Internati<strong>on</strong>al</str<strong>on</strong>g> <str<strong>on</strong>g>C<strong>on</strong>ference</str<strong>on</strong>g> of PhD Students. UM,<br />
Miskolc, pp. 84-90.<br />
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plant available microelements in soil. Acta Agr. Fenn. 123, 223-232.<br />
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Affected by Soil Cadmium C<strong>on</strong>centrati<strong>on</strong>. Soil Sci. Plant Anal. 27. 1765-1777.<br />
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