Joint International Conference on Long-term Experiments ...

<strong>Joint</strong> <strong>International</strong> <strong>Conference</strong> on
Long-term Experiments,
Agricultural Research and
Natural Resources
Debrecen-Nyírlugos
31 st May – 1 st June, 2007

JOINT INTERNATIONAL CONFERENCE ON LONG-TERM EXPERIMENTS,
AGRICULTURAL RESEARCH AND NATURAL RESOURCES
Debrecen-Nyírlugos 31 st May – 1 st June, 2007
Organiser:
Research Institute for Soil Science and Agricultural Chemistry of
the Hungarian Academy of Sciences and
the University of Debrecen Faculty of Agriculture
Co-organisers:
University of Oradea Faculty of Environmental Protection
Town Council Nyírlugos
Editors:
István Láng
János Lazányi
Nicolae Csép
Lectors:
János Lazányi
Nicolae Csép
Published with the financial support of
Westsik Vilmos Foundation for Rural Development in Nyírség Region
4400 Nyíregyháza Westsik Vilmos út 4-6. Hungary
Published by:
University of Debrecen Centre of Agricultural Sciences
4032 Debrecen Böszörményi út 138. Hungary
HU-ISBN: 978-963-473-054-5
Printed in Pentaprint Printing House

CONTENTS
INFLUENCE OF CHEMICAL FERTILIZERS, MANURE AND LIME
APPLIED IN LONG TERM FIELD EXPERIMENTS ON PH-VALUE OF
PRELUVOSOILS FROM NORTH-WEST PART OF ROMANIA
Cristian Hera, Gheorghe Ciobanu, Cornelia Ciobanu,
Cornel Domuţa, A. Vuşcan, Georghe Sarca
EFFECT OF VARIOUS CROP PRODUCTION FACTORS ON THE YIELD
AND YIELD STABILITY OF MAIZE IN A LONG-TERM EXPERIMENT
Zoltán Berzsenyi, Dang Quoc Lap
THE IMPACT OF PLANT BREEDING ON THE IMPROVEMENT OF SEED
OIL CONTENT AND QUALITY IN EVENING PRIMROSE CROPS
Andrew F. Fieldsend
SOME LESSONS LEARNED FROM THE NYÍRLUGOS LONG-TERM FIELD
EXPERIMENT Imre Kádár
ANALYZE OF BIOMASS PRODUCTIVITY BY TIMESERIES
REMOTESENSING DATA IN REGION OF NYÍRLUGOS
János Tamás, Tibor Bíró, Nikolett Szőllősi
RAINFALL, FERTILIZATION AND LIMING RESPONSE ON TRITICALE (X
Triticosecale W.) YIELD IN THE 44 YEAR OLD NYÍRLUGOS FIELD TRIAL
BETWEEN 1999 AND 2006 Márton László
EVALUATION OF THE DATA SET OF THE HUNGARIAN LONG-TERM K-
FERTILIZATION FIELD TRIALS, SET UP BETWEEN 1960 AND 2000
Péter Csathó
LONG-TERM FIELD EXPERIMENTS AS THE BASE OF SITE SPECIFIC
FARMING KESZTHELY LONG-TERM FIELD EXPERIMENTS
Tamás Kismányoky, Zoltán Tóth
LONG-TERM EFFECT OF ORGANIC AND MINERAL FERTILIZATION ON
DIFFERENT SOIL-FERTILITY PARAMETERS
Sándor Hoffmann, Katalin Berecz
GREENHOUSE GAS EMISSION IN LONG-TERM TILLAGE AND
ROTATION EXPERIMENTS
Anita Gál, Péter Hegymegi, Erika Michéli, Tony J. Vyn
INFLUENCES OF AMELIORATIVE FERTILIZATION ON YIELD AND
NUTRITIONAL STATUS OF MAIZE
Kovačević Vlado, Josipovic Marko, Stojić Biserka
3
12
20
29
37
44
51
56
62
68
75
83

YIELD AND QUALITY OF WINTER WHEAT GENOTYPES (Triticum
aestivum L.) Desimir Knezevic, Veselinka Zecevic, Danica Micanovic,
Nevena Djukic, Aleksandar Paunovic, Milomirka Madic, Ivica Djalovic
CROP ROTATION AND GREEN MANURE INFLUENCE ON YIELD AND
WATER USE EFFICIENCY IN WHEAT AND MAIZE FROM CRIS PLAIN
Cornel Domuţa, Gheorghe Ciobanu, Maria Şandor, Alina Samuel, Cornelia
Ciobanu, Nicu Cornel Sabău, Viorel Şcheau, Ioana Borza, Cristian Domuţa,
APPLICATION OF A DIGESTATE AS A NUTRIENT SOURCE AND ITS
EFFECT ON SOME CROPS AND SOIL PROPERTIES Marianna Makádi,
Attila Tomócsik, József Lengyel, Zsolt Bogdányi, Árpád Márton
SOIL TILLAGE SYSTEMS AND NITROGEN FERTILIZATION FOR
WINTER BARLEY AFTER SOYBEAN
Bojan Stipesevic, Danijel Jug, Miro Stosic, Ivan Zugec, Irena Jug
RESEARCHES REGARDING THE INFLUENCE OF THE CROP ROTATION
ON SOME INDICATORS OF THE WHEAT YIELD QUALITY IN THE
CONDITIONS OF THE CRISURILOR PLAIN Gheorghe. Bandici , Cornel.
Domuţa, Ileana Ardelean, Ioana Borza, Cristian Domuţa
CRISANA -THE FIRST ROMANIAN WHEAT VARIETY WITH
TOLERANCE TO ALUMINIUM IONS TOXICITY FROM ACID SOILS
Gheorghe Bunta, Elena Bucurean
EXPERIMENTS WITH PERENNIAL RYE (SECALE CEREANUM) IN
HUNGARY, AT UNIVERSITY OF DEBRECEN Erika Halász, Tamás Sipos
MYCOTOXINS CONTAMINATION IN FRESH AND SILAGE OF ALFALFA
Branislav Gálik, D. Bíro, M. Juráček, M. Šimko, J. Michálková
USAGE OF NATURAL ADDITIVES FOR BROILER CHICKENS
Erika Horniaková, Ladislav Bušta
PERSPECTIVES ON ROMANIAN-HUNGARIAN CROSS-BORDER
COOPERATION. THE BIHOR - HAJDÚ-BIHAR EUROREGION MODEL
Gabriela Popoviciu, M.C. Neacşu
EFFECT OF BORON FERTILIZATION ON ANNUAL FLUCTUATION OF B
IN SWEET CHERRY LEAVES AND FRUIT QUALITY Péter Tamás Nagy,
Sándor Thurzó, Ida Kincses, Zoltán Szabó, József Nyéki
DRY MATTER PRODUCTIVITY AND NITROGEN UPTAKE OF
PERENNIAL RYEGRASS INFLUENCED BY NITROGEN AND WATER
SUPPLY Imre Vágó, János Kátai, Ida Kincses, Andrea Balla Kovács
4
90
95
102
108
113
119
127
132
136
140
149
156

THE EFFECT OF NITROGEN SUPPLY, LIMING AND NICKEL
CONTAMINATION ON THE PRODUCTIVITY OF RYEGRASS (Lolium
perenne L.) IN A GREENHOUSE EXPERIMENT Marianna Sipos, Imre Vágó
TOXIGENIC FUNGI AND MYCOTOXINS IN GRAINS AND BAKERY
PRODUCTS FROM ROMANIA Carmen Puia, Viorel Florian, Alexandra
Suciu, Augusta Lujerdean, Dana Pusta
CORRELATIONS OF SOCIAL AND ECONOMIC FUNCTIONS IN
COMMUNITIES AROUND HORTOBÁGY Erzsébet Csengeri
RIDING TOURISM IN THE NORTHERN GREAT PLAIN REGION
(HUNGARY): STATUS AND POTENTIAL Réka Incze, Gábor Hevessy
TISSUE CULTURE METHODS FOR SCREENING RESISTANCE TO
PECTOBACTERIUM CHRYSANTHEMI IN POTATO
Ildikó Hudák, Mária Hevesi, Judit Dobránszki Katalin Magyar-Tábori
EFFECT OF AUXIN ON CALLUS FORMING CAPACITY OF DIFFERENT
PEA GENOTYPES (PISUM SATIVUM L.) Katalin Magyar-Tábori,
Judit Iszály-Tóth, Judit Dobránszki, Nóra Mendler-Drienyovszki
COMBINING ABILITY STUDIES IN A SEVEN- PARENTAL DIALLEL
CROSS OF DIFFERENT PEAS VARIETES (Pisum sativum L.) Nóra Mendler-
Drienyovszki, Mándi Lajosné, Katalin Magyar-Tábori, Györgyi Csontos
SUSTAINABLE LAND-USE BASED ON WESTSIK CROP ROTATION
EXPERIMENT János Lazányi
THE EFFECT OF LONG-TERM PHOSPHORUS FERTILIZATION ON THE
TRYPTOPHAN AND ZINC CONTENT OF FEED PEAS
Ágnes Elek, Zoltán Győri, Mária Borbély
THE IMPORTANCE OF CALCIUM AND MAGNESIUM SUPPLY ON THE
ACID BROWN FOREST SOILS IN THE NYIRSÉG REGION
Jakab Loch, János Lazányi, Péter Tamás Nagy and Rita Kremper
COMPARATIVE ANALYSIS OF CHEMICAL AND BIOLOGICAL SOIL
EXAMINATION TO DETERMINE THE PLANT AVAILABLE N CONTENT
OF SOIL IN THE NYÍRLUGOS LONG TERM FIELD EXPERIMENTS
Péter Tamás Nagy, János Lazányi, Jakab Loch
EFFECT OF FERTILIZATION SYSTEMS AND LIMING ON MOBILE
PHOSPHORUS CONTENT IN DIFFERENT SOIL TYPES
Augusta Olivia Lujerdean
COMPOSTED SLAUGHTERHOUSE BY-PRODUCTS EFFECT ON CROP
YIELD Péter Ragályi, Imre Kádár
5
163
170
175
181
191
196
201
207
214
220
226
234
239

THE EFFECT OF BENTONITE ON SPECIFIC SOIL PARAMETERS AND
MICROBIAL CHARACTERISTICS OF THE CARBON CYCLE János Kátai,
Magdolna Tállai, János Lazányi, Edina Veres Lukácsné, Zsolt Sándor
INVESTIGATION OF MINERAL CONTENT IN WINTER WHEAT GRAIN
SAMPLES FROM LONG-TERM FIELD EXPERIMENTS
Zoltán Győri, Árpád Tóth, Diana Ungai
COMPARISON OF SOIL-TEST EXTRACTANTS FOR ZINK AND COPPER
Rita Kremper, Sándor Berényi, Jakab Loch, Andrea Balla Kovács
YIELD OF RYE AS AFFECTED BY THE CROPYEAR IN THE WESTSIK`S
CROP ROTATION LONG-TERM FIELD EXPERIMENT
István Henzsel, Gyuláné Györgyi
RESULT OF ALFALFA CROPPING BY THE BASIC OF TIME- AND
VILLAGE TYPE DATA FOR COUNTY OF SZABOLCS SZATMÁR BEREG
László Nagy
THE IMPORTANCE OF LONG-TERM FIELD EXPERIMENTS IN THE
QUALITY ASSURANCE OF CROP PRODUCTION Éva Széles, Zsuzsanna
Szathmáry, Péter Sipos, Dóra Hovánszki, Csilla Uri, Zoltán Győri
MORPHOLOGICAL AND AFLP VARIATION IN SOME GENOTYPES OF
POA ANGUSTIFOLIA L. AND POA HUMILIS EHRH. EX HOFFM Zsuzsa
Lisztes, Ákos Szabó, A. Zubor, Béla Tóthmérész, Mária Papp, József Prokisch
THE EFFECT OF SOME FOLIAR COMPOSITIONS ON THE PRODUCTION,
NUMBER OF INFLORESCENCES AND FRUIT IN THE TECHNOLOGY OF
CULTIVATING TOMATO IN THE FIELD Popa Alina Grigoriţa
METHODS AND MEASURES FOR AN AGROCHEMICAL OPTIMIZATION
OF THE GREENHOUSE SOIL AT THE TOMATOES CULTURE
Popa Alina Grigoriţa
THE INFLUENCE OF THE SEED’ BIOLOGICAL LINKS ON THE WHEAT
PRODUCTION QUALITY Mariana Popovici
THE INFLUENCE OF SOIL OIL POLLUTION ON AGRICULTURAL CROPS
Maria Şandor, Nicu Cornel Sabău, Cornel Domuţa, Cristian Domuţa,
Radu Brejea
EFFECT OF BIOFERTILIZATION ON PARSLEY YIELD AND N CONTENT
ON TWO SOILS
Ida Kincses, Tibor Filep, Andrea B. Kovács, Péter T. Nagy, Imre Vágó
6
247
255
262
269
274
279
284
392
396
300
304
312

THE INFLUENCE OF CHEMICAL AND BIOFERTILIZERS ON THE YIELD
AND NITROGEN CONTENT OF LETTUCE (Lactuca sativa L.)
Andrea Balla Kovács, Ida Kincses, Imre Vágó, Rita Kremper
THE INFLUENCE OF USING BIOLOGICAL ADDITIVES ON THE
NUTRITIONAL QUALITIES OF CORN SILAGE AND ON THE FATTENING
PERFORMANCES OF BULL CALVES
Daniel Mierlita; I. Chereji; Cristina Maerescu
PHYTOPHTORA INFESTANS - ECOLOGICAL CONTROL WITHIN
ROMANIAN (TRANSYLVANIAN PLANE) POTATO CROP
Ioan Oroian, Liviu Holonec, Viorel Florian, Laura Paulette, Antonia Odagiu
THE DYNAMICS OF THE POPULATIONS OF CEREAL PESTS AND
PATHOGENS IN THE ROMANIAN WESTERN PLAIN
Elena Bucurean, Nicolae Csép
THE IMPORTANCE OF CONTROLLING FORMALDEHYDE FUMIGATION
Lucian Bara, Camelia Bara
THE CA AND MG SUPPLY OF THE SOIL EXTRACTED WITH DIFFERENT
SOLVENT SOLUTIONS
István Kocsis, Lajos Szabó, Simándi Péter, László Pásztor
BIOLOGICAL INVESTIGATION ON AN EROSION CATENA
B. Szeder, B. Simon, M. Dombos, T. Szegi
THE PRESENT STATE OF THE ROMANIAN AGRO-ALIMENTARY
CONSUMPTION Anca Monica Brata
FROM FARM TO FORK – EVALUATION OF CROPPING FIELDS WITH
REGARD TO FOOD AND FEED SAFETY AFTER LONG-STANDING
CULTIVATION Péter Sipos, Zsuzsanna Szathmáry, Árpád Tóth, Diána
Ungai, Dóra Hovánszki, Zoltán Győri
MICROCLIMATIC STUDY OF THE MAIN WINE-GROWING REGIONS IN
ROMANIA DURING 2006 Dorin Popa
WHEAT GRAINS PROCESSING INFLUENCE AT QUALLITY
PARAMETERS Timar Adrian
OCEAN DYNAMICS: AIR-SEA INTERFACE GASES EXCHANGE
Speranta Coldea
RESEARCHES ON THE QUICK METHOD OF SUGAR PRODUCTIVITY
EVALUATION Gheorghe Sarca
RESEARCH REGARDING THE CONCENTRATION OF THIN JUICE AND
THICK JUICE Gheorghe Sarca
7
319
327
334
342
347
352
357
364
369
376
382
389
397
402

RESEARCH ON SEEDING AGENTS TO EFFECT THE SUGAR
CRYSTALLIZATION Gheorghe Sarca
WATER DIFFUSION IN BREAD DURING BAKING
Ruska L., Chereji Rodica, Purcărea Cornelia, Timar A.
BREAD PROPERTIES AND CRUMB STRUCTURE
Ruska L., Chereji Rodica, Purcărea Cornelia, Timar A.
THE MATHEMATICAL MODELLING OF THE PRESSURE REGULATOR
AND OF THE ELECTROMAGNETIC INJECTION AT ENGINES WITH THE
REVOLUTION AND THE TEMPERATURE OF THE ENVIRONMENT
SURROUNDINGS Vasile Blaga, Nicolae Chioreanu, Adriana Cătaş
THE VARIATION OF EFFECTIVE POWER AND SPECIFIC EFFECTIVE
FUEL CONSUME OF 106-20 ENGINE WITH THE REVOLUTION AND THE
TEMPERATURE OF THE ENVIRONMENT SURROUNDINGS
Vasile Blaga, Nicolae Chioreanu, Adriana Cătaş
THE EFFECT OF THE GENOTYPE AND EXPLANT UPON THE SOMATIC
EMBRYO INDUCTION IN THREE QUERCUS SPECIES
AI. Timofte, M. Palada,Nicolau, M. Ardelean, D. Pamfil,
COMPARATIVE TESTS FOR SEPARATION METHODS OF
ORGANOCHLORINE PESTICIDES IN MILK Adriana Chiş, Vasile Bara
HIGHLIGHTING THE STATE OF FRESHNESS OF THE FRESH RAW
MATERIAL FISH ON THE BASIS OF THE PHYSICAL-CHEMICAL
PROPERTIES Purgea Ramona
ASSESSMENT OF THE HYGIENIC QUALITY OF THE FISH FRESH RAW
MATERIAL BASED ON THE BACTERIOLOGICAL EXAMINATIONS
Purgea Ramona
FORENSIC ASPECTS REGARDING DISASTER MANAGEMENT
Carmen Radu, Camelia Buhaş, Nicolae Csép, Radu Gabriela, Andrei Csép
ENERGY RECOVERY FROM ANIMAL PRODUCTS László Nagypál 470
THE IMPLICATIONS OF MATERNAL TOXOPLASMOSIS INFECTION IN
REPRODUCTION Sonia-Maria Drăghici, A. Csép, Ildiko Lenard, Adriana
Jarca, Nicoleta Negruţ
BOTULISM: CLINICAL – EPIDEMIOLOGICAL STUDY Ildiko Lenard,
Sonia Draghici, Viorica Coldea, Mirela Indries,Nicoleta Negrut, Andrei Csép,
Timea Lenard
8
407
413
420
427
435
441
453
459
462
465
474
480

STUDIES REGARDING THE INFLUENCE OF SALICYLIC ACID ON THE
GERMINATION OF MAIZE CARYOPSIS (Zea mays) AND ON THE
PEROXIDASE ACTIVITY IN MAIZE PLANTLETS
Cornelia Purcarea, Dorina Cachită-Cosma
THE INFLUENCE OF SALICYLIC ACID ON CONTENT OF
ASSIMILATORY PIGMENTS IN THE PRIMARY LEAVES OF WHEAT
(Triticum aestivum) PLANTLETS Cornelia Purcarea, Dorina Cachiţă-Cosma
TREE GROWTH EQUATIONS IN DIFFERENT SITES FOR MIXED STANDS
OF SESSILE OAK WITH BEECH FROM THE MIDDLE BASIN OF THE
CRISUL REPEDE RIVER Sorin Dorog
THE SIMULATION AND EVALUATION OF A BREAK IN MIXED STANDS
OF BEECH WITH SESSILE OAK Sorin Dorog
THE FREQUENCY OF DAYS WITH DIFFERENT TEMPERATURES IN THE
CRISUL REPEDE HYDROGRAPHICAL BASIN Ana Cornelia Moza
THE RISK OF INUNDATIONS INDUCED BY THE FLOODS ON THE
RIVERS FROM ALMAS-AGRIJ DEPRESSION AND CLUJ - DEJ HILLS
Alina-Daciana Dumitra
RESEARCH REGARDING MAIN CHEMICAL REACTIONS IN
PREDEFECATION OF DIFFUSION JUICE Gheorghe Sarca
DETERMINATION OF NITROGEN-CONTAINING POLYNUCLEAR
AROMATIC HYDROCARBONS IN SMOKED FOODS BY LIQUID
CHROMATOGRAPHY Lucian Bara
THE IMPORTANCE OF VOLATILE N-NITROSAMINES IN FOOD-
PRODUCTS Lucian Bara, Camelia Bara, Vasile Bara, Dana Marele
LUNG CANCER CAUSED BY SMOKING AND CADMIUM AND
CHROMIUM IN HUMAN LUNG TISSUE Camelia Bara
FORMATION OF AFLATOXIN M- TRIFLUOROACETIC ACID
DERIVATIVE IN OPTIMUM CONDITIONS Lucian Bara
DETERMINATION OF “CARCINOGENESIS” BY ALTERNATIVE
BIOASSAYS Camelia Bara, Lucian Bara
CLINICAL AND EPIDEMIOLOGICAL ASPECTS OF HEPATIC HYDATID
CYST Mirela Indrieş, Viorica Coldea, Ciprian Brisc, Ildiko Lenard, Sonia
Drăghici, Andrei Csép, Daciana Sabău, Nicoleta Negruţ
THIN LAYER CHROMATOGRAPHY OF STERIGMATOCYSTIN IN
CHEESE Bara Lucian
9
486
492
497
502
509
516
523
528
531
539
542
546
553
561

TISSUE LEVELS AND BIOLOGICAL EFFECTS OF N-
NITROSODIMETHYLAMINE IN MICE DURING CHRONIC LOW OR HIGH
DOSE EXPOSURE WITH OR WITHOUT ETHANOL Bara Camelia
DETERMINATION OF METALLOTHIONEIN, CADMIUM, COPPER AND
ZINC LEVELS IN HUMAN AND RAT TISSUES Camelia Bara
THE ROLE OF WATER QUALITY IN INFANTILE ALIMENTATION
Andrei Csép, Sonia Draghici, Ildikó Lenard, Mirela Indries, Nicoleta Negrut
SOME ASPECTS OF SHELTER REGENERATION OF QUERCUS SPP.
FOREST STANDS FROM PRODUCTION UNIT II VALEA LUNGA
(FORESTRY DISTRICT, LUGOJ (TIMIS, COUNTY) Ghiţă C. Crainic
WASTEWATER TREATMENT TEHNOLOGIES OF BEER INDUSTRY OF
ROMANIA
Pantea Emilia Valentina, Mirel I., Romocea Tamara, Mitrasca Mihaela
REGIONAL DEVELOPMENT USING CONSTRUCTED WETLANDS FOR
WASTEWATER TREATMENT IN VALEA IERULUI (ÉRMELLÉK) REGION
Botond L. Pete
STUDY OF VERTICAL MOVEMENT OF SOME MICROELEMENTS IN
THE SOIL László Szegedi Lajos Szabó Mária Takács Hájos
Descrierea CIP a Bibliotecii Naţionale a României
INTERNATIONAL CONFERENCE ON LONG-TERM EXPERIMENTS
AGRICULTURAL RESEARCH, NATURAL RESOURCES AND
SUSTAINABLE DEVELOPMENT (5; 2007; Debrecen-Oradea)
<strong>International</strong> <strong>Conference</strong> on Long-term Experiments, Agricultural Research,
Natural Resources and Sustainable Development:
Debrecen-Nyírlugos-Oradea: 31 May-01 June 2007
University of Debrecen, Faculty of Agriculture.
University of Oradea, Faculty for Environmental Protection
Bibliogr.
Index
ISBN 978-973-759-298-9
63(063)
UNIVERSITY OF ORADEA PUBLISHING HOUSE
ACCREDITED BY CNCSIS BUCHAREST, COD 149
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PREFACE
With the general aims to contribute to the exchange of opinions and to
the development of cross border cooperation the <strong>International</strong>
<strong>Conference</strong> on Long-term Experiments, Agricultural Research and
Natural Resources is dedicated to the 45 th anniversary of the
establishment of the Field Experiment at Nyírlugos.
In this regard, one of the most important tasks is to evaluate results
of long-term research sites, test sustainability of water and nutrient
management systems and identify priorities of natural resource
management. For sustainable use of natural resources, we need research
in the principles of multifunctional agriculture and education that will
address the problems of communities. The goal is to bring together
people and resources, to promote an agriculture that is efficient,
profitable, socially acceptable and environmentally sustainable.
The primary objective is to provide a model, where the total
agricultural system and community are taken into account, and
agriculture is not separated from the natural ecosystem of the region. The
most critical challenge is to consider the needs of technological
development and natural ecosystem, and provide an educational
environment for people living in Nyírség region. The international
conference will also help cross border cooperation on the sustainable use
of natural resources. All Hungarian and foreign experts are welcome,
including members and leaders of doctoral schools in the fields of
agriculture, natural and social sciences.
Debrecen-Nyírlugos 31. May 2007
István Láng Tamás Németh
János Nagy János Kátai
Vasile Bara György Hovánszki
11

INFLUENCE OF CHEMICAL FERTILIZERS, MANURE AND LIME APPLIED IN
LONG TERM FIELD EXPERIMENTS ON PH-VALUE OF PRELUVOSOILS
FROM NORTH-WEST PART OF ROMANIA
Cristian Hera*, Gheorghe Ciobanu**, Cornelia Ciobanu**,
Cornel Domuţa**, Adrian Vuşcan***, Georghe Sarca**
* Academy of Agricultural and Forestry Sciences
"GHEORGHE IONESCU-SISESTI" Bucuresti
** University of Oradea, Faculty for Environmental Protection Oradea, Romania
***Agricultural Research and Development Station Oradea
ABSTRACT
In Romania was elaborated since 1968 a stationary long term experiments with fertilizers
and lime in all the Agricultural Research Stations belongs to Research Institute from
Fundulea.
The experiments was set aup using a unitary schem for knowing the evolution of soil
fertilizers and the influence of fertilizers and lime rates and combinations on level and
quality yield of different crops.
The preluvosoil from North-West part of Romania is a medium soil, provide with the
principale nutritive elements, with a weak acid reaction in the ploughing horizon.
In this paper are presented the results regarding the influence of fertilizers and lime
rates and combinations on pH values of preluvosoil from North-West part of Romania.
Long term fertilization with nitrogen determined the decrease of pH values and the
increase of mobile aluminium content up to the phytotoxicity level.
Lime application determined the increase of pH values and the decrease of mobile
aluminium content obtaining positive effects on yield.
INTRODUCTION
In Romania acid ploughing soils are spread on 2,0 millions ha which represent 20% from
total agricol land.
The factors which has a negative influence on growing plants are: high level
concentration by H + and Al 3+ , high level soil content in Fe + and Mn 2+ and low level soil
content in principal nutrients elements, low activity of microorganisms, stagnation of water,
because of unsatisfactory infiltration.
Much research on white luvic soil and preluvosoil conditions (Bedo and Lang, 1977,
Ciobanu and Nagy 1978, Nemeth 1996, Stefanescu 2003) has shown the negative effect of
long-term application of nitrogen as ammonium nitrate on soil reaction, which became more
acidic and led to growth of mobile aluminum and manganese soil content, which can
determine phitotoxicity in the first part of vegetative period, with negative influence on
yield level and quality.
12

For a better knowledge of application effect on time of chemical fertilizers, manure and lime
on soil chemistry was set up in the network of Agricultural Research Stations from
Romania, long-term field experiments in different pedoclimatic conditions.
This paper presents the results regarding the influence of NP chemical fertilizers, manure
and lime on evolution of preluvosoil acidity.
MATERIAL AND METHOD
Experimental site: The research data was obtained at the Agricultural and Development
Research Station Oradea, using a unique design in the all research network of Research
Institute from Fundulea. The investigation has been carried out beginning with the autumn
of 1974 in Oradea, in a flat plain area on the third terrace of the Crisul Repede river, whose
geographical coordinates are: 21 0 56’ Eastern longitude, 47 0 03’ Northern latitude and 136 m
altitude. The solidification rock consists of clay loam. The ground water is located at a depth
of 6-8 m. the soil is a brown one with horizon disposition and the main physical and
chemical characteristics are shown in table 1. The presence of clay migration, B horizon is
to be remarked noticed on the thickness of the soil profile, with high and very high values of
the bulk density and compaction level and low or very low total porosity and hydraulic
conductivity.
The soil reaction is acid in the ploughing A horizon, then slightly acid. The lack of
CaCO3 in the soil profile is underlined. The mobile Al content in the A horizon may cause
poor growth of some crops (clover). The soil is well provided with mobile potassium and
phosphorus. The soil humus medium content may not cause distortions to the neutronic
determination of the soil moisture.
Soil depth
cm
Table 1: The main properties of the brown luvic soil from Oradea – Romania
Sand Silt Clay OC Humus
%
Ca CO3
%
Al mobile
mg/100g
soil
PH
1:2
H2O
N
Total
%
P mobile
ppm
Kmobile
ppm
0 - 5 43,5 28,3 28,2 1,25 2.32 0.00 3.68 6,3 0.12 21.8 83.0
5 - 15 41,8 28,4 29,8 1,12 2.28 0.00 2.32 6,4 0.11 22.7 102.1
15 - 30 40,0 28,5 31,5 1,02 1.91 0.00 0.52 6,3 0.09 5.7 112.1
30 - 60 32,0 28,0 40,0 0,99 1.93 0.00 0.77 6,6 0.09 6.1 117.9
60 - 90 24,1 36,7 39,2 0,29 0.00 0.32 6,6
90 - 150 35,1 27,3 37,6 0,17 0.00 0.59 6,5
The experimental factors in field experiments with NP fertilizers, manure and lime was:
1. Field experiment with NP was set up in 1974 was used from plants in follow crop
rotation-pea-winter wheat-sunflower-maize
P rates ware the same for each plant: 0, 40, 80, 120, 160 kg P2O5 ha -1
N rates were differentiated:
For pea-N0, N25, N50, N75, N100,
13

For winter wheat and sunflower- N0, N40, N80, N120, N160,
For maize- N0, N50, N100, N150, N200,
The chemical NP fertilizers were: ammonium nitrates and superphosphate
2.Field experiment with manure and NP fertilizers was set up in 1974.
Was used a short plant rotation winter wheat-maize
The manure was applied once at four years in autumn for maize using the rates: 0, 20, 40, 60
to/ha.
The NP rates were: N0P0, N50P0, N50P50 and N100P100,
3.Field experiment with lime was set up in 1974 using a crop rotation: pea, winter
wheat, maize, alfalfa.
The lime rates were: 0, 3, 6, 9 to/ha applied once at 6 years.
NPK rates were differentiated:
Pea: N0P80, N30P80, N120P80, N60P80, N90P80, N90P80K80,
Winter wheat: N0P80, N30P80, N120P80, N160P80, N160P80K80,
Maize: N0P80, N80P80, N160P80, N240P80, N240P80K80,
Alfalfa: N0P100, N40P100, N80P100, N120P100, N120P100K80,
Sampling and analytical method: Soil samples from top soil (0-20cm) were collected
from each experiment plot, in august 2000,after wheat harvesting. All samples were taken to
the laboratory and used for routine soil chemical analysis. pH was determined in water
suspension.
RESULTS AND DISCUSSION
Influence of NP fertilizers on preluvosoil reaction(figure 1)
As an effect of systematically applying of NP fertilizers were registered significant
modifications of soil acidity.
The higher influence on soil reaction had the nitrogen fertilizers applied on different P
backgrounds.
In the case of background P0, applying of N fertilizers lead to decreasing of pH values
from 6,20 to 5,15 when N rate is taking values from 0 to 160 kg N/ha.
In the case of all P backgrounds the decreasing trend of pH values due to N fertilizers
applied is obvious. Based on the research data was established the interrelations existing
between pH values and N rates.
The phosphorus fertilizers applied affected more little pH values but it is noticed a low
trend to increase soil acidity once with increasing of P rates, because of depletion of bases
as an effect of higher level of yield obtained.
14

pH values
pH values
8
6
pH values
6.50
6.00
5.50
5.00
4.50
4.00
N rates kg/ha
Background P0
y = 7E-06x 2 - 0.0076x + 6.2034
R 2 = 0.9957
N0 N40 N80 N120 N160
4
0 40 80 120 160
8
6
N rates kg/ha
Background P80
y = -3E-05x 2 - 0.0015x + 5.9257
R 2 = 0.9951
4
0 40 80 120 160
N rates kg/ha
pH values
160
8
6
80
0
P rates kg/ha
Background P40
y = -0.006x + 6.06
R 2 = 0.9965
4
0 40 80
N rates kg/ha
120 160
8
6
Background P160
y = -3E-05x 2 - 0.0009x + 6.0611
R 2 = 0.9982
4
0 40 80 120 160
N rates kg/ha
Figure 1: Influence of nitrogen and phosphorus on preluvosoil reaction from N-W part
of Romania
The lower values of pH was registered in the case of plots fertilized with P80, when was
obtained the higher level of production.
15

Influence of manure applied on different NP backgrounds on preluvosoil reaction
(figure 2)
It is well known that the manure applied in acid soil conditions are increasing the
degree of base saturation in the same time increasing the buffering capacity of the soil. On
this way is possible to avoid unfavorable effect of chemical fertilizers with acid potential.
pH values
7.00
6.00
5.00
Background N0P0
y = 0.0002x 2 - 0.0011x + 6.273
R 2 = 0.9601
4.00
0 20 40 60
pH values
7.00
6.00
5.00
pH values
Manure rates-to/ha
Background N50P50
y = 6E-06x 2 + 0.0061x + 5.8465
R 2 = 0.9971
4.00
0 20 40 60
7
6
5
4
Manure rates -to/ha
60
Manure rates- to/ha
40
20
0
pH values
7.0
6.0
5.0
4.0
pH values
Background N50P0
y = 2E-05x 2 + 0.0052x + 5.609
R 2 = 0.9998
0 20 40 60
7.00
6.00
5.00
Manure rates- to/ha
Background N100P100
y = -4E-05x 2 + 0.0097x + 5.6885
R 2 = 0.9858
4.00
0 20 40 60
Manure rates- to/ha
Figure 2: Influence of manure of preluvosoil reaction from North-West part of Romania
16
N0P0
N50P0
N50P50
N100P100
NP rates kg/ha

In the preluvosoil conditions the manure applied on different NP backgrounds had a
significant positive effect on soil acidity. Applying manure in the rates of 20, 40 and 60
to/ha in the lack of N, P fertilizers the pH values are increasing from 6, 29 to 6,76 units. In
the case of the other NP backgrounds the manure determined an increase of pH values
ranging between 0,4-0,6 units. The negative effect of nitrogen fertilizers application is lower
in the case of manure application.
Neutralization of soil acidity and completion of calcium reserve (and magnesium)
trough lime application is a essential measure for increasing yield capacity of acid soils.
In the case of preluvosoil application of lime in the rate of 3, 6 and 9 to/ha once at six years
lead to increasing of pH values depending on NP background utilized.
When the lime is applied alone pH values are taking values between 6,22 and 7, 08 when
the lime rates are increasing from 0 to 9 to/ha.
Influence of lime application on preluvosoil reaction(figure .3)
pH values
8
6
4
CaCO3 rates
t/ha
Background N0P80
y = 0.0056x 2 + 0.05x + 6.2
R 2 = 1
0 3 6 9
CaCO3 rates - t/ha
9
6
3
0
N0P80
17
4
N160P80K80
N160P80
N90P80
N80P80
pH values
8
6
4
6
8
pH values
NP rates kg/ha
Background N80P80
y = 0.12x + 5.76
R 2 = 0.9529
0 3 6 9
CaCO3 rates - t/ha

pH values
8
6
4
Background N160P80
y = -0.0028x 2 + 0.1683x + 5.155
R 2 = 0.9581
0 3 6 9
CaCO3 rates - t/ha
pH values
8
6
4
Background N160P80K80
y = -0.0028x 2 + 0.195x + 4.885
R 2 = 0.9966
0 3 6 9
CaCO3 rates - t/ha
Figure 3: Influence of lime on preluvosoil reaction from North-West part of Romania
On the other NPK backgrounds the increasing of pH values are taking values between 0,9-
1.42 pH units.
Lime application in preluvosoil conditions is a necessary measure in the case of using NP
fertilizers in high rates.
CONCLUSIONS
The results presented above let us come to the following conclusions:
1. Long term experiments are important tools for examining soil fertility
2. The soil reaction evolution is depends by fertilizers type and by the rates level applied
3. In the case of preluvosoil from North-West Romania by systematic application of
nitrogen fertilizers like as ammonium nitrate leads to a decreasing of pH values from
6.3 to 4.9 as a function of rates level applied
4. Phosphorus fertilizers applied influence not to strong preluvosoil reaction but it can be
observed a slow decreasing of pH values because of depletion of bases due to yields
spores obtained
5. The manure applied alone or associated with NP fertilizers, favorable influenced soil
reaction, pH values increasing with 0.3-0.4 units if the manure rates applied are 40
to/ha respectively 60 to/ha
6. For to avoid the decreasing pH values due to chemical fertilizers applied in preluvosoil
conditions is necessary lime application for acidity neutralization
7. Lime application once at six years in the rate of 9 to/ha maintain pH values between 6.4-
7.0, which ensure optimal growing and developing condition for plants
18

REFERENCES
Bedo, Z.- Lang, L. (1977) - Growing and breeding of quality wheat. Agro-21 Fuzetek,
97/14, 8-28
Ciobanu Gh.. (1999) - Influenta ingrasamintelor organice si minerale asupra productiei si
calitatii graului si porumbului. "Agricultura durabila- performanta". Edit. Agric-
Bucuresti.
Hera Cr., Borlan Z., (1980) – Ghid pentru alcatuirea planurilor de fertilizare. Ed. Ceres
Bucuresti.
Hera Cr., Eliade Gh., Ghinea L., (1984) – Asigurarea azotului necesar culturilor agricole.
Ed. Ceres Bucuresti.
Nemeth, T. (1996) - Environment friendly fertilizer recommendation for sustainable
agriculture. In: Environmental Pollution, Ed. B. Nath et. All ECRP, Queen Mary and
Westfield, college, London, 99-105.
19

EFFECT OF VARIOUS CROP PRODUCTION FACTORS ON THE YIELD
AND YIELD STABILITY OF MAIZE IN A LONG-TERM EXPERIMENT
Zoltán Berzsenyi – Dang Quoc Lap
Agricultural Research Institute of the Hungarian Academy of Sciences, Martonvásár
ABSTRACT
In a long-term experiment set up in 1960 on a medium heavy loamy soil, the effect of
five crop production factors in increasing maize yields was studied in seven treatment
combinations. The factors studied were soil cultivation, fertilisation, plant density,
variety and weed control. All the factors had a minimum and an optimum level. Yield
data recorded over 42 years were evaluated using analysis of variance and stability
analysis. The highest yield (8.59 t ha –1 ) was obtained when all the production factors
were optimum and lowest (2.09 t ha –1 ) when these factors were at a minimum. When
only one factor was at a minimum and all the other factors were optimum the following
yields were obtained (t ha –1 ): soil tillage: 8.32, fertilisation: 5.21, genotype: 4.98, plant
density: 6.31, weed control: 7.01.The crop production factors contributed to the increase
in maize yield in the following ratios (%): fertilisation 30.6, variety 32.6, plant density
20.2, weed control 14.2, soil cultivation 2.4. The highest value of CV%, expressing the
deviation of the yield averages, was obtained when all the production factors were at the
minimum level (45.7%) and when weed control or fertilisation were at a minimum
(36.6% and 34.8%, respectively), while the lowest value was recorded when all the
factors were optimum (19.5%). The significant treatment × year interaction could be
attributed principally to treatments in which weed control, fertilisation, genotype or all
the factors were at a minimum. The regression coefficient of linear regression analysis
provided a satisfactory characterisation of the stability of the treatments in different
environments, while the distance between the straight lines expressed the yield
differences between the treatment pairs. The AMMI (Additive Main Effect and
Multiplicative Interaction) model proved to be a valuable approach for understanding
agronomic treatment x environment interactions and assessing yield stability.
Key words: long-term experiment, maize, production factors, stability analysis, AMMI
model
INTRODUCTION
Long-term experiments are often conducted to compare the long-term effects (e.g.,
sustainability) of various treatments on one or more response variables. Measurements
made on the plots are generally taken each year in the case of crop yield and other plant
measurements. These data are ultimately subjected to some kind of statistical analysis,
often with the goal of understanding something about the potential for different
cumulative effects of treatments over time. As a result, the time x treatment interaction
becomes the focus of an analysis.
It is generally accepted that measurements made in field experiments may be
influenced by uncontrollable environmental factors. Aside from random variation there
is also a predictable element, the long-term average of yield in the experiment. The
20

mean (fixed, i.e. systematic effect) and the variance (random element) are the two main
parameters describing the response pattern of a cropping system. Variations in the
measurements or in the treatment differences across years are simultaneously due to
both the fixed and the random effects, and the two sets are often inseparable from each
other.
The improvement in maize production realized in the fields is the result of the
combined effects of genetic, ecophysiological and technological changes superimposed
on short- and long-term climatological variations. The simple partitioning of yield
improvement into a genetic and an agronomic component assumes the lack of
interaction between the two components. Although the increase has been the result of
both genetic and agronomic-management improvements, most of this improvement is
the result of the genotype x management interaction (Tollenaar and Lee 2002).
The effect of major crop production factors has been investigated in long-term
experiments in Martonvásár since 1960 (Győrffy 1969, Berzsenyi and Győrffy 1995).
The aim of the present work was to carry out a biometric analysis on the 42-year data
series from the experiments in order to determine how the minimum and optimum
levels of five production factors (tillage, fertilisation, plant density, variety, weed
control) influenced the yield of maize, and to use the single-variable and multi-variable
methods of stability analysis to determine the stability of these production factors in
diverse environments.
MATERIAL AND METHODS
The effect of five crop production factors on maize yields has been investigated since
1960 in a long-term experiment set up on medium heavy loam soil. Each factor is
represented by two treatments, one minimum and one optimum. The treatments applied
for each factor were as follows:
Factor Minimum level Optimum level
Tillage * Shallow ploughing Deep ploughing
to a depth of 12–14 cm to a depth of 24–28 cm
Fertilisation ** No fertilisation 60 t ha –1 farmyard manure
every 4 years;
N140P60K60 annually
Plant density ‡ 35,000 plants ha –1 70,000 plants ha –1
Variety Open-pollinated: Hybrid
Aranyözön and
Mindszentpusztai Sárga
Weed control Poor: late thinning and Good: complete freedom
two delayed hoeings from weeds; timely
thinning
* From 1985 onwards the optimum level was deep loosening every four years
** From 1980 onwards the fertiliser rate was doubled
‡ Until 1980 the minimum plant density was 20,000 plants ha –1 and the optimum 40,000
Seven treatment combinations were used in the experiment. In Treatment 1 all the
factors were at the minimum level and in Treatment 2 all were optimum. In Treatments
3–5 one of the crop production factors was at a minimum, while all the others were
21

optimum. The factors at a minimum were tillage in Treatment 3, fertilisation in
Treatment 4, plant density in Treatment 5, variety in Treatment 6 and weed control in
Treatment 7.
The experimental results were evaluated using a variety of methods. The
cumulative yield analysis method (Sváb 1981) designed for the evaluation of long-term
experiments was used to compare the cumulative yield-enhancing effect of individual
production factors compared with the minimum combination. Analysis of variance was
performed by using the “General Analysis of Variance” procedure of the GenStat 9
program. Analysis was conducted separately for each year and pooled together
afterwards. Stability analysis on the experimental treatments was carried out using
single-variable (variance and regression parameters) and multi-variable (AMMI)
methods. Among the variance parameters, the ecovalence (W²), the stability variance
(σ²) and the yield stability (YS) parameters were calculated using the STABLE model
proposed by Kang (1995), where the covariant was the annual rainfall quantity from
April to September. The regression method of stability analysis was applied as
described by Finlay and Wilkinson (1963). The year × treatment interaction was
investigated by means of AMMI analysis, a combination of analysis of variance and
principal component analysis (Crossa 1990).
RESULTS AND DISCUSSION
Annual analysis of variance indicated that the effect of the experimental treatments was
significant at the P=0.1% level every year. Combined ANOVA over the 42 years
demonstrated that both the year effect and the treatment effect were significant, but the
MS values showed that the treatment effect was more than 16 times as strong as the
year effect (Table 1).
Table 1. Analysis of variance of the long-term experiment (1960-2001)
Source df SS MS F-value
Total 1175 8540
Environments (Years) 41 2091 51.0 92.5 ***
Replication 3 16.6 5.54 10.1
Year (replication) 123 67.8 0.55
Treatments 6 5054 842.3 643.9 ***
Residuals 1002 1311 1.31
The cumulative yield-enhancing effect of the production factors compared with the
minimum combination is illustrated for the years 1960–2001 in Figure 1. It is clear from
the figure that under Hungarian conditions deep tillage only had a slight effect, ranging
from 2.4–3.0% over the period in question. A pronounced increase in the fertiliser effect
was observed in the long-term experiment, from 18.6% in the early years to 30.7%. The
effect of an increase in plant density on maize yields changed little over the 42 years,
having values of 19.9–23.3%. Apart from fertilisation, the factor with the greatest
influence on maize yields was the genotype, the effect of which increased from 26.6%
to 32.6% over the years. The effect of weed control on the maize yield ranged from 23–
22

29% in the first 15 years of the experiment, but has declined to 14–18% over the last ten
years.
Cumulative yield increase t ha -1
Years
Basic treatment (1)
Figure 1. Cumulative yield increase in various treatments compared to the minimum
combination of factors (1960-2001)
The effect of the crop production factors in increasing maize yields is depicted in Figure
2, averaged over the 42 years. The greatest difference in the mean yield response was
found between treatments where all the factors were minimum (2.09 t ha –1 ) or optimum
(8.59 t ha –1 ). When only one factor was at a minimum and all the others optimum, the
depth of soil tillage was found to have the least effect on maize yields (mean yield
response: 8.32 t ha –1 ), while both fertilisation and genotype had a substantial influence
on the yield (mean yield response: 5.21 and 4.98 t ha –1 , respectively). The minimum
level of plant density or weed control led to yield reductions of 2.2 and 1.6 t ha –1 (mean
yield responses of 6.36 and 7.01 t ha –1 ). Based on this 42-year data series, the crop
production factors were found to make the following contributions to yield increases in
maize: variety 32.6%, fertilisation 30.6%, plant density 20.2%, weed control 14.2% and
soil tillage 2.4%.
The variance and regression parameters of stability analysis for each experimental
treatment are presented in Table 2. The CV% value expressing the annual deviation in
yield averages was the greatest when all the production factors were at a minimum
(45.7%) and smallest when they were all at the optimum level (19.5%). When one
23

factor was at the minimum level and the remainder were optimum the highest values of
CV% were observed for weed control and fertilisation (36.6% and 34.8%, respectively).
T F D V W
0 0 0 0 0
1 1 1 0 1
1 0 1 1 1
1 1 0 1 1
1 1 1 1 0
0 1 1 1 1
1 1 1 1 1
Treatments
0 1 2 3 4 5 6 7 8 9
Maize grain yield t ha -1
Figure 2. Effect of crop production factors on maize grain yield increments in a longterm
experiment (Martonvásár 1960-2001). Treatments : T: Depth of soil tillage, F:
Fertilisation, D: Plant density, V: Variety, W: Weed control. Each factor at two levels:
0: Minimum level, 1: Optimum level. Treatments designated with the same letter did
not differ significantly according to Duncan’s test.
The magnitude of the ecovalence (W²) and stability variance (σ²) parameters expresses
the contribution of the treatments to the significant year × treatment interaction. It can
be seen that treatments in which weed control, fertilisation, genotype or all the factors
were minimum were chiefly responsible for the interaction. Based on the mean yield
response and the stability, the yield stability (YS) parameters indicated that Treatments
2, 3, 5 and 7 were the most favourable (Table 2).
When the effects of the treatments were investigated in diverse environments by
means of linear regression analysis (Fig. 3), the greatest difference in mean yield
response was observed between the treatments in which all the factors were minimum
or optimum. The yield difference between the two treatments (expressed as the distance
between the straight lines) increased with the environmental mean. The figure shows
that the slight effect of tillage depth could only be detected for a low environmental
mean. When fertilisation was at a minimum the magnitude of the yield reduction was
considerable and increased with the value of the environmental mean. The magnitude
and trend of yield reduction was similar when the genotype was at a minimum. There
was also a similar tendency for the plant density, but the yield reduction was smaller in
this case, particular in the case of a high environmental mean.
24
F
E
E
D
C
B
A

Table 2. Variance and regression parameters of the stability of the production factors
in the long-term experiment (1960-2001)
Stability
parameters
Coefficient of
1 2 3 4 5 6 7
variation (CV%)
Ecovalence
45.7 19.5 21.9 34.8 24.5 25.1 36.6
(W 2 )
Stability variance
117.7 51.0 53.5 219.2 57.1 163.2 325.9
(σ 2 )
Yield stability
3.22 0.94 1.02 6.68 1.15 4.77 10.32
(YS) **
Correlation
+ +
+
+
coefficient (r)
Regression
0.796 0.947 0.962 0.713 0.919 0.690 0.899
coefficient (b)
Mean yield
0.500 1.145 1.236 0.860 1.020 0.580 1.659
response t ha -1
2.094F 8.585A 8.315B 5.205E 6.355D 4.979E 7.014C
Treatments *
* Treatments: 1. Minimum values of all factors, 2. Optimum values of all factors, 3.
Minimum depth of soil cultivation, 4. Minimum level of fertilisation, 5. Minimum level
of plant density, 6. Minimum for variety, 7. Minimum weed control.
** +: variants selected by the STABLE programme.
Table 3. AMMI Analysis for the long-term experiment (1960-2001)
Source df SS MS F-value
Total 1175 8540
Treatments 6 5054 842.3 1969.7 ***
Environments 41 2091 51.0 76.1 ***
Block 126 84 0.67
Interactions 246 988 4.0 9.39 ***
IPCA1 46 467 10.2 23.8 ***
IPCA2 44 238 5.4 12.7 ***
Residuals 156 282 1.8 4.23 ***
Error 756 323 0.43
Compared with the other production factors the weed control minimum exhibited the
opposite environmental dependence, indicating that weed infestation had the greatest
yield-reducing effect at low environmental means and the smallest effect at an
environmental index of 10 t ha –1 . Consequently, it can be stated that Treatments 2, 3 and
5 have average stability (regression coefficients of around 1), while the approximately b
= 0.5 value recorded for Treatments 1 and 6 and the b

Maize grain yield t ha -1
Maize grain yield t ha -1
Maize grain yield t ha -1
18
16
14
12
10
18
16
14
12
10
8
6
4
2
0
8
6
4
2
0
18
16
14
12
10
8
6
4
2
0
Optimum values
of all factors
Minimum
values of all
factors
0 2 4 6 8 10 12
Optimum values
of all factors
0 2 4 6 8 10 12
Optimum values
of all factors
Minimum level
of fertilisation
Minimum for
genotype
0 2 4 6 8 10 12
Environmental index t ha -1
Figure 3. Yield response of maize for minimum levels of production factors vs.
optimum level of all factors in comparision to the environmental index in the longterm
experiment (1960-2001)
26
18
16
14
12
10
8
6
4
2
0
18
16
14
12
10
8
6
4
2
0
18
16
14
12
10
8
6
4
2
0
Optimum values
of all factors
Minimum depth of
soil cultivation
0 2 4 6 8 10 12
Optimum values of
all factors
Minimum plant
density
0 2 4 6 8 10 12
Optimum values
of all factors
Minimum weed
control
0 2 4 6 8 10 12
Environmental index t ha -1

The b = 1.7 value recorded for Treatment 7 can be explained partly by the lower extent
of weed interference in a favourable environment.
The main effects of year and treatments and the interactions were also examined
using AMMI analysis, which combines ANOVA and principal component analysis
(Table 3). The first and second principal components (PCA1, PCA2) explained 71.4%
of the interaction SS. The results of the AMMI analysis are illustrated in Figure 4, with
the yield average on the X axis and the value of the first principal component on the Y
axis, for seven treatments in 42 environments (years). The higher the value of the
principal component, the greater the contribution of the treatment to the interaction, i.e.
the lower the yield stability. It can be seen that Treatments 7, 6, 1 and 4 made the
greatest contributions to the interaction, while Treatments 2, 3 and 5 had the greatest
yield stability.
First principal component scores
3
2
1
0
-1
-2
G1
G7
.
E40
E27
E33 .
E32
.
E26
E42 E3 7
E22 .
E38 G3 E30
E17 E35 E23 .
.
E19 E16
. E39 E10 E41 G2
E34 E14 .
. . . . .
. . . . . G5 E36
. . . . . . E21 . . . . .
. . .
. . . E28 . . . . . . . .
E18
E31
.
E1 E20
.
E4E13
E8
.
E24
E15
.
. E1 2
E9
E11
E5 E7
E25
E6
.
.
E2
E29
E3
.
.
G4
G6
2 4
6
8
Mean yields t ha -1
Figure 4. Plot of the mean yields and first principal component scores of crop
production factors in 42 environments
ACKNOWLEDGEMENTS
This research was supported by a grant from the National Committee for Technological
Development (OMFB-00895/2005).
27
10
12

REFERENCES
Berzsenyi Z., Győrffy B. (1995): Effect of various crop production factors on the yield
and yield stability of maize (Különböző növénytermesztési tényezők hatása a
kukorica termésére és termésstabilitására). Növénytermelés, 44: 507-517.
Crossa, J. (1990): Statistical analyses of multilocation trials. Advances in Agronomy,
44: 55-85.
Finlay, K.W., Wilkinson, G.N. (1963): The analysis of adaptation in a plant breeding
programme. Aust. J. Agric. Res., 14: 742-754.
Győrffy B (1969): Effect of crop production factors on maize yields, Experiment code:
Komplex I (Különböző növénytermesztési tényezők hatása a kukorica termésére,
Komplex I). In: I’só I. (ed.), Maize Production Experiments 1965–1968
(Kukoricatermesztési kísérletek 1965-1968). Akadémiai Kiadó, Budapest.
Kang, M.S. (1995): Simultaneous selection for yield and stability in crop performance
trials: consequences for growers. Agronomy Journal, 85: 754-757.
Sváb J. (1981): Biometrical Methods in Research (Biometriai módszerek a kutatásban).
Mezőgazdasági Kiadó, Budapest.
Tollenaar, M., Lee, E.A. (2002): Yield potential, yield stability and stress tolerance in
maize. Field Crops Research, 75: 161-169.
28

THE IMPACT OF PLANT BREEDING ON SEED OIL CONTENT AND
QUALITY IN EVENING PRIMROSE CROPS
ABSTRACT
Andrew F. Fieldsend
University of Debrecen Centre of Agricultural Sciences
4032 Debrecen, Böszörményi ut 138., Hungary
In recent decades, evening primrose (Oenothera spp.) has been grown as an agricultural
crop in several parts of the world including western and central Europe, North America
and New Zealand. Its value is derived from its seed oil which contains unusually high
levels of gamma linolenic acid (GLA), an uncommon fatty acid which is used in
pharmaceuticals and nutritional supplements. Whilst such improvements have led to a
reduction in the seed price, increases in seed oil content and quality (i.e. percentage of
GLA) can lead to further reductions in the cost of the end product. The breeding
programme of Scotia Pharmaceuticals Ltd, UK was effective in achieving such
increases. For example, in spring-sown crops in New Zealand, seed oil content
increased from around 25% in older cultivars grown in the period 1990-1992 to around
28% in the cultivar “Rigel” grown in the period 1992-1999. Furthermore, GLA content
increased by almost 50%, from around 9.5% in older cultivars to more than 13% in
“Rigel”. It has been estimated that these improvements through plant breeding have led
to an 18% reduction in product costs.
Keywords: Evening primrose; Oenothera spp; breeding; oil content; oil quality
INTRODUCTION
In recent decades, evening primrose (Oenothera spp.) has been grown as an agricultural
crop in several parts of the world including western and central Europe, North America
and New Zealand. Its value is derived from its seed oil which contains unusually high
levels of gamma linolenic acid (GLA), an uncommon fatty acid which is used in
pharmaceuticals and nutritional supplements (Horrobin, 1992). Although found in some
other plant and fungal oils, in evening primrose oil GLA is thought to occur in its most
bioactive state, as part of a triacylglycerol called Enotherol® (Horrobin, 1994). A brief
botanical description of evening primrose was given by Fieldsend (1996a).
Early commercial crops of evening primrose were grown from seed stocks available
through the horticultural trade (Fieldsend, 1996a). One such “traditional” stock, grown
in New Zealand, was simply known as “421”. This and similar lines were unsuitable for
agriculture in many ways, for example in having seed capsules which split as they
matured, meaning that a significant proportion of seed was lost prior to harvest. This
factor simply compounded the problem that the potential seed yield of evening primrose
was already much lower than that of other combinable crops (Nix, 1995).
UK-based Agricultural Holdings, owned by the Balint family, was an important early
seed producer, sourcing much of its seed from Hungary. Its subsidiary, which in time
became Scotia Pharmaceuticals Ltd, a leading manufacturer of products derived from
evening primrose oil, set up an evening primrose breeding programme in the mid-
1970s. Whilst development of cultivars with non-splitting capsules was an important
29

eeding objective, as higher seed yields would lead to a reduction in the price per tonne
of seed, Scotia recognised that of equal importance was an increase in the oil (for
nutritional supplements) and GLA (for pharmaceuticals) contents of the seed. The
difference is that whilst the former are normally consumed by users as a fixed volume
of oil per day, for the latter the daily dose of GLA is the important criterion.
Evening primrose breeding is made difficult by its unusual genetic system (Cleland,
1972). The formation of chromosome rings at meiosis, ordered chromosome movement
and the presence of lethal genes limit segregation (and therefore scope for selecting
improved lines) in the generations following the initial cross. Variable establishment,
coupled with seed dormancy in many genotypes, and the long growing season of the
overwintered crop in temperate climates (approximately 14 months) also present
practical problems (Fieldsend and Morison, 2000a). For unimproved lines, field sowing
would be an unreliable method of trialling, and selection of lines to trial would need to
be made before the previous generation of breeding material had been harvested.
This paper describes the methods developed by Scotia to breed evening primrose and
the improvements in oil content and quality of commercial crops in New Zealand which
have resulted from the use of cultivars developed by the programme.
MATERIALS AND METHODS
The Scotia breeding programme began with the collection, from many sources, of over
2000 individual “accessions” which were numbered in a single series from X1 upwards.
Each accession was evaluated in a single spring-transplanted row of 21 plants. Seeds
were sown in potting compost in 750 mm diameter pots in January and placed in a
refrigerator set at 4ºC for 2-3 weeks to break dormancy, following which the pots were
transferred to a propagator hotbox at a temperature of 27ºC. Soon after emergence 24
seedlings of each accession were pricked out into potting compost in modules and kept
in a heated glasshouse with a mean temperature of 13ºC. The rosettes were moved out
of the glasshouse to acclimate during late April and transplanted into 5 m rows with a
0.55 m row spacing in the field in early May. Assessments on each row were carried out
during the growing season and at maturity representative single plants were hand
harvested and dried using a forced-air drier before the seed was removed from the
capsules by hand-threshing. Following cleaning, seed samples were analysed for oil and
GLA content using methods described by Fieldsend and Morison (2000b).
Eventually, a crossing programme was established to generate new breeding lines.
Crosses were carried out using plants grown in pots in an insect-proof glasshouse. Prior
to the pollen becoming viable, the anthers were removed from flowers of the plants to
be used as female parents and, 24 hours later, the stigmas were hand pollinated using
anthers from the male parent. Reciprocal crosses were carried out as the progeny of
reciprocal crosses within Oenothera can be genetically completely different from each
other (Cleland, 1972). The seed was hand-harvested and stored during the winter.
Subsequent generations were evaluated in breeding rows as described above. In extreme
cases, the F1 population of Oenothera is genetically uniform and contains one complete
genome from each parent. No genetic segregation takes place in subsequent generations.
Even in less extreme cases, only limited segregation occurs. Hence, Scotia adopted a
30

modified pedigree breeding method, entailing a large number of crosses and very small
(21 plant) F2 populations. Unpromising breeding lines could be discarded after the F2
generation whilst in some cases populations were uniform and showed enough
commercial promise for seed multiplication to start.
Clearly, this method does not allow the breeder to evaluate seed yields or the ability of
the breeding line to overwinter. During the 1980s, overwintering ability was assessed on
autumn-transplanted rows of the breeding lines showing the most promise in F3 springsown
rows. Selection of lines for trialling was based on field assessments from the F1,
F2 and (until mid-August when the choice of lines for trialling had to be made) F3
breeding rows plus the oil and GLA content data from the F2 rows. By the 1990s,
however, the establishment ability of the breeding lines had improved to such an extent
that the most commercially promising advanced breeding material could be evaluated in
autumn and spring-sown replicated yield trials (c.f. Fieldsend and Morison, 2000a).
New Zealand is an important source of evening primrose seed, not least because the sixmonth
difference in harvest date compared to the northern hemisphere spreads the
demands of harvest, storage and payment for seed more equably throughout the year.
Overwintered crops are normally sown in February and harvested in March of the
following year, whilst spring-sown crops are sown in September or October and
harvested in April. The husbandry of spring-sown crops is described by Fieldsend
(2003). The total area of crops grown under contract to Scotia in 1990-1997 ranged
between 200 ha and 450 ha, with an approximately equal split between overwintered
and spring-sown crops in the earlier years and a subsequent emphasis on spring
production. This area would be divided between over 20 farmers, who would normally
use field sizes of between 4 and 16 ha (Fieldsend, 2003). After harvest, a seed sample
from each field was analysed for oil and GLA contents and the annual means of these
data (not corrected for differences in field size) are used in this paper.
RESULTS AND DISCUSSION
Oenothera is a very diverse genus and most accessions were immediately rejected as
being unsuitable for commercial seed production after one year of trials. However, three
accessions were “off-type” single plants with non-splitting capsules taken from
commercial crops of the “traditional” stock growing in the UK and Hungary. X444 and
X851 were commercialised as the cultivars “Paul” and “Peter” respectively (similar
accession X852 was not commercialised) and were awarded Plant Variety Rights (PVR)
in several countries including the UK and New Zealand. The reduced risk of seed loss
from “Paul” and “Peter” resulted in an approximately 50% increase in seed yields over
“421” when grown as overwintered crops in New Zealand in 1990 and 1991 (Fig. 1),
but these cultivars were similar to “421” in every other respect. Seed oil contents were
in the region of 26.5% from autumn sowing (Fig. 2) and 25% from spring sowing (Fig.
3) whilst the GLA contents of the oil were around 9% and 9.5% respectively. Higher oil
contents and lower GLA contents from overwintered crops are typical of evening
primrose (Fieldsend, 2000b).
No other accessions were directly commercialised but many were used in the crossing
programme. For example, the cultivar “Merlin” resulted from the cross (X444 x X390).
This was a high yielding, easy to grow cultivar much liked by farmers which was also
31

awarded PVR in several countries. The very high yield in 1990 (Fig. 1) was obtained
from just 2.6 ha so this result must be treated with caution but does show its yield
potential. In 1991-1993 “Merlin” outyielded "Paul” and “Peter” whilst its oil and GLA
contents tended to be higher as well (Figs 2 and 3).
Seed yield (t ha -1 )
2,0
1,6
1,2
0,8
0,4
0,0
1990 1991 1992 1993 1994 1995 1996 1997
Year of harvest
Figure 1: Mean seed yield of crops of evening primrose harvested in New Zealand
from 1990 to 1993. Closed symbols: overwintered; open symbols: spring sown;
circles: 421; triangles up: Paul/Peter; squares: Merlin; triangles down: Rigel.
A series of plants collected from the wild in Ontario, Canada produced seed with high
oil contents and a very high GLA content in the oil (approximately 15%) but were
unsuitable for commercial use. One of these, X945, was crossed with another accession,
X1021, in 1985 and the resulting breeding line (LQB) also produced high quality oil but
the seed yield was low. In turn, in 1988, LQB was crossed with “Merlin” and the result
was the cultivar “Rigel”. This cultivar successfully combined the non-splitting capsules
of “Paul” with the high oil and GLA contents of X945 and a high seed yielding ability.
The great potential of the breeding line was confirmed by the laboratory results from the
F3 breeding row and the unusual step was taken of sending seed from the breeding row
(despite this not having been raised in isolation from other lines) to New Zealand for
trialling. A 1000 m 2 plot was sown on 21 October 1991 and yielded 1121 kg seed ha -1
when harvested in 1992. The oil and GLA contents were 28.5% and 13.3% respectively,
substantially higher than cultivars then in commercial production (Fig. 3).
32

“Rigel” seed multiplication commenced with a 1000 plant, 200 m 2 spring transplanted
plot grown in the UK in 1992 which yielded 25 kg seed. 14.5 ha of “Rigel” crops were
sown in New Zealand in October 1992 and harvested oil and GLA contents were again
much higher than “Merlin”. Unfortunately, yield data after 1990 are not available but
Fieldsend (2003) reported yields of up to 1.87 t ha -1 from spring-sown “Rigel” in New
Zealand in 1998 whilst in 1999 some “Rigel” crops yielded more than 2 t ha -1 (Scotia,
commercial data). This cultivar was again awarded PVR and was the sole cultivar to be
grown commercially for Scotia after 1994.
GLA content of oil (%)
Seed oil content (%)
14
13
12
11
10
9
8
29
28
27
26
25
24
1990 1991 1992 1993 1994 1995 1996 1997
Year of harvest
Figure 2: Seed oil content and GLA content of oil of overwintered crops of evening
primrose harvested in New Zealand from 1990 to 1997. Cultivars are designated
thus: circles: 421; triangles up: Paul/Peter; squares: Merlin; triangles down: Rigel.
33

Thus, by the late 1990s, oil and GLA contents of overwintered crops in New Zealand
averaged around 28.5% and 12.5% respectively whilst the equivalent data from springsown
crops were 28% and 13.5%. The impact on the GLA content of the seed was an
increase from around 2.5% in 1990 to over 3.5% by 1999, with spring-sown crops
normally giving the highest results (Fig. 4). The big difference between sowing dates in
1994 is due to the fact that overwintered crops were almost exclusively “Paul” and
“Merlin” whilst all spring-sown crops were “Rigel”. The Scotia breeding programme
closed in 1999 but “Rigel” continues to be commercially grown.
GLA content of oil (%)
Seed oil content (%)
14
13
12
11
10
9
8
29
28
27
26
25
24
1990 1991 1992 1993 1994 1995 1996 1997
Year of harvest
Figure 3: Seed oil content and GLA content of oil of spring sown crops of evening
primrose harvested in New Zealand in from 1990 to 1997. Cultivars are designated
thus: circles: 421; triangles up: Paul/Peter; squares: Merlin; triangles down: Rigel.
34

The benefits of the cultivars bred by Scotia can be seen in different ways. For
nutritional supplements, the daily dose is normally specified as the number of (gelatine)
capsules to be taken but a higher GLA content in the oil gives the retailer a marketing
benefit. For pharmaceuticals, where the dose normally much higher and is calculated as
daily GLA intake, the number of capsules taken can be reduced. This has the twin
benefits of improving patient compliance and also, owing to savings along the entire
manufacturing chain, reducing product costs. It has been estimated (Scotia, commercial
data) that the change to “Rigel” oil led to an 18% reduction in costs.
Seed GLA content (%)
4,0
3,5
3,0
2,5
2,0
1990 1991 1992 1993 1994 1995 1996 1997
Year of harvest
Figure 4: Mean seed GLA content of crops of evening primrose harvested in New
Zealand from 1990 to 1997. Closed symbols: overwintered; open symbols: spring
sown.
The oil and GLA contents achieved with “Rigel” are close to the upper limit observed in
the genetic material evaluated by Scotia. However, the high linoleic acid content of the
seed oil (approximately 75% in “Merlin” (Fieldsend and Morison, 2000b)) is a substrate
for potentially much higher GLA contents which could possibly be achieved through
genetic engineering. Borage (Borago officinalis L.) oil, for example, is more complex
and contains approximately 37% linoleic acid and 21% GLA, i.e. a ratio of less than 2:1
(Senanayake and Shahidi, 2000). de Gyves (2004) demonstrated the concept by using
an Agrobacterium-mediated transformation procedure to deliver a cDNA encoding a 6desaturase
from borage under the control of a cauliflower mosaic virus (CaMV) 35S
35

promoter in “Rigel”. Analysis of the transformed plants demonstrated an altered profile
of polyunsaturated fatty acids, with an increase in GLA and octadecatetraenoic acid in
leaf tissues when compared with control lines. The fatty acid profile of the seed oil was
virtually unaltered, but this is not surprising as CaMV 35S is not a seed-specific
promoter.
Fieldsend (1996b) coined the term “factories in the field” to describe the cultivation of
evening primrose as a source of GLA. The farmer is the “factory manager”. Agricultural
research provides him with the correct “equipment” in the form of cultivars adapted to
his growing conditions, and the know-how, or “operating manual”, in the form of
husbandry guidance. Almost ten years later, the concept has been adopted as part of the
stakeholder proposal for the “Plants for the Future” technology platform (Anon, 2005).
In evening primrose, the advances envisaged by the technology programme have
already been achieved in practice.
REFERENCES
Anon. (2005): Plants for the Future. Stakeholder Proposal for a Strategic Research
Agenda 2025; Part 1: Summary, EPSO, 28 p.
Cleland, R.E. (1972): Oenothera, cytogenetics and evolution. Academic Press, London.
Fieldsend, A.F. (1996a): Evening primrose - from garden flower to oilseed crop. The
Horticulturist, 5: (3): 2-5 p.
Fieldsend, A.F. (1996b): Factories in the Field. Green Futures, 2: 20-21 p.
Fieldsend, A.F. (2003): Agronomic factors affecting the yield of crops of evening
primrose (Oenothera spp.) Buletinul USAMV-CN, A-H, 59/2003: 58-63 p. ISSN
1454-2382.
Fieldsend, A.F. and Morison, J.I.L. (2000a): Contrasting growth and dry matter
partitioning in winter and spring evening primrose crops (Oenothera spp.) Field
Crops Research, 68: 9-20 p.
Fieldsend, A.F. and Morison, J.I.L. (2000b): Climatic conditions during seed growth
significantly influence oil content and quality in winter and spring evening
primrose crops (Oenothera spp.) Industrial Crops and Products, 12: 137-147 p.
de Gyves E.M., Sparks C.A., Sayanova O., Lazzeri P., Napier J.A. and Jones H.D.
(2004): Genetic manipulation of γ-linolenic acid (GLA) synthesis in a commercial
variety of evening primrose (Oenothera sp.) Plant Biotechnology Journal, 2: (4):
351-357 p.
Horrobin, D. F. (1992): Nutritional and medical importance of gamma-linolenic acid.
Progress in Lipid Research, 31: 163-194 p.
Horrobin, D.F. (1994): Natural ≠ Safe. Pharmaceutical Technology Europe, December
1994, 2 pp.
Nix, J. (1995): Farm management pocketbook. 26 th edition. Ashford, Kent: Wye
College, University of London
Senanayake, S.P.J.N. and Shahidi, F. (2000): Lipid components of borage (Borago
officinalis L.) seeds and their changes during germination. Journal of the American
Oil Chemists' Society, 77: (1): 55-61 p.
36

ABSTRACT
SOME LESSONS LEARNED FROM THE NYÍRLUGOS
LONG-TERM FIELD EXPERIMENT
Imre Kádár
Research Institute for Soil Science and Agricultural Chemistry
H-1022, Budapest, Herman Ottó Str. 15, Hungary. E-mail: kadar@rissac.hu
The paper reports results achieved in a 44-year-old field trial set up on acid sandy
brown forest soil in the Nyírség region. Characteristics of the site: pH (KCl) 4.3-4.6,
humus 0.5-0.7%, CEC 3-4 meq/100 g in the ploughed layer. The topsoil was poor in all
five macronutrients (N, P, K, Ca, Mg) and the groundwater depth was 2-3 m. Fertilizers
applied in form of Ca-ammonium nitrate, superphosphate, potassium chloride,
powdered limestone and dolomite. The following conclusions could be drawn:
1. As the result of the regular N fertilization the pH (KCl) value in the topsoil dropped
to 3.5 (control plots 4.3), while it rose to 6.4 after addition of 1 t/ha/yr CaCO3. As a
function of liming, the exchangeable Ca 2+ enhanced from 0.13 to 2.18 meq/100 g,
while Al 3+ decreased from 0.68 to 0.40, Fe 2+ from 0.43 to 0.15 meq/100 g; CEC
increased from 3.3 to 3.6-3.8, S-value (sum of bases) from 0.4 to 2.5 meq/100 g, base
saturation (V) from 12% to 69%.
2. The NH4-acetate+EDTA soluble (Lakanen and Erviö 1971) element contents in
plowlayer also mirrored the changes caused by liming and fertilization. So, Ca lifted
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
to 0.53, Ni from 0.10 to 0.19 mg/kg.
3. Fertilizer responses were time dependent. In the 1 st decade of the experiment N alone
increased the potato and rye yields. In the 2 nd decade, the yield on N-plots declined
near to control. In the studied last few years, the yields on N-plots were negligible,
the soil lost its fertility and became leached and very acid. To maintain or increase
the yields of crops NPKCaMg fertilization was needed. Fertile plots have a pH (KCl)
5.5-6.0, 120-150 mg/kg ammoniumlactate-soluble P2O5 and K2O in plowlayer and
require the application of about 150 kg N and 1 t/ha ground dolomite yearly.
Keywords: fertilization, liming, time dependence, responses, long-term field
experiment
INTRODUCTION
There is a nearly universal acceptance of the fact that most long-term ecological
research is inherently interdisciplinary in nature. The successful performance of longterm
field experiments requires the special recognition of certain key elements in the
work. The scientific questions and objectives must be clearly defined and stated because
they must guide research for several generations of scientists, administrators and
funding agencies.
37

The people involved in the long-term field experiments must possess a shared
philosophy, appropriate training, the acceptance of a team leadership, and mutual trust
and respect. The site at which long-term field experiments are performed must be
representative of the biological and ecological (soil, weather, type of cultivation etc.)
setting. Both the site and research programme must have solid financial support and
continuity, and they must have a strong group of people associated with them. Of
course, the actual application and implementation of long-term field experiments may
vary from place to place and time to time.
Every human use of the natural ecosystem results in the removal of material and
embodied energy which are later not returned or broken down at the same place. This
means that human uses change natural ecosystems through different forms of matter and
energy transports, involving removal or supply. Long-term field experiments are vital in
order to represent cumulative effects and unusual events caused by human use,
agricultural practice, such as cultivation, manuring etc.
Long-term field experiments make visible processes and events often invisible in most
short-term experiments. As Magnuson et al. (1983) stated: "Because we are unable to
directly sense these slow changes and because we are even more limited in our abilities
to interpret cause and effect relations for these slow changes, processes acting over
decades are hidden and reside in the invisible present." This is the time scale of acid
deposition, the introduction of synthetic chemicals, acidification of soils, air borne
pollution and climate changes made by man.
Slow phenomena, or accumulative changes in soil properties may be captured only in
long-term studies. Limited duration might miss important results, or worse, cause the
results to be misinterpreted. Experiments in different years yield different results
because of weather changes. Long-term experiments are one of the sure ways to
determine slow processes.
The various experiments at Rothamsted, including both the classic and modern,
symbolize the value of long-term studies in general. The original goals of several of
these experiments have long since been fulfilled. However, they continue to be of value
as demonstrations, and as sources of continuing insight into agricultural practice
(Johnston 1989). In Hungary, most long-term field experiments are less than 40 years
old, though this number has been increasing since the late 1980’s, as the regional
research stations and institutes were established in the late 1950’s and 1960’s.
MATERIALS AND METHODS
The trial was established in autumn 1962 on brown forest soil, acid sand with thin
interstratified layers of colloid and sesquioxide accumulation called “kovárvány”. The
soil has a particle-size distribution in plow-layer as follows: sand over 0.05 mm 70-
85%, loam 0.05-0.002 mm 8-20%, clay under 0.002 mm 3-6%. Clay in colloid
accumulation layers makes up to 10-18%. The saturation percentage was 25-30, pH
(H2O) 5.4, pH (KCl) 4.3, humus 0.5-0.8%, CEC 3-5 meq/100 g. The site is situated
38

about 100 m above sea-level with yearly precipitation of 550-600 mm and sunny hours
1900-2000 h/year. The min/max temperature was about –25 Cº and +35 Cº interval, the
watertable level found at a depth of 2-3 m. The site is extremely drought sensitive.
The trial has 32 treatments x 4 replication = 128 plots with 5 x 10 = 50 m 2 plot size and
randomized block design. The forms of fertilizers applied were Ca-ammonium nitrate,
superphosphate, muriate of potash, powdered limestone and dolomite. The crop
sequence was potato-rye in the first 10 years, than potato-winter wheat (8 years), than
followed different crops the next 10 years (like white lupine, sunflower, grasses, spring
barley, tobacco). In 1991, the 29 th year of the trial, a triticale monoculture was
established which is now 16 years old. The nutrients applied in the trial are shown in
table 1.
Table 1: Fertilization and liming applied in the experiment, kg/ha/year (Brown forest
soil, acid sand, Nyírlugos, Nyírség region)
Nutrient
Applied nutrients, kg/ha/year
levels N P2O5 K2O CaCO3 MgCO3
0 0 0 0 0 0
1 50 60 60 250 140
2 100 120 120 500 280
3 150 180 180 1000 -
Remark: Ca-ammonium-nitrate, muriate of potash, powdered limestone and dolomite
The plant material was analysed after wet ashing with cc.HNO3 + cc.H2O2, for the N
determination with cc.H2SO4 + cc.H2O2 treatment. The soil samples were analysed
using NH4-acetate + EDTA method (Lakanen and Erviö 1971), basic soil properties
according to Baranyai et al. (1987), adsorptive soil characteristics according to
Bascomb (1964). All the mineral elements were measured using ICP technics. The
composite soil samples per plots were made of 20 subsamples, the composite plant
samples were made of 1 m 2 plant material per plot. Material and methods and the main
results of the experiment were summarized elsewhere (Láng 1963, 1973, 1984; Kádár
és Szemes 1990, 1994; Kádár et al. 1999; Márton 2002 a, b, c).
RESULTS AND DISCUSSIONS
As we can see in table 2 the value of pH (H2O) dropped down from 5.4 (control) to 4.6
on the N3 plot, while lifted up to 6.8 on the limed plot. The pH (KCl) values showed
analoge picture between 3.5 minimum and 6.4 maximum. The hydrolytic acidity
followed the movement of pH within a 4-14 interval. The humus content did not change
significantly in the ploughed layer as a function of fertilization and liming. The NH4acetate+EDTA
soluble phosphorus and potassium mirrored the P-and K-application.
The P-supply of soil turned to a satisfactory, while the K-supply of soil still remained
under a satisfactory level in the topsoil. The downward movement of K-fertilizer
probably enriched partly the subsoil.
39

Table 2: Effect of fertilization and liming on the soil chemical characteristics in 2006,
in the 44 th year of the trial. Ploughed layer.
Treatment
code
pH
(H2O)
pH
(KCl)
Hydrolytic
acidity,y1
Humus
%
NH4-acetate+EDTA
soluble
P2O5 mg/kg K2O mg/kg
Control 5.4 4.3 7.6 0.6 92 67
N1 5.0 4.2 9.6 0.8 85 43
N2 4.7 3.6 12.6 0.8 105 41
N3 4.6 3.5 13.6 0.7 89 44
N2P1 4.9 3.8 10.6 0.8 135 43
N2P2 4.6 3.9 11.6 0.9 158 50
N2P3 4.7 3.9 11.6 0.8 191 41
N2K1 4.7 3.7 11.4 0.8 101 56
N2K2 4.9 3.8 10.4 0.7 92 78
N2K3 4.8 3.7 11.1 0.8 99 73
N2P2K2 4.8 3.8 11.6 0.8 163 67
N2P2K2Ca3 6.8 6.4 3.6 0.7 225 62
N2P2K2Mg2 6.3 6.0 4.8 0.8 198 69
N2P2K2Ca2Mg2 6.7 6.2 3.8 0.7 220 65
LSD5% 0.3 0.5 2.1 0.3 41 12
Mean 5.2 4.3 9.5 0.8 139 57
Table 3 gives an overview about the effect of some treatments on soil chemical
properties in the topsoil as a function of pH and the total grain yields of triticale during
the last two 4-years periods. The close positive correlations among the pH (H2O), pH
(KCl) and hydrolytic acidity values are unambiguous. The exchangeable cations Al 3+
and Fe 2+ dropped down with increasing pH, while Ca 2+ and Mg 2+ ions lifting up when
Ca or Mg was applied. Exchangeable cations expressed in % the S-value (sum of bases)
show relative relationships among these elements.
On the leached and very acid soil Ca 2+ amounted altogether to 32%, while on the limed
one to 88%. In case of Mg 2+ , the N3 plot soil had 8%, while the fertilized with dolomite
one 22% relative weight. The K + made out 22% in the very acid soil, while 6% in the
limed soil. Considering the adsorption characteristics, one can state that the cation
exchange capacity shows increasing trend. The sum of bases and the % of base
saturation enhanced 5-6 times in the limed soils compared to that of the N3 plot (Table
3).
40

Table 3: Effect of treatments on soil chemical properties in ploughed layer in 2006
and on the cumulative yield of triticale. 2003-2006.
Measured and
Treatments code
LSD5%
calculated characteristics N3 N2 N1 Control NPKMg NPKCa
pH(H2O) 4.6 4.7 5.0 5.4 6.3 6.8 0.3
pH (KCl) 3.5 3.6 4.2 4.3 6.0 6.4 0.5
Hidrolytic acidity (y1) 13.6 12.6 9.6 7.6 4.8 3.6 2.1
Exchangeable cations, meq/100 g
Al 3+ 0.68 0.72 0.67 0.63 0.48 0.40 0.08
Fe 2+ 0.43 0.40 0.32 0.25 0.20 0.15 0.07
Ca 2+ 0.13 0.25 0.41 0.85 1.51 2.18 0.28
Mg 2+ 0.03 0.04 0.08 0.11 0.49 0.05 0.06
Adsorption characteristics
CEC, meq/100 g 3.3 3.4 3.4 3.4 3.8 3.6 0.4
S-value, meq/100 g 0.4 0.5 0.7 1.2 2.2 2.5 0.3
Base saturation, % 12 15 21 35 58 69 8
Exchangeable cations as % of sum of bases (S-value)
Ca 2+ 32 50 57 71 68 88 11
Na + 28 22 14 9 5 4 7
K + 22 16 13 12 6 6 7
Mg 2+ 8 8 11 9 22 2 5
NH4-acetate+EDTA soluble, mg/kg
Fe 152 158 124 104 121 111 25
Al 134 146 123 103 100 103 15
Ca 87 117 161 238 456 767 88
Mg 18 19 25 28 97 22 12
Mn 8 16 28 35 26 36 9
Sr 0.40 0.78 1.20 2.22 2.88 2.76 0.56
Co 0.15 0.30 0.47 0.47 0.35 0.53 0.13
Ni 0.10 0.16 0.16 0.20 0.18 0.19 0.04
Grain yield, t/ha/ 4 yrs* 6.1 9.3 8.6 6.1 18.0 18.5 3.2
Grain yield, t/ha/ 4 yrs** 1.9 4.8 5.5 3.9 10.0 10.6 2.6
*Sum of the 1999-2002. years
**Sum of the 2003-2006. years.
The NH4-acetate+EDTA soluble element contents change also as a function of the pH
(KCl). The content of Fe and Al diminished significantly with the increased pH (KCl)
values while the content of Ca, Mg and Mn enhanced. The limed soil was enriched also
41

with microelements Sr, Co and Ni. According to our analysis, the used powdered
limestone had 0.3% Mg, 70 mg/kg Mn, 158 mg/kg Sr, 1.3 mg/kg Ni and 0.2 mg/kg Co.
The used dolomit powder contained around 24% Ca, 12% Mg, 6-7 mg/kg Mn, 75 mg/kg
Sr and 0.4-0.5 mg/kg Ni. The content of Co was under detection limit of 0.08 mg/kg.
The Fe made out 730 mg/kg in limestone and 150 mg/kg in dolomite, while Al was 410
mg/kg in limestone and 140 mg/kg in dolomite powder. In spite of the Fe and Al input,
the NH4-acetate+EDTA soluble Fe and Al content in the ploughed layer diminished. So,
the solubility of these elements lessened, assessed with this method. The Sr is usually
moving in line with Ca both in rocks and soils which can be followed here. The same
time, the very low pool of Mn, Sr, Co and Ni in the extremely acid N3 soil might be a
consequence of leaching of these elements.
Table 4: Effect of fertilization and liming on the yield of triticale in the 41 st – 44 th
years of the experiment. (The 12 th – 16 th years of the triticale monoculture)
Treatments 2003 2004 2005 2006
code Grain Straw Grain Straw Grain Straw Grain Straw
Control 1.2 1.8 1.8 1.8 0.3 2.3 0.6 0.7
N1 1.2 1.8 3.6 3.9 0.3 2.7 0.4 0.6
N2 1.1 1.6 3.0 3.5 0.4 2.9 0.3 0.3
N3 0.6 1.0 1.1 1.5 0.1 1.2 0.1 0.2
N2P1 1.1 1.7 4.0 4.7 0.5 4.3 0.3 0.4
N2P2 1.8 2.5 4.6 5.3 0.4 3.2 0.3 0.2
N2P3 1.7 2.1 5.0 5.3 0.3 3.2 0.3 0.4
N2K1 1.1 1.5 2.2 3.0 0.2 2.1 0.2 0.2
N2K2 1.7 2.9 3.2 4.1 0.3 3.0 0.3 0.4
N2K3 1.0 1.6 2.3 2.6 0.3 3.0 0.4 0.3
N2P2K2 1.7 2.4 4.2 4.5 0.5 4.8 0.4 0.6
N2P2K2Ca3 1.8 2.7 5.6 5.8 0.9 9.1 2.1 1.8
N2P2K2Mg2 2.2 2.7 5.7 6.1 0.8 7.0 1.3 2.1
N2P2K2Ca2Mg2 2.1 3.3 6.7 6.7 0.9 8.1 1.9 2.5
LSD5% 0.6 0.8 1.5 1.4 0.2 1.8 0.7 0.6
Mean 1.5 2.1 3.8 4.2 0.4 4.1 0.6 0.8
The yield of triticale was depressed on the acidified soil. Fertilizer responses were time
dependent (Table 4). In the 1 st decade of the experiment N alone increased the potato
and rye yields. In the 2 nd decade, the yield on N-plots declined near to control. In the
studied last few years, the yields on N-plots were negligible, the soil lost its fertility,
became leached and very acid. To maintain or increase the yields of crops NPKCaMg
fertilization was needed. Fertile plots have a pH (KCl) 5.5-6.0, 120-150 mg/kg
42

ammoniumlactate-soluble P2O5 and K2O in plowlayer and require the application of
about 150 kg N and 1 t/ha ground dolomite yearly.
REFERENCES
Baranyai F.- Fekete A.- Kovács I. (1987): A magyarországi talaj tápanyagvizsgálatok
eredményei. Mezőgazdasági Kiadó. Budapest.
Bascomb, C. (1964): Rapid method for the determination of cation exchange capacity
of calcareous and noncalcareous soils. J. Soil Sci. Food Agri. 15:821-823.
Johnston, A.E. (1989): The value of long-term experiments, a personal view. In: Longterm
studies in ecology; approaches and alternatives. Ed: Linkens, E.G. 175-179.
Springer Verlag. New York. USA.
Kádár I.- Szemes I. (1990): Műtrágyázás és meszezés tartamhatásának vizsgálata
savanyú homoktalajon. Növénytermelés. 39:147-155.
Kádár I. – Szemes I. (1994): A nyírlugosi tartamkísérlet 30 éve. MTA Talajtani és
Agrokémiai Kutató Intézete. Budapest.
Kádár I. (1999): A hazai homoktalajok műtrágyaigényéről. Agrokémia és Talajtan.
48:217-223.
Lakanen, E. & Erviö, R. (1971): A comparison of eight extractants for the
determination of plant available micronutrients in soil. Acta Agr. Fennica.
123:223-232.
Láng I. (1963): A homoktalajok műtrágyázásának kérdései. MTA Agrártud. Oszt.
Közl. 22:431-434.
Láng I. (1973): Műtrágyázási tartamkísérletek homoktalajokon. Akad. Doktori
Disszertáció. Kézirat. MTA. Budapest.
Láng I. (1984): Homoktalajok termőképességének fokozása. Agrokémia és Talajtan.
14:75-87.
Magnuson, J.J., Bowser, C.J. & Beckel, A.L. (1983): The invisible present, long-term
ecological research on lakes. L and S Magazine. College of Letters and Science.
Univ. Wisconsin-Madison. Fall. 3-6.
Márton L. (2002a): A csapadék és a tápanyagellátás és az őszi búza (Triticum
aestivum L) termése közötti kapcsolat. Növénytermelés. 51:529-542.
Márton L. (2002b): A csapadék és tápanyagellátottság hatásának vizsgálata a tritikale
termésére tartamkísérletben. Növénytermelés. 51:687-701.
Márton L. (2002c): Az éghajlatingadozás és a N-műtrágyázás hatása a rozs (Secale
cereale L.) termésére. Növénytermelés. 51:199-210.
43

ABSTRACT
ANALYZE OF BIOMASS PRODUCTIVITY BY TIMESERIES
REMOTESENSING DATA IN REGION OF NYÍRLUGOS
János Tamás – Tibor Bíró – Nikolett Szőllősi
University of Debrecen Agricultural Sciences,
Faculty of Agronomy,
Department of Water and Environmental Management
The biomass growth can be described in regional space and time by continuous
equations. Effective calculation of biomass was not available because there was not
large scale resolution punctual data. We have introduced a math treatment first time in
Hungarian biomass research which is made by dissociation of 6 years time series from
earth satellite multispectral remote sensing data in region of Nyírlugos. This model
consists of linear trend, periodic, autoregressive and random component which were
successfully transformed forward. Result of trend analyze filtered out long term effect.
This means that the affects of biomass increase and decrease like wet or drought years
were detached. Long term climate effects forecast was difficult but periodic and
autoregressive component in short-term and in middle term forecast provided more
responsible values. Finished and practical method is more fitting than foregoing to
determinate biomass potential.
Keywords: biomass, time series, multispectral, remote sensing
INTRODUCTION
According to Csete and Láng sustainable competitiveness is accomplished through
sustainable farming methods, whose key motives are sustainable production,
adaptability, quality to any extent and favourable investments levels, consequently this
kind of competitiveness is altogether different from any old practice.
The U.S. Department of Agriculture (USDA) and the National Aeronautics and Space
Administration (NASA) signed a Memorandum of Understanding (MOU) to strengthen
future collaboration. In support of this collaboration, NASA and the USDA Foreign
Agricultural Service (FAS) jointly funded a new project to assimilate NASA's Moderate
Resolution Imaging Spectroradiometer (MODIS) data and products into an existing
decision support system (DSS) operated by the Production Estimates and Crop
Assessment Division. It produces objective, timely and regular assessments of global
agricultural production outlook and the conditions affecting food security. In monitoring
crop conditions for a specific region, remotely sensed vegetation index data are used to
track the evolution of the growing season compared to reference long-term mean
conditions (Tucker, 1985). A global normalized difference vegetation index (NDVI) is
produced from MODIS data, and is referred to as the "continuity index" similar to the
existing archive of NOAA-AVHRR derived NDVI . SPOT Vegetation NDVI data and
44

MODIS NDVI data from the Terra and Aqua platforms represent improvements in the
ability to monitor land photosynthetic capacity . The croplands are highly variable both
temporally and spatially. Croplands vary from year to year due to events such as
drought and fallow periods, and they vastly differ across the globe in accordance with
characteristics such as cropping intensity and field size (Anyamba, et al., 1998). To
describe this temporally heterogeneity a global NDVI time-series database, with a
spatial resolution of 250 meters has been assembled using a 16-day compositing period,
allowing for interannual comparisons of growing season dynamics (Tamás - Németh,
2005).
MATERIAL AND METHOD
Basic data source is made after dual average by MODIS Terra remote sensing data. This
MODIS NDVI dataset is reprojected and mosaicked to suit the Nyírlúgos region.
There was made maps in Nyírlugos which is after mean of territorial matrix of 10x10
km with 250 meter rescission. These data are monthly frequency data of 16 day period
which had respect for in 6 years duration. The analyzed areas are in black square in Fig
1. Bright fields signify higher biomass intensity.
RESULTS AND DISCUSSIONS
The progress of biomass process is a climate dependent periodicity in time so it is
manifest to analyze the observation of time series by time steps methods.
Forecast, operation, simulation are solvable this way. Basic data source consists of dual
averaging NDVI values counted by MODIS Terra remote sensing data. There was made
maps in Nyírlugos which is originated from means of territorial matrix of 10x10 km
with 250 meter resolution. These data are monthly frequency covering 16 day periods
which had respected for in 6 years duration (Figure 2.).
45

Figure 1. NDVI time series of Nyírlugos in 2006
(January-December from to left by lines)
46

NDVI
0,8
0,75
0,7
0,65
0,6
0,55
0,5
0,45
0,4
0,35
0,3
1 5 9 13 17 21 25 29 33 37 41 45 49 53 57 61 65 69
Time (m)
Figure 2. The time series of NDVI data sources (2001-2006, n=72)
Region of Nyírlúgos
Lineáris (Region of
Nyírlúgos)
The mean of presented time series of NDVI data sources in region of Nyírlugos is
0.536; standard deviation: 0.136; maximum: 0.759; while minimum: 0.316.
Discrete time steps in time describe phenomenon of biomass growth. The components
of time series model were determined in four steps. The applied theoretical model were
described by Kontur et al., (1993), used for another phenomenon (groundwater
hydrograph) which was continuous in time and space. This adopted time series analysis
based on the next formula:
Y = T + P + A + V
(eqv. 1.)
i
i
i
i
i
Where Ti trend, Pi periodic, Ai autoregressive and Vi contingent component. In addition
data sources in Figure 1. were terminated by equation 1.
First of all the trend was detached from the series for term 2001-2006. Due to this
process the long term effect was eliminated from the data source. This means the effect
of serial of wet and drought years resulted of biomass production changing was filtered
out.
The detachment of linear trend component:
Ti = d0 + d1* i (eqv. 2.)
i= 1, 2, …., N (timesteps)
47

With proper replacement the element of parameter vectors d0 and d1, independent
variable i=1, 2,…N, dependent variable Y1, Y2,… YN and errors y1, y2,…yN can be
obtained the series without trend.
d
2(
N − 2)
− 3
∗Y
− i ∗Yi
= 3
2(
N + 2)
− 3 N 1
−
3 2
0 +
where d0 = constant of trend function
d1 = coefficient of trend function
N = number of element
i = i th element
Y = mean of original time series
P i
= a
0
2π
2π
+ acos
∗i
+ bsin
∗i
r r
d
1
N + 1
− ∗Y
+ i ∗Yi
=
2
2
N −1
12
(eqv. 3.)
Calculated result of Equation 2. is the following: Ti=0.5113+0.0007i.
The Pi periodic component of time steps of annual biomass intensity is exists in
temperate climate. As next step the periodic component was detached by using the
model. In that case when the periodic time of periodic component is known, the
determination of the period amplitude has to be done only. We suppose there is a 12
months periodic in biomass change. The calculation of one time period is the following:
where:
(eqv. 4.)
a0 = constant of period
a és b = coefficient of function
The relevant period time is r = 12 months. The value of i can be counted in month. The
a0, a and b parameters were determined. In this case y1 time step without trend with Pi
periodic component was converged (Eqv. 5.):
yi = Pi + p (eqv.5.)
The value of pi is: pi = yi - Pi
The determination “a” and “b” (Eqv. 6.) is needed to count Pi.
N 2 2π
a = yi
cos i
N 12
∑
i=
1
N 2 2π
b = yi
sin i
N 12
48
∑
i=
1
(eqv. 6.)

The a*cos(2π*i/12) periodic component value was 0,000131776, while b*sin(2 π
i*i/12) periodic component value was 2,80205E-05 between 2001-2006. Pi periodic
component and substances are demonstrated by Figure 3.
c
0,2
0,15
0,1
0,05
0
-0,05
-0,1
-0,15
-0,2
1 5 9 13 17 21 25 29 33 37 41 45 49 53 57 61 65 69
p ′ ′ + V ′
′ i = c1
∗ pi−1
1
N −1
1
N
N −1
∑
i=
1
′ 1 = r1
=
N
∑
i=
1
p
p
i
2
i
p
i−1
i
a*cos(2π*i/12)
b*sin(2π*i/12)
Figure 3. Time steps of biomass production periodic component and substances
After the isolation of the periodic component from the data source the autoregressive
and random components are remained.
During the interpretation of the prepared model the autoregressive component expressed
the connecting effect of biomass periods (year by year) which are written in plant
production practice as year effect while random component determines the uncertainty
of model.
Then analyzes was continued with determination of pi random component supposing the
trend and periodic component effects are not exist anymore. So there is only
autocorrelation between time series values. The one step autoregressive sub-model is
the following:
Pi
(eqv. 7.)
where t refers time step and Vi is autoregressive component, the c1is constant of the
above function.
c1’equal to autocorrelation factor, which is rj (Eqv.8):
49
(eqv. 8.)

Value of c1’ is 0,274262498 while mean of Vi value is 0,04 the minimum 0,08 the
maximum 0,11. When taking into consideration the all regularity these values mean the
precision of the determination of biomass production. The random component depends
on the climate and cultivation condition mainly. But only that part of it which is not
affected by the periodicity and the trend. Higher random factor can be occur anywhere
(e.g. extreme year) but it is more likely on irregular increasing interval of biomass-curve
(e.g. indeterminate sow conditions).
SUMMARY
We have introduced a math treatment first time in Hungarian biomass research which is
made by dissociation of 6 years time series from earth satellite multispectral remote
sensing data in region of Nyírlugos. This model consists of linear trend, periodic,
autoregressive and random components which were successfully transformed forward.
Result of trend analyze filtered out long term effect. This means that the affects of
biomass increase and decrease like wet or drought years were detached.
With detaching periodic component sinus changing was separated from data source.
Autoregressive analyses quantify statistical rules which resulted from dependence of
sequential years.
Random component demonstrated the uncertainty of model. This is the factor which
makes biomass production characteristics undetermined in an added point or in
investigational space. This means by taking into consideration the all well-known
influential factor divergent value can be appeared according to the expected value.
After adjusting model component successfully backward transformation was carried out
which procedure proved the applicability clearly. This confirms the fact that significant
regularity can be assumed in biomass growth in such a dynamic model was set in this
study.
REFERENCES
Anyamba, A., J. Eastman R., Tucker, C. J. (1998). Warm Enso Event of 1997/98: NDVI
Precursors and Drought Pattern Prediction for Southern Africa. Greenbelt, NASA pp. 1-
40.
Csete, L., Láng, I.(2005) A fenntartható agrárgazdaság és vidékfejlesztés. Magyarország
az ezredfordulón.MTA Társadalomkutató Központ. Marosi-Print Kiadó. Budapest. 1-
313.
Kontur I., Koris K., Winter J. (1993). Hidrológiai számítások. Akadémiai Kiadó,
Budapest. pp. 143-184.
Tamás J., Németh T. (szerk.) (2005): Agrárkörnyezetvédelmi indikátorok elmélete és
gyakorlati alkalmazásai. Debreceni Egyetem, Debrecen, p. 138
Tucker, C. J., Vanpraet, C. L., Sharman, M. J , van Ittersum, G. (1985) Satellite remote
sensing of total herbaceous biomass production in the Senegalese Sahel: 1980–1984,”
Remote Sens. Environ., vol. 17, pp. 233–249,
50

RAINFALL, FERTILIZATION AND LIMING RESPONSE ON TRITICALE
(X Triticosecale W.) YIELD IN THE 44 YEAR OLD NYÍRLUGOS FIELD TRIAL
BETWEEN 1999 AND 2006
Márton László
Research Institute for Soil Science and Agricultural Chemistry of the Hungarian
Academy of Sciences (RISSAC-HAS)
1022 Budapest, Herman O. u. 15. Hungary, e-mail: marton@rissac.hu
ABSTRACT
Precipitation amount, distribution and nitrogen (N)-, phosphorus (P2O5)-, potassium
(K2O)-, calcium (CaO)-, and magnesium (MgO) fertilization interaction effects were
studied on a sandy acidic lessivated brown forest soil; WRB: Haplic Luvisol in the 44
year old Nyírlugos Field Trial (NYFT) in a Hungarian fragile agro-ecosystem in
Nyírség region (N: 47 0 41’ 60’’ and E: 22 0 2’ 80’’) on triticale (X Triticosecale W.)
yield from 1999 to 2006. At the trial set up in 1962, the soil had the following
agrochemical properties: pH (H2O) 5.9, pH (KCl) 4.7, hydrolytic acidity 8.4, hy1 0.3,
humus 0.7%, total N 34 mg kg -1 , ammonlactate (AL) soluble-P2O5 43 mg kg -1 , AL-K2O
60 mg kg -1 in the plowed (0-25 cm) layer. From 1980 to 2006, the experiment consisted
of 32x4=128 plots in randomised block design. The gross plot size was 10x5=50 m 2 .
The average fertilizer rates in kg ha -1 year -1 were nitrogen 75, phosphorus 90 (P2O5),
potassium 90 (K2O), calcium 437.5 (CaCO3) and magnesium 140 (MgCO3). The
groundwater table has had at a depth of 2-3 m below the surface. The main results are as
follows. During drought conditions the respective yield of the control areas was -25%
less than for average years. The application of N alone, or of NP and NK treatments, led
to yield reduction of -19.7%, while that of NPK, NPKCa, NPKMg or NPKCaMg caused
an -28.3% drop during these types of years. In the wet years, the yield decreased by -
22.2% on the unfertilized soils; in the case of N, NP, or NK nutrition with an -14.1%;
and increased at 13.8% on NPK, NPKCa, NPKMg and NPKCaMg treated plots. In the
very wettest year, the yields were dropped -43.1% on control soils, -39.3% of N, NP, or
NK loadings and -35.8% on NPK, NPKCa, NPKMg and NPKCaMg treatments to those
in the average year. The relationships between rainfall quantitiy during the vegetation
period N, P, K, Ca and Mg nutrition and yield were characterised by polynomial
correlations (control: R= 0.7212***, N: R = 0.7410***, NP: R = 0.6452***, NK: R =
0.6998***, NPK: R = 0.5555***, NPKCa: R = 0.5578***, NPKMg: R = 0.4869**,
NPKCaMg: R = 0.4341**). However, the total regression coefficients ranged from 0.43
to 0.74 in depence on the different nutrient application. Maximum yields of 5.8-6.0 t .
ha -1 were achieved in the rainfall range of 580-620 mm. At values above and below this
domain of the precipitation the grain yield reduced quadratically. So, it can be stated
that both, drought and excess rainfall conditions resulted dramatically in significant
negative effects between fertilization (N, P, K, Ca, Mg) and triticale yield.
Keywords: precipitation, fertilization, liming, triticale, yield
51

INTRODUCTION
The hazards associated with climate change are depend on the interaction of several
systems with many variables (Johnston, 2000). Accummulation of carbon dioxide,
methane, water vapor, ozone, nitrous oxide, sulfur hexafluoride, hydrofluorocarbons,
perfluorocarbons, chlorofluorocarbons (build-up of greenhouse gases) in the atmosphere
and trends in their emissions suggest that we can expect significant environmental
changes in the 21th century (Cynthia and Ana, 2006; Eric 2006). However, a recent
consensus has emerged that between the greenhouse gases rising of atmospheric
concentrations of carbon dioxide could become the more dangerous bacause it causing
the global warming (Láng, 2005).
Today, most researchers believe that higher temperature, drought and rainfall
excess caused by climate change will depress crop yields in many places in the coming
decades (Kádár et al., 1999; Jolánkai, 2005). Thus, in the last decades many agricultural
investigations focused on understanding the relation between mean climate change and
crop production (Runge, 1968; Várallyay, 1992). Changes in weather patterns were
observed thoughout Europe including Hungary as early as 1850. Among the natural
consequences of changing weather patterns, years of drought (rainfall deficit) and wet
(rainfall excess) conditions, resulted in problems among plant nutrition and field crop
production (EU, 2003). Triticale (Kádár et al., 1999; Márton, 2002) is a crop of
worldwide importance, limited research exists about the effects of climate change on
these crop. The crop is sensitive to the prevailing weather conditions (such as rainfall)
and, for this reason, understanding the effects of anthropogenic climate change on it’s
production is important (Márton, 2006). In addition to growing season triticale
conditions (e.g., soil agrochemical properties, fertilization, liming) affect the growth and
yield of crop (Lobell and Asner, 2003) and cause yield variations. Understanding the
fertilization, liming and rainfall effects have been a continuous endeavor toward
improving farming technology and management strategy to reduce the negative impacts
of fertilization, liming and rainfall and to increase crop yield (Kádár and Szemes 1994;
Várallyay 1994; among others).
Our main objective of this research it was study and clarify the precipitation
amount and distribution and nitrogen (N)-, phosphorus (P2O5)-, potassium (K2O)-,
calcium (CaO)-, and magnesium (MgO) fertilization interaction effects on a sandy
acidic lessivated brown forest soil; WRB: Haplic Luvisol in the 44 year old Nyírlugos
field trial in a Hungarian fragile agro-ecosystem in Nyírség region on triticale (X
Triticosecale W.) yield from 1999 to 2006. Furthermore, it was our intent to emphasize
that the net effect of multiple environmental changes is far more important than the
effect of a single factor on the crop.
MATERIALS AND METHODS
The net-influence of rainfall (quantity, distribution) and mineral fertilization (N, P2O5,
K2O, CaO, MgO) were studied in a long term field experiment established at the
Research Institute for Soil Science and Agricultural Chemistry of the Hungarian
Academy of Sciences Experiment Station (RISSAC-HAS ET) in Hungary on a Haplic
Luvisol (sandy acidic lessivated brown forest soil) with triticale (X Triticosecale W.)
indicator crop under fragile agro-ecological circumstances at Nyírlugos for 8 years from
52

1999 to 2006. The main experiment’s soil agrochemical characteristics in the plowed
(0-25 cm) layer are presented in Table 1. at the experimental set up in 1962 (Láng,
1973). From 1980 to 2006 the experiment consisted of 32x4=128 plots in randomized
block designs. The gross plot size has been having 10x5=50 m 2 . The experimental
treatments and combinations are shown in Table 2.
The fertilizers were applied in the form of 25% calcium ammonium nitrate, 18%
superphosphate, 40% potassium chloride, calcium carbonate and magnesium sulphate.
The groundwater table has had at a depth of 2-3 m below the surface. The plant samples
had had taken by manually at the harvest time. Rainfall amounts (deviation in rainfall
from the average over many years: dry year -10 - -20%, drought year -20% over, wet
year +10 - +20%, year with excess rainfall +20% over) and other related data
determined on traditional Hungarian (Harnos, 1993) and Research Institute for Soil
Science and Agricultural Chemistry of the Hungarian Academy of Sciences (Márton,
2004) standards, and MANOVA (Multivariate Analysis of Variance) by SPSS test
(SPSS Inc., 2000).
Table 1: The main soil agrochemical properties in the plowed (0-25 cm) layer at the
experiment set up. (Brown forest soil, acid sand; WRB: Haplic Luvisol, Nyírség
region Nyírlugos 1962)
Content PH HA* Hy1 Humus Total AL**
(H2O) (KCl) nitrogen P2O5 K2O
5.9 4.7 8.4 0.3
% 0.7
mg . kg -1 34 43 60
* Hydrolytic acidity, ** ammoniumlactate (AL) soluble
Table 2: Fertilization and liming treatments in the experiments, kg ha -1 year -1
(Brown forest soil, acid sand; WRB: Haplic Luvisol, Nyírség region Nyírlugos)
Applied kg ha -1 yr -1
Treatment
Levels N P2O5 K2O CaCO3 MgCO3
0 0 0 0 0 0
1 50 60 60 250 140
2 100 120 120 500 280
3 150 180 180 1000 -
Note: In the form of Ca-ammonium-nitrate, Superphosphate, potassium cloride,
powdered limestone and dolomite.
RESULTS AND DISCUSSION
During drought conditions, in conformity with Adams et al. (1995), Rosenzweig and
Tubiello (1997) and McMaster (1999) the respective yield of the control areas was -
25% less than for average years. The application of N alone, or of NP and NK
treatments, led to yield losses of -19.6%, while that of NPK, NPKCa, NPKMg or
53

NPKCaMg caused an -28.3% drop during these types of years. In the wet years, the
yield decreased by -22.2% in the unfertilized plots; in the case of N, NP, or NK
nutrition with an -14.1%; and increased at 13.8% on NPK, NPKCa, NPKMg and
NPKCaMg treatments. In the very wettest year, the yields were dropped -43.1% on
control soils, -39.3% of N, NP, or NK loadings and -35.8% on NPK, NPKCa, NPKMg
and NPKCaMg treatments to those in the average year, reverse of Asbjorn et al. (2004).
The relationships between rainfall quantitiy during the vegetation period N, P, K, Ca
and Mg nutrition and yield were characterised by polynomial correlations (control: R=
0.7212***, N: R = 0.7410***, NP: R = 0.6452***, NK: R = 0.6998***, NPK: R =
0.5555***, NPKCa: R = 0.5578***, NPKMg: R = 0.4869**, NPKCaMg: R =
0.4341**). However, the total regression coefficients ranged from 0.43 to 0.74 in
depence on the different nutrient application. Maximum yields of 5.8-6.0 t . ha -1 were
achieved in the rainfall range of 580-620 mm. At values above and below this range the
grain yield reduced quadratically.
To sum up we can say climate change will gradually and, at some point, be even
abruptly affects Hungary and Europa agriculture. Warming temperatures and a greather
incidence and intensity of extreme weather events possible lead to significant reductions
in triticale yield. Expanded ranges of crop agrochemicals and altered transmission
dynamics of different irrigation solutions might exacarbate these reductions. Since
farmers’ strategies grow out of experience, they can find that the past will be a less
reliable predictor of the future.
ACKNOWLEDGEMENTS
This research was supported by Hungarian Academy of Sciences, H-Budapest and the
Hungarian and Spanish Intergovernmental S & T Cooperation Project of E-2/04-
OMFB-00112/2005 and Hungarian and Indian Intergovernmental S & T Cooperation
Project of IND-3/03/2006.
REFERENCES
Adams, R. M., Fleming, R. A., Chang, C. C., McCarl, B. A. and Rosenzweig, C.
(1995): A reassessment of the economic effects of global climate change on U.S.
agriculture. Climatic Change, Vol. 30, pp. 147-167.
Asbjorn, T., Michelle, T. and Bárd, R. (2004): Climate Change Impacts on
Agricultural Productivity in Norway. CICERO. Oslo
Cynthia, R. & Ana, I. (2006): Potential impact of climate change on world food
supply. Data sets from a major crop modeling study. Socioeconomic Data and
Applications Center. Columbia University. New York
Eric La F. (2006): Adapting crops for climate change. UBC Botanical Garden and
Centre for Plant Researches.
EU (European Union). (2003): Drought costs EU farmers euro of 11 billion. European
Report, Brussels
Harnos, Zs. (1993): Weather and weather-yield interaction analysis. (In Hungarian) In:
Aszály 1983 (Szerk.: Baráth Cs-né., Győrffy B., Harnos Zs.). KÉE. Budapest
Johnston, A. E. (2000): Some aspects of nitrogen use efficiency in arable agriculture.
K. Scogs-o. Lantbr. Akad. Tidskr. 139:8.
54

Jolánkai M. (2005): Effect of climate change on plant cultivation. (In Hungarian) In:
„AGRO-21” Füzetek. 41. 47–58.
Kádár I., Németh T., Szemes I. (1999): Triticale trágyareakciója a nyírlugosi
tartamkísérletben. Növénytermelés. 48:647-661.
Kádár I., Szemes I. (1994): A nyírlugosi tartamkísérlet 30 éve. MTA Talajtani és
Agrokémiai Kutató Intézete. Budapest.
Láng I. (1973): Műtrágyázási tartamkísérletek homoktalajokon. Akad. Doktori
Disszertáció. Kézirat. MTA. Budapest.
Láng I. (2005): Weather and climate change: change-effect-response. (In Hungarian)
In: „AGRO-21” Füzetek. 43. 3–10.
Lobell, D. B., Asner, G. P. (2003): Climate and management contributions to recent
trends in U.S. agricultural yield. Science. 299. 1032.
Márton L. (2002): A csapadék és tápanyagellátottság hatásának vizsgálata a tritikale
termésére tartamkísérletben. Növénytermelés. 51:687-701.
Márton L. (2004): Rainfall and fertilization effects on crops yield in a global climate
change. In: Proc. 4 th Agroenviron Symposium. Role of Multipurpose Agriculture
in Sustaining Global Environment-AGROENVIRON 2004 (Udine, 20–24 Oct.,
2004). Part 3. 451–456. DPVTA. Udine
Márton L. (2006): Ecological changes of rainfall and artificial fertilization on crop
yield formation. ESA, Memphis, Tenesse
McMaster, H. J. (1999): The potential impact of global warming on Hail Losses to
winter crops in New South Wales. Climatic Change, Vol. 43, No. 2, October, pp.
455-476.
Rosenzweig, C. P. and F. N. Tubiello.( 1997): Impacts of global climate change on
Mediterranean agriculture: Current methodologies and future directions: An
introductory essay. Mitigation and Adaptation Strategies for Global Change, Vol.
1, pp. 219-232.
Runge, E. C. (1968): Effect of rainfall and temperature interaction during the growing
season on corn yield. Agron. J, 60:503-507.
SPSS. (2000): SigmaPlot for Windows. Ver. 3.2, Chicago, III.: SPSS, Inc
Várallyay, Gy. (1992): Globális klímaváltozások hatása a talajra. Effect of Global
Climate Change to soil. Magyar Tudomány, 9:1071-1076.
Várallyay Gy. (1994): A nyírlugosi tartamkísérlet talajszelvényeinek leírása és
laborvizs-gálati eredményei. In: A nyírlugosi tartamkísérlet 30 éve. 216-226.
(Kádár I.- Szemes I.) MTA Talajtani és Agrokémiai Kutató Intézete. Budapest.
55

EVALUATION OF THE DATA SET OF THE HUNGARIAN LONG-TERM
K-FERTILIZATION FIELD TRIALS, SET UP BETWEEN 1960 AND 2000
Péter Csathó
Research Institute for Soil Science and Agricultural Chemistry of the Hungarian
Academy of Sciences, H-1022 Budapest, Herman O. út 15, Hungary.
ABSTRACT
Potassium is considered to be the third most important macronutrient in Hungary
following nitrogen and phosphorus. The magnitude of responses to K application is
effected not only by the available K levels in the soil, but by the crops, too.
Evaluating the results of Hungarian 1- to 10-years-old long-term potassium fertilization
field experiments found in the literature between 1960 and 2000, conclusions were
made as follows:
The connection between the AL-K content of K-control (NP) plots and the
responses to K, expressed in relative yields (100Yield in NP plot/Yield in NPK plot,%)
could be described by a Mitscherlich-like equation, modified by Bray (1944): / Y'= 100
(1-10 -cx ) /, where x = AL-K content in the K-control (NP) plots; Y' = relative yield in
the trials, having "x" AL-K contents in the K-control plots; and c = proportional
constant (Mitscherlich's "working factor").
As a result of K application, surplusses in maize were higher than in winter wheat
or alfalfa. Average surplusses in maize grain yields varied between 0.0 and 1.9 t/ha, in
alfalfa between 0.0 and 1.6 t/ha and in winter wheat between 0.1 and 0.4 t/ha, resp.
The AL- (ammonium-lactate) (Egner, Riehm and Domingo 1960) soluble K indicated
the natural K- supplying power of the different soils adequately.
Keywords: field trials, potassium, soil texture, Bray-Mitscherlich approach, new
fertiliser recommendation system
INTRODUCTION
In the early 60's, when intensive (NP) K fertilization had not been introduced, about
half of the arable land in Hungary was poorly or moderately supplied with K. For that
reason the synthesis of the national field trial series of the above two crops is especially
important (Corey 1987). Alfalfa is also among the most important crops produced in the
country. As a first attempt, the data set of K fertilization trials with winter wheat, maize
and alfalfa has been established.
MATERIALS AND METHODS
A database of the results of 1- to 10-year-old Hungarian potassium field experiments
with maize, winter wheat and alfalfa, found in literature was compiled. A large number
of these trials belong to the National Long-term Field Trial Series with Fertilizers
(OMTK), supervised by the Pannon Agricultural University, Keszthely. In all trials,
by-products were removed from the fields. Soil characteristic data of NP (K-control)
treatments, grain yield data of NP and also NP+K treatments giving maximum
economic yields (about 95% of the maximum yields) were collected. The results were
56

classified according to the soil texture groups. The correlation between soil test (AL-K)
values and responses to K, using Bray's relative index method, was investigated. The
AL-method has been widely used in the Scandinavian countries, in the Netherlands, in
Portugal and in some Central European countries, and is still the official soil PK test
method in many of these countries.
RESULTS AND DISCUSSIONS
Evaluating the results of Hungarian 1- to 10-years-old potassium field experiments with
maize, winter wheat, and alfalfa found in literature between 1960 and 2000, conclusions
were made as follows:
As a result of previous NPK experiment series in Hungary with cereals and row
crops, N-responses were highest in both groups, which was followed by the Kresponses
in row crops and P-responses in cereals. N-responses were twice as high as
K- or P-responses (Várallyay sr 1950, Denke 1974a and 1974b, Debreczeni and
Debreczeni 1994).
Regarding the texture of Hungarian soils, 16% are sands, 10% sandy loams, 43%
loams, 19% clay loams and 7% are clayey soils.
The soils of the experimental fields did not receive previous significant potassium
doses so the results of the trials helped in estimating the original K supplying power of
the soils with different soil texture.
Soils with higher clay contents provided a better original (native) K-supplying power.
Among clay minerals, usually smectite was dominant in heavy soils, and illite in light
ones.
For estimating the extractable K-contents of the soils in Hungary, the AL-method
(0.1 mol/L ammonium lactate + 0.4 mol/L acetic acid, pH = 3.7, according to Egnér,
Riehm and Domingo (1960) has been official since the 60's.
On the basis of K field experiments with winter wheat, maize and alfalfa, carried
out between 1960 and 2000, the AL-method indicated the native K-supplying power of
the soils quite accurately (Figures 1 to 3). By this method probably not only the K
adsorbed and in the soil solution, but also some parts of fixed K was extracted.
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.
The connection between the AL-K content of K-control (NP) plots and the
responses to K, expressed in relative yields (100Yield in NP plot/ Yield in NPK plot,%)
could be described by a Mitscherlich-like equation, modified by Bray (1944): / Y'= 100
(1-10 -cx ) /, where x = AL-K content in the K-control (NP) plots; Y' = relative yield in
the trials, having "x" AL-K contents in the K-control plots; and c = proportional
constant (Mitscherlich's "working factor").
The "c" values - similarly to the findings of Bray (1944) - were different for maize
(0.0083 and for the other crops: alfalfa (0.0109) and for winter wheat (0.0153),
respectively.
The determination coefficient (R2) for maize and alfalfa trials was higher (0.77*** and
0.80***) than winter wheat trials (0.67***), respectively.
The connection between the AL-K contents of K-controls and surplusses (yield in
NPK-NP, t/ha) for the same crops could be described by a hyperbolic curve.
57

Relative yield (100 NP/NPK), %
Relative yield (100 NP/NPK), %
1.A. Winter wheat 1.B. Winter wheat
Y’ = 100 (1-10 –cx ); n = 59; Y’ = 1/(ax+b); n = 59
c: = 0.0153; r = 0.67
4.0
a = 0.04; b = -0.29; r = 0.72
100
80
60
40
20
0
0 100 200 300 400 500
AL-K2O, mg/kg
Grain increases (NPK-NP), t/ha
3.0
2.0
1.0
0.0
0 100 200 300 400 500
AL-K2O, mg/kg
2..A. Maize 2.B. Maize
Y’ = 100 (1-10 –cx ); n = 86; Y’ = 1/(ax+b); n = 86
c: = 0.0083; r = 0.77 a = 0.01; b = -0.02; r = 0.70
100
4.0
80
60
40
20
0
0 100 200 300 400 500 0 100 200 300 400 500
AL-K2O, mg/kg
AL-K2O, mg/kg
Figures 1 and 2. Relationship between the AL-K2O contents of K control (NP) plots and
responses of winter wheat and corn to K fertilization, calculated from the database of
Hungarian field trials on K fertilization, 1960–2000, using the Bray-Mitscherlich approach
Responses to K fertilization were eliminated when soil K test values exceeded 160-170
mg/kg for winter wheat, 170-180 mg/kg for alfalfa and 200 mg/kg for maize (Figure 1-
3.).
58
Grain increases (NPK-NP), t/ha
3.0
2.0
1.0
0.0

Relatív termés (100 NP/NPK), %
3.A. Alfalfa 3.B. Alfalfa
Y’ = 100 (1-10 –cx ); n = 23; Y’ = 1/(ax+b); n = 23
c: = 0.0109; r = 0.80 a = 0.03; b = 0.94; r = 0.74
100
4.0
80
60
40
20
0
0 100 200 300 400 500
AL-K2O, mg/kg
Terméstöbblet (NPK-NP), t/ha
3.0
2.0
1.0
0.0
0 100 200 300 400 500
AL-K2O, mg/kg
Figure 3. Relationship between the AL-K 2O contents of K control (NP) plots and responses of
alfalfa to K fertilization, calculated from the database of Hungarian field trials on K
fertilization, 1960–2000, using the Bray-Mitscherlich approach
The AL- (ammonium-lactate) (Egner, Riehm and Domingo 1960) soluble K indicated
the natural K- supplying power of the different soils adequately. With higher soil clay
contents in the trials, the original K supplying capacity of the soils was usually better.
Maize (and wheat) responses to K showed a better correlation with the soil clay
contents, than with the clay mineral compositions, resp.
Table 1. The effect of soil texture on responses to K application in Hungarian field trials with
maize, set up between 1960 and 2000
Soil
texture
groups
Number
of trials
(n)
homok h. vályog vályog a. vályog agyag
K A * AL-K 2 O
mg/kg
(NP)
K 2 O dose
kg/ha**
(K-control)
Grain yield
on NP plots
(NP/NPK)
Relative
yield,%
(NPK-NP)
Surplus,
t/ha
(NPK-NP)
Sands 13 29 80 120 4,27 71 1,91
S. loams 8 36 128 100 5,54 88 0,71
Loams 20 39 159 54 6,17 93 0,43
C. loams 27 45 187 33 6,26 97 0,19
Clay soils 5 55 206 0 7,81 100 0,00
Mean 73 40 155 59 5,90 90 0,61
*Upper limit of plasticity, according to Arany (similar to the US plasticity index).
** K 2 O doses needed for maximum economic yields (~ 95% of maximum yield)
59

Table 2. The effect of soil texture on responses to K application in Hungarian field trials with
winter wheat, set up between 1960 and 2000
Soil
texture
groups
Number
of trials
(n)
K A * AL-K 2 O
mg/kg
(NP)
K 2 O dose
kg/ha**
(K-control)
Grain yield
on NP plots
(NP/NPK)
Relative
yield,%
(NPK-NP)
Surplus,
t/ha
(NPK-NP)
Sands 5 28 75 112 3,08 89 0,41
S. loams 7 35 120 66 4,22 94 0,32
Loams 17 39 160 15 4,15 98 0,06
C. loams 20 46 210 7 3,98 98 0,08
Clay soils 5 56 218 0 4,41 100 0,00
Mean 54 42 171 26 4,02 97 0,13
*Upper limit of plasticity, according to Arany (similar to the US plasticity index).
** K 2 O doses needed for maximum economic yields (~ 95% of maximum yield)
Table 3. The effect of soil texture on responses to K application in Hungarian field trials with
alfalfa, set up between 1960 and 2000
Soil
texture
groups
Number
of trials
(n)
K A * AL-K 2 O
mg/kg
(NP)
K 2 O dose
kg/ha**
(K-control)
Grain yield
on NP plots
(NP/NPK)
Relative
yield,%
(NPK-NP)
Surplus,
t/ha
(NPK-NP)
Sands 2 27 60 120 5.55 78 1.60
S. loams 3 35 163 13 8.42 98 0.26
Loams 2 42 205 46 8.95 90 0.74
C. loams 12 45 172 16 7.74 98 0.18
Clay soils 3 55 150 0 8.50 100 0.00
Mean 22 40 175 20 8,92 96 0,33
*Upper limit of plasticity, according to Arany (similar to the US plasticity index).
** K 2 O doses needed for maximum economic yields (~ 95% of maximum yield)
On the basis of these data sets, new AL-K 2 O supply categories were elaborated for
maize, winter wheat and alfalfa, according to their different responses to K fertilization.
The new AL-K 2 O supply categories for clay loam and clay soils are much lower
than those in former fertilizer advisory systems.
CONCLUSIONS
For evaluating the connection between AL-K2O values and responses to K fertilization
in Hungarian field trials with maize, alfalfa and winter wheat, Bray's relative index
method was a useful tool. The difference between the responses of maize, winter wheat
and alfalfa to K fertilization was also indicated in the different "c" values. AL- K values
indicated the K-supply of the different soils adequately.
60

ACKNOWLEDGEMENT
This study was financially supported by the Hungarian National Scientific Research
Fund (OTKA), under grant N.o. T 06511.
REFERENCES
Bray, R. H. (1944) Soil-plant relations: I. The quantitative relation of exchangeable
potassium to crop yields and to crop response to potash additions. Soil Sci. 58,
305-324.
Corey, R.B. (1987) Soil test procedures: Correlation. In: Soil testing: Sampling,
correlation, calibration, and interpretation (ed.: Brown, J.R.) 15-22. SSSA Special
Publ. N.O. 21. Madison, Wisc. USA.
Csathó, P. (1994) Database of Hungarian K-fertilization field trials with maize and
winter wheat, 1960-1990. In: (Eds.: Borin, M. and Sattin, M.) Proc. 3rd ESA
Congress, Abano-Padova, Italy. 464-465.
Denke, J. (1974a) (Ed.) Results of fertilization researches. 2. Maize. KAE Keszthely,
Hungary. 72 p. (In Hungarian)
Denke, J. (1974b) (Ed.) Results of fertilization researches. 5. Alfalfa. KAE Keszthely,
Hungary. 48 p. (In Hungarian)
Debreczeni, B. and Debreczeni, K. (eds.). (1994) Fertilization Researches, 1960-1990.
Akadémiai Kiadó, Budapest. (In Hungarian)
Egner, H., et al. (1960) Untersuchungen über die chemische Bodenanalyse als
Grundlage für die Beurteilung des Nahrstoffzustandes der Böden. II. Kungl.
Lantbrukshögskolans Annaler, 26: 199-215.
Sárdi, K. and Németh, T. (1993) Studies on the plant available K-content of different
soils at constant moisture. In "Proc. Symp. on Soil Resilience and Sustainable
Land Use" (Ed. Szabolcs I) Agrokémia és Talajtan 42,183-194.
Várallyay, Gy. sr. (1950) Experiments and analyses directing fertilizer use. Agrokémia
2. 287-302.
61

LONG-TERM FIELD EXPERIMENTS AS THE BASE OF SITE SPECIFIC
FARMING
KESZTHELY LONG-TERM FIELD EXPERIMENTS
Tamas Kismányoky – Zoltan Toth
University of Pannonia Georgikon Faculty of Agriculture
Long-term field experiments existed, exist and will exist at least in places, where their
importance is recognised. Long-term field experiments are the source of such iformation
from which we can learn a lot about the factors that influence soil fertility and its
sustainability. In most cases effects and interactions can be understood only from longterm
data even if different soil and climatical conditions are compared to each other.
These information are valuable for farmers, extension consultants, decision makers as
well as scientists either in local or wide scale general concern.
Long-term field experiments can be considered such living laboratories that let us
study the physical and biological parameters of the agricultural environment reliably,
further give scientists and policy decesion makers basic information in order to follow
and control environmental mechanisms.
The survey and the register of the European long-term field experiments were
published by Körschens (1994, 1997, 1999, 2000). Valuable publications were also
made in this topic by Mazur et. al (1993), Debreczeni B. (1994), Leigh and Johnston
(1994), Merbach et. al (1999), Smith et. al (2001), Láng et. al (2002), Debreczeni K and
Körschens, M. (2003).
Most of the long-term field experiments were set up to study the effects and
interactions of crop rotation, continuous cropping, organic and mineral fertilizers,
microelements, liming, soil tillage, water management, plant protection, nutrient
respond of new varieties and hi-breds, etc. Similar goals were the reason for the
estabilishment of the long-term field experiments in Keszthely, Hungary.
The long-term field experiments of the World and Hungary are listed in Table 1.,
Table 2. and Table 3.
Table 1. Long-term field experiments of the World
(Debreczeni K., Körschens, M. 2003)
Continent 10-20 20-50 50-100 > 100 year Total
Europe 24 292 81 20 417
Africa 32 37 69
Asia 28 7 35
Australia 7 14 21
North-America 1 41 21 5 68
South-America 9 1 10
World’s total 25 409 161 25 620
62

Table 2. The oldest and most important long-term field experiments of the World
(Körschens et. al., 1994)
Place Country Year of estabilishment
Rothamsted England 1843
Grignon French 1875
Illionis USA 1876
Halle/Saale Germany 1878
Columbia USA 1888
Dakota USA 1892
Askov Denmark 1894
Auburn USA 1896
Bad Lauchstädt Germany 1902
Dikopshof Germany 1904
Table 3. The oldest long-term field experiments of Hungary
Place Year of estabilishment
MTA TAKI 1959
Putnok 1963
Kompolt 1961
Szarvas 1989
Kecskemét 1959/60
Gödöllő 1970
Karcag 1977
Debrecen 1978
GKI 1979
Gyöngyös 1994
Westsik 1929
Nyirlugos 1962
OMTK 1967
Keszthely 1963
Martonvásár 1959/61
The crop production research of Europe and Hungary has several common aspects
either in the fields of research, or in the structure of the research organisations or in the
principle of funding research. On the other hand the process of implementation is not
uniform. In the western European countries the crop production research is made by the
universities and by regional research institutes. The research institutes are financed by
the administration, or by private organisations. A part of the agrotechnical research is
non profit type (eg. plant nutrition, crop rotation, soil management), while others like
breeding or pesticide research are profit oriented ones.
Obviously on those fields, where the experimental results has a direct return (new
species, or hibred, new pesticide or machine etc.) the financial background of the
research is evident. There is a completely opposit situation in case of those researches,
63

that have higher costs than their direct incomes, since the reliable results can be reached
and published after several replication and besides this publications are available for
everyone wihout any compensation, but on the other hand they are important for the
society.
The results of the applied agrotechnical research can be transfered to the practice
by extension effectively, if the research institutes are located in that particular region.
One of the prominent examples for this is the National Long-term Field Experiment
Network.
A scientific school was formed since 1945 in Keszthely, Hungary, based on the
successful research activity of several decades in the field of soil fertility and nutrient
management. Dr. Kolbai Károly, Dr. Kemenesy Ernő, Dr. Nyéki Jenő, Dr. Láng Géza,
Dr. Kováts András professors had an unexegerateble role in this development.
At the beginning research was aimed at organic (farmyard) manure treatments and
application, delivering useful results for the practice.
Since the 1960-es, when the mineral fertilizer application rate increased
dramatically, the research was focused on the effective application of fertilizers in order
to utilize the genetical potential of the different species and hibreds.
Although mineral fertilizer application had been playing the main role in the
maintainance of soil fertility, the new questions of organic matter and organic manures
had to be studed as well. In this period the research work of the department included the
study of the utilization of agricultural by products like straw (incorporation or burning)
as well as slurry.
The studies covered the specifications of technologies according to the different
requirements of the species, varieties and hibreeds (plant nutrition, sowing date, crop
rotation, plant density, tillage methods, etc.) as well as the methods of physical and
chemical improvement of soils.
The researh activity of the department focused mainly on the yield improvement
and cost effective cropping methodes of the leader field crops as wheat, maize, barley,
potato, sunflower, rape, sugarbeet, and others (soybeen, fababeen).
This studies and trials were conducted in small and large plot field experiments on
30 ha arable land in three different sites (Keszthely, Szentgyörgyvölgy,
Homokszentgyörgy-Mariettapuszta) and also in pot experiments with a well equipped
lab. The three different experimental site had different type of forest soils. Ramann-type
brown forest soil (Keszthely), Pseudogleyic brown forest soil (Szentgyörgyvölgy) and
leached sandy soil (Homokszentgyörgy-Mariettapuszta). The results from this different
study sites can be applied for the most part of the Transdanubian region, where the
different classes of the forest soils are typical.
The half of the long-term field experiments were set up in 1963-1970 (Láng Géza,
Kemenesy Ernő, Kováts András, Nyéki Jenő, Cseh Ervin, Németh István). This trials
represent high scientific values. From the 2-3-4 decade long database reliable
conclusions can be drawn about the alteration of soil fertility as well as about the effects
of the different crop years in these different agroecological regions.
The great increase in the volumen of mineral fertilizer production and application –
beside the previously started experiments – required a country scale study of the
application of fertilizers in order to provide a fertilization advisory system based on
uniform basic principles. A board was estabilished for this purpose and worked out the
64

plan of the uniform countrywide field trial network in 1965. In this board the staff of the
Department of Plant Production in Keszthely played significant role (Láng Géza,
Mártonffy Tamás). Then in 1966 the administration allocated funds for the
estabilishment of this trial network and field experiments have been set up in 18 sites
operated by 9 research organisations (institutes and universities). In the next step the
number of the experimental sites rised up to 26, from which 3 (Keszthely, Lábod-
Vadaspuszta, Nagykanizsa) belonged to the Department of Plant Production in
Keszthely.
As a recognation of the research work of the department the Ministry of
Agriculture and Food Industry estabilished an operational board in 1968 in order to
improve and help the new uniform field trial network which was chaired by Dr. Láng
Géza, and the secretary was Dr. Mártonffy Tamás. The expenditures of the experiments
were covered by different funding programs (MÉM MÜFA fund, OMFB, Chemical
Industry Union SZP-2 program and G-9 program since 1986). The Department of Plant
Production was in charge of the operation of this new National Field Trial (NFT)
Network until 1985, then it was taken over by the Department of Agrochemistry under
Dr. Debreczeni Béla’s and Dr. Debreczeni Béláné’s leadership. In 2002 the operation of
the network got back to the Department of Plant Production and Soil Science.
Nowadays the number of the experimental sites of the NFT network has reduced to 9
and the financial sources run out of supplies.
The success of the research activity of the department was recognised by the
following facts (it was considered as a school and a scientific workshop).
The Ministry of Agriculture and Food Industry launched different middle-term
operative research programs in 1970, from which the program of fertilization
(„Research on Soil Power Management”) was run by the department with great
experience in this field with academician Láng Géza’s leadership. He was also in charge
of the domestic and international public relations, as well as of the coordination of the
research in this field by making research plans and comprehensive reports.
In 1971 the ministry tended to rise the research focusing on the improvement of
soil fertility as the basic factor of the increase of production and technological progress
to the priority of the country scale research programs. Dr. Géza Láng was the person in
charge of the preliminary arrangements as a head of the operating board.
In 1972 the ministry decided to start the country scale research program: K-9
„Improvement of soil fertility…”. The program had several chapters as fertilization, soil
improvement, soil tillage and soil conservation. The coordinator of the research
program was Dr. Láng Géza professor the head of the operating board and senior
researcher of the Keszthely university of Agriculture. The subprogram of soil fertility
was operated directly also by Keszthely. This program worked until 1980. Since 1981
the subprograms continued as independent programs. The soil fertility subprogram was
continued by the department under the title of „ SZP-2 Agrochemical research”. Since
1986 it was runed as the project No. 6 of the G-9 Research Program.
The department research activity covered the field of the „Study of soil
acidification as a result of intensive plant production” connecting to the soil fertility
research program between 1982-1986. In the frame of the project the reasons for soil
acidification and the methods of their compensation were studied on different soil types
65

and in field and pot experiments. Farm scale data are also were processed for this
purpose.
The next important work (1986-1990) in this field was connected to the project No.
OTKA-513 called „Factors influencing the utilization of yield potential of the new
varieties, the dinamics of plant growth affected by the environmental conditions in case
of some arable crop species and varieties.” The research was carried out either in longterm
or in annual field experiments in Keszthely, Hungary.
In the following years the centralized systematic funding of these research projects
by the administration came to an end and the R+D sources of the institutions decreased
dramatically. In the following years the funding of long-term field experiments have
been based on short-term projects (OTKA, FVM, NKFP, GVOP, etc.) and their
existence is quite uncertain nowadays.
Since the ’90-es the long-term field experiments have been run by the today’s
generation of researchers. The main long-term field experiments of the department are
as follows:
• Comparative study of organic and mineral fertilizer application (1960-63)
• Minimum tillage experiment in wheat-maize diculture (1972)
• Mineral and organic fertilization in different crop rotations (1963)
• Nitrogen fertilization experiment with the study of residual effect (1965 Keszthely,
1965 Szentgyörgyvölgy)
• Phosphorus fertilization experiment with the study of residual effect (1963
Keszthely, 1965 Szentgyörgyvölgy)
• Potassium fertilization experiment with the study of residual effect (1963
Keszthely, 1965 Szentgyörgyvölgy)
• Mineral fertilization in continuous maize cropping experiment (1969)
• <strong>International</strong> organic and mineral nitrogen fertilization experiment (1983)
• National Long-term Field Experiment (OMTK) Network trial (1967-69)
The results of the long-term field experiments are published in several journals,
conference proceedings and books. The aim of our field research is to develop such
systems of fertilization and agrotechnics that resulted in the greatest yields in given
agroecological condition without environmental hazards.
During the history of the agricultural research the aim of the trials was always to
clarify the limiting factors. While in the ’60-es farmers had to be persuaded about the
effects of mineral fertilizer application now beside the clarification of particular
questions the integrated polyfactorial and ecosystem oriented approach of research is
the typical.
SUMMARY
The importance and necessity of long-term field experiments:
• Long-term effects can only be studed after several decades reliably.
• The agrotechnical development (chemical, biological, etc.) of the last decades can
be measured by long-term data and the plans for the future should also be based on
this data base.
• Nutrient balances can be estimated reliably.
66

• The effect of climate change can be estimated by the comparison of the long-term
data of the different study sites.
• Long-term database is the background of the computer models.
• Long-term field experiments are the references of the the sustainable development
of agriculture and environment.
REFERNCES
Debreczeni, B., Debreczeni, Bné. (1994) Trágyázási kutatások 1960-1990. Akadémiai
Kiadó Budapest, p. 411.
Debreczeni, K., Körschens, M. (2003) Long-Term Field Experiments of the World
Archives of Agronomy and Soil Science, October 2003, Vol. 49, pp. 465-483.
Körschens, M. (2000) IOSDV <strong>International</strong>e organische
Stickstoffdauerdüngungsversuche, UFZ-Bericht No./15/2000. Leipzig-Halle
GMBH. ISSN 0948-9452, p. 192.
Körschens, M. (1999) Experimentelle Möglichkeiten zur Ableitung optimaler Corg-
Gehalt in Ackerboden., UFZ-Bericht No./9/1999. Beziehungen Zwischenb
organischer Bodensubstanz und bodenmikrobiologischen Prozessen. Leipzig-Halle
GMBH. ISSN 0948-9452, pp. 75-94.
Körschens, M. (1997) Die Wichtigsten Dauerfeldversuche der Welt-Übersicht,
Bedeutung, Ergebnisse. Arch. Acker- Pfl. Boden., Vol. 42, pp. 157-168.
Körschens, M. (1994) Der Statische Düngungsversuch Bad Lauchstädt nach 90 Jahren.
B.G. Teubner Verlagsgesellschaft Stuttgart-Leipzig, p. 179.
Láng, I., Lazányi, J. and Németh, T. (2002) Tartamkísérletek, tájtermesztés,
vidékfejlesztés. Nemzetközi Konferencia I., Debrecen, Nyírlugos, Nyíregyháza,
Livada Sárköz, Románia 2002. június 6-8. ISBN 963-472-654-20 p. 418.
Leigh, R. A. and Johnston, A. E. (1994) Long-term Experiments in Agricultural and
Ecological Sciences. Proceedings of a conference to celebrate the 150 th
Anniversary of rothamsted Experimental Station, held at Rothamsted, 14-17 July
1993, p. 428.
Mazur, K., Filipek, J. and Mazur, B. (1993) Proceedings of the <strong>International</strong>
Symposium ’Long-term static fertilizer experiments’ June 15-18, 1993. Warszawa-
Kraków, Poland. 253.
Merbach, W., Schmidt, L., Garz, J., Stumpe, H. und Schliephake, W. (1999) Die
Dauerdüngungsversuche in Halle-ein Überblick. UFZ-Bericht No.24/1999
Dauerdüngungsversuche als Grundlage für nachhaltige Landnutzung und
Quantifizierung von Stoffkreisläufen, ISSN 0948-9452.
Smith, P., Fallon, P., Smith, J. U. and Powlson, D. S. (2001) GCTE Global Change
and Terrestrial Ecosystems. Report No 7. 2 nd edn. Soil Organic Matter Network
(SOMNET): Model and Experomental Metadata. Published by the CGTE Focus 3
Office Wallingford, UK, p. 223.
67

LONG-TERM EFFECT OF ORGANIC AND MINERAL FERTILIZATION ON
DIFFERENT SOIL-FERTILITY PARAMETERS
ABSTRACT
Sándor Hoffmann and Katalin Berecz
University of Pannonia, Georgikon Faculty of Agricultural Sciences
Deák F. str. 16, H-8360 Keszthely/Hungary
Eleven variations of farmyard manuring (FYM) and mineral fertilization treatments of a
long-term field trial have been selected for the present investigations. Attributes of
changes in soil fertility were investigated. FYM and equivalent NPK fertilization
resulted in similar pHH2O values, while FYM increased TOC content of the soil by 10%
as compared to equivalent NPK fertilizer. FYM increased K, while equivalent doses of
NPK fertilizer increased P content of soil to a higher degree, as compared to nil plot.
Yearly 14 t ha -1 FYM or equivalent NPK fertilizer kept soil P and K contents at the
range of “medium-well-supplied”. However, not even yearly 21 t ha -1 of FYM resulted
in maximum yields in 1998-2006. 35; 70 and 105 t FYM ha -1 5 year -1 increased grain
yields to a lesser extent (by 0.66, 1.44 and 0.85 t ha -1 , respectively), as compared to
equivalent NPK doses, which was mainly due to the better utilization of mineral
fertilizer. After harvesting all the plots contained relatively high amounts of nitrate in
the 0-300 cm soil layer. Nitrate content of the non-treated plot was considerably high as
well. Despite the low yield response to FYM-treatments, less nitrate was found in the
soil, than with mineral fertilization.
INTRODUCTION
Field experiments are essential means for studying the changes in soil fertility as
affected by different fertilization and farming systems. With changing economical and
social conditions and farming systems, also the judgement of the role of farmyard
manuring (FYM) kept changing. The effect of FYM on soil fertility is very different.
However, the nutrient content of FYM have the most direct and highest effect, which
can be compared to the effect of equivalent NPK fertilizer doses. This effectiveness is
influenced by various factors and it is generally lower, than in case of fertilizers with
the same active ingredients and equivalent doses. Nevertheless, this tendency may turn
with the increase of clay content in the soil (Sarkadi, 1993). The NPK fertilizer
equivalency of FYM can be determined not only in experiments comparing the effect of
FYM and NPK fertilizers of equivalent doses, but also in combined polyfactorial
experiments (Baldock and Musgrave, 1980; Bouldin et al., 1984), and the results are
nearly similar (Hoffmann, 1999). Many authors detected that the nutrient effect of FYM
can be fully substituted for fertilization (Stumpe et al., 2000), others founded that the
effect of fertilization is higher except for the case of very heavy soils (Árendás and
Csathó, 1994). On the other hand, the secondary effect of FYM, namely the increase in
general soil fertility is very important and it manifests itself in the structure, organic
matter composition and microbial activity of soil, and this effect can not be substituted
for mineral N-fertilization in many instances (Albert, 2001).
68

MATERIALS AND METHODS
An organic and mineral fertilization field experiment was established with a randomized
block design, four replications, and two crop rotations on Keszthely in 1963. The initial
soil (Eutric Cambisol) parameters of the long-term experiment were as follows: (KA =
37; pHH2O = 7.3, TOC = 0.87; AL-P2O5 = 27; AL-K2O = 135 mg kg -1 ). The plot size was
98 m 2 . In the present study, only the results of crop rotation potato - maize - maize -
winter wheat - winter wheat and 11 of the 15 treatments of the experiment (35, 70, 105 t
ha -1 FYM or mineral fertilizer with equivalent NPK doses or FYM+NPK combinations
as shown in Table 1) are discussed. In case of simple, double and triple doses, FYM was
distributed before sowing of the potato for 5 years, timed half-and-half to the potato for
5 years, and to the maize (in the autumn of the previous year) in the 3 rd year,
respectively. Grain yields in cereal units [CU] (potato tuber yields were multiplied by
0.3.) in 1998-2006, pHH2O, organic carbon (TOC), P, K and mineral N-content of the 0-
300 cm soil layer (in increments of 30 cm, 4 sample per plot), as attributes of changes in
soil fertility were evaluated.
Composite soil samples were taken from the 0-20 cm soil layer after harvest, 10
samples for each plot. P and K content of the soil was determined applying ammonium
lactate extractant, which is used in Hungary for routine soil tests (Egner-Rhiem-
Domingo, 1960). TOC contents in the ploughed soil layer were measured by the
classical methods of Turin. Mineral-N was extracted with 1% KCl then NH4-N and
NO3-N were analysed by steam distillation and titration procedure.
Treatment
No.
Table 1. Selected treatments of the experiment
Codes of treatments kg NPK ha -1 yr -1
1. Control -
2. 1 FYM N44P38K49
3. 2 FYM N88P76K98
4. 3 FYM N132P114K147
5. 1 eqv N44P38K49
6. 2 eqv N88P76K98
7. 3 eqv N132P114K147
8. 1 FYM + 3 eqv N176P152K196
9. 4 eqv N176P152K196
10. 1 FYM + NPK N172P110K181
11. 1 eqv + NPK N172P110K181
Legends: 1 FYM= 35 t ha -1 farmyard manure in 5 years
1 eqv = mineral NPK equivalent to 35 t ha -1 FYM in 5 years
NPK = N640P360K660 in 5 years
69

RESULTS AND DISCUSSION
Soil pHH20 in the control plot was 7.02, which is 0.3 pHH2O lesser than the initial value
(7.3). Very similar pH values were measured with the different treatments and the
different fertilizer type or doses practically did not change the soil acidity. On an
average of the doses, 40 years long-term FYM and equivalent NPK treatments resulted
in pHH20 values of 6.94 and 6.99, respectively (Figure 1). Not even the treatments with
the highest NPK content caused lower pH values.
pH H2O
7,6
7,4
7,2
7
6,8
6,6
6,4
6,2
6
5,8
5,6
5,4
5,2
5
control
ab
1FYM
ab
1eqv
ab
2FYM
ab
2eqv
a
3FYM
ab
3eqv
ab
1FYM+3eqv
ab
Figure 1: pHH2O values of soil
4eqv
b
1FYM+NPK
ab
1eqv+NPK
TOC content of soil was evaluated in two sampling dates, in the 20 th and 40 th years of
the field experiment. The type of the fertilizer influenced the organic matter content of
the soil definitely (Figure 2) FYM treatments alone increased TOC by 17% on the
average, while equivalent NPK doses only by 7%, as compared to the control plot. A
clear- cut conclusion about the effect of combined treatments on the TOC content can
not be drawn. No definite change in the TOC content could be detected as an affect of
20- or 40-years organic and/or mineral fertilization.
70
ab

TOC %
1,2
1
0,8
0,6
0,4
0,2
0
bc b c c
bc bc
ab c
bcab
ab
ab ab
ab
ab ab
a aab
a
a
Cont
1FYM
2FYM
3FYM
1 eqv
2 eqv
3 eqv
1FYM+3eqv
4 eqv
Figure 2: TOC content of soil
1FYM+NPK
1eqv+NPK
Figures 3 and 4 show data about the long-term impact of organic or mineral
fertilization on the phosphorus and potassium content. FYM and equivalent NPK
influenced soil P and K content differently. While mineral NPK fertilization resulted in
22 mg kg -1 more P2O5 in the soil than FYM, K2O content of the soil in the FYM treated
plots was 26 mg kg -1 higher than in case of mineral fertilization.
In spite of the differences between the effect of the organic and mineral fertilizers,
it can be stated, that yearly14 t ha -1 FYM (70 t ha -1 5 yr -1 ) or equivalent PK fertilization
can keep soil P and K contents at the range of “medium-well-supplied”, taking into
consideration the absolute soil P and K levels.
P 2O 5
mg kg
250
-1
200
150
100
50
0
Cont
1FYM
2FYM
3FYM
1 eqv
2 eqv
3 eqv
1FYM+3eqv
4 eqv
Figure 3: AL-P2O5 content of soil
71
1FYM+NPK
1eqv+NPK

K 2O mg kg -1
300
250
200
150
100
50
0
Cont
1FYM
2FYM
3FYM
1 eqv
2 eqv
3 eqv
1FYM+3eqv
4 eqv
1FYM+NPK
Figure 4: AL-K2O content of soil
1eqv+NPK
Figure 5 shows the crop yields expressed in cereal units and in the average of 1998-
2006.
t ha
8
7
6
5
4
3
2
1
0
-1
control
a
1FYM
b
2FYM
b
3FYM
c
Crop yields 1998-2006
1eqv
c
2eqv
d
3eqv
de
1FYM+3eqv
de
4eqv
de
1FYM+NPK
Figure 5: Crop yields (CU) t ha -1 1998-2006
e
1eqv+NPK
Despite the relative high soil P and K levels, neither 14 t ha -1 yr -1 nor 21 t ha -1 yr -1
FYM resulted in maximum yields. The organic fertilizer doses, (35; 70 and 105 t
FYM ha -1 5 year -1 ) increased grain yields to a lesser extent (by 0.66, 1.44 and 0.85 t ha -
1 , respectively), as compared to equivalent NPK doses. The average crop yield
amounted to 5.103 t ha -1 with FYM, while to 6.154 t ha -1 with equivalent NPK. The
increase was 28% as compared to the control, which was mainly due to the better
utilization of fertilizer.
72
e

The much better performance of the mineral nutrient form can be the result of the better
utilization, especially that of N.
The higher TOC contents as a result of the FYM treatments suggest, that higher N
quantities were incorporated into the humic substances from the organic fertilizer,
which resulted in a lesser N supply and lower crop yields. The highest yields could be
achieved with the combined treatments with high NPK content
NO3-N (mg/kg)
0 20
0
30
60
90
120
150
180
210
240
270
300
Control
NO3-N (mg/kg)
1 FYM
NO3-N (mg/kg)
1 eqv FYM
NO3 – N mgkg -1
NO3-N (mg/kg)
2 FYM
NO3-N (mg/kg)
2 eqv FYM
NO3-N (mg/kg)
Figure 6 :Nitrate-N content of soil (0-300cm)mgkg -1
After harvest all the plots contained relatively high amounts of nitrate in the 0-300 cm
soil layer (Figure 6). Nitrate content of the non-treated plot was considerably high as
well. Despite the low yield response to FYM-treatments, less nitrate was found in the
soil, than with mineral fertilization. The lower nitrate reserves in the soil with FYM
treatments, in parallel with the lower crop yields, also confirms our suggestion that
higher N quantities were incorporated into the humic substances from the organic than
from the mineral fertilizers.
The amount of ammonium was higher, but it did not show any correlation with
fertilization.
73
3 FYM
NO3-N (mg/kg)
3 eqv FYM
NO3-N (mg/kg)
4 eqv FYM

REFERENCES
Albert, E. (2001): Effect of long-term different mineral and organic fertilization on
yields, humus content, netN-mineralization and N-balance. Arch. Acker-Pfl.
Boden., Vol. 46,. 187-213 p.
Árendás T., Csathó P. (1994): Azonos NPK – hatóanyagú szerves- és műtrágyázás
hatása a talajtulajdonságok függvényében Agrokémia és Talajtan 43: 399-408 p.
Baldock, J.O.,Musgrave, R.B. (1980): Manure and mineral fertilizer effect in
continous and rotational crop sequences in Central New York. Agron. J. 72, 511-
518 p.
Bouldin, D.R., Klauser, S.D., Ried; W.S. (1984): Use of nitrogen from manure.
Nitrogen in crop production, Madison Wisconsin USA.. 221-243 p.
Egner, H., Rhiem, H., Domingo. W.R. (1960): Untersuchungen über die chemische
Bodenanalyse als Grundlage für die Beurteilung des Nährstoffzustandes der
Boden. II. K. Lantbr. Hogsk. Ann. 26:199 p.
Hoffmann, S. (1999): Az istállótrágya N-egyenértékének megállapítása nem
hatóanyagazonosságra alapozott tartamkísérletekben. „Növénytermesztés és
környezetvédelem”. Szerk.: Ruzsányi-Pepó. MTA Agrártudományok Osztály
Kiadványa. Budapest, 97-101 p.
Sarkadi, J. (1993): Szerves- és műtrágyák tápelemtartalmának érvényesülése
tartamkísérletekben. I. Nitrogénforgalom. Agrokémia és Talajtan 42: 293-308 p.
Stumpe, H. et al. (2000): Effect of humus content, farmyard manuring, and mineral-N
fertilization on yields and soil properties in a long-term trial. J. Plant Nutr. Soil.
Sci. 163: 657-662 p.
74

GREENHOUSE GAS EMISSION IN LONG-TERM TILLAGE AND
ROTATION EXPERIMENTS
1 Anita Gál, 1 Péter Hegymegi, 1 Erika Michéli, 2 Tony J. Vyn
1 Szent István University, Soil Science and Agrochemistry Department, Gödöllő, HU
2 Purdue University, Department of Agronomy, West Lafayette, Indiana, USA
ABSTRACT
Greenhouse gases such as carbon-dioxide (CO2), and nitrous oxide (N2O) are strongly
linked to global warming. Soil organic matter is influenced by tillage and rotation, and
through its decomposition and accumulation, soils have a major effect on the gas
composition of the atmosphere. Tillage systems vary substantially in the intensity and
depth of soil disturbance, in crop residue placement, and in the resulting depths of soil
carbon gains or losses. The actual effect of different tillage practices on soil carbon
storage is highly dependent on the crops produced in the field. Several studies have
been conducted to determine the quantity and distribution of organic carbon and total
nitrogen under different tillage and rotation systems in long-term studies but less is
known about the relationship of OC and N stored in the soil and the emission of CO2,
and N2O from the soil over the crop production cycle. In our research we have
compared the long-term effects of conservation (no-till) and conventional (chisel plow
and moldboard plow) tillage systems on soil CO2, and N2O emission, in continuous corn
and soybean-corn rotation. Our results show that both tillage and rotation systems
influence significantly gas emission. Carbon-dioxide and nitrous-oxide emission
declines in the growing season with time. Nitrous-oxide emission increases after
application of nitrogen fertilizer and after heavy rain events. Corn monoculture gives
the highest emission of carbon-dioxide, and also for nitrous-oxide, rotations including
corn or corn monoculture have the highest emission. As for tillage treatments, chisel
plowing results in the highest emission of nitrous-oxide.
Keywords: organic matter, carbon sequestration, no-tillage, gas emission, rotation
INTRODUCTION
Greenhouse gases such as carbon-dioxide (CO2), and nitrous oxide (N2O) are strongly
linked to global warming because of their effects on radiation (Sparks, 2003). Organic
matter is a major soil component which is influenced by tillage and rotation, and
through its decomposition and accumulation, soils have a major effect on the gas
composition of the atmosphere (Eswaran et al., 1993, Kimble et al., 1990).
Tillage systems vary substantially in the intensity and depth of soil disturbance, in
crop residue placement, and in the resulting depths of soil carbon gains or losses. In notill,
which is the ultimate conservation tillage system, soil disturbance is limited to the
opening of a small slot for seed and fertilizer placement. In contrast, moldboard plowing
inverts the soil and most surface residue to a depth between 20-25 cm (Phillips et al.,
1980). Chisel plowing loosens soil to similar depths but does not invert the soil profile,
so it distributes residues more evenly and at shallower depths than moldboard plow.
After several years, soils which have undergone no-till have a higher carbon
75

concentration in the surface layer than moldboard plowed or chiseled plots. On the other
hand, carbon levels at lower depths are generally similar in both systems, or higher
under moldboard plow (Angers et al. 1997, Gal, 2005).
The actual effect of different tillage practices on soil carbon storage is highly
dependent on the crops produced in the field. Crop rotation contributes to increased
yield (Mitchell et al., 1991) and nitrogen contribution by legumes can be very
important. Crop rotation is known to increase soil microbial activity, which has the
direct effect of breaking down organic matter by the microbes. Residue quantity is the
main factor in determining soil organic carbon, for example lower soil organic carbon
level corresponds with sequences including soybean, but organic carbon increases as
soybean is not present or corn is present in a sequence according to Studdert et al.
(2000). However, the effect of residue amount is conditioned by several other factors.
Several studies have been conducted to determine the quantity and distribution of
organic carbon and total nitrogen under different tillage and rotation systems in longterm
studies (Kern and Johnson, 1993; Angers et al., 1997; VandenBygaart et al., 2002).
Gal et al. (2007) have found on the same experimental plots that while no-till clearly
resulted in more organic carbon and nitrogen accumulation in the surface 15 cm than
moldboard plow, the relative no-till advantage declined sharply witd depth. Indeed,
moldboard plow resulted in more organic carbon and total nitrogen int he 30-50 cm
depth interval. However, less is known about the relationship of OC and N stored in the
soil and the emission of CO2, and N2O from the soil over the crop production cycle. In
order to understand the effect of different tillage and rotation systems on carbon
sequestration - which is the long-term storage of carbon in soil and can help to stop the
increase of greenhouse gases in the atmosphere - we have to examine these processes
more in details. Extended knowledge of the factors influencing soil capacity to be a net
sink or source of greenhouse gases in the atmosphere is one of the most relevant and
urgent needs in agricultural policy making and also in extension.
The objective of the research was to compare the long-term effects of conservation
(no-till) and conventional (chisel plow and moldboard plow) tillage systems on soil
CO2, and N2O emission, in continuous corn and soybean-corn rotation.
MATERIALS AND METHODS
Research area
The sampling site chosen for the study is located at Purdue University (West Lafayette,
IN, USA), at the Agronomy Center for Research and Education. The soil is a poorly
drained Chalmers silty clay loam (Typic Haplaquoll according to the US Taxonomy)
dark, with approximately 4 % organic matter in the surface 30 cm, developed under
prairie vegetation (Vyn et al., 2000). The Long-Term tillage experiment has been
conducted for 30 years. Moldboard plow, chisel plow and no-till tillage treatments were
compared in continuous corn and soybean-corn rotation.
The experiment is a randomized complete block in a split-plot (small plots) design
with treatments replicated four times. Crop rotations are the main units, and the tillage
treatments the subunits. In the course of this experiment, starter N (36 kg N ha -1 as 34-0-
0) and side-dressed N (222 kg N ha -1 as 28% urea ammonium nitrate, UAN) were
applied.
76

Carbon-dioxide and nitrous-oxide sampling
Carbon-dioxide and nitrous-oxide fluxes were measured by the vented flux chamber
method. In each measurement plot, duplicate anchors measuring 73.7 X 35.4 X 12.0 cm
capped with U-shaped channels (1.8 cm wide by 1.9 cm deep) welded to the outer-edge
were driven about 10 cm into the soil. Within each plot, anchors were placed 10 m
apart and a carpenter’s level was used to ensure that the anchors were level with the soil
surface. Gas fluxes were measured by placing chambers that measured 75.8 X 38 X 13
cm over the anchors to cover the soil surface. To ensure that there was no gas exchange
between chamber and atmosphere during sampling, the U-shaped channel of the anchor
was filled with water to form an air-tight seal before sampling began. Samples were
then collected from the chambers through a rubber septum at regular intervals of 0, 5,
10, and 15 min after deployment using a 20 mL polypropylene syringe and pressurized
into 12 mL pre-evacuated vials. Gas sampling was done 14 times ont he following days:
June 15, 19, 20, 22, 27, 29; July 6, 10, 14, 21, 26; and August 4, 9, 16. CO2, and N2O
concentration of the gas samples was determined using Varian CP 3800 Gas
Chromatograph.
Soil temperature and soil moisture data
Soil moisture content (%) and soil temperature (°C) measurement was done together
with gas sampling on the same sampling points. Soil moisture was determined with a
TRIME-TDR equipment at depth of 12 cm. Soil temperature was measured with a
thermometer at 10 cm depth close to the gas samling chambers. Daily precipitation
(mm) and average daily air temperature (°C) data were compiled from the
meteorological station of the experimental farm (http://agmetx.agry.purdue.edu).
Statistical analysis
Statistical analysis was performed with SAS Version 8.2 program (SAS Institute, Inc.,
2002). Multivariate linear regression analysis was carried out to determine the effect of
soil moisture content, soil temperature and day of sampling (after application of
nitrogen fertilizer), besides the effect of tillage and rotation treatments for carbondioxide
and nitrous-oxide emission. Predicted values of gas emission were calculated
using the regression equations.
RESULTS AND DISCUSSION
Carbon-dioxide emission
Carbon-dioxide emission was significantly (P=0,05) influenced by soil temperature and
sampling day. Carbon-dioxide emission was the highest in no-till , although the
emission was higher than 6000 mg/m 2 /hr in all treatments; among crop rotations corn
monoculture showed the highest emission values. Figure 1 shows the predicted values
of CO2 emission influenced by soil temperature and sampling day.
77

CO 2 flux (mg/m 2 /hr)
CO 2 emission (mg/m 2 /hr) BC-CH+NT vs BC-PL vs CC-CH vs CC-NT vs CC-PL
12000
10000
8000
6000
4000
2000
60
50
40
30
Day
20
Day vs Temp vs Pred BC CH+NT CO 2
Day vs Temp vs Pred BC PL CO 2
Day vs Temp vs Pred CC CH CO 2
Day vs Temp vs Pred CC NT CO 2
Day vs Temp vs Pred CC PL CO 2
10
20
22
24
26
0
Soil temperature ( o C)
Figure 1 Predicted values of CO2 emission in soybean-corn rotation and corn
monoculture
Regression equations (R 2 = 0,330973):
BC CH+NT CO2 = 3016,09 + 179,75 Temp – 1,78 Temp * Day
BC PL CO2 = 1996,81 + 179,75 Temp – 1,78 Temp * Day
CC CH CO2 = 4674,25 + 179,75 Temp – 2,92 Temp * Day
CC NT CO2 = 5177,25 + 179,75 Temp – 2,92 Temp * Day
CC PL CO2 = 5347,96 + 179,75 Temp – 2,92 Temp * Day
78
28
30

Nitrous-oxide emission
Nitrous-oxide emission was influenced significantly (P=0,05) by soil temperature and
soil moisture content. Among the three tillage treatments chisel plowing resulted in the
highest nitrous-oxide emission, and there was no significant difference between
moldboard plowing and no-till, so these latter treatments were considered as one group
on the figures. Among crop rotations, corn monoculture had higher values than
soybean-corn rotation, however these differences were not significant. Figure 2 shows
the predicted and actual values of N2O emission influenced by soil temperature and soil
moisture content in soybean-corn rotation and in corn monoculture.
N 2 O flux (mg/m 2 /hr)
70
60
50
40
30
20
10
0
-10
20
22
24
26
28
30
Soil temperature ( o C)
N 2 O emission (mg/m 2 /hr) CH vs PL+NT
80
70
60
50
Soil moisture (%)
40
30
20
Pred Moist vs Pred Temp vs CH Pred
Pred Moist vs Pred Temp vs PL-NT Pred
CH Moist vs CH Temp vs CH N2O
PL-NT Moist vs PL-NT Temp vs PL-NT N2O
Figure 2. Actual and predicted values of N2O emission in soybean-corn rotation and
in corn monoculture (effect of tillage treatments)
Regression equations (R 2 = 0,282134):
79

CH N2O = - 8,856 + 0,701 * Moist – 0,012 Moist * Temp
PL + NT N2O = - 8,856 + 0,607 * Moist – 0,012 Moist * Temp
Although there was no significant difference between crop rotations, Figure 3. shows
the predicted values in soybean-corn rotation and in corn monoculture.
N 2 O flux (mg/m 2 /hr)
40
30
20
10
0
Soil temperature ( o C)
-10
20
2224262830
N 2 O emission (mg/m 2 /hr) CH-BC vs CH-CC vs PL+NT-BC vs PL+NT-CC
80
70
60
50
Soil moisture (%)
40
Moist vs Temp vs CH-BC Pred N2O
Moist vs Temp vs CH-CC Pred N2O
Moist vs Temp vs PL+NT-BC Pred N2O
Moist vs Temp vs PL+NT-CC Pred N2O
Figure 3. Predicted values of N2O emission in soybean-corn rotation and in corn
monoculture (effect of tillage and rotation)
Regression equations (R 2 = 0,287105):
80
30
2

CH BC N2O = - 8,839 + 0,6904 * Moist – 0,0125 Moist * Temp
CH CC N2O = - 8,839 + 0,6904 * Moist – 0,0114 Moist * Temp
PL + NT BC N2O = - 8,839 + 0,5961 * Moist – 0,0125 Moist * Temp
PL + NT CC N2O = - 8,839 + 0,5961 * Moist – 0,0114 Moist * Temp
Carbon-dioxide and nitrous-oxide emission declined further in the growing season,
mainly, because the amount of nutrients necessary for microbial activity
(undecomposed crop residues and nitrogen fertilizer) has declined in the soil with time.
Nitrous-oxide emission increased after application of nitrogen fertilizer and after heavy
rain events (approximately 30-40 mm). Corn monoculture seems to give the highest
emission of carbon-dioxide, and also for nitrous-oxide, rotations including corn or corn
monoculture have the highest emission. As for tillage treatments, chisel plowing results
in the highest emission of nitrous-oxide.
Based on the results of our study from 2002-2004 we concluded that tillage effects
are more important in organic carbon and nitrogen storage than crop rotation effects.
We have found, that despite the 30 % higher total crop residue returned in continuous
corn, the soybean-corn and continuous corn rotations resulted in similar organic carbon
and nitrogen levels. The greater CO2 and N2O emission in continuous corn relative to
soybean-corn rotation explains the similar organic carbon and nitrogen storage under
the two crop sequences.
REFERENCES
Angers, D.A., M.A. Bolinder, M.R. Carter, E.G. Gregorich, C.F. Drury, B.C.
Liang, P.R. Voroney, R.R. Simard, R.G. Donald, R.P. Beyaert, and J. Martel.
1997. Impact of tillage practices on organic carbon and nitrogen storage in cool,
humid soils of eastern Canada. Soil and Tillage Research 41:191-201.
Eswaran, H., E. Van Den Berg, and P. Reich. 1993. Organic Carbon in Soils of the
World. Soil Science Society of America Journal 57:192-194.
Gál, A. 2005. Depth dependency of soil carbon changes with long-term tillage and
rotation systems. MSc thesis, Purdue University
Gál A., T. J. Vyn, E. Michéli, E. J. Kladivko, William W. McFee. 2007. Soil carbon
and nitrogen accumulation with long-term no-till versus moldboard plowing
overestimated with tilled-zone sampling depths. Soil and Tillage Research. (in
press, doi 10.1016 j.still.2007.02.007)
Kimble, J., T. Cook, and H. Eswaran. 1990. Organic matter in soils of the tropics .
p.250-258. In Proc. Symp. Characterization and role of organic matter in different
soils. Int. Congr. Soil sci. 14 th, Kyoto, Japan. 12-18 Aug. 1990. ISSS,
Wageningen, the Netherlands.
Kern, J.S., and M.G. Johnson. 1993. Conservation tillage impacts on national soil and
atmospheric carbon levels. Soil Science Society of America Journal 57:200-210.
Mitchell, C.C., R.L. Westerman, J.R. Brown, and T.R. Peck. 1991. Overview of
long-term agronomic research. Agronomy Journal 83:24.
Omonode, R.A., Gál, A., Stott, D.E., Abney, T.S., Vyn, T.J. Short-term versus
continuous chisel and no-till effects on soil carbon and nitrogen. Soil Science
Society of Agronomy Journal 70:419-425.
81

Phillips, R.E., R.L. Blevins, G.W. Thomas, W.W. Frye, and S.H. Phillips. 1980. Notillage
agriculture. Science 208:1108-1113.
Sparks, D.L. 2003. Chemistry of soil organic matter, p. 75-112 Environmental Soil
Chemistry, Second ed. Academic Press (Elsevier Science).
Studdert, G.A., and H.E. Echeverria. 2000. Crop rotations and nitrogen fertilization
to manage soil organic carbon dynamics. Soil Science Society of America Journal
64:1496-1503.
VandenBygaart, A.J., X.M. Yang, B.D. Kay, and J.D. Aspinall. 2002. Variability in
carbon sequestration potential in no-till soil landscapes of southern Ontario. Soil
and Tillage Research 65:231-241.
ACKNOWLEDGEMENT
Anita Gál received a Hungarian State Eötvös Scholarship in 2006 for the duration of the
study.
82

INFLUENCES OF AMELIORATIVE FERTILIZATION ON YIELD
AND NUTRITIONAL STATUS OF MAIZE
Kovačević Vlado , Josipovic Marko 2 , Stojić Biserka 3
1 University J. J. Strossmayer, Faculty of Agriculture HR-31000 Osijek, Croatia
2 Agricultural Institute, Juzno predgradje 17, HR-31000 Osiejk, Croatia
3 Petrokemija d.d. Fertilizer Factory HR-44320 Kutina, Croatia
ABSTRACT
Suboptimal levels of plant available nutrients are oft limiting factor of field crop yields
under environmental conditions, especially in middle and north-western part of Croatia.
Farmers mainly have not basic information about their soils (for example, their
agrochemical characteristics). In frame of education of farmers in Djulovac (Bjelovar-
Bilogora County), Pakrac (Pozega-Slavonia County) and Okucani (Brod-Posavina
County) municipalities during March of 2003 total 108 soil samples (0-30 cm depth)
were taken form unfertilized fields (total 279 ha) and soil testing was made (soil pH,
plant available phosphorus and potassium according AL-method). About 60% of tested
soil samples are acid ( pH in KCl less than 5). Also, about 40% of soil samples are poor
or moderate supplied with phosphorus (P), while potassium (K) status is better because
only 13% samples are in this range of K availability. Based on soil test, choice of plots
for fertilization experiment was made. Aim of this study was testing of ameliorative
fertilization (spring 2003) with P and K in form of NPK fertilizer on yield and
nutritional status of maize (the growing seasons 2003-2005). In the second and third
year of testing only standard fertilization of the experiment was applied. Maize
responded to applied fertilization by moderate yield increases up to 10% only. We
presume that Al-method is not suitable solution for prediction of plant available P status
in this soil. Ameliorative fertilization resulted by significant influences on increased
plant available P, K and Fe status in the soil, while differences of the other tested
elements (S, Ca, Mg, Mn, Zn, Cu and Cd) were in level of statistical error. The applied
fertilization had the most influences on maize nutritional status in the first year of
testing (2003) because significant differences for P, K, Mn (increasing trend) Ca, Mg,
Mn and Zn (decreasing trend) were found. In the second year (2004) non-significant
differences were found, while in the third year (2005) significant differences were found
for P, K and Mg only. It is important that cadmium status in maize plant was
independent on applied fertilization because there are examples in literature that
application of the high P rates is mainly in connection with increases of Cd
concentrations in plant. Ameliorative fertilization had mainly low influences on maize
grain composition because only in the third year of testing significant differences for K,
Mg (increases) and Zn (decrease) status were found. Cd concentrations in maize grain
were under detectable range of the applied method (

humanity organization, in cooperation with Croatian Co-operative Association and
Croatian Agricultural Extension Service, in frame of ECRA project – revitalization of
war affected communities- which are supported by USAID agency, began education of
farmers in Djulovac (Bjelovar-Bilogora County), Pakrac (Pozega-Slavonia County) and
Okucani (Brod-Posavina County) municipalities. During March of 2003 total 108 soil
samples (0-30 cm depth) were taken form unfertilized fields (total 279 ha) and soil
testing was made (soil pH, plant available phosphorus and potassium according ALmethod).
About 60% of tested soil samples (total 65) were acid ( pH in KCl less than 5)
and liming could be useful. Also, about 40% (total 42) of soil samples were poor or
moderate supplied with phosphorus, while potassium status is better because only 13%
samples (total 14) were in this range of K availability. These results were in detail
elaborated in the previous study (Kovacevic et al., 2005). Soil test was criterion for
choice of the plots for field experiments with increased fertilization with P and K
including Korenicani experiment in Bjelovar-Bilogora County. Grain yields of maize in
this experiment for three-year period were elaborated in detail by the previous study
(Kovacevic et al., 2006). In this study, influences of the fertilization on leaf and grain
composition were shown.
MATERIAL AND METHODS
The field experiment: Four fertilization treatments by application of NPK 7:20:30
fertilizer (the product of Petrokemija Fertilizer Factory Kutina, Croatia: a = 0 = standard
fertilization, b = a+1250, c = a+2500 and d = a+3750 kg ha -1 ) were applied on
Korenicani soil (municipality Djulovac, Bjelovar-Bilogora County). Different amounts
of nitrogen (N) for the a-d treatments were equalized by addition of CAN (calcium
ammonium nitrate) in the amounts 974, 649, 325 and 0 kg ha-1, for the a, b, c and d,
respectively. Only standard fertilization of the experiment for the 2004 and the 2005
growing seasons were applied. The experiment was conducted in four replicates by
randomized block design. Gross of the basic plot was 92,4 m 2 (11.0 m x 8.4 m). Maize
hybrids OsSK552 (the growing seasons 2003 and 2004) and OsSK499 (the growing
season 2005) were sown by pneumatic sowing machine in terms end of April/beginning
May. Planned (theoretical) plant densities (TPD: plants ha -1 ) were 51948, 54 946 and
57143, for 2003, 2004 and 2005, respectively. Maize was harvested manually (four
rows from each plot). Grain yiels were calculated on 14% grain moisture basis and plant
densities as follows: 90% TPD (2003), 70% TPD (2004) and 95% TPD (2005),
respectively.
Sampling and chemical analyses: Soil samples were taken after maize harvest at end
October 2005. The samples were taken by auger to 30 cm of depth (15 individual
sampling in the mean sample). The ear-leaf of maize was taken at beginning of silking
(middle of July; 25 leaves in the mean sample). Soil, leaf and grain sampling were made
from three replicates of the trial.
Nutritional status of soil (mobile fraction of the elements) was predicted by
extractions with NH4Acetate + EDTA (pH 4.65) according Lakanen and Ervio (1971)
and their determination by the ICP-AES procedure. For general soil test AL-extraction
was used (Egner et al., 1960). Soil reaction and organic matter were determined
according to ISO (1994, 1998). The total amounts of the elements in the leaf and grain
84

samples were measured by the ICP-AES technique after their microwave digestion
using concentrated HNO3+H2O2. The soil and plant analyses were done in the
laboratory of the Research Institute for Soil Science and Agricultural Chemistry
(RISSAC), Budapest, Hungary.
The soil characteristics: By soil test (end of March 2003) acid reaction (pH in 1nKCl =
4.98), low organic matter contents (1.86%) and moderate levels of mobile phosphorus
and potassium (5.20 mg P2O5 and 8.13 mg K2O 100 g of soil -1 according the ALmethod)
were found.
RESULTS AND DISCUSSION
In general, in spite of relative low status of plant available P were found by the
preliminary soil test, response of maize to ameliorative fertilization was low because
yield were increased up to 10% only (Table 1). We presume that Al-method is not
suitable solution for prediction of plant available P status in this soil. Ameliorative
fertilization resulted by significant influences on increased plant available phosphorus,
potassium and iron status only, while differences of the other tested elements was in
level of statistical error. Applied fertilization influenced on moderate increases of soil
reaction (pH value), while humus contents were similar for all fertilization treatments.
Also, values of plant available P and K which were determined by two extraction
methods are similar (Table 2).
Table 1. Realized plant density and grain yield of maize as affected by ameliorative
fertilization - the experiment Korenicani (Kovacevic et al., 2006)
a)
b)
c)
d)
Fertilization* Realized plant density (RPD in %TPD=theoretical plant
(17. 04. 2003.) density) and grain yield of maize**
kg ha -1 Maize hybrid OsSK 552 Hybrid OsSK 499
2003 2004 2005
RPD Yield RPD Yield RPD Yield
NPK CAN % t ha -1 % t ha -1 % t ha -1
0 974 87.3 12.38 73.7 10.12 98.9 11.78
1250 649 85.9 13.33 72.4 10.87 94.0 11.92
2500 325 89.4 13.52 70.1 11.17 94.6 11.88
3750 0 84.6 13.41 72.7 10.82 96.1 11.89
Mean 86.8 13.16 72.2 10.75 95.9 11.87
TPD (plants ha -1 ) 51948 54946 57243
LSD 5% 0.54 0.17 n.s.
* for 2004 and 2005 standard fertilization of the a-d treatments; **yield calculations on
14% moisture basis and TPD 90% (2003), 70% (2004) and 95% (2005), respectively.
85

NPK
7:20:30
Table 2. Influences of fertilization (spring 2003) on soil characteristics
The experiment Korenicani: 0-30 cm of soil depth
(sampling 20 October 2005)
NH4Acetate-EDTA extraction (pH 4.65): concentrations in mg kg -1 of soil
kg ha -1 P2O5 K2O Ca Mg S Fe Mn Zn Cu Cd
a) 0
b) 1250
c) 2500
d) 3750
64.7
62.7
154.3
155.0
100.4
109.7
147.0
153.3
1835
1764
2100
2067
299
298
365
357
7.9
7.8
9.2
8.6
448
421
597
590
261
245
269
267
2.68
2.65
3.19
3.23
3.59
4.49
4.39
4.21
0.077
0.073
0.089
0.091
LSD5% 74.9 34.0 ns ns ns 78 ns ns ns ns
Mean 109.2 127.6 1942 330 8.4 2.94 261 514 4.17 0.082
General agrochemical soil test
pH AL-method: mg g -1 %
a) 0
b) 1250
c) 2500
d) 3750
H2O KCl P2O5 K2O Humus
6.45
6.46
6.87
6.82
5.12
5.12
5.64
5.53
84.7
96.3
164.7
172.0
85.0
99.3
149.7
176.7
1.94
1.92
1.99
1.94
LSD5% 0.23 0.36 28.6 24.0 n.s.
Mean 6.65 5.35 129.4 127.7 1.95
The applied fertilization had the most influences on maize nutritional status in the first
year of testing (2003) because significant differences for P, K, Mn (increasing trend)
Ca, Mg, Mn and Zn as well (decreasing trend) were found. In the second year (2004)
non-significant differences were found, while in the third year (2005) significant
differences were found for P, K and Mg only (Table 3). It is important that cadmium
status in maize plant was independent on applied fertilization because there are
examples in literature that application of the high P rates are mainly in conncetion with
increases of Cd concentrations in plant (Bergmann, 1992; Mengel and Kirkby, 2001).
Ameliorative fertilization had mainly low influences on maize grain composition
because only in the third year of testing significant differences for K, Mg (increases)
and Zn (decrease) status were found. Cd concentrations in maize grain were under
detectable range of the applied analytical procedure (Table 4).
Loncaric et al. (2005) applied 500 and 1000 kg ha -1 P2O5 and K2O alone and in
their combination on standard fertilization. Although low levels of P and K were found
by the soil test (AL-method), maize yields were similar for applied treatments in both
years. For this reason, in question is scientific application of AL-method in
interpretation of soil nutritional status for this and similar soil types. Probably acid
reaction of soil could be responsible for low influences of ameliorative fertilization on
maize yield in our study and in the study Loncaric et al. (2005).
Also, with aim of more precise prediction of soil nutritional status it is need
inclusion soil testing by application of different extraction methods.
86

Table 3. Influences of ameliorative fertilization on nutritional status of maize
(composition of the ear-leaf at silking stage)
Korenicani field experiment: Influences of fertilization in spring 2003 (NPK 7:20:30 on
standard fertilization) on nutritional status of maize
NPK Concentrations in dry matter of ear-leaf of maize at silking stage
7:20:30 Percent (%) mg kg -1
kg ha -1 P K Ca Mg S Fe Mn Zn Cu Cd
0
1250
2500
3750
The growing season 2003
0.29
0.31
0.30
0.32
2.14
2.55
2.50
2.50
0.71
0.65
0.63
0.60
0.44
0.30
0.25
0.24
0.19
0.20
0.18
0.19
122
127
129
135
83
108
105
110
66.4
63.7
56.4
56.4
16.5
17.4
15.3
17.0
0.08
0.08
0.09
0.11
LSD5% 0.02 0.21 0.09 0.03 n.s. n.s. 18 10.1 n.s. n.s.
Mean 0.30 0.24 0.65 0.31 0.19 128 101 60.7 16.5 0.09
0
1250
2500
3750
The growing season 2004
0.38
0.38
0.39
0.38
2.29
2.25
2.37
2.29
0.75
0.72
0.73
0.72
0.22
0.22
0.21
0.22
0.21
0.21
0.21
0.20
190
190
196
202
81
77
87
84
55.3
58.8
60.5
57.3
17.6
18.7
19.0
17.5
0.06
0.06
0.07
0.07
LSD5% n.s. n.s. n.s. n.s. n.s. n.s. n.s. n.s. n.s. n.s.
Mean 0.38 0.23 0.73 0.22 0.21 194 82 58.0 18.2 0.07
0
1250
2500
3750
The growing season 2005
0.36
0.38
0.41
0.41
2.06
2.19
2.23
2.47
0.66
0.63
0.66
0.65
0.27
0.23
0.23
0.22
0.21
0.21
0.22
0.21
155
191
189
171
97
100
85
96
42.7
46.5
49.6
48.3
15.0
16.1
16.8
16.1
0.02
0.04
0.03
0.02
LSD5% 0.03 0.30 n.s. 0.04 n.s. n.s. n.s. n.s. n.s. n.s.
Mean 0.39 2.18 0.65 0.24 0.21 177 94 46.8 16.0 0.03
In general, based on our field experiment experiences in the last decade period, liming
was more influencing factor of field crops yield compared to ameliorative fertilization
with P and K. For example, Kovacevic et al. (2006) applied carbocalk (waste of sugar
factory, 39% CaO) in five rates up to 90 t/ha. Liming with carbokalk resulted with
considerable increases of field crop yields (maize-maize-sunflower-barley rotation )as
follows: up to 50% and 36% (maize for 2001 and 2002, respectively), up to 49%
(sunflower 2003) and up to 30% (barley 2004). In general, by application of carbocalk
in level of 90 t/ha, yields drastically decreased mainly to level of control as affected by
overliming. Using 15 t ha -1 of carbocalk on Djakovo pseudogley (the growing season
1993) yields of maize hybrids (total 10) were increased depended on hybrids in range
from 6% to 45% (Banaj and Kovacevic, 2000). Liming with carbocalk (30 t ha -1 ) was
especially useful for maize under drought stress of the 2000 growing season on Slatina
pseudogley because yield was doubled compared to the control. At the same time,
contribution of female-sterile plants was decreased from 42% to 7% only (Antunovic et
al., 2001).
87

Table 4. Influences of ameliorative fertilization on nutritional status of maize
(composition of grain at maturity stage for the growing seasons 2004 and 2005)
Korenicani experiment: influences of NPK fertilization on grain composition of maize
NPK Concentrations in dry matter of grain at maturity stage*
7:20:30 Percent (%) mg kg -1
kg ha -1 P K Ca Mg S Fe Mn Zn Cu
The growing season 2004
0 0.307 3.63 0.005 0.099 0.098 21.9 4.60 21.2 1.52
3750 0.295 3.61 0.007 0.092 0.084 18.4 4.43 19.5 1.31
LSD5% n.s. n.s. n.s. n.s. n.s. n.s. n.s. n.s. n.s.
Mean 0.301 3.62 0.006 0.095 0.091 20.1 4.52 20.3 1.42
The growing season 2005
0 311 3.51 0.002 0.087 0.087 22.7 4.81 30.3 1.66
1250 308 3.52 0.002 0.086 0.088 23.1 5.04 22.7 1.69
2500 319 3.83 0.002 0.089 0.083 25.6 5.09 18.0 1.71
3750 312 3.74 0.002 0.096 0.086 22.1 4.97 18.2 1.70
LSD5% n.s. 0.12 n.s. 0.004 n.s. n.s. n.s. 9.1 n.s.
LSD1% n.s. 0.18 0.006 n.s. n.s. n.s. n.s.
Mean 0.312 3.65 0.002 0.089 0.086 23.4 4.98 22.3 1.69
Cd concentrations in grain were under detectable range Cd (

Egner, H., Riehm, H., Domingo, W.R. (1960): Untersuchungen uber die chemische
Bodenanalyse als Grundlage fur die Beurteilung des Nahrstoffzustandes der Boden
II. Chemische Extractionsmethoden zu Phosphor- und Kaliumbestimmung. K.
Lantbr. Hogsk. Annlr. W.R. 1960, 26, 199-215.
ISO (1994): Soil quality. Determination of pH. ISO 10390:1994(E)
ISO (1998): Soil quality. Determination of organic carbon by sulfochromic oxidation.
ISO 14235:1998(E).
Kovacevic V., Banaj D., Kovacevic J., Lalic A., Jurkovic Z., Krizmanic M. (2006):
Influences of liming on maize, sunflower and barley. Cereal Res. Comm. 34 (1):
553–556.
Kovačević V., Lončarić Z., Rastija M. (2005): Analize tla i gnojidba u funkciji
povećanja prinosa kukuruza. U: Zbornik radova, XL. Znanstveni skup hrvatskih
agronoma s međunarodnim sudjelovanjem (Kovačević V. i Jovanovac Sonja, ur.),
Opatija, 15-18. veljače 2005. str. 457-458.
Kovačević V., Stojić Biserka, Rastija Mirta, Bukvić Gordana, Antunović Manda
(2006): Reakcija kukuruza na melioracijsku gnojidbu fosforom i kalijem. In:
Proceednings, 41 Croatian & 1st <strong>International</strong> Symposium on Agriculture, (S.
Jovanovac and V. Kovacevic Eds.), 13-17. February 2006, Opatija, Croatia,
Faculty of Agriculture, University J. J. Strossmayer in Osijek, p. 423-424.
Lončarić Z., Kovačević V., Šeput Miranda, Šimić B. and Stojic Biserka (2005):
Influences of fertilization on yield and nutritional status of maize. Cereal Research
Communications 33 (1):259-262.
Mengel K., Kirkby E. A. (2001): Principles of plant nutrition. Kluwer Academic
Publishers Dordrecht/Boston/London.
89

YIELD AND QUALITY OF WINTER WHEAT GENOTYPES
(Triticum aestivum L.)
Desimir Knezevic 1 , Veselinka Zecevic 2 , Danica Micanovic 2 , Nevena Djukic 3 ,
Aleksandar Paunovic 4 , Milomirka Madic 4 , Ivica Djalovic 4
1 Faculty of Agriculture, Zubin Potok, University of Pristina, Serbia,
2 Center for Small Grains Kragujevac, S. Kovacevica 31, 34000 Kragujevac, Serbia
3 Faculty of Natural Science, Department of Biology, University of Kragujevac, Serbia,
4 Agricultural Faculty, Cacak, Cara Dusana 32, 32000, Cacak, Yugoslavia
ABSTRACT
Variability of yield and quality parameters were studied in 11 wheat lines and cultivars.
Wheat cultivars were grown in the experimental field in 5 replication. part of The
differences in average values for all the studied parameters among investigated cultivars
were determined. High variability of investigated traits was established. The high
variability for analyzed yield and quality components were.
Keywords: wheat, yield, quality, protein sedimentation, gluten
INTRODUCTION
Increasing of yield in wheat has been an important task of breeders. The breeders in the
last six decades have been investigated impact of genetic, physiological, biochemical,
ecological parameters for increases in yield and quality improvement. Numerous data of
investigation indicated that levels of water, fertilizer, pesticide and herbicide application
have influence to impovement of wheat yield. In successful breeding program wheat
breeders has produced a series of cultivars with improved resistance to pests and
diseases, wider adaptation to adverse soil and climatic conditions and better quality
(Bedo et al., 2005). Genetic yield potential may be defined as the yield of a cultivar
grown in environments to which it is adapted, with nutrients and water non-limiting,
and with pests, diseases, weeds, lodging and other stresses effectively controlled. The
genetic yield potential of a wheat cultivar may be dependent on favorable conditions
and good agronomy for its expression. The ideal cultivar for high grain yield or for any
other desirable traits need to express genetic potential in different environment with low
value of variance in different environmental factors of growing (Joshi et al. 2002). The
grain yield of wheat is variable traits, which depends on numerous yield components
and environmental factors (Kraljevic-Balalic et al., 2001). The characters as height of
plants, length of spike, number of spikelets per spike are in positive correlation with
grain yield (Zecevic, 2005). The yield variability is less studied than yield itself.
The wheat grain and its product are very important in human nutrition. The great
nutritional importance of wheat have stimilated the investigation of quality parameters.
During the recent few decades significant advanced have be made in the biochemical,
genetic and molecular biology studies of protein components. Proteins play major role
in determination of quality of grain, flour, dough and bread. A number of genes
encoding proteins were isolated and sequenced.
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To resolve genetics of grain quality in wheat the main investigations were focused on
protein sedimentation volume, protein and gluten contents, grain hardness and
farinograph characteristics, are genetically controlled. A combination of those
parameters and the flour protein content can be used as a base for estimation of the
technological quality of selection materials. For improving of wheat grain quality is
especially important to identify the biotype polymorphisms with numerous favorable
traits and gene donors controlling high protein content with high grain yield and create
high quality cultivars (Bebyakin, 1985, Knezevic et al., 2006).
The aim of this paper is study of variability of yield, height of plants, thousand
grain mass, hectoliter mass and technological quality parameters in genetically
divergent wheat genotypes.
MATERIALS AND METHODS
The variability of yield and and qualyty parameters (thousand grain mass, hectoliter
mass, protein contents, protein sedimentation volume, wet and dry gluten contents,
water absorption) were investigated in 11 genetically divergent wheat lines and cultivars
(KG-35265-1/94, KG-52, KG-3625/97, KG-3089/97, KG-3062/1-93, KG-4, KG-
3617/98, KG-35171/94, KG-3419-3/97, Pobeda and Partizanka). The experiment was
performed in randomized block design in five replication on the experimental field of
Center for Small Grains, Kragujevac, Serbia. The seeds of cultivars were sown on basic
plot 5 m 2 . The technological quality of 11 winter wheat cultivars was estimated by
standardized laboratory methods. Standard milling was performed with a Buller type
MLU-202 experimental mill (Buller, Uzwill, Switzerland). Sedimentation value was
analyzed by Zeleny modified in which 2 or 5 g of wheat could be evaluated
satisfactorily. Rheological quality were analyzed with micro-Brabender farinograph
using 10 g flour sample.
RESULTS AND DISCUSSIONS
The analysis of yield and technological quality parameters showed differences among
cultivars. Sedimentation volume varied between 39 ml (KG-3089/97) and 68 ml
(Partizanka). According to sedimentation volume wheat lines which had sedimentation
volume higher than 40 ml are belonging into the 1 st quality group. Wheat lines which
had less than 40 ml sedimentation volume are clasified into 2 nd quality group (Table 1).
Sedimentation volumes of analyzed cultivars have been different.
High sedimentation protein values indicate high protein content and good gluten quality
(Menkovska et al., 1995). This was established in wheat line KG-3419-3/97. Protein
sedimentation volume, an important quality component, has a positive correlation with
other quality parameters such as gluten quality and loaf volume (Zuoji et al., 1989).
High values of protein sedimentation volume mainly depends from protein
contents as well from amount of gliadin and glutenin and gliadin/glutenin ratio,
respectively. Glutenin content is directly related to protein contents and therefore
influences sedimentation volume (Knezevic et al., 2000). Protein contents varied from
11.90% (KG-35265-1/94) to 17.21% (KG-3419-3/97) Table 1. Glutenin content had a
positive genetic correlation with technological quality of flour, sedimentation volume
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whereas gliadin had a negative correlation (Sasek et al., 1987; Pogna et al., 1990;
Menkovska et al., 2002).
Table1. Technological qualityparameters of Kragujevac's winter wheat lines
Cultivar Sv (ml) Wg % Dg % Wac Qsg Qn Pc
KG-35265-1/94 63 43.23 16.55 62.4 A1 91.2 11.90
KG-52 58 38.00 13.28 58.4 A2 89.5 14.02
KG-3625/97 42 37.26 11.25 63.4 C1 40.2 14.36
KG-3089/97 39 39.31 14.45 63.4 A2 76.7 15.22
KG-3062/1-93 47 37.95 12.06 62.6 B1 56.6 14.08
KG-4 40 38.89 12.52 64.0 B2 46.7 15.10
KG-3617/98 41 43.68 13.59 63.8 B1 55.4 15.22
KG-35171/94 50 39.26 13.34 62.4 B1 66.4 15.45
KG-3419-3/97 62 47.84 17.28 66.2 A2 71.6 17.21
Pobeda 56 41.21 13.51 62.8 A2 72.8 14.02
Partizanka 68 45.91 14.59 64.2 A2 80.0 13.15
Sv=Sedimentation value (ml); Wg=Wet gluten %; Dg=Dry gluten %; Wac=Water
absorption capacity ; Qsg=Quality subgroup;Qn= Quality number; Pc=Protein contents
In this study different value of gluten content in wheat cultivars were established. In
average the highest gluten content 47.84% (KG-3419-3/97), and the lowest 37.26% in
the KG-3625/97 line. The ratio of variation of gluten contents is in agreement with data
(Pavlovic et al., 1994) which found that gluten content varies from 16 to 52% (crude)
and 5-20% (dry). Grain consistency, temperature and moisture conditions during
maturation have influence to gluten quality. High temperature and low moisture
produce strong gluten with less extensibility and the opposite conditions produce weak
glutens and extensible doughs (Kodanev, 1976). Gluten is formed in the early stages of
ripening (milk) but its quality changes during maturity to the final characteristics.
Gluten structure has a great influence on flour rheological properties and quality.
Gluten quality depends on the ability of dough to keep CO2 from fermentation in gas
cells in order to provide loaf volume and bread crumb.
All the varieties in study belonged to semi-dwarf wheat class expressing the plant
height from x= 63.4 cm (partizanka) and x = 76.3 cm (kg-52), according to mean
values over studied period (table 2). This semi-dwarf ccultivars have advantages in
relation to dwarf and tall cultivars as a source of desirable genes in the future breeding
programs. The similar results reported Dimitrijevic et al. (1996) that indicated that
genotype had higher influence than environment on expression of plant height. the
proportion of total mass to grain yield in mostly modern cultivars is more than 2:1
which indicates excessive consumption of nutrients for straw formation instead of grain.
This is one reason why breeders have concept of decreasing vegetative part of wheat
and realize optimal proportion in order to increase the harvest index to more than 50%
(Zecevic at al., 2005).
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Table 2. Average values of analyzed traits of Kragujevac's winter wheat lines
Cultivar Plant height Hectoliter TGW Yield
(ml) mass (kg) (g) (kg)
KG-35265-1/94 67.8 82.30 38.28 4265
KG-52 76.3 80.90 40.19 4556
KG-3625/97 71.6 82.50 37.52 4756
KG-3089/97 68.8 80.10 37.52 4321
KG-3062/1-93 64.3 81.30 36.96 3882
KG-4 67.0 81.30 37.29 4327
KG-3617/98 69.0 82.10 38.98 3718
KG-35171/94 64.4 82.10 39.21 3890
KG-3419-3/97 63.5 80.75 36.96 2875
Pobeda 67.0 81.31 39.36 4753
Partizanka 63.4 82.10 41.47 3962
Decreasing of stem height gave higher possibility of efficient utilization mineral
elements for the head development. Also translocation of mineral elements is more
efficient in semidwarf cultivars from the vegetative organs to grain.
During investigated periods the highest average value of of thousand grain weight had
Partiyanka cultivar (x=41.47) the lowest average value of thousand grain weight
expressed KG-3062/1-93 cultivar (x=30.62) Table 2. Number of grain per spike and
thousand grain weight have strong influence on spike productivity. Yield of analyzed
wheat genotypes varied between 2875 kg ha -1 (KG-3419-3/97) and 4756 kg ha -1 (KG-
3625/97) Table 2. Yield is in high relationships with the number of stems per m 2 a nd the
percentage of fertile spikes, number of kernels per spike, thousand grain weight. The
genetic controls of stem height and other yield components are very important, but
environment has influence on expression of analyzed traits. Generally speaking,
genotypes reacted similarly to environmental variation for plant height, and number of
spikiest per spike, number of kernels per spike, thousand grain weight, differing in main
effect. On the contrary, genotype reaction for yield components in interaction making
phenotypic expression for those trait more unpredictable. The remobilization of reserve
into harvested organs varies among the cultivars. Yield is very important agroeconomic
traits which will in the focus of investigation in the future.
REFERENCES
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combinative ability of the varieties of spring common wheat for grain quality
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aestivum L) for different adaptation types in multifunctional agricultural
production. Turk. Jagric.For., 29, 151-156.
Dimitrijevic, M., Knezevic, D., Petrovic Sofija, and Zecevic Veselinka (2000): Stability
of yield components in wheat (Triticum aestivum L.). EUCARPIA, XI th Meeting of
the Section Biometrics in Plant Breeding, pp.105-106. Paris/France.
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Joshi, S.K., Sharma, S.N., Singhania, D.L., Sain, R.S. (2002): Genetic analysis of
quantitative and quality traits under varying environmental conditions in bread
wheat. Wheat Information Service, 95, 5-10.
Knezevic, D., Zecevic Veselinka, Urosevic, D., Pavlovic, M., Marinkovic Ivana,
Micanovic Danica and Urosevic Vesna (2000): Quality Parameters of Yugoslav
Common Winter Wheat Cultivars (Triticum aestivum L.). Annual Wheat
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Jadranka (2006): Yield and quality parameters of winter wheat lines (Triticum
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Montenegro. 20-24 Septembar, Kotor. pp.423-429
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Variability and gene effect in wheat. In: Monograph Genetic and Breeding of
Small Grains. (eds. S.Quarrie et all) pp. 9-49
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Quality of Some Macedonian Bread Wheat Varieties: I. Relation With the
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Macedonia,14,1,31-34.
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725.
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protein composition of durum wheat lines containing the Gli-D1/Gli-D3 loci. In:
CW Wrigley ed. Gluten 96, Cereal Chemistry Devision, Royal Australian
Chemical Institute, North Melbourne, Sidney, pp. 18-22.
Sasek, A., Kubanek, J. & Cerny, J. (1987) Gliadin and glutenin polymorphism of some
cultivars – Populations of common wheat (T. aestivum L.). Scientia Agriculturae
Bohemoslovaca 19, 2, 93-99.
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Correlation between bread making quality and other quality parameters and yield
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(2005): the inheritance of plant height in winter wheat (Triticum aestivum L.).
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Genetika, 37, 173-179.
94

CROP ROTATION AND GREEN MANURE INFLUENCE ON YIELD AND
WATER USE EFFICIENCY IN WHEAT AND MAIZE FROM CRIS PLAIN
C. Domuţa, Gh. Ciobanu, Maria Şandor, Alina Samuel, Cornelia Ciobanu,
N.C. Sabău, V. Şcheau, Ioana Borza, Cr. Domuţa
University of Oradea, Oradea, Romania
ABSTRACT
The paper is based on the results researches obtained during 2004 – 2006 in Oradea. In
the crop rotation of 4 years with clover in comparison with wheat – maize crop rotation,
the yield gains statistically assured were obtained both in the variants with organic
fertilization and in the variant with organic + chemical fertilization. The use of the
mixture lupin + oat in this crop rotation determined to obtain yield gains in comparison
with variants with lupin, pure crop as green manure; yield obtained using lupin + oat
were close to the yield from variant with manure, 25 t/ha. Meliorative crop rotation and
fertilization determined the improve of the water use efficiency in comparison with the
controls; water use efficiency from variants with lupin + oat was bigger than water use
efficiency obtained in the variant with lupin, too.
Key words: crop rotation, green manure, yield, wheat, maize.
INTRODUCTION
In the sustainable agriculture system, the crop rotation is the central pivot and organic
fertilization is one of the most components (Budoi and Penescu, 1998; Gus and al,
1998; Pakurar and Nagy, 2005, Pepo, 2005, Domuţa, 2006). Green manure can
occupied a very important place between the organic fertilizer types but the correct use
are need consist of the mixture between lupin and oat (rye) and rape. (Domuţa, 1999,
2005). Due the small C/N rapport the use of the lupin in pure crop determines explosive
microbiological processes and intense humus mineralization and finally the decrease of
the soil humus reserve. (Eliade and al, 1983, Samuel and al, 2006).
MATHERIAL AND METHOD
The researches were carried out in Oradea, Western Romania, on erosioned preluvosoil
with slope of 8˚. On ploughing land the pH value is of 6.2, humus content is of 2.1,
mobile phosphorum is of 34.1 ppm and mobile potassium content is of 209.2 ppm.
Structure degree is of 55.8% and field capacity (24.3%) and wilting point (9.1%) have
median value.
Research period was 2004 – 2006 but the experiment is in the second cycle of the
researches. Experimental dispozitive includes three factors:
Factor A: crop rotation
a1 : wheat – maize
a2 : oat + clover – clover – wheat – maize
Factor C: annual fertilization
c1: N0P
c2: N90P60K60
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Factor B: organic fertilization
b1 : control
b2 : manure, 25 t/ha
b3 : manure, 50 t/ha
b4 : lupin
b5 : lupin + oat

Number of repetition used: 4. Surface of the experiment plot: 300 m 2 .Total surface of
the experiment: 6000 m 2 .Green manure was produced in 2001, 2002 and 2003 like main
crop. Sowing rate: 200 kg/ha in lupin pure crop and lupin 100 kg/ha and oat 80 kg/ha in
the mixture. The harvesting was made in the flowering stage of the lupin. After
harvesting, the green manure was kept like mulch on the soil surface. After 10 days the
plough land of 25 cm depth was made. The maize was cropped in the first year after
organic fertilization. Water use efficiency was established dividing the wheat and maize
yield to water consumption. Wheat and maize water consumption was established using
the soil water balance method; balance depth used was 0 – 150 cm.
RESULTS AND DISCUSSION
Influence of crop rotation on wheat yield
The average of the yields on the period 2004 – 2006 emphasizes bigger yields in the
crop rotation with clover both in the background N0P0 (4944 kg/ha vs. 3944 kg/ha,
25.3%) and in the background N90P60K60 (6092 kg/ha vs. 5562 kg/ha). The relative
differences between yield registered in the researched crop rotation during the years
were of 18.6% (N0P0) and of 12.4% (N90P60K60) in 2004, of 30.0% (N0P0) and of 9.0%
(N90P60K60) in 2005, of 27.3% (N0P0) and of 7.5% (N90P60K60) in 2006 (table 1).
Influence of crop rotation and green manure on maize yield
All the organic fertilization variants determined to obtain very significant yield gain in
comparison with control both in the crop rotation wheat – maize and in the crop rotation
oat + clover – clover – wheat – maize. The biggest yields were obtained in the variant
with manure 50 t/ha.
Table 1: Influence of the crop rotation and fertilization on wheat yield,
Oradea 2004 – 2006
Organic fertilization
N0P0
Chemical fertilization
N90P60K60
2004 2005 2006 Average 2004 2005 2006 Average
Wheat - maize
1. Control 2740 3010 3580 3110 4010 4690 5010 4570
2. Manure 25 t/ha 3890 3850 4320 4020 5320 5720 6190 5740
3. Manure 50 t/ha 5020 4580 5030 4880 6280 6440 6760 6490
4. Lupin 3690 3420 3840 3650 4960 5130 5530 5210
5. Lupin + oat 4080 3810 4290 4060 5620 5700 6080 5800
Average 3884 3734 4212 3944 5238 5536 5914 5562
Oat + clover – clover – wheat – maize
1. Control 3640 4010 4520 4060 5020 5330 5840 5400
2. Manure 25 t/ha 4760 5030 5570 5120 5980 6090 6460 6180
3. Manure 50 t/ha 5620 5710 6240 5860 6390 6890 7100 6790
4. Lupin 4210 4540 5010 4590 5720 5680 6010 5800
5. Lupin + oat 4800 4990 5480 5090 6320 6170 6390 6290
Average 4606 4856 5364 4944 5886 6032 6360 6092
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Organic Chemical Chemical fertilization x Organic fertilization x
fertilization fertilization Organic fertilization Chemical fertilization
a1 a2 a1 a2 a1 a2 a1 a2
LSD 5% 140 150 160 110 230 210 200 190
LSD 1% 250 290 230 208 310 420 350 360
LSD 0.1% 590 470 310 390 420 730 560 580
In the variant with lupin + oat, an yields bigger than yields obtained in the variants with
lupin pure crop were obtained 410 kg/ha in the crop rotation wheat – maize and 500
kg/ha in the crop rotation oat + clover – clover – wheat – maize. The yield obtained in
the variants with lupin + oat are very closed to the yields obtained in the variant with
manure 25 t/ha.
Chemical fertilization applied on organic background determined the increase of
the yields with 28.3% in crop rotation wheat – maize and with 23.3% in crop rotation
oat + clover – clover – wheat – maize.
Table 2: Maize yield obtained under the influence of fertilization and crop rotation,
Oradea 2004 – 2006
Chemical fertilization
Organic fertilization
N0P0
N90P60K60
2004 2005 2006 Average 2004 2005 2006 Average
Wheat - maize
1. Control 5170 5020 4860 5020 6420 6340 6110 6290
2. Manure 25 t/ha 5990 5620 5520 5710 7030 6830 6820 6890
3. Manure 50 t/ha 6710 6100 6130 6310 7620 7320 7200 7380
4. Lupin 6820 6140 6100 6350 7710 7300 7240 7416
5. Lupin + oat 8010 7320 7930 7750 8760 8520 8410 8560
Oat + clover – clover – wheat – maize
1. Control 5990 5880 5710 5860 7230 6820 7010 7020
2. Manure 25 t/ha 6810 6400 6320 6510 8010 7350 7560 7640
3. Manure 50 t/ha 7520 6820 6860 7067 8780 7810 7900 8163
4. Lupin 7500 6900 6990 7130 8790 7900 7960 8220
5. Lupin + oat 8410 8020 7910 8113 9220 9000 9020 9080
Organic Chemical Chemical fertilization x Organic fertilization x
fertilization fertilization Organic fertilization Chemical fertilization
a1 a2 a1 a2 a1 a2 a1 a2
LSD 5% 180 150 120 110 230 210 208 190
LSD1% 310 290 240 208 410 420 310 360
LSD 0.1% 520 470 410 390 710 730 530 580
Both in the crop rotation wheat-maize and in the crop rotation oat + clover-clover-wheat
+ maize, the lowest maize yields values were obtained in the variant with lupin; the
differences in comparison with the control were of 13.7% (N0P0K0) and of 9.5%
(N90P60K60) in the crop rotation wheat-maize and of 11.1% (N0P0K0) and of 8.8%
(N90P60K60) in the crop rotation with clover (table 2).
In the variant with lupin + oat, the maize yields registered were bigger than in the
variants with lupin, pure crop; overall on the studied period the differences registered
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were of 600 kg/ha (N90P60K60) and of 490 kg/ha (N90P60K60) in the crop rotation wheatmaize;
in the crop rotation with clover the differences were of 557 kg/ha (N0P0K0) and
of 523 kg/ha (N90P60K60)).
Generally, in the variant with manure 25 t/ha the yield registered were closed to the
yield obtained in the variant with lupin + oat; the biggest yields was registered in the
variant with manure 50 t/ha in all cases.
Influence of the crop rotation and organic fertilization on wheat water consumption
All the years, in the crop rotation with clover, a bigger quantity of rainfall was stored
during the cold period. The same phenomenon was registered in the variant with organic
fertilization in comparison with the control. In these conditions the values of the wheat
water consumption are bigger in the variants from crop rotation with clover and from
organic fertilization. (Table 3)
Table 3: Influence of the crop rotation and organic fertilization on wheat water
consumption, Oradea 2004 – 2006
Variant of organic
Crop rotation
fertilization
Wheat - maize Oat + clover – clover – wheat - maize
2004 2005 2006 2004 2005 2006
1. Control 3670 3820 4250 4020 4210 4520
2. Manure 25 t/ha 3810 3980 4420 4200 4400 4660
3. Manure 50 t/ha 3960 4210 4760 4420 4580 4880
4. Lupin 3680 3850 4310 4100 4290 4530
5. Lupin + oat 3780 3930 4400 4180 4350 4700
Average 3780 3958 4436 4184 4366 4658
Influence of the crop rotation and green manure on maize water consumption
In the variants with green manure and manure bigger quantities of rainfall were stored
during the cold period leading to values of the maize water consumption higher than the
control. In the variant with lupin + oat the values of the maize’ water consumption were
bigger than in maize’ water consumption in the variant with lupin pure crop. The
highest value of the maize’ water consumption was registered in the variant with
manure 50 t/ha. In the crop rotation with clover bigger quantities of rainfall were stored
in the cold period determining values of the water consumption higher than the values
registered in the wheat-maize crop rotation (table 4).
Table 4: Maize water consumption obtained under the influence of the organic
fertilization and crop rotation, Oradea 2004 – 2006
Variant of organic
fertilization
2004
Wheat - maize
2005
Crop rotation
Oat + clover – clover – wheat - maize
2006 2004 2005 2006
1. Control 4910 4760 5370 5360 5110 5620
2. Manure 25 t/ha 4970 4820 5460 5420 5200 5710
3. Manure 50 t/ha 5100 4830 5510 5510 5320 5790
4. Lupin 5120 4900 5500 5560 5300 5600
5. Lupin + oat 5320 5020 5610 5720 5420 5810
Average 5084 4866 5490 5514 5270 5706
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Influence of the crop rotation and organic fertilization on water use efficiency in wheat
In average on the studied period, in the background N0P0, in the crop rotation oat +
clover – clover – wheat – maize the wheat quantity obtained on 1 m 3 was bigger than
the value obtained in wheat – maize crop rotation with 44.3% (1.40 kg/m 3 vs. 0.97
kg/m 3 ). The same situation was registered in the background N90P60K60, (1.38 kg/m 3 vs.
1.25 kg/m 3 ) but the relative difference (10.4%) is smaller.
The green manure represented by mixture lupin and oat determined to obtain a water
use efficiency bigger than the values obtained in the variant with lupin both in crop
rotation wheat – maize and in the crop rotation oat + clover – clover – wheat – maize;
the same situation was registered in the background N0P0 and N90P60K60. Generally, the
water use efficiency obtained in the variant with lupin + oat like green manure was very
closed to water use efficiency obtained in the variant with manure, 25 t/ha.
Table 5: Influence of the crop rotation and fertilization on water use efficiency in
Variant of organic
fertilization
wheat, Oradea 2004 – 2006
Crop rotation
Wheat - maize Oat + clover – clover – wheat – maize
2004 2005 2006 Average 2004 2005 2006 Average
1. Control 0.75 0.79 0.84
N0P0
0.79 1.25 1.31 1.39 1.32
2. Manure 25 t/ha 1.12 0.97 0.98 0.99 1.42 1.38 1.47 1.42
3. Manure 50 t/ha 1.27 1.09 1.06 1.13 1.45 1.50 1.45 1.47
4. Lupin 1.00 0.89 0.89 0.93 1.40 1.32 1.33 1.35
5. Lupin + oat 1.08 0.97 0.98 1.01 1.51 1.42 1.36 1.43
Average 1.02 0.94 0.97 0.97 1.41 1.39 1.40 1.40
N90P60K60
1. Control 1.00 1.11 1.11 1.07 1.25 1.27 1.29 1.27
2. Manure 25 t/ha 1.27 1.30 1.33 1.30 1.42 1.38 1.39 1.40
3. Manure 50 t/ha 1.32 1.41 1.39 1.37 1.45 1.50 1.45 1.47
4. Lupin 1.15 1.20 1.22 1.19 1.40 1.32 1.33 1.35
5. Lupin + oat 1.28 1.31 1.29 1.29 1.51 1.42 1.36 1.43
Average 1.20 1.27 1.27 1.25 1.41 1.38 1.36 1.38
Influence of the crop rotation and organic fertilization on water use efficiency in maize
In comparison with the control, in the variant with lupin pure crop the lowest values of
the water use efficiency were determined both in the crop rotation wheat-maize (1.08
kg/m 3 in the N0P0K0 background and 1.36 kg/m 3 in the N90P60K60 background) and in
the crop rotation with clover (1.3 kg/m 3 in the N0P0K0 and 1.45 kg/m 3 in the N90P60K60
background). In the variants with lupin + oat, the water use efficiency was better than in
the variant with lupin pure crop. The values registered in the variants with manure 25
t/ha were the same or a little bigger than the values obtained in the variant with lupin +
oat; the highest value of the water use efficiency was registered in the variant with
manure 50 t/ha (table 6)
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Table 6: Water use efficiency (kg/m 3 ) in maize under the influence of the organic
fertilization and crop rotation, Oradea 2004 – 2006
Variant of organic
fertilization
Wheat - maize
Crop rotation
Oat + clover – clover – wheat – maize
2004 2005 2006 Average 2004 2005 2006 Average
N0P0
1. Control 1.05 1.05 0.91 1.00 1.31 1.33 1.14 1.26
2. Manure 25 t/ha 1.12 1.17 0.96 1.08 1.41 1.42 1.25 1.36
3. Manure 50 t/ha 1.32 1.26 1.11 1.23 1.49 1.52 1.31 1.44
4. Lupin 1.33 1.25 1.11 1.23 1.51 1.49 1.32 1.44
5. Lupin + oat 1.51 1.46 1.41 1.46 1.65 1.75 1.50 1.63
Average 1.27 1.24 1.10 1.20 1.47 1.50 1.30 1.42
N90P60K60
1. Control 1.31 1.33 1.14 1.26 1.35 1.33 1.37 1.35
2. Manure 25 t/ha 1.41 1.42 1.25 1.36 1.48 1.41 1.45 1.45
3. Manure 50 t/ha 1.49 1.52 1.31 1.44 1.59 1.47 1.48 1.51
4. Lupin 1.51 1.49 1.32 1.44 1.58 1.49 1.50 1.52
5. Lupin + oat 1.65 1.70 1.50 1.62 1.61 1.66 1.66 1.64
Average 1.47 1.49 1.53 1.42 1.52 1.47 1.45 1.49
CONCLUSIONS
1. The yields wheat obtained in the crop rotation oat + clover – clover – wheat –
maize were bigger than yields obtained in the wheat – maoze crop rotation both in
N0P0 background and in N90P60K60 background.
2. In the variant with lupin+oat, the maize yield obtained was bigger than the maize
yield obtained in the variant with lupin pure crop both in the wheat-maize crop
rotation and in the crop rotation with clover. The same situation was registered both
in the N0P0K0 background and in the N90P60K60 background.
3. Organic fertilization (manure 25 t/ha, manure 50 t/ha, lupin, lupin + oat) but
especially organic fertilization associated with annual background N90P60K60
determined to obtained very significant yield gains in comparison with control.
4. The use of the green manure composed by lupin + oat (applied for maize)
determined yield gains bigger than green manure composed by lupin like pure crop.
The yields gains were registered in the twice crop rotation studied.
5. In the crop rotation with clover the rainfall storages in the cold period were bigger
than in the crop rotation wheat – maize; the same situation, was registered in the
variants with organic fertilization. Due the values of wheat and maize water
consumption increased.
6. Both in wheat and maize the water use efficiency values obtained in the crop
rotation with clover were bigger than in the crop rotation wheat – maize; the values
registered in the background N90P60K60 were bigger than the values obtained in the
background N0P0. In the variant with lupin + oat like green manure the water use
efficincy had bigger values than in the variant with lupin pure crop.
7. The researches emphasizes the importance of the crop rotation in wheat and maize
and recomand to use of the green manure like mixture beetween lupin and oat and
not like lupin pure crop.
100

REFERENCES
1. Budoi Gh., Penescu A. (1996) – Agrotehnica. Ed. Ceres, 427 – 432 p.
2. Domuţa C., (1999) – Ameliorarea fertilităţii solurilor erodate pe terenurile în pantă
din vestul ţării. Cereale şi plante tehnice nr. 7.
3. Domuţa C. (2005) – Agrotehnica terenurilor în pantă din nord – vestul României.
Ed. Universităţii din Oradea, 96 – 117 p.
4. Domuţa C. (2006) – Agrotehnica diferenţiată. Ed. Universităţii din Oradea, 377 –
442 p.
5. Eliade Gh., Ghinea L., Ştefanic Gh., (1983) – Bazele biologice ale fertilităţii
solului. Ed. Ceres, 127 – 130 p.
6. Guş P., Lăzureanu A., Săndoiu D., Jităreanu G., Stancu I. (1998) – Agrotehnica.
Ed. Risoprint, 496 – 499 p.
7. Pakurar M., Nagy I. (2005) - Effect of different tillage treatments on the yield of
maize. <strong>International</strong> Symposium A.R.D.S. Oradea, 7 – 8 July, 158 – 165 p.
8. Pepo P. (2005) – New issues in sustainable wheat production. <strong>International</strong>
Symposium A.R.D.S. Oradea, 7 – 8 July, 17 – 28 p.
9. Samuel D., Drăgan – Bularda M., Domuţa C (2006) – The effect of green
manure on enzymatic activities in a brown luvic soil. Studia Universitas Babeş –
Bolyai, Biologia, L I, 83 – 93 p.
101

APPLICATION OF A DIGESTATE AS A NUTRIENT SOURCE AND ITS
EFFECT ON SOME SELECTED CROPS AND SOIL PROPERTIES 1
ABSTRACT
Marianna Makádi 1 , Attila Tomócsik 1 , József Lengyel 2 ,
Zsolt Bogdányi 1 , Árpád Márton 1
1 Research Centre of CAS of University of Debrecen
H-4432 Nyíregyháza, Westsik Vilmos u. 4-6.
2 Bátortrade Ltd. H-4300 Nyírbátor, Árpád u. 156/A.
The largest biogas plant of Hungary is operated in the city of Nyírbátor, NE Hungary. A
substantial amount, some 95.000 m 3 of digestate is originating every year, as the
residual material of biogas production. A study was conducted on the utilization of this
material as a nutrient source for different crops. In this paper we present the results
obtained with winter wheat and triticale. The use of the digestate has a more
pronounced effect on triticale than on winter wheat. On the average, the yield of winter
wheat and triticale were increased by 20.8 % and 51.42 %, respectively, and the grain to
straw ratio was also increased by the treatments with the digestate. As for soil
properties, the treatments caused positive changes in the case of humus content, but they
increased the Na content of the soil.
Keywords: biogas, digestate, nutrient supply, winter wheat, triticale
INTRODUCTION
According to the regulations of Law No. XLIII./2000 on waste management, by 2014
the amount of biologically degradable waste types disposed in landfills should be
reduced to 35 % of the total amount of the waste landfilled in 1995. A significant
amount of organic waste can be composted and used as natural fertilizer and soil
improving material. The other possible treatment of organic wastes is their utilization in
biogas plants. The main products of this treatment are biogas (methane) and heat, while
by-product is digestate, which can also be utilized as source of nutrients in agriculture.
This anaerobic treatment is environmentally friendly because it utilises renewable
energy sources such as plants, and destroys and utilises organic wastes such as offal.
The fermentation residue – the digestate – can substitute for the artificial fertilizers
(Kovács, 2007).
Numerous papers deal with the importance of quality and quantity of basic
materials in biogas production. But very few papers are available on the utilization of
the digestate as a plant nutrient source. Qi et al. (2005) examined in North China the
effect of fermented waste as organic manure in cucumber and tomato production. They
have found increasing yield and vitamin C-content of about 18 %. Banik and Nandi
(2004) investigated biogas residual slurry manure on the yield, protein and mineralcontents
of oyster mushroom. They have found positive effects.
Besides the examination of the effect of the digestate on the crop yields of different
plants, it is also very important to study its effects on the soil properties.
1 This project is supported by the grant GVOP-3.1.1.-2004-05-0220/3.0.
102

MATERIALS AND METHODS
The experiment was carried out in the Nyírség region, the NE Hungary. The biogas
plant is a part of a big agricultural system with crop production, livestock (beef and
diary cattle), slaughterhouse and processing plant. The biogas plant works with
mesophylic and thermophylic systems, the basic materials are livestock manure, plant
crop and offal. The total capacity of thermophylic and mesophylic digesters are 7,600
m 3 and 9,400 m 3 , respectively. The remaining digestate after anaerobic digestion can be
utilized in crop production as a nutrient supply.
No. of
treatment
Table 1: Applied treatments in the experiments with digestate.
Phenological stage at the time
of treatment
Proportion
Quantity of
digestate
Quantity of water
1. Control 0 0 0
2. after sowing 100% mean 0
3. after sowing 100% +30% 0
4.
5.
6.
7.
after sowing
plants with 6-7 leafs
after sowing
plants with 6-7 leafs
after sowing
plants with 3-4 leafs
plants with 6-7 leafs
after sowing
plants with 3-4 leafs
plants with 6-7 leafs
50%
50%
50%
50%
25%
50%
25%
25%
50%
25%
mean 0
+30% 0
mean 0
+30% 0
8. after sowing 100% 0 +30%
9. after sowing 100% 0 mean
10.
after sowing
plants with 6-7 leafs
50%
50%
0 mean
11.
after sowing
plants with 6-7 leafs
50%
50%
0 +30%
after sowing
25%
12.
plants with 3-4 leafs
50%
0 +30%
plants with 6-7 leafs
25%
after sowing
25%
13.
plants with 3-4 leafs
50%
0 mean
plants with 6-7 leafs
25%
The rates of the digestate to be applied was determined by its actual N-content, because
N is one of the main macronutrients for plants, but it could be a serious environmental
pollution if leached to the soil water and accumulates in it. The main parameters of the
digestate are the followings: pH 8.025, density 1025.75 kg m 3 , dry matter content
1.1824 m/m% (row matter), total-N 0.376 m/m% (row matter), Mg 32.7 mg kg -1 (r.m.),
Na 272.5 mg kg -1 (r.m.).
103

In the experiments (Table 1.) we applied the “mean” quantity of digestate, which means
the quantity of it equals the average N-demand of the crop plants, while the high dose
was the digestate containes “mean+30%” nitrogen. The digestate contains much water,
therefore we applied the same quantity of water in the experiment with winter wheat. In
the triticale-experiment we could not use water. No additional fertilizer was applied.
The sowing time was 15.10.2005. for winter wheat cv. ‘Sixtus’ (Triticum aestivum
ssp. vulgare L.) and 05.11.2005. for triticale cv. ‘Versus’ (X Triticosecale Wittm.). The
winter wheat was harvested on 15.07.2006, the triticale on 19.08.2006.
We measured the yield of the crop, physical, chemical and biological parameters of the
soils. One-way analysis of variance, Tukey’s test and T-test were used to determine
treatment effects by SPSS statistical program.
RESULTS AND DISCUSSIONS
1./ Winter wheat: Winter wheat is the main cereal crop in Hungary. It is mainly
cultivated on chernozem soils. In the Nyírség region the main cereal crop is rye, which
produces satisfactory yield on sandy soils. Nevertheless, some better sandy soils are
also appropriate for winter wheat production with the subsequent delivery of nutrients
in the large-scale farming systems (Bocz (ed.), 1992). In our experiment the nutrient
supply of the winter wheat was ensured by the digestate. The doses of digestate were
calculated on the N-demand of the crop, based on the literature, and the calculated
amounts were applied to the surface of soil and plants in full, or split in two or three
portions. In the case of good nutrient supply the crop yield of winter wheat is generally
around 6-7 t ha -1 on chernozems and 4-5 t ha -1 on sands. In our experiments the average
crop yield was 3.72 t ha -1 , which is less than the average on sandy soils. The results of
treatments can be seen in Figure 1.
t*ha -1
6
5
4
3
2
1
0
Winter wheat crop yield
1 2 3 4 5 6 7 8 9 10 11 12 13
No. of treatments
Figure 1: Winter wheat crop yield as affected by the treatments of digestate and
water. Treatments are numbered according to Table 1.
We have not found any significant difference among the treatments on the p

average of watered treatments resulted in a 3.35 % yield increase, while the average of
digestate-treatments increased the yield by 20.8 %, compared to the control. The
positive effect of digestate benchmarked against water based on its nutrient content and
its stimulating effect on soil microorganisms.
We have to take into account, that if the digestate is applied in full after sowing,
not all the nutrients can be utilized by plants, part of them will move down to lower soil
layers and may pollute soil water. If the digestate is split into two or three portions at
the time when the plant needs it, the utilization of the digestate by plants will be more
efficient, the application will be environmentally friendly. Nowadays in Hungary the
yearly amount of the digestate can be utilized only before sowing (except our
experiments), because the digestate, as a category, is not defined in Hungarian law. The
digestate is handled as sewage sludge. On the basis of our results, and also considering
the results from tests on other crops, the application of the digestate, based on Ndemand
of plants, split in two or three portions is the right way both in economic and
environmental respects. We computed the grain to straw ratio from the averages of the
water and digestate treatments (Table 2).
Table 2: Grain to straw ratio of winter wheat in the experiment.
Control
Digestate
Water
Mean + Std. Dev.
0,819 + 0,037
0,907 + 0,110
0,857 + 0,075
The digestate treatments increased this ratio, which means that digestate resulted less
straw and more yield in the treated plots.
2./ Triticale: Triticale is the typical crop of marginal soils. It has higher yield than rye
has, that is why triticale displaced rye from the better sandy soils. The average yield of
this plant on the field plots of Bátortrade Ltd, with the use of fertilizers, is 2.8-3.1 t ha -1 .
In our small-plot experiment the samples were collected from sampling plots of 1 m 2 in
size, and after threshing the grain and the straw yield was taken. The yield was higher
than the average as a result of the manual harvesting and handling.
For technical reasons, this experiment was only treated with the digestate. The
results obtained from the treatments 1-7 can be seen in Figure 2.
Table 3: Grain to straw ratio of triticale in the experiment.
Control
Digestate
Mean + Std.Dev.
0.702+ 0.074
0.820 + 0.079
Comparing these results to those obtained with winter wheat, treatments were more
efficient since we found significant treatment effect (p

compared to the control. It is important to conduct experiments with a large variety of
different crop plants, because they respond differently to the digestate treatment.
g*m 2
450
400
350
300
250
200
150
100
50
0
a
a
b
Triticale crop yield
a
b
a
b
1 2 3 4 5 6 7
No. of treatments
Figure 2: Triticale crop yield as affected by the treatments of digestate. Treatments
are numbered according to Table 1. Signs a,b means significance groups according to
Tukey’s test (p

Table 4: Changes in soil properties under the winter wheat after 2 years of digestate
application. Treatments are numbered according to Table 1.
Treatment pH H2O pH KCl y1 Humus %
NO3-N (mg kg -1 P2O5 ) (mg kg -1 )
K2O (mg kg -1 Na
) (mg kg -1 )
Mg
(mg kg -1 )
Before treatment 5,49 4,52 2,34 1,038 33,834 348 221 17,5 78,4
1 5,89 4,94 8,50 0,867 0,171 138 54 12,8 3,8
2 4,80 3,88 11,00 0,696 7,068 85 95 26,1 10,3
3 5,36 4,40 12,75 0,585 9,013 104 102 14,5 9,7
4 6,35 5,81 8,75 0,978 8,868 98 88 18,8 13,8
5 5,06 4,36 11,25 1,018 21,044 109 132 33,4 15,8
6 4,86 3,90 11,25 1,301 5,509 133 76 32,0 10,5
7 4,76 4,04 11,75 0,897 9,922 194 155 29,7 21,0
8 5,64 4,25 8,50 0,897 0,216 173 52 16,0 3,6
9 6,03 4,98 6,50 0,827 0,184 200 54 14,6 4,9
10 5,63 4,72 10,25 1,139 0,195 197 47 17,7 2,3
11 5,65 4,95 10,25 0,887 0,622 170 46 20,9 7,1
12 5,83 4,46 8,25 0,847 0,480 197 63 19,6 7,1
13 5,66 4,39 12,75 0,948 0,329 192 66 20,1 6,9
Table 5: Changes in soil properties under the triticale after 1 year of digestate
application. Treatments are numbered according to Table 1.
Treatment pH H2O pH KCl y1 Humus %
NO3-N (mg kg -1 P2O5 ) (mg kg -1 )
K2O (mg kg -1 Na
) (mg kg -1 )
Mg
(mg kg -1 )
Before treatment 5,46 3,88 9,65 0,44 4,688 245 179 11,5 60,3
1 5,42 3,92 12,75 0,68 0,190 242 131 76,8 3,7
2 5,74 4,29 12,25 0,83 1,322 264 209 67,5 13,0
3 5,57 4,18 11,75 0,76 0,232 173 125 89,8 13,4
4 5,71 4,66 12,25 0,84 0,205 253 210 86,3 12,5
5 5,38 4,01 12,75 0,92 1,433 258 155 49,6 12,8
6 5,26 4,04 8,75 0,79 0,342 258 157 49,5 11,8
7 5,39 4,47 12,50 0,50 2,863 205 136 44,5 3,7
NO3-N and Mg-content of soils decreased in both treatments, while Na-content was
increased. Increased Na-content indicates unfavourable changes in the soils. We
indicated this process for the operators, and the decreasing of Na-content of the
digestate was successful.
REFERENCES
Banik, S., Nandi, R. (2004): Effect of supplementation of rice straw with biogas
residual slurry manure on the yield, protein and mineral contents of oyster
mushroom. Industrial Crops and Products, 20: 311-319.
Bocz, E. (ed.) (1992): Szántóföldi növénytermesztés. Mezőgazda Kiadó, Budapest.
Kovács, K. (2007): Biogas in Hungary: the state of the art in 2007, and future
opportunities. Waste, 2: 11-16.
Qi, X., Zhang, S., Wang, Y., Wang, R. (2005): Advantages of the integrated pigbiogas-vegetable
greenhouse system in North China. Ecological Engineering,
24:177-185.
107

ABSTRACT
SOIL TILLAGE SYSTEMS AND NITROGEN FERTILIZATION
FOR WINTER BARLEY AFTER SOYBEAN 1
Stipesevic, Bojan, Jug, Danijel, Stosic, Miro, Zugec, Ivan, Jug, Irena
Faculty of Agriculture Osijek, Trg Sv. Trojstva 3, 31000 Osijek, Croatia
The winter barley crop production is not adequately researched regarding soil tillage
systems, especially in crop rotation with the soybean, both crops gaining importance as
food for the animals. The research at experimental site Boksic (Croatia), during the
years 2005 and 2006, showed no difference in yields from conventional tillage, based
on ploughing, and reduced tillage, based on diskharrowing, in each of six nitrogen
fertilization levels (0, 30, 60, 90, 120 and 150 kg N/ha). Regarding N fertilization, yield
increase was not significantly higher after applied 90 kg N/ha.
Keywords: winter barley, soybean, conventional tillage, reduced tillage, nitrogen
INTRODUCTION
The soil tillage systems for winter barley production had been reconsidered during the
last decade, especially in the light of the Croatian needs for more affordable and high
quality cattle fodder. This process is a result of worldwide trends and research results
about tillage simplifications for higher sustainability of the agriculture, in which the
environment protection and decreases of tillage costs is especially emphasised (Karlen
et al., 1994). In the Slavonia, the most agricultural region of the Republic of Croatia,
various systems of reduced tillage for different crops have been already tested
(Stipesevic et al., 1997, 2000; Zugec et al., 2000; Filipovic et al., 2006; Jug et al., 2006),
with main goals to decrease the costs of production, maintain agrosphere sustainability
and to preserve high yield (characteristic for this region) despite the reduction of applied
agritechniques. Along with the introduction of reduced tillage systems, the awareness
has been raised of different approach toward fertilization, soil compaction, weed control
and other problems connected with lesser soil agitation. The simplified soil tillage
particularly raised the question of efficiency of fertilizers, especially nitrogen, in
interaction with the tillage systems.
MATERIALS AND METHODS
This research was conducted near Boksic in Eastern Croatia, for the winter barley
(Hordeum vulgare L.) in a crop rotation after soybeans (Glycine max L.) for crop
seasons 2004/05-2005/2006. The site's soil type was determined as a eutric cambisol,
with loamy clay texture, total porosity between 32.2-44.7%, bulk density from 1.30 to
1.70 kg dm -3 , neutral reaction (pH in KCl 6.8), with rather high content of humus (4.%),
and with poor fertility (6.6 mg P2O5 and 6.8 mg K2O per 100 g of soil, 2.8 % of CaCO3)
in 0-30 cm depth. The main experimental set-up was a split-plot design in three
1 This research has been sponsored by Croatian Ministry for Agriculture, Forestry and
Water Management (VIP Project V-29-7/04) and family farm "Oto Kovacevic" from
Boksic, Croatia, on whose land this experimental site had been established.
108

epetitions, where the main treatment was soil tillage with two steps: CT=conventional
tillage (autumn ploughing up to 25 cm depth, spring diskharrowing, followed by
seedbed preparation with rototiller and standard sowing) and DS=autumn diskharrowing
up to 20 cm depth, seedbed preparation with rototiller in spring and standard sowing.
The sub-treatment of the nitrogen fertilization consisted of six steps of nitrogen
fertilization: N1=0, N2=30, N3=60, N4=90, N5=120 and N6=150 kg N ha -1 (see Table
1 for the nitrogen fertilization distribution), with the same amount of phosphorus (83 kg
P2O5 ha -1 ) and potassium (124 kg K2O ha -1 ) each season. The phosphorus and potassium
amounts were determined by soil analyses and planned crop uptake recommendations.
The basic experimental plot size was 5 m wide and 30 m long (total area of 150 m 2 ).
The winter barley cultivar "Trenk" was sown, the creation of the Agricultural Institute
Osijek, Croatia, in recommended plant density of 450 plants m -2 , within the optimal
sowing dates (31. October 2004 and 20. October 2005). During the harvest time, plots
were harvested one by one and complete grain mass from each plot was weighted on
portable electronic scale, whereas moisture content was determined by "Dickey John
GAC 2000" grain moisture meter, from ten subsamples taken during the harvest and
preserved in the plastic bags. Soil cone penetrologger "Eikelkamp" model SN with cone
diameter 2.00 cm and 60° cone angle was used for recoding soil resilience up to 60 cm
depth in 1 cm steps with 10 measures for each basic experimental plot, read once in
May 2005 and May 2006. The split-split-plot ANOVA was performed by SAS statistic
package (V 8.02, SAS Institute, Cary, NC, USA, 1999) with Year as the main level,
Tillage as sub-level and added N as sub-sub-level for the winter barley yields, and with
Year as the main level, Tillage as the sub-level and Depth as the sub-sub-level for the
soil resilience. The Fisher protected LSD means comparisons were performed for
P=0.05 significance levels.
Table 1: Fertilization scheme for the Boksic experimental trial
Fertilization levels
Autumn fertilization
0PK NPK Urea
1st
Sidedressing
KAN
2nd
Sidedressing
KAN
0:20:30 8:22:33 46% N 27% N 27% N
N1: 0 kg N ha -1 413 - - - -
N2: 30 kg N ha -1 - 375 - - -
N3: 60 kg N ha -1 - 375 - 111 -
N4: 90 kg N ha -1 - 375 - 111 111
N5: 120 kg N ha -1 - 375 - 167 167
N6: 150 kg N ha -1 - 375 65 167 167
RESULTS AND DISCUSSION
1. Soil resilience: Differences between two tillage treatments are presented in the
Figure 1, where DS treatment formed compacted pan shallower than the CT, which was
emphasised by Birkas et al. (2002) as "diskharrowing pan" and "ploughing pan".
Previous deeper soil tillage residual looseness below "diskharrowing pan" for DS
treatment is also clearly visible between 13-16 cm depth. Lower depth of soil
109

disturbance by reduced tillage was clearly notable through statistically significant
differences between two soil tillage treatments at 10-25 cm soil depth. In any case, the
detected soil resilience values did not present limiting factor for normal growth and
development of the winter barley, although some authors noticed that more compacted
0-30 cm layer under reduced soil tillage can contribute toward lower grain yield of these
treatments (Izumi et al., 2004).
Figure 1: Soil resiliance for the site "Boksic", average for 2005 and 2006
2. Winter barley yield: The rather favourable weather condition in both years managed
rather high yields for winter barley under both tillage systems with adequate available
nitrogen, with no statistical differences between Tillage treatments for all Nitrogen
treatment levels (Table 2), although DS tended to be slightly higher, especially at lower
Nitrogen levels (N1 through N3). This effect was probably the result of higher
concentration of incorporated soybean residues in shallower upper soil layers. At the
fertilization levels higher than 60 kg N ha -1 (N4 through N6), this difference tended to
decrease due to the sufficient nitrogen supply in both Tillage treatments. These results
were in accordance with some authors whose trials included ploughing and continuous
diskharrowing, who reported that, under favourable agroecological conditions, yields
did not differ among conventional tillage and other reduced tillage systems (Halvorson
et al., 1999). However, some other authors with similar soil tillage treatments recorded
110

that usually conventional tillage had better results than reduced soil tillage systems
(Varvel et al., 1989; Birkas et al., 2002), although some of them pointed out (Varvel et
al., 1989) that different placement and higher amount of nitrogen could alleviate
negative effects of reduced tillage and thus unfavourable soil preparation for cereals
growth at the yield level achieved with the conventional tillage based on ploughing.
Along with the increase of N amount, the winter barley responded with significant
yield growth between N2 to N3 (2.69 to 3.35 t ha -1 , respectively) and between N3 to N4
(from 3.35 to 3.85 t ha -1 ), with better crop reaction between N2 to N3 (for 663 kg ha -1 or
increase of 25% in comparison with the yield at N2 level), than between N3 to N4 (for
498 kg ha -1 or 15% if compared with N3). These yield leaps are corresponding with the
Mitscherlich's theory of lowering yield's increase, and roughly determining Nitrogen
levels N3 and N4 as Baule 2 and Baule 3 values of added nutrient, respectively.
Subsequently, nitrogen rates higher than 90 kg N ha -1 did not produce significantly
higher winter barley yields, which is leading toward the conclusion that, for given
agroecological conditions and observed cultivar of winter barley, there is ineffective to
fertilize with more than 90 kg N -1 in crop rotation after the soybean.
Table 2: Winter barley grain yield (kg ha -1 ) at 13% grain moistur, Boksic site,
Croatia, average for both years ( 2005 and 2006)
N level
CT
(Ploughing)
DS
(Diskharrowing)
Average (N)
N1 (0 kg N ha -1 ) 2380 a † 2571 a 2475 A ‡
N2 (30 kg N ha -1 ) 2625 a 2752 a 2688 A
N3 (60 kg N ha -1 ) 3224 b 3479 b 3351 B
N4 (90 kg N ha -1 ) 3841 c 3858 bc 3849 C
N5 (120 kg N ha -1 ) 3923 c 3928 bc 3925 C
N6 (150 kg N ha -1 ) 4036 c 4107 c 4072 C
average (Tillage) 3338 3449
†
The winter barley yields within the same Tillage level and labeled with the same
lowercase letter are not different at the P=0.01 significance level
‡
The winter barley yields labeled with the same uppercase letter are not different at the
P=0.01 significance level
ACKNOWLEDGEMENTS
This research has been sponsored by Croatian Ministry for Agriculture, Forestry and
Water Management (VIP Project V-29-7/04) and family farm "Oto Kovacevic" from
Boksic, Croatia, on whose land this experimental site had been established. Authors
would like to express their deep gratitude to Oto and Pavao Kovacevic for their
contribution during this project. Gratitude should also been given to the "Arvi" ltd.,
Croatian branch, for their contribution in fertilizers used for this experiment.
111

REFERENCES
Birkas, M., Szalai, T., Gyuricza, C., Gecse, M., Bordas, K. (2002): Effect of disk
tillage on soil. condition, crop yield and weed infestation. Rostlinna Vyroba, 48(1),
20-26 p.
Filipovic, D, Husnjak, S, Kosutic, S., Gospodaric, Z (2006) Effects of tillage systems
on compaction and crop yield of Albic Luvisol in Croatia. J. of Terramechanics, 43
(2): 177-189 p.
Halvorson, A.D., Black, A.L., Krupinsky, J.M., Merrill, S.D., Tanaka, D.L. (1999):
Dryland winter wheat response to tillage and N within an annual cropping system.
Agron. J. 91 (4), 637-642 p.
Izumi, Y., Uchida, K., Iijima, M. (2004), Crop production in successive wheatsoybean
rotation with no-tillage practice in relation to the root system
development. Plant Prod Sci 7 (3), 329-336 p.
Jug, D., Stipesevic, B., Zugec, I., Horvat, D., Josipovic, M (2006): Reduced soil
tillage systems for crop rotations improving nutritional value of grain crops. Cereal
Res. Comm., 34 (1): 521-524 p.
Karlen, D.L., Wollenhaupt, D.C. Erbach, E.C. Berry, J.B. Swan, N.S. Eash, J.L.
Jordahl. (1994): Long-term tillage effects on soil quality. Soil Tillage Res 32:
313-327 p.
Stipesevic, B., Zugec, I., Juric, I., Petrac, B. (1997): Possibility of reduced soil tillage
for winter wheat in East-Croatia conditions. Fragmenta Agronomica, 2B/97, 613-
616 p.
Stipesevic, B., Zugec, I., Josipovic, M. (2000): Investigation of Rational Soil Tillage
for Maize (Zea mays L.) in Eastern Croatia. ISTRO 15th Proc. Fort Worth, Texas,
USA (CD-ROM)
Varvel, G.E., Havlin, J.L., Peterson, T.A. (1989): Nitrogen placement evaluation for
winter wheat in three fallow tillage systems, SSSA J. 53 (1), 288-292 p.
Zugec, I., Stipesevic, B., Kelava, I. (2000): Rational Soil Tillage for Cereals (Winter
Wheat - Triticum aestivum L. And Spring Barley - Hordeum vulgare L.) in Eastern
Croatia. Proceedings of the 15th <strong>Conference</strong> of the <strong>International</strong> Soil Tillage
Research Organization (ISTRO), CD-ROM, 2-7 VII 2000., Fort Worth, Texas,
SAD
112

RESEARCHES REGARDING THE INFLUENCE OF THE CROP ROTATION ON
SOME INDICATORS OF THE WHEAT YIELD QUALITY IN THE CONDITIONS
OF THE CRISURILOR PLAIN
Gh. Bandici , C. Domuţa, Ileana Ardelean , Ioana Borza , Cr. Domuţa
University of Oradea, Environmental Protection Faculty, Oradea, Romania
ABSTRACT
The paper sustain the importance of the crop rotation on quality of the wheat yield and is
based on the results carried out during 2003-2006 in a long term trial out placed on the
preluvosoil from Oradea in 1990. Both in nonirrigated and irrigated conditions the smallest
values of the protein, wet gluten and dry gluten were obtained in wheat monocrop; the
values increased in the crop rotation wheat maize and the biggest values were registered in
the crop rotation wheat-maize-soybean.
INTRODUCTION
The quality of the wheat yield is influenced by many factors. Protein acumulation in the
grains is influenced by wheat type, cultivar, climate conditions, natural fertility of the soil,
nitrogen doses used, irrigation (Muntean L.S. et al, 2003, Domuţa C., 2005, Ardelean
Ileana, 2006). Gluten content of the wheat grain is influenced first of all by climate
conditions (Bîlteanu Gh., and Bîrnaure V., 1979, Bandici Gh., 1997).
The influence of the crop rotation and irrigation on the protein and gluten content is
presented in the paper.
MATHERIAL AND METHODS
The paper is based on the research obtained in the long term trial with crop rotation placed
in 1990 in Oradea on preluvosoil. On ploughing depth, the soil is low acid (pH= 6,8), humus
content is low (1,75%), phosphorus (22,0 ppm) and potassium (845,4 ppm) have medium
values; macroagregates hydrostability (47,5%) is high and bulk density (1,44 g/cm 3 ) is high,
too. The experiment dispositive includes:
Factor A: crop rotation
a1 = wheat, monocrop
a2 = wheat-maize
a3 = wheat-maize-soybean
Factor B: water regime
b1 = nonirrigated
b2 = irrigated
The surface of the experiment parcele = 50 m 2 . Number of repetition = 4. Place methods =
blocks method. Cultivar used: Dropia
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In the irrigated variant soil water reserve on 0-50 cm was maintained between easily
available water content and field capacity determining the soil moisture fifteen to fifteen
days and using the irrigation when the situation required. (Domuta C., 2005).
Dry gluten and wet gluten were determined by usually methods.
Gross protein was determined using the following formula: Nt x 5,7; when Nt = total
nitrogen. The rainfall registered during the vegetation period of the wheat from harvesting
were of 110,7 mm in 2003, 177,6 mm in 2004, 223,0 mm in 2005 and 287,2 mm in 2006.
RESULTS AND DISCUSSION
Crop rotation influence on protein content of the wheat grains
Both nonirrigated and irrigated conditions, crop rotations influenced the protein content of
the wheat yield. There were specifical situation for every year studied.
Protein content of the wheat grains determined in the wheat-monocrop in 2003 was of 9,1%
in nonirrigated conditions and of 9,0% in irrigated conditions. The values determined in the
wheat-maize crop rotation, 11,0% and 10,9%, were significant statistically bigger than
values from wheat monocrop. The biggest values of the protein content were registered in
the wheat-maize-soybean crop rotation, 13,8% and 13,7%; the differences in comparison
with monocrop, 4,7% both in nonirrigated and irrigated conditions is very significant
statistically.
In the year 2006, the smallest values of the protein content were registered in the
monocrop of wheat, too: 71% in nonirrigated and 6,9% in irrigated conditions. In the wheatmaize
crop rotation the values increased with 45% and 46% and in the wheat-maizesoybean
crop rotation with 73% and 77% respectively.
In average on the researched period, the smallest values of the protein content of the wheat
grains were registered in monocrop, 7,98% in nonirrigated conditions and 7,73% in irrigated
conditions. In the wheat-maize crop rotation the values of the protein content (10,7% and
10,45%) increased distingue significant in comparison with monocrop. The biggest values
of the protein content was obtained in the wheat-maize-soybean crop rotation, 13,02% in
nonirrigated and 12,93% in irrigated conditions.
Table 1: Crop rotation and irrigation influence on protein content of the wheat
grain, Oradea 2003 – 2006
Crop rotation
Water regime
Nonirrigated Irrigated
Protein
% % % %
Average on
the crop
rotation
1. Wheat- monocrop 7,98 100 7,73 100 7,86 Mt
2. Wheat-maize 10,7 135 10,45 135 10,56 **
3. Wheat-maize-soybean 13,02 164 12,93 167 12,98 ***
Average on the water regime 10,57 Mt
100 9,73 -
98,1 -
114

Crop rotation Water regime
Water regime x
Crop rotation
Crop rotation x
Water regime
LSD 5% 1,17 0,73 1,4 1,43
LSD 1% 2,16 1,46 2,6 2,73
LSD 0,1% 3,96 2,96 4,8 4,43
Crop rotation influence on wet gluten content of the wheat grains
Crop rotation influenced very strong the wet gluten content of the wheat grain. Every year
the smallest contents were obtained in wheat monocrop both nonirrigated and irrigated
condition.
The year 2003 was the year with the biggest drught and values of the wet gluten were
the biggest too.In wheat monocro, the values of the wet gluten were of 22,6% in
nonirrrigated conditions and of 21,9% in irrigated conditions. The values registered in the
whet-maize crop rotation (29,9% and 29%) and in the wheat-maize-soybean crop rotation
(36,1% and 33,8%) were very significant statistically bigger than the values registered in the
wheat-monocrop. (table 1).
The values of wet content registered in 2004 in whet-monocrop were of 20,4 % in
nonirrigated conditions and of 19,6% in irrigated conditions. There were very significant
differences in the wheat-maize and wheat-maize-soybean crop rotation; relative defferences
were of 36% and 61% in nonirrigated conditions and of 38% and of 63% in irrigated
conditions. This year were registered the biggest values of the wet gluten of the studied
period.
In 2005 in wheat-monocrop, the content of the wet gluten from grains were of 21,3 in
nonirrigated conditions and of 21% in irrigated conditions. Diferences registered in the
wheat-maize and wheat-maize soybean crop rotation were very significant statistically, 31%
and 61% in nonirrigated conditions, 30% and 57% in irrigated conditions respectivelly.
In the year 2006, the smallest values of the wet gluten were registered in the wheat
monocrop, 19,9% in nonirrigated conditions and 19,5% in irrigated conditions; in the wheatmaize
crop rotation the values increased with 36% and 37% and in the wheat-maizesoybean
crop rotation with 59% and 62%.
The average data of the period 2003-2006 show that the smallest content of the grain
wet gluten was registered in monocrop. In wheat-maize and in wheat-maize-soybean crop
rotation were registered the differences very significant statistically in comparison with
wheat-monocrop: 34% and 60% in nonirrigated conditions, 34% and 55% in irrigated
conditions, respectivelly. (table 2).
115

Table 2: Crop rotation and irrigation influence on wet gluten content of the wheat
grains, Oradea 2003 – 2006
Water regime
Average on
Crop rotation
Nonirrigated Irrigated the crop
% % % % rotation
1. Wheat- monocrop 21,1 100 20,5 100 20,8 Mt
2. Wheat-maize 28,2 134 27,5 134 27,85 ***
3. Wheat-maize-soybean 33,7 160 32,6 159 33,15 ***
Average on the water regime 27,7 Mt
100 26,9 o
96,9 -
Crop rotation Water regime
Water regime x
Crop rotation
Crop rotation x
Water regime
LSD 5% 1,43 0,75 1,70 1,63
LSD 1% 2,40 1,45 3,03 2,96
LSD 0,1% 4,46 3,41 5,24 5,05
Crop rotation influence on dry gluten content of the wheat grains
In 2003 the values of the dry gluten content from wheat grain for monocrop were of 10,8%
in nonirrigated and of 10,3% in irrigated conditions. The differences registered in wheatmaize
crop rotation were significant statistically, 19% in nonirrigated conditions and 17% in
irrigated conditions. In the wheat-maize-soybean crop rotation were distingue significant:
35% in nonirrigated conditions and 39% in irrigated conditions. (table 2).
The dry gluten content of the wheat grains in 2004 in the monocrop were of 9,8% in
nonirrigated conditions aof 9,3% in irrigated conditions. The statistically significant of the
differences vs. wheat-monocrop registered in the wheat-maize-soybean crop rotation have
similar statistically significant with the differences registered in 2003: significant and
distingue significant; the biggest values, 13,7% in nonirrigated conditions and 13,0% in
irrigated conditions, were registered in wheat-maize-soybean crop rotation. (table 3).
In 2005, the smallest values of the dry gluten were registered in wheat-monocrop, too:
10,2% in nonirrigated conditions and 9,4% in irrigated conditions. A similar situation with
2003 regarding statistically significant of the differences in comparison wheat monocrop
was registered in 2005, too. The biggest values of the dry gluten, 14,0% in nonirrigated
conditions and 13,3% in irrigated conditions, were registered in the wheat-maize-soybean.
In the year 2006, the smallest values of the dry gluten were registered in the wheat
monocrop, 9,5% in nonirrigated variant and 9,3% in irrigated variant. In the wheat-maize
crop rotation the values of the dry gluten increased with 23% both irrigated and nonirrigated
conditions and in the wheat-maize-soybean crop rotation with 38% and 39% respectivelly.
In average on the studied period, the values of the dry gluten content of the grains wheat
from monocrop were of 10,01% in nonirrigated conditions and of 9,58% in irrigated
116

conditions. The values, registered in wheat-maize crop rotation were significant statistically
bigger: (12,20% and 11,53%) and in wheat-maize-soybean were registered the biggest
values (13,88% and 13,38%) and differences distingue significant in comparison with
wheat-monocrop. (table 3).
Table 3: Crop rotation and irrigation influence on dry gluten content of the wheat
grains, Oradea 2003 – 2006
Water regime
Average on
Crop rotation
Nonirrigated Irrigated the crop
% % % % rotation
1. Wheat- monocrop 10,01 100 9,58 100 9,80 Mt
2. Wheat-maize 12,20 122 11,53 120 11,87 **
3. Wheat-maize-soybean 13,88 139 13,38 140 13,59 ***
Average on the water regime 12,03 Mt
100 11,49 -
95,6 -
Crop rotation Water regime
Water regime x
Crop rotation
Crop rotation x
Water regime
LSD 5% 0,91 0,65 1,18 1,14
LSD 1% 1,56 1,16 2,12 1,90
LSD 0,1% 2,49 2,14 3,95 3,48
CONCLUSIONS
The results obtained in a long term trial (1990-2006) emphasized the importance of the crop
rotation in the protein, wet gluten and dry gluten of the yield wheat.
During 2003-2006 both nonirrigated and irrigated conditions the smallest values of the
protein, wet gluten and dry gluten were obtained in wheat monocrop.
In comparison with wheat monocrop, in the wheat-maize crop rotation the values of the
protein, wet and dry gluten increased significant statistically. The differences very
significant statistically in comparison with the wheat monocrop were registered every year
in the wheat-maize-soybean crop rotation in all three parameters of the wheat yield quality
analyzed.
Irrigation determined to obtain smaller values of the protein, wet and dry gluten in the
wheat grains in comparison with nonirrigated variants from all the crop rotations.
117

REFERENCES
1. Ardelean Ileana (2006) - Contributions in the known and modification of the crop
rotation influence on quantity and quality of the winter wheat yield croped on the acid
soils from North-Western Romania. Thesis degree, USAMV Cluj-Napoca
2. Bandici Gh. (1997) – Contributions to establishing the influence of forreruner plant
and fertilisation level upon dinamics acumulation of wheat biomass cultivated on soils
with excessive moisture in centre of North-Western Romania. Thesis degree, USAMV
Cluj-Napoca.
3. Bîlteanu Gh., Bîrnaure V. (1979) – Phytotechny. Publishing house Ceres Bucureşti
4. Domuţa C., 2005 – Irrigation crops. Publishing house University of Oradea
5. Muntean L.S., Roman Gh..V., Borcean I., Axinte I., (2003) – Phytotechny. Ed. Ion
Ionescu de la Brad Iaşi
118

CRISANA -THE FIRST ROMANIAN WHEAT VARIETY WITH TOLERANCE
TO ALUMINIUM IONS TOXICITY FROM ACID SOILS
Gh. Bunta*, Elena Bucurean**
*Agricultural Research and Development Station Oradea,
**University of Oradea, Oradea, Romania
ABSTRACT
The paper present the new wheat variety registered in 2005 in Romania, Crisana. This
one was created in Agricultural Research and Development Station Oradea by
hybridization and repeated genealogical selection, especial for the acid soils from
Transilvania and other parts of Romania. The acid soils from hill zones have low pH
and aluminum ions toxicity, but Crisana variety is very tolerant to this. In the same
time, this new variety has very good backing qualities, being superior to the presents
Romanian wheat variety cultivated in the north part of Romania. In present, the variety
is in a multiplication process in the fields of Agricultural Research and Development
Station Oradea and it will be cultivated on acid soils from west and north of Romania.
Keywords: aluminum toxicity, variety, wheat, quality.
INTRODUCTION
In 1960, Neenan described for the first time the differential reaction of genotypes to
aluminum ion toxicity from acid soils. After this finding, the tolerance of plants to
aluminum ion toxicity was approach by many researchers. At the beginning, the
problem was studied in genetically point of whew in Brasilia and CIMMYT –Mexico
(Camargo and Felicio, 1988) and after this like a breeding objective (Aniol and
Gustafson, 1990).
In Romania, for the first time the problem was studied in 1987 (Bunta et al.), when
started a breeding program in cooperation with National Institute for Agricultural
Research and Development Fundulea. The study begun with elaboration of an original
method of testing in laboratory conditions, in hydroponics solutions. The researches
continued with the study of a large collection of genotypes to identify some genetically
sources of tolerance to ions toxicity. In the same idea, we collected many local wheat
populations and old cultivates (Bunta, 1997). After this, we initiated studies regarding
heredity of wheat tolerance to aluminum ions toxicity (Bunta, 1999/a; Bunta, 1999/b).
The new variety Crişana, which is for the moment the single Romanian wheat
variety with high tolerance to ions toxicity, has another qualities too, like a good
backing quality and high productive potential.
The studies regarding the breeding for tolerance to aluminum toxicity proceeding
for triticale in National Institute for Agricultural Research and Development Fundulea
(Ittu and Săulescu, 1988) and for wheat Agricultural Research and Development Station
Albota, too (Voica and Dumitrascu 2002).
MATERIALS AND METHODS
Th Crisana variety was created by genealogical repeted selection from hybrid
combination Fundulea 4 / Atlas 66.
119

The romanian Fundulea 4 variety vas utilized like a genitor becouse in 1986 it was the
most productive wheat variety in our country. It was too the most addapted to our zone,
an ideotype for north-west of Romania, an wet and colder geografic areal.
Atlas 66, an old variety from United States of America (South Carolina), has a lot
of defficiences, like any old cultivar: sensibility to diseases, tall, sensitive to falling and
too tardive. Hovewer, this variety have a very high content in protein and high tolerance
to aluminum ions toxicity, beeing an etalon in all genetical study.
The hibridation Fundulea 4/Atlas 66 was efectuated in 1986 and F1 hybrid vas
tested in 1987. During the period 1988 - 1990 (F2 – F4 generations) we selected every
year plantes with aluminum tolerance, in solution with growing concentration of
aluminum in laboratory and in a testing fild near Sudrigiu, in Beius Depression.
Beginning with the year 1991 (F5 generation) and finishing with 1996 (hybrid
generation F10) the selection has agronomical objective, in two experimental fields,
Sânmartin and Oradea. During 1997 – 1998 (D1 and D2), the breeding line Oradea 133
G vas tested in comparative micro-trials, simultaneous in all three fields. In the period
1999 – 2001 we proceded to testing of breeding line 133 G 41113124-S353 in paralel in 2
comparative experiments, in Oradea and Sânmartin. Because of it performances during
of all this years, during 2002 – 2004 the breeding line Oradea 133 was tested in network
of State Institute for Variety Testing and Registration (S.I.V.T.R.). Finally, in 2005 this
breeding line vas registred like variety with the name Crisana.
Simultaneoussly, we started the multiplication of seeds, than today the new variety
is cultivated on more than 200 ha in Agricultural Research Station Oradea fields.
To caracterize the genotipes regarding the tolerance to aluminium, we used an original
method (Bunta, 1996) and another two method addapted by proffesor dr. Gallia Butnaru
from the University of Agricultural Sciences and Veterinary Medicine of Banat county
(USAMVB) from Timişoara, the laboratory of genetics.
The qualities analyses were effectuated in the laboratory of National Institute for
Agricultural Researches and Development Fundulea and State Institute for Variety
Testing and Registration Bucureşti.
RESULTS AND DISCUSSIONS
The actuals Romanian cultivars do not have tolerance to aluminium toxicity or here
tolerance level is reduced (Albota, Fundulea 133, Arieşan, Trivale). The cause consists
in the fact that the Romanian breeding programs did not fallow this objective.
The breeding program for tolerance to aluminum ions toxicity started at
Agricultural Research and Development Station Oradea in 1982, like a program
regarding creation of varieties well adapted to local condition of hill zone from western
Romania.
The first stage look to transfer aluminum tolerance from spring wheat varieties
originated from Brazil and another country in local winter wheat genotypes. In the same
time, we look to associate aluminum tolerance with different another characters, like:
plant height, precocity, spike characteristics, the position and color of lives, etc.
To can realize the main objective of our program, it was necessary to create a
specific breeding scheme. These consist in the classic method (recombination and
selection), with some adaptations (fig. 1).
120

In addition, we used recurent selection to broke some correlations of aluminum
tolerance with: plant heigh, falling susceptibility, disease sensibility, bad coocking
quality and another indesirable phisiological or morphological characters.
SUDRIGIU SÂNMARTIN ORADEA
IIIIIIIIIIIIIIIIII
IIIIIIIIIIIIIIIIII
OIIIOIIIIOIIIIO
OIIIOIIIIOIIIIO
IIIIIIIIIIIIIIIIIII
IIIIIIIIIIIIIIIIIII
SIVTR
cecking
IIIIIIIIIIIIIIIIIIIIII
XXXXXXXXXX
00000000000000
00000000000000
**************
**************
OIIIOIIIIOIIIIO
OIIIOIIIIOIIIIO
IIIIIIIIIIIIIIIIIIIIII
IIIIIIIIIIIIIIIIIIIIII
IIIIIIIIIIIIIIIIIIIIII
IIIIIIIIIIIIIIIIIIIIII
Study of initial material
Hybridizations
Hybrids F1
Individual selection in fild
and laboratory, F2-Fn
Micro comparative trials
( control field)
Orientative trials
IIIIIIIIIIIIIII Comparative
IIIIIIIIIIIIIII trials
IIIIIIIIIII Preliminary
multiplication
Seed
multiplication
Figure 1: The original breeding scheme for creation of wheat variety with aluminum
tolerance
Finally, the best breeding lines are promovated to be tested in the State Institute for
Variety Testing and Registration network. Simultaneously the seed of promissing
breeding lines are multiplicated to reduce the period of extension in the large fields.
121

Class Genotype
Table 1: Regenerating capacity of some wheat breeding lines
48 ours later exposure to high Al 3+ rates
Root elongation (mm)
10 ppp 20 ppm 40 ppm
Al 3+
Al 3+
Al 3+
1 32 B 822 31,0±1,4 15,2±2,4 4,6±2,5
2 14 I 323 11,1±1,1 12,5±5,0 8,7±1,2
3 81 J 475 12,5±2,8 6,0±2,0 0,4±0,3
4 CRISANA 12,0±1,3 4,9±1,0 0,4±0,3
5 138 C 412 17,3±2,9 0 0
6 5 E 122 15,2±6,4 0 0
7 4 K 464 9,2±1,4 0,4±0,2 0
8 88 I 159 5,2±1,4 0 0
9 21 H 454 4,2±1,4 1,4±1,7 0
10 8 J 1010 4,5±1,5 1,5±0,5 0
11 10 K 487 1,4±1,2 0,4±0,4 0
12 29 D 125 1,8±0,9 0 0
13 19 I 197 1,6±0,3 2,9±0,9 0
14 93 J 222 0,3±0,3 0 0
15 10 F 414 0 0 0
16 32 F 132 0 0 0
17 35 F 133 0 0 0
18 76 J 545 0 0 0
19 128 G 128 0 0 0
20 FUNDULEA 29 0 0 0
Estimation
Very tolerant
Tolerant
Susceptible
From a large list of breeding lines created to have a high tolerance to aluminum ions
toxicity, some of them were evidentiated everytime, in any method of testing. Thus,
Oradea 133 line (the futhure variety Crisana) restarted his elongation of roots, even at
40 ppm Al 3+ in solution (table 1).
Another method are based on maximal roots length of plants at 4 ppm Al 3+ , after
10 days of exposure (table 2). The hydroponic solution consist only in distiled water and
aluminium ions, assured from AlCl 3, withouth any other mineral, at 4.5 pH, the
nutritive elements being assured by reserve elements from seed. This method is an
original one, adapted by us for a large series of breeding individuals in segregating
populations.
122

Table 2: The root length of some wheat genotypes at 4 ppm Al 3+ concentration
Class Genotype Roots length Differ. Differ. Estimation
(mm) % (mm) signiff.
1 Crisana 112,3 154,8 +39,7 *** Tolerant
2 Oradea 30 110,1 151,9 +37,6 ***
3 Oradea 33 93,4 128,8 +20,9 ** Medium
4 Fundulea 472 79,9 110,2 +7,4
tolerant
5 Arieşan 76,0 104,7 +3,5
Experimental average 72,5 100,0 0 - -
6 Fundulea 4 40,4 55,7 -32,1 000
7 Lovrin 34 35,8 49,4 -36,7 000 Susceptible
8 Fundulea 29 32,2 44,4 -40,3 000
LSD5%= 10,4 mm; LSD1%= 14,8 mm; LSD0,1%= 21,4 mm.
By total locations and years average (table 3), the variety Crisana produced a yield
output of 355 kg/ha comparative to check variety Fundulea 4 (106,1%), respective 610
kg/ha comparative to Apullum (110,5%). Comparative to the two-check average, the
yield output was 482,5 kg/ha (108,3%).
Location
Table 3: The yields of Crişana variety in SIVTR network
2002 2003 2004 Location averages
Kg/ha %
% check
averages
Kg/ha
Fundulea Apullum
4
Satu Mare 8596 3213 9064 6958 107,6 102,8 105,2
Huedin /Luduş 4785 3785 6848 5139 97,0 134,2 115,6
Galda de Jos 5268 4188 5952 5136 108,5 104,2 106,4
Sibiu 6546 5121 7678 6448 105,8 111,0 108,4
Hărman/Tg.Secuiesc 7083 1572 8036 5564 118,5 112,1 115,3
Rădăuţi 4705 4677 7527 5636 99,3 98,6 99,0
YEARS
AVERAGES
6164 3759 7518 5814 106,1 110,5 108,3
A simple estimation of economical efficiency demonstrate that, accepting an price of
0,11 dollars/kg, only in the west of Romania (more than 500.000 ha cultivated with
wheat), the total profit possible to be realized by cultivation with this new variety
exceed 25 million dollars/year.
123

Table 4: Quality indicators of Crisana variety comparative to check varieties utilized
in SIVTR network
Quality indicators Estim.
Variety
unit. Fundulea 4 Arieşan Apullum Crisana
Falling number sec.
Schrot analise
238 236 272 299
Total proteine % 14,12
Flour analise
13,94 14,53 14,71
Wet gluten % 26,35 26,72 27,50 27,28
Dry gluten % 9,75 9,82 10,67 10,70
Glutenic index - 75,09 75,76 84,21 87,56
Sedimentation index Ml 50,2
Flour graphic
52,5 57,5 60,5
Hidratation capacity % 67,0 65,5 66,1 68,5
Elasticity UV 100 95 92,5 95
Elongation UV 60 55 52,5 55
Flour strength - 61,2 64,7 63,2 62,5
Valuable index - 65,1 67,5 68,4 68,0
Estimating groupe - B1-A2 B1-A2 B1-A2 A2-B1
Crişana variety have a high yelding pottential (with a maximum of 9.064 kg/ha at Satu
Mare, in 2004), and a high milling and backing quality, demonstrated by the results
showed in table number 4. Quality analizes were dune in SIVTR Bucureşti laboratories
of chemistry on wheat sample collected from all the testing locations. Thus, the results
prove the quality stability of this new variety in different climatically conditions.
Falling number and total protein is higher even than Apullum, considered like a superior
quality variety.
The analises of flour (especially glutenic index) proved the superior quality of
Crisana variety. These index and flour graphic analize were utilized to calculate
valuable index and classiffied Crisana variety, by this index, in the groupe A2-B1 (in the
rate of 3:1). Thus, this variety is a very good one, having the capacity to ameliorate the
flour quality of another variety, by mixture.
In 2006, the new variety Crisana was tested for yield capacity and backing quality
comparative to another 15 ones (table 5). Crisana has a good yielding potential and
quality indicators, compatible with the best Hungarian varieties created in Martonvasár
Institute. In adition to the high yielding pottential, the tolerance to soil acidity and
aluminium ions toxicity, the superior milling and backing quality, another characteristics
are very good in case of Crisana variety: the good or very good resistance to disease,
winter frost and falling
124

Table 5: Productions and quality results realized by the new wheat variety
Crisana in the demonstrative field. Biharia, 2006
Class. Variety
Yield Quality index
kg/ha % Wet Total Zeleny
gluten % protein % index
1. Toborzó 7148 118.4 30 13.9 46
2. Pálma 7003 116.0 27 13.7 51
3. Emese 6861 113.6 30 14.1 52
4. Marsall 6787 112.4 30 14.9 54
5. Romulus 6370 105.5 30 14.5 53
6. Süveges 6348 105.1 31 15.7 59
7. Crisana 6165 102.1 31 14.2 54
8. Capo 6086 100.8 29 14.1 58
9. Palotás 6067 100.5 32 15.5 61
Experimental
average
6038 100.0 29.9 14.6 57.1
10. Kristina 5890 97.5 23 12.5 52
11. Magdaléna 5653 93.6 32 15.7 64
12. Csárdás 5369 88.9 32 16.0 63
13. Magvas 5326 88.2 33 15.4 61
14. Verbunkos 5285 87.5 31 15.4 58
15. Joszef 5189 85.9 31 15.0 73
16. Apache 5059 83.8 27 13.6 54
Acording with all this characteristics, the State Institute for Variety Testing and
Registration recommended in his Official Catalogue (2005) that the new variety Crisana
to be cultivated in Transilvania, on acid soils from hill zone.
REFERENCES
Aniol, A., Gustafson, J. P., (1990) - Genetics of tolerance in agronomic plants. Heavy
metal tolerance in plants: evolutionary aspects. Ed. A. J. Shaw, Boca Raton, USA,
255-267 p.
Bunta, Gh., Bunta, Anuţa, Negruţiu, I., (1987) – Toleranţa la aciditate şi exces de
ioni de aluminiu din sol. Principalele obiective de ameliorare a grâului pentru zona
colinară şi de deal din vestul ţării. Probl. genet. teor. aplic., XIX, (4), 173-188 p.
Bunta, Gh. (1996) - Cercetări privind efectul ionilor de aluminiu asupra lungimii
rădăcinilor câtorva soiuri de grâu. Analele Universitatii Oradea, fasc. Agric., II,
121 - 130 p.
Bunta, Gh., (1997) - Identificarea unor surse genetice de toleranţă la aluminiu în cadrul
populaţiilor locale de grâu. Probl. genet. teor. aplic., XXIX, (1-2), 11-21 p.
Bunta, Gh., (1999/a) - Results regarding the genetic control of tolerance to aluminum
ion toxicity in wheat. Romanian Agricultural Research, (11-12), 1-8 p.
125

Bunta, Gh., (1999/b) - Ereditatea nucleo-citoplasmatică a toleranţei grâului la
toxicitatea ionilor de aluminiu. Probl. genet. teor. aplic., XXXI, (1-2), 1-12 p.
Camargo, C., E., Felicio, J., C., (1988) – Wheat breeding at the Campinas Agronomic
Institute. Wheat breeding for acid soils; review of Brazilian/CIMMYT
collaboration, 1974 - 1986. Mexico, D., F., CIMMYT, Ed. Kohli, M., M. and
Rajaram, S., 39-49 p.
Ittu,Gh., Săulescu, N., (1988) – Cercetări privind crearea de soiuri de grâu de toamnă
tolerante la toxicitatea produsă de ionii de aluminiu. Probl. Genet. Teor. Aplic.,
XX, 3: 139 – 149 p.
Neenan, M., (1960) – The effects of soil acidity on the growth of cereals with particular
reference to the differential reaction of varieties thereto. Plant Soil, (12), 324 -338
p.
Voica, Maria, Dumitraşcu, Navara, (2002) – Toleranţa unor genotipuri de grâu de
toamnă la toxicitatea ionilor de aluminiu. An. ICCPT Fundulea, LXIX : 51 – 63 p.
*** 2005 – Catalogul oficial al soiurilor de plante de cultură din România. Bucureşti,
110 – 111p.
126

EXPERIMENTS WITH PERENNIAL RYE (SECALE CEREANUM) IN HUNGARY,
AT UNIVERSITY OF DEBRECEN
ABSTRACT
Erika Halász – Tamás Sipos
Research Centre of Nyíregyháza, University of Debrecen
Perennial rye as a new culture crop has some problems which make more difficult the
growth technology. The most important from these is that germination of seeds is compared
to winter rye very few (50-70). We found correlation between the rev and the germination
ability, but it wasn’t any correlation between harvesting in one step and two steps.
In the Research Centre of Nyíregyháza we started a development of a new forage
mixture with perennial rye and perennial papilionaceae (alfalfa, bird’s-foot trefoil). The two
plants with different fodder value in the mixture are complementary to each other from the
point of view of dietetics; the mixture produce great mass fodder with digestible fiber and
high quantity of crude protein. In second year perennial rye produced 30-35 t/ha green
mass, but the mixed stand yielded 78-85 t/ha green mass. We didn’t find relevant
differences between total green yields or dry-matter products of mixed treatments, but the
proportion of two components differed remarkably.
Keywords: perennial rye, germination, forage mixture
INTRODUCTION
Perennial rye (Secale cereanum) is a stable hybrid of the winter rye (Secale cereale L.) and
the perennial wild rye (Secale montanum Guss.). Crossings in the genus Secale were made
already at the end of the XIX century to study crossability, relationship, influence on
cytological parameters, etc. (Kotvics, 1963). Later the objectives of plant breeding (e.g.
increasing the winter and drought resistance, developing resistance to biotic and abiotic
factors, increasing the protein content) get more importance. Many researchers found it
interesting to make a new perennial rye crop. The aim was to develop a hybrid, which all
over its botanical importance unites the best qualities of winter rye and perennial rye in
form of a perennial, highly productive, highly tolerant crop with good fodder value, which
can be grown also on weak habitats. The hybrid was developed in several countries.
Kotvics G. made the first plants in Hungary, which were useful for the plant breeding, in
the 1950’s (Kotvics, 1970.).
Kruppa J. developed the variety “Kriszta” from this material at the Research Centre of
Nyíregyháza (University of Debrecen), which got the state allowance in 1998. Another
perennial rye variety was created from the same origin in Hungary, at Research Institute in
Kompolt. Diploid and tetraploid varieties were made in Germany from similar crossings
(Reimann-Philipp, 1995).
Perennial rye is a new crop of the disadvantageous fields, and it has some problems of
its growth technology: the germination of the seeds is often not good, about 50-70% (likely
because of their form they are sensitive for trash), the spikes are often fragile, etc. These
problems can be solved on the one hand through plant breeding, on the other hand through
improvement the growth technology – and this latter way seems to be faster.
There aren’t too much possibilities of utilization of weak acid sandy soils. In our
opinion we can get from the same area on these soils more valuable fodder if we produce
perennial rye mixed with perennial papilionaceae.
127

MATERIALS AND METHODS
Perennial rye variety of our experiments was ‘Kriszta’. Owing to its high green
productivity, low seed productivity and perennial type it is utilized as a green fodder, as
grazing land or grass-land. This variety can be in culture 3-5 years, after this time the stand
grows thinner and thinner, and get weedy. Its seed-production is about 500-1000 kg pro
year. The plant is 150-180 cm high, thousand-kernel weight: 12-15 g
It can be sown with 3-3.5 million seeds in September and in the springtime in March, too.
Its drought resistance is very good, this perennial crop adapts well to the disadvantageous
soil and weather conditions, so it can be sown also on weak habitats, loose sandy soils. It
covers the fields the whole year. Attributable to this, to the fast shooting and excellent
stooling (the number of its shoots can reach the 100) perennial rye is instrumental in
protecting the loose sandy soils from erosion and deflation – it fixes the surface of the land.
Expectedly its role will extend in the game farming too, because it is exceedingly suitable
to winter pasture: it can be grazed also in winter and it tillers fast early in spring.
In one of our experiments we analyzed the relationship between the germination and
some harvest and thresh methods: we studied the effect of the set of threshing machine and
combine (rev) on germination ability. We harvested perennial rye in two ways: one-stepharvesting
and two-steps-harvesting (threshing one weak after mowing). Several standards
were threshed by hand. The germination was carried out accordingly to the cereal standard.
In our other experiment we started the development of a new forage mixture with two
components (lucerne (Medicago sativa L.) or bird’s-foot trefoil (Lotus corniculatus L.) and
perennial rye (Secale cereanum)). According to our previous experiments we established
that for this mixture lucerne is suitable, bird’s-foot trefoil is suppressed by perennial rye.
In a forage mix experiment we prepared 7 different mixes with perennial rye (variety
Kriszta) and alfalfa (variety Klaudia) and we compared the mixtures with two standard
treatments (perennial rye alone in 1,8 and 2,7 million plant per hectare doses). According to
previous experiments, we choose perennial rye doses of 0.4, 0.7, 0.8, 1.4 million plant/ha,
and 2.5, 3.2, 5.6 million alfalfa plant/ha. The small parcels of this experiment were sown on
acid sandy soil (pHKCl=4,35, KA=28, H%=0,78) in Kisvárda in March of 2002. We mowed
the parcels four times a year and weighed the green and dry mass product. We compared
the products of the components by mowing and the results of the mixed sowing and
separately–rows sowing.
RESULTS
The results of germinating study are shown in Figure 1. In cases of either threshing
methods we found correlation between the rev and the germination ability. By the plot
combine threshing and also by the single ear thresher we found that the lowest rev
eventuates the best results (82,3 and 92,5 %), which don’t differ significantly from the
control threshed by hand (89,0 %). The disadvantage of low rev is that after threshing a lot
of seeds stay in the ears. The difference between the two mechanical methods follows
probably from the length of threshing-time: the ears stay much shorter in the threshing
drum of plot combine than in the single ear thresher. Between the one step and two steps
harvesting methods on the score of germination we didn’t find any significant differences.
In the forage mix experience the productivity was considerably low in the year of sow,
the green yield suffice for grazing. From the second year the relation of perennial rye and
papilionaceae changed as follows: in the first growth the perennial rye was presented by 40-
60 % in the green mass, this value later decrease and the fourth growth we could harvest
only alfalfa. Because of the frequent mowing technology (which adapts to alfalfa) the
forage mix can utilize for three years, then the perennial rye get thinner. After this the area
can utilized as alfalfa seed producer field. In second year perennial rye produced 30-35 t/ha
128

green mass (6,8-7 t/ha/year dry mass), but the mixed stand yielded 78-85 t/ha green mass
(15,7-16,5 t/ha/year dry mass – Table 1.).
Germination %
100
90
80
70
60
50
40
30
20
10
0
one-step-harvesting
two-steps-harvesting
Rev 1/min 1000 1500 1800 1000 1500 1800
Threshed by: Plot combine Single ear thresher Manually
Figure 1: Effects of harvest methods on germination ability of perennial rye
Table 1: Yield in the second year of forage mix experiment, Kisvárda 2004.
Treatments
Sowing
method
Product of green yield Product of dry-matter
t/ha
t/ha
P. rye Alfalfa Total P. rye Alfalfa Total
1. P. rye 1,8 Mp/ha 30,93 - 30,93 6,87 - 6,87
2. P. rye 2,7 Mp/ha 33,42 - 33,42 7,07 - 7,07
3.
4.
5.
6.
7.
8.
9.
P. rye 1,4 +
Alfalfa 3,2 Mp/ha
P. rye 1,4 +
Alfalfa 5.6 Mp/ha
P. rye 0,7 +
Alfalfa 3,2 Mp/ha
P. rye 0,7 +
Alfalfa 5,6 Mp/ha
P. rye 0,4 +
Alfalfa 3,2 Mp/ha
P. rye 0,4 +
Alfalfa 5,6 Mp/ha
P. rye 0,8 +
Alfalfa 2,5 Mp/ha
mixed 15,18 69,34 84,52 2,88 13,41 16,30
mixed 17,42 64,28 81,70 3,31 12,73 16,03
mixed 9,07 72,88 81,95 1,74 13,41 15,15
mixed 11,25 72,88 84,13 2,04 14,02 16,07
mixed 11,10 73,57 84,67 2,16 14,20 16,36
mixed 8,00 78,90 86,90 1,57 14,95 16,52
separated
rows
21,99 56,78 78,77 4,45 11,30 15,74
129

We didn’t find relevant differences between total green yield or dry-matter products of
mixed treatments, but the proportion of two components differed remarkably (product of
perennial rye was 17,96 % in the 3. treatment and 9,2 % in the 8. treatment).
Our experiment proved that perennial rye is adapted to mixing with alfalfa. The new
forage mixture is able to produce considerable green yield from the second year either on
acid sandy soil. The two forage crop gave more yield in the experiment when they mixed
than they sowed in separately rows (which is hardly can be implemented in the practice).
DISCUSSION
To bring this new plant species into use it’s essential to solve the problem of seedproduction.
The primary method of this could be the right set of thrashing parameters of
combine-harvester.
Perennial rye can play a role in forage-mixture, mixed with perennial papilionaceae. The
new forage mixture has the follow environmental and economic advantages:
• The two plants with different fodder value in the mixture are complementary to each
other from the point of view of dietetics; the mixture produce great mass fodder with
digestible fiber and high quantity of crude protein.
• Because of the lack of fodder on pastures in summer it is necessary to eat hay or apply
additional feeding at this time. The mixture with perennial rye can produce pasturable
green mass from spring to autumn.
• If the mixture is utilized by grazing, the surplus of green yield in spring and autumn
can be utilized by making hay or silage, which can make use of feeding in winter.
• The influence of this new mixture from papilionaceae and gramineae is advantageous
for the soil. Alfalfa has deep taproot, that reduces the compactness of soil and it can
collect nitrogen from the air. The fibrous root system of perennial rye contributes to
development of favorable structure of surface soil.
• It is advantageous from the point of water balance of soil, that perennial rye and alfalfa
utilize the water from different bed owing to they different root-system and they don’t
enter into competition with each other.
• The forage mixture can profitably be utilized for three-five years, it depends on the
soil and intensity of utilization, so the expense of planting is divided among some
years and the maintenance cost is low.
• This mixture can play great part in soil protective agriculture, because the perennial
culture overgrow the surface of the soil during the whole year and it affords possibility
of ecological farming.
• The new forage crop association makes the sustainable farming possible for a long
time, which is given financial assistance by the European Union (Sipos 2006/B.).
130

REFERENCES
Kotvics G. (1963): A Secale cereale L., Secale montanum Guss. és hibridjeinek
morfológiai, citogenetikai és fejlődés-élettani tulajdonságainak vizsgálata, kandidátusi
értekezés, Gödöllő.
Kotvics G. (1970): Investigations os increasing the protein content of Secale cereale L. In
Ed. Bálint A. : Protein Growth by Plant Breeding 89-98 Akadémiai Kiadó, Budapest.
Reimann-Philipp R. (1995): Breeding Perennial Rye. In. Ed. Jules Janick 0-471-.57343-4
Plant Breeding Reviews, Volume 13.
Sipos T., Kruppa J. (2006/A): A Kriszta évelő rozs – alternatív energianövény a gyenge
termékenységű homoktalajokra. Őstermelő, 10. évf. 3. sz.
Sipos T., Halász E. (2006/B): A „Kriszta” évelő rozs fajta szerepe a homoktalajok
védelmében. Előadás, V. Alföldi Tudományos Tájgazdálkodási Napok, Mezőtúr 2006.
Lektorált kiadvány 117. o.
131

MYCOTOXINS CONTAMINATION IN FRESH AND SILAGE
OF ALFALFA (Medicago sativa)
B. Gálik, D. Bíro, M. Juráček, M. Šimko, J. Michálková
Slovak University of Agriculture, Department of Animal Nutrition, Nitra,
Slovak Republic
ABSTRACT
The aim of this work was detection of mycotoxin contamination in fresh alfalfa (dry
matter: 228 g.kg -1 ) and in alfalfa silage. Ensiled matter was wilted at six hours. Dry
matter content was 277.7 g.kg -1 in silage. Concentrations of mycotoxins
(deoxynivalenol 5/5, fumonisin, T-2 toxin, aflatoxin and ochratoxin) were determined
in fresh forage and silage of alfalfa. This was used immuno-enzymatic analysis by
ELISA Reader (Noack, Slovak Republic) for investigation amount of mycotoxins.
Concentrations of mycotoxins were higher in silage in comparison with concentrations
of mycotoxins in fresh matter.
The value of deoxynivalenol (DON) was 100 μg.kg -1 in fresh matter and 233.3
μg.kg -1 (average concentration) after conservation. We found 61.9 μg.kg -1 of fumonisin
after conservation and 51.2 μg.kg -1 in fresh matter. The content of T-2 toxin was 21.9
μg.kg -1 before ensiling and 88.5 μg.kg -1 in silage of alfalfa (minimum was 72.3 μg.kg -1
and maximum was 99.2 μg.kg -1 ). Contents of mycotoxins were produced by storage
fungi (aflatoxin and ochratoxin) were higher in silage than numbers of mycotoxins in
fresh alfalfa. Concentrations of all detected mycotoxins were lower than critical values.
Keywords: mycotoxins, contamination, fresh matter, silage, alfalfa
INTRODUCTION
Mycotoxins are toxic secondary metabolites produced by fungi. There are hundreds of
mycotoxins known, but few have been extensively researched and even few have good
methods of analysis available (Nelson et al., 1993, Nedělník, Moravcová, 2006).
Mycotoxins exhibit a variety of biological effects in animals: liver and kidney toxicity,
central nervous system effects and estrogenic effects, to name a few (Whitlow et al.
1998). Worldwide, approximately 25% of crops are affected by mycotoxins annually.
Many different mycotoxins have been found to occur on forages either in the field or in
storage as hay or silage (Lancey, 1991). The limiting factor for fungi growth in hay is
moisture, in silage pH (Kalac, Woolford, 1982). At present nearly 400 mycotoxins
produced by a wide spectrum of fungal pathogens are described but the occurrence of
only the most frequent and harmful ones is monitored on a regular basis (Lew et al.
2001). Fungi and their mycotoxins identified in feeds (silage) may occur either through
their deleterious effects on nutrient and hygienic quality (Tapia et al. 2005).
Aflatoxins are a family of extremely toxic, mutagenic and carcinogenic
compounds produced by Aspergillus flavus and Aspergillus parasiticus (Deiner et al.
1987). Zearalenone is estrogenic metabolite of several species of Fusarium.
Zearalenone has been reported to occur in corn, other grains and silage in many areas
of the world (Hagler et al. 1984). Trichothecenes, T-2 toxin and deoxynivalenol too, are
commonly found in agricultural commodities (Desjardins et al. 1993). Fumonisins has
132

een shown to cause leucoencephalomalacia in horses (Marasas et al. 1988), pulmonary
edema in swine (Harrison et al. 1990) and hepatoxicity in rats (Geldebrom et al. 1991).
Unlike foodstuffs, ultimate concentrations of individual mycotoxins are not for
individual categories of animals and feeds in many European countries. Therefore
detected results were compared to recommended limits published in the United States
of America. The critical concentration of zearalenone in all feeds and categories of
animals is 0.5 mg.kg 1 , the limiting concentration of T-2 toxin is the same. The critical
concentration of fumonisin is 5 mg.kg -1 . The limiting concentration of deoxynivalenol
is specific for cattle 10 mg.kg -1 , the critical amount of deoxynivalenol is 5 mg.kg -1 for
pigs.
The intention of the present experiment was to establish mycotoxins contamination
in fresh forage and silage of alfalfa.
MATERIAL AND METHODS
Second cut alfalfa (Medicago sativa) was harvested at the blooming stage, variety
Palava. After wilting (6 hours), alfalfa was chopper by chopper harvester to 15 mm
particle length. Wilted matter of alfalfa was conserved (281.6 g.kg -1 of dry matter)
without additives in silage units (capacity 15 dm 3 ). Silage units were opened after two
months. Samples of fresh alfalfa and silage were analyzed for mycotoxin
contamination. Fresh matter and alfalfa silage were tested on a number of mycotoxin in
two replicates for each mycotoxin. The following mycotoxins were monitored:
zearalenone, deoxynivalenol 5/5, fumonisin, aflatoxin and ochratoxin. We used
immuno-enzymatic method for determination amounts of mycotoxins by ELISA reader
(Noack, Slovak Republic) with veratox tests (quantitative test). Principle of the test is
determination of concentration in extract of feedstuffs by spectrophotometric method
(650 nm). Before extraction, samples had to been homogenous. Extraction was
performed with distilled water (DON 5/5), 70% solution of methanol (V/V) for
fumonisin, zearelenone and aflatoxin or 50% solution of methanol (V/V) for ochratoxin
and T-2 toxin.
RESULTS AND DISCUSSION
Many different mycotoxins have been found to occur on forages either in the field, or in
storage as hay or silage (Lacey, 1991). Silage contained higher numbers of mycotoxins
in comparison to amounts of mycotoxins in fresh matter (Table 1). Content of
deoxynivalenol was range from 200 to 300 μg.kg -1 it was 100 μg.kg -1 in fresh matter.
The highest concentration of fumonisin was 71.9 μg.kg -1 . Fumonisin has been shown to
reduce milk production in dairy cattle at 100 mg.kg -1 (Diaz et al., 2000). We found
similar results of zearalenone like Nedělník and Moravcová (2006), range from 635.8 to
703.1 μg.kg -1 . In Slovak climatic conditions, mycotoxins (aflatoxin and ochratoxin)
have produced by storage fungi (Leibetseder, 1995). We found aflatoxin with toxic
mutagenic and carcinogenic characteristics (Deiner et al., 1987) range from 10.4 μg.kg -1
to 11.9 μg.kg -1 , lower than reported Kalac and Woolford (1982). The concentration of
ochratoxin was 12.5 μg.kg -1 in fresh matter, it was range from 8.2 to 14.5 μg.kg -1 in
silage.
133

Table 1: Concentrations of tested mycotoxins in fresh matter and alfalfa silage
Indicators of
DON ZON T-2 FUM AFL OT
contamination µg.kg -1
fresh
matter
alfalfa
silage
n=2 100 210 21.9 51.2 5.2 12.5
Min. 200 635.8 72.3 52.8 10.4 8.2
Max. 300 703.1 99.2 71.9 11.9 14.5
n=6 Average 233.3 451.4 88.5 61.9 7.4 7.6
DON: deoxynivalenol, ZON: zearalenone, T-2 : T-2 toxin, FUM : fumonisin,
AFL : alfatoxins, OT : ochratoxin
REFERENCES
Deiner, U.L., Cole, R.J. et al. 1987. Epidemiology of aflatoxin formation by Aspergillus
flavus. In: Annual Review of Phytopathology, vol. 25, 1987, p. 240-270.
Desjardins, A.E., Hohn, T.M., McCormick, S.P. 1993. Trichothecene biosyntesis in
Fusarium sp.: chemistry, genetics and significance. In: Microbiology Reviews,
vol. 57, 1993, p. 594-604.
Diaz, D.E., Hopkins, L.M., et al. 2000. Effect of fumonisin on lactating dairy cattle. In:
Journal of Dairy Science, vol. 83, 2000, p. 1171 (abstr.).
Geldebrom, W.C.A, Kreik, N.P.J., et al. 1991. Toxicity and carcinogenicity of the
Fusarium moniliforme metabolite, fumonisin B1, in rats. In: Carcinogenesis, vol.,
12, 1991, p. 1247-1251.
Hagler, W.M., Jones, F.T., Bowman, D.T. 1984. Simultaneous occurrence of
deoxynivalenol, zearalenone and aflatoxin in 1982 scabby wheat from the
Midwestern United States. In: Applied of Enviromental Microbiology, vol. 47,
1984, p. 151-154.
Harrison, L.R., Colvin, B.M., et al. 1990. Plumonary edema and hydrothax in swine
produced by fumonisin B1, a toxic metabolite of Fusarium moniliforme. In:
Journal of Veterinary Diagn. Invest., vol., 2, 1990, p. 217-221.
Kalac, P., Woolford, M.K. 1982. A review of some aspects of possible associations
between the feeding of silage and animal health. In: British Veterinary Journal,
vol. 138, 1982, p. 305-320.
Leibetseder, J. 1995. The European perspective on mycotoxins. In: Lyons, T.P.,
Jacques, K.A.: Biotechnology in the Feed Industry, Nottingham University Press:
Notthingam, 1. vyd., 496 s., ISBN 18-97-67656-5, s. 65-75.
Lancey, J. 1991. Natural occurrence of mycotoxins in growing and conserved forage
crops. In: J.E. Smith, R.E. Henderson: Mycotoxins and Animal Foods. 1991, 1.
edt., Boca Raton Florida: CRC Press, p. 363-379.
Lew, H. et al. 2001. Occurrence of toxigenic funghi and related mycotoxins in plants,
food and feed in Austria. In: Occurrence of toxigenic funghi, Cost Action 835,
European Commision, 2001, s. 3-12.
134

Marasas, W.F.O., Kellerman, W.C.A, et al. 1988. Leucoencephalomalacia in a horse
induced by fumonisin B1 isolated from Fusarium moniliforme. In: Onderstepoort
Journal of Veterinary Research, vol., 55, 1988, p. 197-203.
Nedělník, J., Moravcová, H. 2006. Mycotoxins and forage crops problems of the
occurrence of mycotoxins in animal feeds. In: 12 th <strong>International</strong> Symposium:
Forage Conservation, VFU: Brno, 2006, p. 15-26.
Nelson, P.E., Desjardins, A.E., Plattner, R.D. 1993. Fumonisins, mycotoxins produced
by Fusarium species : Biology, chemistry and significance. In: Annual Review of
Phytopathologie, vol. 31, 1993, p. 233-249.
Tapia, M.O., Stern, M.D., Soraci, A.L. et al. 2005. Patulin-producing molds in corn
silage and high moisture corn and effects of patulin on fermentation by ruminal
microbes in continuous culture. In: Animal Feed Science and Technology, vol.
119, 2005, p. 247-258.
Whitlow, L.W., Hagler, W.M., Hopkins, B.A. 1998. Mycotoxins occurrence in farmer
submitted samples of North Carolina feedstuffs 1989-1997. In: Journal of Dairy
Science, vol. 81, 1998, p. 1189.
135

USAGE OF NATURAL ADDITIVES FOR BROILER CHICKENS
Erika Horniaková, Ladislav Bušta
Department of Animal Nutrition, Faculty of Agrobiology and Food Resource,
Slovak University of Agriculture - Nitra, Slovakia
ABSTRACT
The goal of this experiment was an additional verification of the natural additive
preparation Clinoptilolit of sedimentary origin with an addition of microelements and
citric acid, the influence of this APC feed mixture on feed consumption, feed
conversion, live weight and weight gain in broiler chickens.
The trial included broiler chickens of the ROSS 308 type which were divided into a
control group (C) and 2 experimental groups (T1 and T2), each containing 100 chickens
with four repetitions. The feed mixtures differed in concentration according to crude
protein and clinoptilolit preparation. The feeding periods were determined by the total
consumption of 15 kg HYD-01/1, alternatively 65 kg HYD-01/2, further 100 kg of
HYD-02 and finally 170 kg of HYD-03. The highest concentration of crude protein was
offered to be 230 g.kg-1 in the control group, a lower value of 215 g.kg-1 in the first
trial group and 200 g.kg-1 in the second group as starter food (HYD-01/1). During
fattening the protein concentrations were slowly decreased to 220.0, 210.0 and 200.0
g.kg-1 in HYD-01/2, 200.0, 190.0 and 180.0 g.kg-1 in HYD-02 and for finishing the
feed mixture HYD-03 contained 190.0, 180.0 and 170.0 g.kg-1 . In the groups T1 and
T2 the additive clinoptilolit was added on a 0.2% concentration level.
The highest total feed intake per chicken per day was in the C group (101.39 g) as
compared with the experimental groups, which consumed 93.65 % and 96.92 % of feed
respectively. A similar tendency was found in the intake of dry matter, crude protein
and energy. The live weights were 1956 g, 2137 g and 2001 g in the same order of the
considered groups. The differences were statistically significant on the level P

minerals (Čuvanová et al. 2006). The usage of zeolits on an 0.2% dose along with
nicarbazin and narazin positively affected daily weight gain (P

weight and weight gain. These parameters were determined individually in the trial
group and were compared with the parameters determined in the control group.
Statistical evaluations were carried out with the program
RESULTS AND DISCUSSION
In evaluating the average feed consumption for the starter period (HYD-01/1) for one
broiler chicken per day (table 3) feed consumption in the control group was 24.27 g and
in the experimental groups the observed values were 23.55 g for group T1 and 23.85 g
for group T2. The relative values were 2.97 % less in T1 and 1.73 % less in T2 less
compared to the control group. For the prefattening period (HYD-01/2) feed
consumption in the control group was highest (67.90 g) in comparison with T1 and T2,
which resulted in smaller values to be 6.42 % and 4.14 % less. The same parameters in
the groups were evaluated also for the fattening period (HYD-02) and finishing period
(HYD-03). For the total feed consumption for one broiler chicken per day, feed
consumption in the control group was 101.39 g, 94.95 g in the first trial group and 96.92
g in the second trial group. Average feed consumption per kg live weight showed the
same tendency within the considered groups as the other parameters. Values for average
feed conversion were observed to be highest in the control group (1.906 kg), while in
the experimental groups values were lower, in the trial groups T1 (1.840 kg) and T2
(1.873 kg) during the whole experiment.
Table 2: Analytical analysis and nutritional value of FM
FN group CP MEN Ca P
HYD-01/1
HYD-01/2
HYD-02
HYD-03
C 233.4 12.9 13.7 7.8
T1 217.9 12.8 10.2 5.2
T2 206.2 12.8 10.1 5.2
C 227.9 12.9 12.5 7.1
T1 209.1 12.8 8.9 4.9
T2 198.4 12.8 8.0 4.7
C 207.0 12.8 9.8 7.6
T1 192.6 13.0 8.4 5.3
T2 186.9 13.0 7.7 5.2
C 191.3 13.1 10.2 7.1
T1 187.9 13.0 7.5 5.1
T2 178.3 13.1 7.7 6.4
These data of decreasing feed consumption per chicken and day and also per live weight
were confirmed by Gezen et al. (2002), and by Agrar Production and Consulting (2005,
2003) in their experiments with applications of zeolit or APC preparations as additives
to feed mixtures.
The average live weight (table 4) of broiler chickens was compared with the
control group in the individual periods starter, prefattening, fattening and finishing
(0.155, 0.494, 1.001 and 1.825 kg). In the experimental groups a decrease of live weight
was observed only in the first two periods (starter HYD-01/1 and prefattening HYD-
138

01/2). For the latest periods (fattening and finishing) where we used the HYD-02 and
HYD-03 feed mixtures, the values in live weight increased. Within the values of live
weight significant and highly significant differences were noticed. The smaller content
of crude protein is responsible for the delayed growing up at the beginning of fattening.
The additives with clinoptilolit result in a better utilization of protein and thus a lower
required supply along with a higher live weight and weight gain at the end of fattening.
Reduction of nitrogen in the feed mixture resulted in better utilization and a reduction in
nitric manure.
Table 3: Feed consumption and feed conversion
FM
Feed consumption Feed conversion
C T1 T2 C T1 T2
HYD-01/1 24.27 23.55 23.85 0.944 1.017 1.018
HYD-01/2 67.90 63.54 65.09 1.852 1.879 1.953
HYD-02 113.95 109.94 111.49 1.968 1.828 1.842
HYD-03 166.05 154.76 159.59 2.069 1.974 2.005
Total 101.39 94.95 96.92 1.906 1.840 1.873
Table 4: Live weight and weight gain
Period C T1 T2 C T1 T2
starter 0.155 0.144 0.142
prefattening 0.494 0.491 0.475 0.340 0.347 0.333
fattening 1.001 1.033 1.020 0.508 0.542 0.545
finishing 1.825 1.897 1.876 0.824 0.864 0.856
REFERENCES
1. Agrar Production and Consulting, 2003,2005 available on : http://www.apcaustria.com/deutsch/produkte/index.htm
2. Reháková, M., Čuvanová, S., Gavaľová, Z., Rimár, J. 2003: Využitie prírodného
zeolitu typu klinoptilolitu v agrochémii a v poľnohospodárstve. Chem. listy 97,
(2003), s. 260-264,.
3. Čuvanová,S., Reháková, M., Rimár, J., Dzivák, M.2006: Klinoptilolit ako nosič,
regulátor a stabilizátor uvoľňovania živín. Acta Montanistica Slovaca Ročník 11
,(2006) , mimoriadne číslo 2, 285-289
4. Skalická, M., Koreneková, B., Makóová, Z., Naď, P.: Vplyv aditív na vybrané
produkčné ukazovatele hydiny. V. Dni výživy a veterinárnej dietetiky. Zborník z
medzinárodnej vedeckej konferencie, Hrádok, (2002), 237–239.
5. Gezen S.S, Mustafa, E. 2002: Broyler Rasyonlazma Katalin Narasin ve
Nikarbazinin, Zeolit Yle Etkilisimin Besi Perfornabsu Userine Etkileri. Uludag,
Univ. Y. Fac. Vet. Med, 21,(2002), p. 95-101
139

PERSPECTIVES ON ROMANIAN-HUNGARIAN CROSS-BORDER
COOPERATION. THE BIHOR - HAJDÚ-BIHAR EUROREGION MODEL
Gabriela Popoviciu 1 , M.C.Neacşu 2
1 University of Oradea, 2 Academy of Economic Studies Bucharest - Romania
ABSTRACT:
The promotion of regional cooperation, including cross-border initiatives, represents a
new concept for international relations between two or more neighbouring countries,
which have the same interest in interregional research and development cooperation in
order to stimulate investments in the private sector and lead to a general improvement
of living standards. On the other hand, the regional development policy promoted some
measures by local public and central administration, in partnership with the private and
public sector, and volunteers’ community also, for ensuring an economic, dynamic and
sustainable growth through the efficient valorisation of the local and regional potential
aiming.
This is one of the main goals of cooperation between Romania and Hungary.
Particularly, in this case study we will make notifications about regional development
between the two countries, in connection with their natural resources and their ways of
maintaining them in good quality for future generations. For this reason, both countries
have identified cross-border cooperation as a priority area and have made further steps
to enhance it.
Key words: Development, cooperation, cross-border, perspectives, regional
INTRODUCTION
In the past few years, the cross-border relations and cooperation between states have
started to follow a new tendency. These countries have begun to develop new crossborder
cooperation systems, the new one consisting of Euroregions.
In this context, we carried out a research on the topic of Cross-border Cooperation
in one of the newest Association Systems between two neighbouring countries,
Romania and Hungary. We will mention here the Bihor-Hajdú-Bihar Euroregion, which
exists in this configuration since October 2002.
The Bihor-Hajdú-Bihar Euroregion has its origins in the Carpathian Euroregion
(composed by Hungary, Poland, Ukraine, Slovakia and Romania), the first cross-border
cooperation form between Central and Eastern European countries. However, over the
years, this Euroregion has had various problems with transposing the entire countries’
programmes for a common working. This moment was the starting point for the Bihor-
Hajdú-Bihar Euroregion. It consisted of two cross-border counties, Bihor (RO) and
Hajdú-Bihar (HU). The official agreement’s act of partnership was signed by the
Bihor’s County Council and Self-government’s Hajdú-Bihar along with consolidating
cooperation among authorities, other public or private organisations and, last but not
least, between the population from those counties.
140

1. GENERAL APPROACH TO EUROREGIONAL PHENOMENA
The moment worth to be symbolically named “1989” and which marks the beginning of
the communist system wane in Europe, is followed by two opposite tendencies:
fragmentation and integration (Neguţ S., 2005). On the one hand the territorial
disintegration is obvious, peaceful (the dismemberment of Czechoslovakia, 1993) or
conflicting (e.g. Yugoslavia, 1991-1992, former USSR, 1991). On the other hand, an
economic integration tendency through successive waves of integration in European
Union (2004, 2007) manifested.
One of the dimensions of integration was the regionation or euroregionation,
which has not entailed a fragmentation, but vicinity rather, through the effort to recede
economic, social, cultural discrepancies between territorial units spatially close
together, but which belonged to different states, in an attempt to make the most of
geographical complementarity.
The euroregional model implies the association between local territorial
communities, from two or more neighbouring states, creating a unitary region,
characterized by some common particularities and interests. Euroregions concretize
geographical solidarities, despite state frontiers, making new spaces of decentralized
cooperation.
The territories of these close cross-border spaces could be characterized as
territorial units of a relative homogeneity with regard to the natural background of the
two or more states from the respective Euroregion, which are similar from the point of
view of their features, traditions, activities and preoccupations.
Thus, the collaboration phenomenon within Euroregions consists of establishing
some direct relationships between the regions and communities on both sides of a
state’s frontier, by virtue of the local authorities’ competences, as they are defined in
every country’s national legislation.
The prerequisites for an optimal development of one of these Euroregions,
conditions that the two counties hereof (Bihor – Romania and Hajdú-Bihar - Hungary)
seem to have successfully accomplished, are: a minimal economic equilibrium, relief,
language and culture unity, similar administrative structure and, why not, the same
historical heritage, given the fact that every state has minorities settled in the
neighbouring state. In fact, euroregions 1 (Neguţ S., 1999) represent a factor of regional
stability and security at the frontier, which aims to outrun a negative historical heritage
in the context of reinstating an economic equilibrium able to promote a high living
standard and an improved quality of life.
1 On the occasion of the meeting from Praga, 1998, concerning the matter of regional
cooperation formulas in Central and South-East Europe, dr. Sabathil, as member of the
European Commission Delegation, emphasized: the most traditional and efective form
for cooperation between cross-border regions and (…) a relatively flexible instrument
for collaboration in these frontier zones.
141

2. THE BIHOR-HAJDÚ-BIHAR EUROREGION– A CROSS-BORDER DEVELOPMENT
MODEL
2.1. Geographical, human and economic complementarities
An illustrative example for all we have noted above, which meets all the necessary
conditions and elements for cross-border cooperation, is the Bihor - Hajdú-Bihar
Euroregion. This is the newest cross-border Euroregional structure 2 and it is situated in
the North-Western part of Romania, in the Romanian-Hungarian cross-border area (fig.
1).
Fig.1. The Bihor-Hajdú-Bihar Euroregion
The Bihor – Hajdú-Bihar
Euroregion overlaps on three
essential geographical units, which
are the West Plain, the West Hills
and the Apuseni Mountains (on the
Romanian side) while only the plain
is characteristic to the Hajdú-Bihar
County.
The maximum altitude of the
Euroregion reaches almost 2 000 m
(Cucurbăta Mare Peak, 1 849 m)
while the minimum altitude does not
surpass 100 m (Hortobágyi Region).
The studied Euroregion also has a plentiful hydrological network (Barcău, Crişul
Repede, Crişul Negru, Crişul Alb, Tisa, Körös, Keleti and Hortobágy) and important
natural resources.
Similar to other cross-border cooperation structures (Ilieş Al., 2004) formed by
including the state frontier, the Bihor-Hajdú-Bihar Euroregion shares some of their
general characteristics (such as the ones regarding organisation and working mode) but
also has a few specific features, which we will present as follows.
The Bihor-Hajdú-Bihar Euroregion represents an open cross-border association,
which does not possess its own juridical personality (art.4 from Euroregion Statute).
Within the cooperation process, the parts have identical rights and obligations. Through
the present cooperation, the parts have expressed their accord that their actions be based
on mutual trust, spirit of collaboration and good neighbouring relations (art.5 from
Euroregion Statute).
From a structural point of view, the Bihor-Hajdú-Bihar Euroregion is an
association of administrative-territorial units of the same rank (Bihor county and Hajdú-
Bihar county), on one hand, and other individual or in association inferior rank units,
(Bihor’s Frontier Localities Association and Hajdú-Bihar’s Frontier Localities
2 Romania and Hungary also participate together to the following euroregions of the
cross-border type: the Carpathian Euroregion (alongside Poland, Slovakia and Ukraine,
with a 148 095 km² surface and over 15 mil. inhabitants; actually, the Bihor-Hajdú-
Bihar Euroregion was created within this macro-region) and the Danube-Cris-Mures-
Tisa Euroregion (alongside Serbia; this euroregional structure extends over 77 243 km²
and has about 6 mil. inhabitants).
142

Association – fig. 2), on the other hand. As we can see, the cooperation within our
Euroregion is extending both horizontally and vertically 3 .
This structure was organised through bilateral participation, which endows it with a
high rate of functionality and an increased focus on specific problems that derive from
the interfacing position of the two areas.
The favourable cross-border cooperation premises in the Bihor - Hajdú-Bihar
Euroregion also derive from the fact that, especially during the past few years, the
majority of the economic and decisional factors have accepted the perspective of crossborder
development. The following aspects determine the complexity of the crossborder
dimension in our region:
- Dimensionally speaking, the Euroregion is a structure of medium value because
of its surface (13 755
km 2 ) and its number
of inhabitants (about
1.1 mil.), which
means that it
represents a substate
structure.
- With regard to
the shape of the
Euroregion, it is
similar to that of a
highly functional
system, thus ensuring
a balanced
distribution of
localities in the
territory in relation to
the county’s residence
(Oradea and
Debrecen) as well as
in relation to the other
cities.
- The Euroregion
represents a crossborder
cooperation
structure (Ilieş Al.,
2003) situated in
Crişurilor’s and Tisa’s
Fig.2. The Frontier Localities Associations from Bihor -
Hajdú-Bihar Euroregion
hydrographical basins, with an in tread morphology of the main units of relief and an
3 Reference is making to the international agreement concerning the partnership
principle extended on both a horizontal and vertical level. This one refers to the
collaboration for mutual support and a good neighbouring policy between nations,
states, localities, authorities etc.
143

accent on complementarity which benefits by a determinative compatibility in the
administrative-territorial system.
- The Euroregion benefits from a communication network favourable to
complementarity as a basis for economic development.
In the area of the Euroregion the infrastructure of 16 “potential” cross-border roads
exists, out of which three are operative in the international regime (Salonta - Mehkerek,
Borş - Artand and Valea lui Mihai - Nyirabrany) and two are meant for occasional
traffic (Toboliu - Körösnagyhomorog and Săcuieni - Letavertes). Four railroad systems
can be added to these, out of which three are active in the regime of international traffic
(Salonta - Kötegyan, Episcopia Bihor - Biharkeresztes and Valea lui Mihai -
Nyirabrany) (Stamate, G., 1997).
- The Euroregion has a main Romanian-Hungarian ethnic base with a balanced
share of the two ethnic
groups. This ethnic base
is characterised by a
powerful tradition of
cohabitation between the
majority and the local
minorities. Other coinhabiting
nationalities
(fig. 3) can be added, in a
larger or smaller number
on both sides of the
frontier.
- The Euroregion evinces
a higher level of
economic development
as compared to the
average in the Eastern
part of Europe.
- Currently, the
Euroregion includes only
one state frontier sector
(which has a military
Fig. 3. The ethnic structure in the Bihor – Hajdú-
Bihar Euroregion (the values are in %)
function determined by
the NATO membership
of both countries 4 , and a
control function of the people and merchandise fluxes).
- The Romanian-Hungarian border length on the Bihor County’s side is of approx.
166 km, out of which approx. 160 km are on land and 6 km on water. These totalize
about 37% of the entire length of the Romanian-Hungarian border (Stamate G., 1997).
Due to the land support of this frontier sector, there is an active potential for
4 Starting with 2007, once with Romania’s adhesion to the EU, this sector has
becomeone of its internal frontiers
144

communication between the two associated territorial systems, which ensures a crossborder
flux similar to the one in other European countries.
- The local administrative organisms of the Euroregion established as priorities
those actions that refer to the consolidation of administrative-institutional capacities,
which will ensure their practical role, but also the creation of an informational
framework regarding the potential of the cooperation between Romanians and
Hungarians.
- On both counties’ territory, there are important natural resources and anthropic
edifices, marked by landscape diversity and a natural complementary background. This
provides a special potential for tourism within the Euroregion. For this purpose, “The
Romanian-Hungarian Corridor for Conservation of Biodiversity” was created in the
wetland ponds’ area of Cefa, in the Natural Zoological Reservation from Rădvani
(Romania), as well as in the Körös-Máros National Park (Hungary), locations that are
actually situated “in mirror position” from one another.
The variety of
geographical conditions
is essential for a unitary
development of the
Euroregion, because it
can lead to multiple
possibilities of land
usage. In other words,
the Bihor – Hajdú-Bihar
Euroregion develops
according to its natural
conditions (industrial
activities in the
mountainous zone,
agricultural activities
and services in the other
areas) (fig. 4).
If we compare the
two counties, we can
see that the industrial
sector still plays the
most important role,
Fig. 4. The population structure on the activities sectors
in Bihor - Hajdú-Bihar Euroregion
especially in the economy of Bihor County, unlike in the Hajdú - Bihar County, where
the services segment is dominant. At the same time, the general trend is to diminish the
importance of agriculture, in favour of increasing the one of services.
145

2.2. Oradea-Debrecen the next urban cross-border agglomeration 5
Although the historical evolution of the two cities has been different, the
complementarity of these two future attraction poles is obvious: on one hand, Oradea is
mentioned in a 1113 document as a fortress set in a marshy area and it evolves
spatiotemporally under the ruling of its political-administrative function, and Debrecen
on the other hand, of a similar age, renowned economical centre.
After 1920, along with the delimitation of state frontiers, the two cities suffered a
slight syncope in the evolution of their interdependence relations, each of them
belonging to its own decisional centre. The functionality of the urban system revitalized
after 1990 through a cultural convergence (university exchange programs, both cities
endowed with renowned and very dynamic Universities) as well as a business related
one.
Nowadays the two centres represent the pillars of a future cross-border urban axis
grafted on the two metropolitan areas: Oradea (the city of Oradea and the administrative
territory of the adjoining villages – Biharia, Bors, Cetariu, Paleu, Nojorid, Oşorhei,
Sânmartin and Sântandrei), with a population of over 250 000 inhabitants (out of which
the largest part is located in Oradea – about 87%) and Debrecen (with a population just
over 200 000 inhabitants), which would spread over about 75 kms on the Oradea –
Biharkeresytes – Berettyóújfalú – Derecske – Debrecen axis (Săgeată R. et. al., 2004,
Ilieş Al., 2004).
3. THE JURIDICAL-INSTITUTIONAL BACKGROUND REGARDING REGIONAL
DEVELOPMENT IN THE BIHOR – HAJDÚ-BIHAR EUROREGION. OBJECTIVES
Because Euroregions represent frontier cooperation structures, which contribute not to
regional isolation but to regional development (Ghidul U.E., 2004), both states have
adopted the Convention of Madrid (May 21, 1980) regarding cross-border cooperation
between territorial communities and authorities. Moreover, they are both part of the
European Charter of Local Self-Government (Strasbourg, Oct. 15, 1985). In order to
render functional the settlements comprised in these two European acts, the legislative
step made by both countries was the adoption of two national laws concerning local
administration (Law no. 215/2001 – Romania and Law no. XXI/1996 - Hungary).
With regard to the legislative and statutory aspects, the two laws are compatible, in
the sense that they both designate the authorities, communities and organisms playing a
role in exercising regional functions as the local and county’s Council (for the
Romanian side) and the county’s and town hall’s local self-government (for the
Hungarian side). We must mention here that only the typology of the two laws is
different, as the understanding of the settlement is identical.
In accordance with Article 2, paragraph 1 of the Euroregion’s Constitution
Agreement, “the cross-border cooperation is exercised by the qualified territorial
collectivities and authorities, as they are defined in the internal law.”
5 This model is currently in the phase of research project initiated by the “Forum”
Foundation from Oradea, with the participation of the University of Oradea and
University of Debrecen, Thinking the future together. The Debrecen-Oradea Crossborder
Agglomeration (2020).
146

The juridical settlement of the Bihor – Hajdú-Bihar Euroregion was created by a private
law contract, whose settlements and provisions are available only on the Euroregion’s
territory. According to the Statute and Agreement of Association of the two counties,
the main objectives of the partnership between the two territorial-administrative
authorities are:
• Encouraging the plan for unitary development of the rural communities in both areas;
• Encouraging the development of communities both locally and regionally, based on
the existing natural and human resources;
• Stimulating the native efforts for community’s development and promoting local
initiatives;
• Reinforcing the partnership and creating a network between different regional and
local non-governmental organizations, local administrations and the business sector;
• Reinforcing the managerial capacities of local and regional non-governmental
organizations, as well as those of local administrations.
4. CONCLUSIONS
The Bihor – Hajdú-Bihar Euroregion has a common natural and cultural heritage. For
the conservation of those common values, concrete forms of cross-border cooperation
are necessary.
The European project of Euroregions and cross-border cooperation between
neighbouring communities is built on the common patrimony of European societies, on
specific national principles, values and regulations, which may be identified in the two
counties reunited into the Bihor – Hajdú-Bihar Euroregion.
The consideration of all these aspects will open new long-term perspectives for a
sure and durable development of the Bihor – Hajdú-Bihar Euroregion and for the
inclusion of its inhabitants into the world development circuit.
In accordance to all of the above, the Bihor – Hajdú-Bihar Euroregion maintains a
high functionality, which we believe to be accentuated by the position of the two
associated counties at the border between two different political entities, with common
collaboration and cooperation premises, which render viable the existence of the studied
Euroregion.
REFERENCES
1. Ilieş, Alexandru (2003), România între milenii. Frontiere, areale frontaliere şi
cooperare transfrontalieră (I),. Universităţii din Oradea Publishing House;
2. Ilieş, Alexandru (2004), România. Euroregiuni, Universităţii din Oradea Publishing
House;
3. Neguţ, Silviu (1999), Les Euroregions, in „Revue Roumaine de Geographie”,
Academiei Publishing House, Bucharest;.
4. Neguţ, Silviu (2005), Europa între fragmentare şi integrare, in Introducere în
geopolitică, Meteor Press Publishing House, Bucharest, p.161-166.
5. Săgeată, Radu (coord.) (2004), Soluţii de optimizare a organizării administrativteritoriale
a României în perspectiva aderării la Uniunea Europeană, Ars Docendi
Publishing House, Bucharest, p.43.
147

6. Stamate, Grigore (1997), Frontiera de stat a României, Militară Publishing House,
Bucharest;
7. * * * European Institute of Romania (2004), Ghidul U.E., Aquis comunitar nr. 1
series, 2 nd edition, coord. Tamara Ferluşcă.
8. * * * (1996), Legea nr. XXI din 1996 privind administraţia publică locală a
Ungariei.
9. * * * (2001), Legea nr. 215/2001 privind administraţia publică locală, în M. Of. al
României nr. 204 din 23 aprilie 2001, Bucureşti.
10. * * * Kozponti Statisztikai Hivatal (2002), Hajdú-Bihar megyei Igazgatósága.
11. * * * National Institute of Statistics (2003), Date statistice după recensământul
populaţiei din 2002.
12. * * * (2002), The Euroregion’s Constitution Accord and The Statute of The
Euroregion Bihor – Hajdú-Bihar.
13. Web sites: www.euroregiuni.ro, www.freeweb.hu/romanul, www.mie.ro
148

EFFECT OF BORON FERTILIZATION ON ANNUAL FLUCTUATION OF B
IN SWEET CHERRY LEAVES AND FRUIT QUALITY
Péter Tamás Nagy 1 - Sándor Thurzó 2 - Ida Kincses 1 - Zoltán Szabó 2 - József Nyéki 2
1 Department of Agricultural Chemistry and Soil Science, University of Debrecen,
Centre of Agricultural Sciences, Faculty of Agronomy
2 Institute for Extension and Development, University of Debrecen, Centre of
Agricultural Sciences
ABSTRACT
The goal of the study was to examine response of sweet cherries (Prunus avium L.) to
boron (B) fertilization. The experiment was conducted during 2005–2006 in West
Hungary on mature cv. ‘Germersdorfi 3’ grafted on Prunus mahaleb rootstock.
Sweet cherry trees planted on a calcareous chernozem soil. Trees were foliar-fertilized
with B. Foliar B sprays were performed: (1) in the spring, at the stage of white bud,
beginning of flowering, and (2) repeated 5 weeks after full bloom. In each of spring
spray treatments, B was applied at a rate of 0.15 kg ha −1 . Trees untreated with B served
as a control.
The results showed that B fertilization had effect on B concentration in leaf tissues,
mostly after ripening. Mean fruit weight was slightly increased by B fertilization. Fruit
sensitivity to cracking was not influenced by B fertilization. Nevertheless, from our data
it can be conclude that the sensitivity of fruit to cracking is improved when the fruit is
riper, the fruit density and fruit weight are higher. The soluble solids varied between
15.0 and 15.9% according to the treatments. Our results for the monosaccharides
investigated varied between 7.6 and 8.0 as glucose and fructose as well.
It is concluded that under conditions of this experiment, B fertilization can be
recommended in sweet cherry culture to improve fruit quality and their appearance.
Keywords cherry, foliar nutrition, boron fertilization, fruit quality
INTRODUCTION
However the role of Boron (B) is well-known in the plant nutrition, especially fruit
nutrition, there are very little information about its application and use in Hungarian
orchards.
Boron is an essential micronutrient in plants that is often deficient in most soils
because most of the boron in the soil is adsorbed to clay minerals, hydrous metal oxides,
and organic matter in soils. In addition, boron can be co-precipitated with calcium
carbonate making it unavailable to the roots. Moreover the B uptake is hindered by very
wet or very dry soils, increased leaching and cold soil temperatures.
The major role of B in fruit trees involves fruit set (Faust, 1989). Apple, pear and
cherry flowers are very high in B. The B needed in the flower is transported mainly
from the reserves in the adjacent branches and not from the roots during the
development flower. It is essential for reproduction, aids in the formation of pollen
germination and pollen tube growth. Boron aids in the metabolism of hormones and in
the translocation of calcium, sugars and growth regulators, required for protein
synthesis. In addition B is important for early growth, flowering and fruit set (Kamali
149

and Childers, 1970), maintains balance between sugar and starch, aids in auxin
regulation and of course it is necessary for cell division and differentiation, and root tip
development.
Therefore, the close attention to B levels is important because both low and high
concentrations cause poor fruit quality. Low B results in short storage life with the fruit
having a higher susceptibly to storage breakdown and fruit deformities. High B results
in a higher incidence of internal disorders such as watercore and internal breakdown.
MATERIALS AND METHODS
The study was conducted during 2005–2006 in West Hungary on cv. ‘Germersdorfi 3’
grafted on Prunus mahaleb rootstock. Trees were planted in the spring of 1999. Trees
spaced 7 x 5 m, and growing in a calcareous chernozem soil at Siófok in West-Hungary.
Orchard was not irrigated in 2005 and 2006. For the purpose of the experiment, 20 trees
were randomly selected from a population of trees with uniform characteristics. The
applied foliar applications are presented in Table 1.
For spraying, Solubor (Disodium Octaborate Tetrahydrate - 20.9% B) was applied.
Table 1: Applied foliar fertilization system
Applied nutrient Dose (g/L) Time of applications Code of
treatment
Control - - C
B as Na2B8O13·4H2O 0.5
at full bloom
5 weeks after full bloom
1. Soil sampling and preparation
Two soil samples were collected from three layers (0-20, 20-40 and 40-60 cm) of each
treatment by using manual soil sampling equipment following the Hungarian sampling
guidelines and according to Nagy et al. (2006). Sampling was performed before
treatments, at the beginning of the vegetation period in March 2005. Sample preparation
of the soil samples was performed according to Hungarian guideline (MSZ
20135:1999). The following parameters were measured: pH, KA, content of humus and
AL soluble P and K according to Hungarian guidelines.
2. Plant sampling and preparation
Plant (leaf) samples were taken, from May to September (14 May, 30 May, 30 June and
10 September) in 2005and 2006. Leaves were taken from all trees according to
international conception and Hungarian sampling guidelines (Stiles and Reid 1966; MI-
08 0468-81). For leaf analysis 1 g plant sample was ashed in a muffle furnace at 450 °C.
Ash was dissolved in 5 ml of a 1M HCl at room temperature, mixed, and measured by
photometric method (Azomethin-H method).
150
B1
B2

3. Examinations of fruit quality
For fruit assessments, 50 fruits were examined.
3.1. Fruit cracking (%)
It was calculated as a ratio of the number of cracked fruits and the number of total
fruits.
3.2. Fruit weight (g)
It was measured with a digital analytical scale with 0.1 g punctuality.
3.3. Fruit density (1-9)
This parameter was given a number on a subjective scale from 1 to 9 (determined at
10 trees). Where, 1 means the tree without fruit and 9 means the tree fully covered
fruits. It was established two weeks before harvest.
3.4. Maturity (1-9)
This parameter was given a number on a subjective scale from 1 to 9 (determined at
10 trees). Where, 1 means the unripe fruit and 9 means the totally ripe fruit. It was
established two weeks before harvest.
3.5. Soluble solids and sugars
The sugars (fructose and glucose) and soluble solids (Brix) were studied at harvest
in the sweet cherries (Prunus avium L.) cv. ‘Germersdorfi 3’ in 2005 and 2006. All
fruit samples had been picked at the optimal ripening time. For examination 12
fruits were performed. Fruits were pressed and the obtained juice was filtered
through (FILTRAK Qual. grade:132) folded filters. Soluble solids and sugars were
determined in the juice by refractometer (ATAGO PAL series) at 20 ºC.
RESULTS AND DISCUSSIONS
1. Soil analysis
The orchard soil type is calcareous chernozem soil. The upper layer of soil (0-60 cm)
contained 1.7 % humus, 178 mg/kg and 372 mg/kg AL-soluble P and K. The plasticity
index according to Arany (KA) was 39. According to our results the soil is slightly
alkaline (pH(H2O)=7.65) loamy soil and calcareous in deeper layers.
2. Plant analysis
Boron concentration was varied between 35 and 65 mg/kg in leaves, during examined
vegetation period. It continuously increased during examined period in every treatment
till ripening. Then, in the control, it significantly decreased while in B treatments its
amount increased continuously (Fig. 1). So although shoot leaves from all treatments
collected in summer had similar boron concentrations, B was present in different
content in leaf tissues late (after ripening) in the season.
This effect is very important for the next year´s growth. Because, the higher foliar B
content plays efficient role in transport processes from the leaves into storage tissues for
the next year´s growth. Moreover, we conclude that leaf B after harvest is translocated
to above-ground storage organs before leaf fall and is then used the following season.
Based on the data of ripening, the leaf B values varied between 59 and 61.5. In all
treatments, these values were higher than the optimal range (Mills and Jones 1986;
Papp, 2004). Obtained results can be explained by the soil properties and additional
boron application.
151

B mg/kg in dry matter
70
60
50
40
30
20
10
0
May 14 May 30 June 30 Sept. 10
Sampling time
control
Figure 1: Effect of applied foliar treatments on annual fluctuation of B in leaves
Obtained results confirmed that earlier findings that timing of B maintenance sprays is
not critical for cherry trees if the trees already contain adequate amounts of B and do
not show visual evidence of B insufficiency (Peryea, 1994; Peryea et al., 2003).
3. Quality factors and fruit analysis
Some investigated fruit quality factors are represented in Figure 2.
Fruit sensitivity to cracking was influenced differently by B fertilization. B1 treatment
decreased, while B2 increased the number of cracked fruits.
Mean fruit weight was significantly increased by both B fertilizations.
Quality factors (relative scale)
20
16
12
8
4
0
LSD 0.05 =1.96
Fruit cracking
(%)
LSD 0.05 =0.83
LSD 0.05 =0.57
B1
B2
Control
B1
B2
LSD 0.05 =0.33
Fruit weight (g) Fruit density Maturity
Figure 2: Effect of applied foliar treatments on some quality factors
152

Fruit density was increased applying foliar boron treatments, but only the B2 treatment
had significant effect on it. Maturity was not effected by boron treatments. The lowest
value was founded in the B1 treatment and there was not statistical difference between
B2 treatment and control (Fig. 2).
From our data it can be conclude that the sensitivity of fruit to cracking is
improved when the fruit is riper, the fruit density and fruit weight are higher.
Figure 3 shows the soluble solids varied between 15.0 and 15.9% according to the
treatments. Our results are similar to those reviewed by Kaack et al. (1996) and Predieri
et al. (2004). Our results were pointed out that B applications influenced the soluble
solids content. Result of B1 treatment may be explained by the lower value of maturity.
Moreover the soluble solids content is connection with maturity and fruit weight as
Blažková et al. (2002) reported.
%
18
16
14
12
10
8
6
4
2
0
LSD0.05=0.52
LSD0.05=0.27
Control
B1
B2
LSD0.05=0.26
Brix Fructose Glucose
Figure 3: Effect of applied foliar treatments on contents of soluble solids (%) and
sugars (g/100g of fresh weight)
Our results for the monosaccharides investigated are similar to those reviewed by
Wrolstad and Schallenberger (1981) who reported mean values for a large variety of
cherries as glucose 7.78 g/100 g; fructose 7.09 g/100 g and Gardiner et al. (1993) who
reported the content of glucose is 7.025 g/100 g and fructose is 6.7 g/100 g
approximately. Our values are also comparable to those obtained by Wills et al. (1987)
for Australian cherries and Dolenc and Štampar (1998) in Slovenia. Sucrose was not
detected in the unprocessed cherries, which is consistent with the findings of Wrolstad
& Schallenberger (1981) who reported the absence of sucrose in some cultivars (e.g.
cherries) and only low concentrations in others. No attempt was made to inactivate
invertase in the unprocessed cherries as it is unlikely that this would be done in a
commercial situation.
153

Furthermore, our results confirmed those earlier results as the content of glucose and
fructose is present in approximately equal amounts in most nectar (Van Handel et al.
(1972)).
It is concluded that under conditions of this experiment, B fertilization can be
recommended in sweet cherry culture to improve fruit quality and their appearance.
REFERENCES
J. Blažková - I. Hlušičková and J. Blažek (2002): Fruit weight, firmness and soluble
solids content during ripening of Karešova cv. sweet cherry. Hort. Sci. (Prague),
29, 2002 (3): 92–98
Dolenc K. and F. Štampar (1998): Determining the quality of different cherry
cultivars using the hplc method. Acta Hort. (ISHS) 468:705-712
Faust M. (1989): Physiology of temperate zone fruit trees. Wiley, New York.
Gardiner M. A. - R. Beyer - L. D. Melton (1993): Sugar and anthocyanidin content of
two processing-grade sweet cherry cultivars and cherry products. New Zealand
Journal of Crop and Horticultural Science, 1993, Vol. 21: 213-218.
Kaack K. – Spayd S.E. – Drake S.R. (1996): Cherry processing. In: Cherries: Crop
Physiology, Production and Uses. Eds. By Webster A. D. and Looney N. E. CAB
<strong>International</strong>, Wallingford. 471-485 pp.
Kamali A.R. and N.F. Childers (1970): Growth and fruiting of peach in sand culture
as affected by boron and fritted form of trace elements. J. Amer. Soc. Hort. Sci.
95:652–656.
Mills H.A. and J.B. Jones (1996): Plant Analysis Handbook II. MicroMacro
Publishing, Inc.
MI-08 0468-81: Plant analyses. Orchards. Sampling, preparation of samples, storing of
samples. Hungarian Standards Institution. Ministry of Agriculture. Budapest (in
Hungarian)
MSZ 20135:1999: Determination of the soluble nutrient element content of the soil.
Hungarian Standards Institution. Budapest (in Hungarian)
Nagy, P. T. - Gonda I. - Dremák P. and Holb I. (2006): Study on the micronutrient
content of soil and leaf o fan organic apple orchard in Eastern Hungary.
<strong>International</strong>Papp J. (2004): Fruit Production 1st Ed.; Mezőgazda Kiadó:
Budapest, Hungary, 686 pp. (in Hungarian)
Peryea F.J. (1994): Boron nutrition in deciduous tree fruit, p. 95–99. In: A.B. Peterson
and R.G. Stevens (eds.). Tree fruit nutrition. Good Fruit Grower, Yakima, Wash.
Peryea F.J. - D. Nielsen and G. Neilsen (2003): Boron maintenance sprays for apple:
Early-season applications and tank-mixing with calcium chloride. HortScience
38:542–546.
Predieri, S. - R. Dris and F. Rapparini (2004): Influence of growing conditions on
yield and quality of cherry: II. Fruit quality. Food, Agriculture & Environment
Vol.2 (1) : 307-309.
Stiles W.C., and Reid W.S. (1966): Orchard nutrition management. Cornell
Cooperative Extension. Information Bulletin 219.
Van Handel E. – J. S. Haeger and C. W. Hansen (1972): The sugars of some Florida nectars.
Amer. J. Bot. 59(10): 1030-1032.
154

Wills, R. B. H. - Lim, J. S. K. - Greenfield, H. (1987): Composition of Australian foods. 40.
Temperate fruits. Food technology in Australia 39: 520-521,530.
Wrolstad, R. E. - Schallenberger (1981): Free sugars and sorbitol in fruits—acompilation from
the literature. Journal of the Association of Official Analytical Chemists 64: 91-103.
155

DRY MATTER PRODUCTIVITY AND NITROGEN UPTAKE OF PERENNIAL
RYEGRASS INFLUENCED BY NITROGEN AND WATER SUPPLY
ABSTRACT
Imre Vágó – János Kátai – Ida Kincses – Andrea Balla Kovács
University of Debrecen, Faculty of Agriculture
Department of Agricultural Chemistry and Soil Science
H-4015 Debrecen, P.O.Box 36.
Besides the genetically coded productivity of the cropped plants, other factors also
determine the quality and the quantity of yields. Among these factors the nitrogen and
water supply is very important. Investigations were made in greenhouse pot
experiments to reckon if these factors influence the effects of each other. The same
nitrogen and water supply has different effect in soil types. That is why vegetation pot
experiment on three extremely different soil types of Hungary, near of Debrecen was
carried out.
Perennial ryegrass (Lolium perenne L.) was used as a test plant. The effect of
nitrogen (0, 40, and 80 mg kg -1 ) and water (50 and 75% of the total water capacity)
supply on the dry matter production of ryegrass was examined. The nitrogen
concentration of the shoot was measured and the nitrogen uptake of plants was
calculated.
The dry matter production of plants reduced during the experimental period from
4.02 g per pot (first cut) to 1.78 g per pot (third cut). The reason is the nutrient
deficiency of small soil mass in the pots and the ageing of grasses. At cuts according to
the mean of treatments the calcareous chernozem was the most productive, while that of
marshy meadow and brown forest soils was similar to each other. The 80 mg kg -1
nitrogen supply doubled the yield in the brown forest soil and calcareous black soil,
while in marshy meadow soil abounding in organic matters the increase of crop mass
was smaller. On the calcareous chernozem and marshy meadow soil, the water supply
increased the yield but on the sand the maximum yield at the 50% water capacity was
detected.
During the three cuts summarized extracted nitrogen amount depending on the
soil type (P=0.1%). The calcareous chernozem soil and the marshy meadow soil
supplied similar (nearby 200 mg per pot) amount of nitrogen for the ryegrass. This fact
can be explained that the higher shoot production of ryegrass on calcareous black soil
and the higher nitrogen content of ryegrass on marshy meadow soil equalise each other.
From the brown forest soil only 110 mg nitrogen per pot in average was extracted by
plant shoots, because of the smaller plant production and its lower concentration of
nitrogen.
Keywords: Nitrogen and water supply, productivity and nitrogen uptake of ryegrass
INTRODUCTION
The productivity of the plants is influenced by a lot of different factors. The most
important of them is the genetic potential, but the environmental and the technological
factors also influence the quantity and the quality of the yields (Mengel, 1976; Loch and
156

Nosticzius, 1992). Among these, soil parameters, nitrogen and water supply are the
most decisive ones. The nitrogen and the water supply significantly modified the
dynamics of nutrient uptake at the different crops (Borchmann, 1963). It should be
noted, however, that there are significant differences in the plant species’ ability to take
up the nutrients (Eichler et al., 1997; Eichler and Köppen, 2001; Eichler, 2004).
The identical nitrogen and water supply has a different effect on the soil types. To
investigate this phenomenon, a vegetative pot experiment was set up on three extremely
different soil types from the near of Debrecen. The advantage of the pot experiment is
that several factors (e.g. water supply) can be controlled and the same conditions can be
ensured for all the treatment combinations. In addition, the amounts of added nutrients
are exactly known due to the size of the pot and the roots of the plant interwove the
whole pot.
Perennial ryegrass (Lolium perenne L.) was used as a test plant. The advantage of
this plant is that it endures well the conditions of the greenhouse and it can be cut
several times during the vegetation period, therefore, the dynamics of the processes can
be concluded too.
In the greenhouse experiment, the effect of different nitrogen and water supply on
the plant’s dry matter production and its dynamics was examined. The nitrogen
concentration of the yielded shoot dry matter was measured and the amount of nitrogen
extracted from the soil by the plant was calculated for the different cuts and for the
whole period of the experiment.
MATERIALS AND METHODS
In the Table 1 are included the basic initial parameters of the three untreated soils used
in the pot experiment. The initial total carbon and nitrogen content of soils was
measured by Nagy, P. T. (2000). Calculated for air-dry state, 2.5 kg of brown forest
soil, and of chernozem soil and 2.0 kg of marshy meadow soil were put separately into
plastic pots of 2.0 dm 3 volume.
The standard background treatment of each experimental pots was a joint 10 cm 3
solution of 40 mg kg -1 P2O5 and 40 mg kg -1 K2O, as pro anal. compounds of KH2PO4
and K2SO4 in distilled water.
The treatment combinations applied on the three soils are shown in Table 2. Each of
them was carried out in four replications to ensure the possibility of statistical
evaluations.
The transpirated and evaporated water was supplemented by irrigating the soils
with distilled water. In the treatment plan given water supply level set up was ensured
by daily irrigation of the vegetation pots to a calculated specific weight. At high
vegetative mass, and also on the hot days plants were irrigated in the afternoon to avoid
the threat of wilting.
Plants were cut three times, first time five weeks after the sowing, the 2 nd and the 3 rd
times in three weeks intervals. The grass shoot yield was first air-dried and after that
dried at 60 ºC until reaching constant mass in a desiccator. The dry matter production
per pot was measured by analytical balance.
157

Table 1: Parameters of untreated soils
Soil parameter
Soil site
Pallag Látókép Dombos
Soil type Brown forest soil Calcareous Marshy
chernozem meadow soil
Soil texture Sand Loam Clayes loam
KA 28 46 58
pH-H2O 5.38 6.13 7.60
pH-KCl 4.04 5.45 7.27
pH-CaCl2 4.62 5.91 6.98
Hydrolytic acidity y1 8.16 7.85 -
CaCO3 content (%) - - 26.7
Salt content (%) 0.002 0.039 0.022
Total C content (%) 0.34 1.93 7.20
Total N content (%) 0.043 0.206 0.473
C/N ratio 7.97 9.35 15.24
Organic C (%) 0.26 0.97 1.75
Organic/total C ratio 76.5 50.3 24.3
0.01 M dm -3 CaCl2 extractable (mg kg -1 )
NO3 - -N 0.51 4.95 3.44
NH4 + -N 0.31 3.67 0.56
Inorganic N 0.82 8.62 4.00
Organic N 0.14 0.59 1.46
Total soluble N 0.96 9.21 5.46
The nitrogen content of the dry matter was measured on the principle of the Kjeldahl
method with a Kjel-Foss analyzer. From the mass of the shoot and the nitrogen
concentration of it, the amount of nitrogen extracted from the soil per cutting was
determined. Finally, the total amount of nitrogen extracted was calculated. The obtained
results were evaluated by a three-variate analysis of variance.
Table 2: Combinations of treatments
Combination N treatment
code (mg kg -1 Water supply
soil) (% of water capacity)
1. 0 50
2. 40 50
3. 80 50
4. 0 75
5. 40 75
6. 80 75
In the analysis of variance, F and SD 5 % values were calculated for each factor and for
each double and triple interaction, then the significancy levels were determined.
158

RESULTS AND DISCUSSIONS
The dry matter production of the perennial ryegrass reduced with time (main average:
1 st cut: 4.02; 2 nd cut 2.04; 3 rd cut: 1.78 g per pot). The reasons for the reduction of dry
matter production are the nutrient-deficiency originating from low soil mass and the
ageing of grasses. The nutrient supply of soils heavily influenced the dry matter
production of plants at all cuts. In the average of treatments, the calcareous chernozem
soil was the most productive (significant at P = 0.1 % level), while the productivity of
the brown forest and the marshy meadow soil was similar (Table 3).
The dry matter production increase due to the nitrogen treatment varied for the different
soils. This is supported by the fact, that the interaction of factors A*B (N dose*soil
type) is highly significant (mostly P = 0,1 %). The application of 80 mg kg -1 nitrogen
doubled the yield on the brown forest and the calcareous chernozem soil, while on the
marshy meadow soil with high organic matter content the increase was also significant,
but smaller (30 %).
Table 3: Dry matter production of ryegrass (g pot -1 ) Sum of 1 st , 2 nd and 3 rd cuts
W.c. (%) Soil site N supply (mg kg -1 ) (A)
(C) (B) 0 40 80 Average
Pallag 4.38 7.43 10.53 7.44
50 Látókép 6.92 8.75 11.57 9.08
Dombos 5.10 6.60 7.27 6.32
Average 5.47 7.59 9.79 7.62
Pallag 4.40 5.80 6.73 5.64
75 Látókép 7.33 10.82 14.43 10.86
Dombos 6.48 7.70 8.85 7.68
Average 6.07 8.11 10.00 8.06
Pallag 4.39 6.61 8.63 6.54
Average Látókép 7.13 9.79 13.00 9.97
Dombos 5.79 7.15 8.06 7.00
Average 5.77 7.85 9.90 7.84
Table of variance
Factor FG MQ F SD 5%
A 2 102.30 117.6 *** 0.54
B 2 83.18 95.6 *** 0.54
A*B 4 6.68 7.7 *** 0.94
C 1 3.51 4.0 * 0.44
A*C 2 0.26 0.3 0.77
B*C 2 22.88 26.3 *** 0.77
A*B*C 4 5.18 6.0 *** 1.33
Error 51 0.87
*** Significant at P=0.1 % level
* Significant at P=5.0 % level
The higher level of water supply increased the perennial ryegrass dry matter production
at every cut. The significance of the B*C (soil*water capacity) interaction (P=0,1 %)
159

proves that the highest yield can be achieved by 75 % water supply on calcareous
chernozem and marshy meadow soil and by 50 % water supply level on brown forest
soil. It can also be stated that the effect of nitrogen fertilization was not modified by the
different water supply. The positive effect of nitrogen treatments was detectable in the
dry matter production of plants at each cuts (significant at P = 0.1 % level).
The nitrogen content of plants is the highest (2.81 %) on marshy meadow soil
(P = 0.1 %), while on brown forest soil and calcareous chernozem the nitrogen content
of ryegrass was lower, 1.34 % and 1.91 %, respectively (Table 4). The tendency is in
accordance with the original total nitrogen contents of the experiment soils (Table 1).
The higher water supply decreased the nitrogen content of plants in almost every
treatment and soil combination (P = 1.0 % or 5.0 %).
In the mean of the experiment, the amount of nitrogen extracted per pot reduced with
time (1st cut: 87.7; 2 nd cut 42.8 and 3 rd cut: 37.2 mg N per pot). The total amount of
nitrogen extracted by the three cuts is also considerable with 167.7 mg per pot. The
decrease in the amount of nitrogen extracted can be explained by the gradually
decreasing yield.
Table 4: Nitrogen content (%) of the ryegrass 1 st cut
W.c. (%) Soil site N supply (mg kg -1 ) (A)
(C) (B) 0 40 80 Average
Pallag 1.13 1.27 1.83 1.41
50 Látókép 1.38 2.00 2.52 1.97
Dombos 1.85 2.90 3.72 2.83
Average 1.45 2.06 2.69 2.07
Pallag 1.00 1.27 1.52 1.27
75 Látókép 1.35 1.78 2.45 1.86
Dombos 2.10 2.75 3.55 2.80
Average 1.48 1.93 2.51 1.98
Pallag 1.06 1.27 1.67 1.34
Average Látókép 1.36 1.89 2.49 1.91
Dombos 1.97 2.83 3.64 2.81
Average 1.47 2.00 2.60 2.02
Table of variance
Factor FG MQ F SD 5%
A 2 7.72 291.5 *** 0.09
B 2 13.27 501.0 *** 0.09
A*B 4 0.56 21.1 *** 0.16
C 1 0.15 5.71 * 0.08
A*C 2 0.08 2.85 + 0.13
B*C 2 0.02 0.82 0.13
A*B*C 4 0.05 1.99 0.23
Error 51 0.03
*** Significant at P=0.1 % level; * Significant at P=5.0 % level
+ Significant at P=10.0 % level
160

The amount of nitrogen uptake by the plants corresponds with the increasing nitrogen
supply. The amount of the nitrogen extracted from the soils was 97.2; 159.4 and
246.5 mg N per pot in the average of control treatments, at 40 mg kg -1 and at 80 mg kg -1
nitrogen doses, respectively (Table 5).
The amount of nitrogen extracted was dependent on the soil (P = 0,1 %). The
calcareous chernozem and the marshy meadow soil supplied the grass with
approximately the same amount of nitrogen, which can be explained by the fact that the
higher grass yield on the chernozem soil and the higher nitrogen content of plants on
marshy meadow soil offset each other. However, the ryegrass plants extracted only 110
mg nitrogen per pot from the brown forest soil.
Table 5: Nitrogen uptake of ryegrass (g pot -1 ) Sum of 1 st , 2 nd and 3 rd cuts
W.c. (%) Soil site N supply (mg kg -1 ) (A)
(C) (B) 0 40 80 Average
Pallag 61.6 113.3 206.2 127.0
50 Látókép 113.8 169.4 276.4 186.5
Dombos 112.8 189.7 248.9 183.8
Average 96.1 157.5 243.8 165.8
Pallag 60.0 95.7 125.0 93.6
75 Látókép 98.7 186.2 330.5 205.1
Dombos 136.4 202.2 292.1 210.2
Average 98.3 161.4 249.2 169.9
Pallag 60.8 104.5 165.6 110.3
Average Látókép 106.3 177.8 303.4 195.8
Dombos 124.6 196.0 270.5 197.0
Average 97.2 159.4 246.5 167.7
Table of variance
Factor FG MQ F SD 5%
A 2 134992.9 295.5 *** 12.40
B 2 59348.8 129.9 *** 12.40
A*B 4 4742.3 10.4 *** 21.48
C 1 265.3 0.6 10.13
A*C 2 14.1 0.0 17.54
B*C 2 6361.1 13.9 *** 17.54
A*B*C 4 3207.4 7.0 *** 30.38
Error 51 344.0
*** Significant at P=0.1 % level
In spite of the fact, that water dose had an effect both on yield quantity and the nitrogen
content of plants, the level of water supply did not influence the amount of nitrogen
uptake significantly. The reason for this was that while yield increased paralelly with
the water supply, the nitrogen content decreased. These two counter-effects balanced
each other in the nitrogen uptake of plants.
161

ACKNOWLEDGEMENTS
The research work was carried out with the financial support of the Hungarian Scientific
Research Fund (OTKA T 032343).
REFERENCES
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Futterpflanzen von der Höhe der Wasserversorgung. III. Zeitschrift für
Landwirtsch. Versuchs- und Untersuchungswesen, 9: 173-198. p.
Eichler, B. (2004): Möglichkeiten zur Einflussnahme auf Phosphorkreisläufe für die
Gestaltung nachhaltiger Bodennutzungssysteme. Habilitationsschrift. Rostock.
Eichler, B. – Köppen, D. (2001): Die Nutzung ausgewählter Zwischenfrüchte zur
Verringerung von Phosphatverlusten in der Landwirtschaft. Mitt. Gesellschaft für
Pflanzenbauwiss., 13: 166-167 p.
Eichler, B. – Köppen, D. – Grüner, A. (1997): Nutzung von Phosphorvorräten durch
Zwischenfrüchte im Gefäßversuch. 41. Jahrestagung vom 24.-27. 09. 1997 in Kiel,
Tagungsband S. 309
Loch J. – Nosticzius Á. (1992): Agrokémia és növényvédelmi kémia. Mezőgazda
Kiadó, Budapest
Mengel, K. (1972): Ernährung und Stoffwechsel der Pflanze. VEB Gustav Fischer
Verlag, Jena
Nagy, P.T. (2000): Égetéses elven működő elemanalizátor alkalmazhatósága talaj- és
növényvizsgálatokhoz. Agrokémia és Talajtan, 49: 521-534 p.
162

ABSTRACT
THE EFFECT OF NITROGEN SUPPLY, LIMING AND NICKEL
CONTAMINATION ON THE PRODUCTIVITY OF RYEGRASS
(Lolium perenne L.) IN A GREENHOUSE EXPERIMENT
Marianna Sipos, Imre Vágó
University of Debrecen, Faculty of Agriculture
Department of Agricultural Chemistry and Soil Science
H-4015 Debrecen, P.O.Box 36.
It is well known that the dry matter production of plants is determined by the nutrient
and water supply and calcium carbonate content of soils and the amount of toxic
materials (besides their genetic potential). However, it is not known how the
anthropogenic pollutions (e. g. the heavy metal ion content of sewage) modify the effect
of these factors.
For studying this phenomenon, experiments were carried out on two extremely
different soils (leached calcareous chernozem with good fertility and acidic shifting
sand soil with low buffering capacity and humus content). In the pot experiment, the
effect of four factors (soil, nitrogen supply, nickel loading and CaCO3 treatment) on the
dry matter production of perennial ryegrass (Lolium perenne L.) was studied.
In the experiment, the dry matter yield of plants was measured in the different
treatments and treatment combinations.
On chernozem soil, nitrogen had the greatest effect on yield. In the sum of cuts,
lime had a yield-increasing effect, while nickel had no significant effect on yielding
capacity.
On shifting sand soil significant differences were found in the effect of treatments
on yield: nitrogen and lime, and nickel ion have exerted their positive and negative
effect, respectively; under high Ni-loading chlorotic symptoms could be observed
visually on plants. The yield-reducing effect of nickel was striking mainly in the
treatments without lime, while in limed variants the yield-depressing effect of Ni
contamination was more moderate.
Calcium carbonate had a stronger influencing effect on dry matter formation than
nitrogen treatments. These results indicate that improving the pH value of acidic soils is
important since in addition to improving yielding-capacity, it also limits the depressing
effect of toxic elements.
Keywords: Nitrogen, calcium carbonate, nickel contamination, productivity of ryegrass
INTRODUCTION
Nowadays, environmental considerations, study of environmental pollution and
mitigation of damages are becoming increasingly important. Anthropogenic pollution
represents a serious burden for the living and non-living environment. It might happen
that our striving to protect the environment and reduce the pollution will not be efficient
enough without satisfactory knowledge on the sources of danger. A good example of
this is the heavy metal pollution from sewage sludge applications, which can limit the
favourable yield-increasing effect of sewage or can even reduce yields.
163

Fertilizers, liming materials and manure have a low nickel content, therefore, it is not
probable that they would cause serious nickel contamination (Simon, 1999).
In a low exterior concentration, nickel has a favourable effect on the germination
of plant germination and growth. Nickel has only one physiological function in plant
life: it has a role in building the urease enzyme and its functioning. For the functioning
of the enzyme, 6-8 Ni 2+ ions are necessary per molecule, therefore, it is an essential
element for plants (Láng, 1998).
Luckily, nickel content is so low in most soils, that no toxic effect should be
expected: the average Ni 2+ concentration in non-contaminated soils is between 5 and 50
mg kg -1 (Stefanovits et al., 1999). However, in case of its accumulation, it hinders
growth and can result in chlorosis or necrosis. It influences the uptake of ions with two
positive chargings, primarily that of iron and it hinders the translocation of iron into the
shoot. This has a disadvantageous effect on the water management of the plant and
results in an oxidative stress. The human health risks of nickel are significant, it was
proven that nickel accumulating in the human body has a primary carcinogenic effect.
In the 1950s, coating of seeds with heavy metal compounds was a common practice also
in Hungary. In large-scale companies, coating with NiCl2 solution was also used among
others against rust and smut diseases of wheat. The improper use of the remaining
solutions at the place of coating resulted in punctual contaminations (Csathó, 1994).
In recent decades, it became a general practice to use sewage for fertilization after
composting or directly. By the application of sewage, valuable nutrients are provided
for soils (Balláné and Filep, 2000).
However, large amounts of heavy metal can enter the soil with the application of
sewage, even if the concentration of toxic elements is under the official limit since high
volumes are applied. Although the amount of toxic elements that can be applied to the
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
kg ha -1 , the danger of accidental overdosing and the fact that repeating applications
result in accumulation of pollutants in the soil should be considered.
A significant part of the contamination in the soil fortunately becomes immobile
and only a fraction of it enters the plants. The heavy metal binding capacity of the soils
varies greatly. It should also be considered that environmental and production factors
(e.g. nutrient supply, changes in soil pH, precipitation) can modify the mobility of
nutrient elements (Kincses et al., 2005) and pollutants.
All the above facts indicate that it is important to study the effects and interactions
of factors influencing nickel-uptake with special regards to plant yields.
MATERIALS AND METHODS
The objective of the experiment was to reveal the interactions between and direct effects
of nickel contamination and factors having a favourable effect on plant development
(fertilization, CaCO3 application).
For studying the interaction between nickel contamination, nitrogen supply and
CaCO3 application, a pot experiment was set up at the Department of Agricultural
Chemistry in May 2006. The advantage of the pot experiment is that several factors
(e.g. water supply) can be controlled and the same conditions can be ensured for all the
treatment combinations. In addition, the amount of nutrients, pollutants and CaCO3 are
164

exactly known due to the size of the pot and the roots of the plant interwove the whole
pot. Perennial ryegrass (Lolium perenne L.) was used as a test plant. The advantage of
this plant is that it endures well the conditions of the greenhouse and it can be cut
several times during the vegetation period, therefore, the dynamics of the processes can
be concluded too.
The effect of four factors (soil, nitrogen supply, calcium carbonate application and
nickel loading) on plants was studied.
As it is already known, the basic soil characteristics such as soil type, humus
content, pH etc. have a major effect on plant development. Accordingly, the experiment
was carried out on two extremely different soils. One of the soils was calcareous
chernozem with high fertility and buffering capacity from Debrecen-Látókép, while the
other was an acidic sandy soil with weak fertility from Újfehértó.
The basic parameters of the untreated soils used in the greenhouse experiment are
included in Table 1. Calculated for air-dry state, 1.0 kg of each soil was put separately
into the vegetation pots.
Table 1: Parameters of untreated soils
Soil site Debrecen-Látókép Újfehértó
Soil type Calcareous chernozem Shifting sand
Soil texture Loam Sand
Hygroscopicity acc. to Arany (KA) 39 26
pH-H2O 6.05 5.01
pH-KCl 5.41 3.98
pH-CaCl2 5.73 4.19
Hydrolytic acidity (y1) 9.07 7.06
Total C content (%) 1.89 0.27
Mass volume (kg dm -3 ) 1.19 1.49
Calculated lime requirement (t ha -1 ) 6.16 3.19
Calculated lime requirement (g kg -1 ) 2.07 0.86
The initial total carbon content of soils was measured by Nagy, P. T. (2000). The
standard treatment of pots was a joint solution of 100 mg kg -1 P and 160 mg kg -1 K, in
10 cm 3 KH2PO4 and K2SO4. The combinations of treatments applied on the soils are
shown in Table 2.
Nitrogen and nickel were added to the soils as NH4NO3 and NiSO4 solution,
respectively. Lime requirement of the soils was calculated according to the method of
Filep (1988) as
CaCO3 (kg ha -1 ) = 17.4 * KA * y1
then was added to the soil as fine powder. Each treatment was carried out in four
replications.
The water supply was ensured by daily irrigation to a specific weight. The
transpirated and evaporated water was supplemented by irrigating the chernozem and
sandy soil with distilled water to 75% and 60% of the field water capacity, respectively.
At high vegetative mass, plants were irrigated in the afternoon also on hot days to avoid
wilting.
165

Table 2: Combinations of treatments
Number of
combination
N treatment
(mg kg -1 )
Ni treatment
(mg kg -1 )
CaCO3
treatment code (!)
1. 80 0 0
2. 80 50 0
3. 80 100 0
4. 80 0 1
5. 80 50 1
6. 80 100 1
7. 160 0 0
8. 160 50 0
9. 160 100 0
10. 160 0 1
11. 160 50 1
12. 160 100 1
(!) 0 = without CaCO3 1 = CaCO3 dose according to calculated lime requirement
Plants were cut two times, 33 and 61 days after sowing, the grass yield was dried at
60ºC until reaching constant mass in a desiccator and the dry matter production per pot
was measured by analytical balance.
RESULTS AND DISCUSSIONS
The obtained results were evaluated by a three-variate analysis of variance on the basis
of Sváb’s method (1981). In the analysis of variance, F and SD 5 % values were
calculated for each factor and for each double and triple interaction, then the
significance levels were determined.
Summarized yields of the two cuts and their statistical evaluation on chernozem
and sandy soils are shown in Tables 3 and 4.
On the chernozem soil at Látókép, it can be stated when considering the sum of dry
matter production of two cuts that nitrogen had a significant effect on yield (P = 0.1 %).
Nitrogen supply had a different effect in the different lime and nickel combinations.
This is indicated by the fact that the N*Ni*CaCO3 triple interaction was significant at
P = 5.0 % level, meaning that factors (in our case nitrogen) exert their effect depending
upon the other factors.
In combinations not treated with calcium carbonate, the yield-increasing effect of
nitrogen could be observed at all three nickel dosages, however this effect was
decreasing with increasing nickel dosages. Whereas in limed treatments, the increasing
nickel dosages do not limit the yield-increasing effect of nitrogen.
The yield-reducing effect of Ni is not significant on this soil in the sum of the two cuts
as confirmed by the statistical analysis.
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Table 3: Dry matter production of perennial ryegrass (g pot -1 )
Látókép chernozem, 1st+2nd cut
Factors Unlimed Limed Average
N=80
Ni=0 7.53 8.02 7.77
Ni=50 7.65 7.55 7.60
Ni=100 7.68 7.68 7.68
Average 7.62 7.75 7.68
N=160
Ni=0 10.20 9.18 9.69
Ni=50 9.93 10.05 9.99
Ni=100 9.67 9.60 9.64
Average 9.93 9.61 9.77
N average
Ni=0 8.86 8.60 8.73
Ni=50 8.79 8.80 8.79
Ni=100 8.67 8.64 8.66
Average 8.78 8.68 8.73
Table of variance
Factor MQ DF F SD5%
N 52.292 1 234.9 *** 0.28
Ni 0.076 2 0.34 0.38
N*Ni 0.273 2 1.22 0.48
CaCO3 0.11 1 0.49 0.28
N*CaCO3 0.63 1 2.83 0.398
Ni*CaCO3 0.86 2 0.39 0.48
N*Ni*CaCO3 0.876 2 3.94 * 0.68
*** Significant at P=0.1 % level * Significant at P=5.0 % level
No considerable differences were found in dry matter yield of the first and second cuts.
This suggests that the amount of nutrients applied to 1 kg chernozem soil and the
natural nutrient-supplying capacity of the soil were satisfactory for plant development.
Regarding the summarized yield of the two cuts on shifting sandy soil, it can be stated
that all three treatment factors had a significant effect.
Increasing nitrogen dosages increase yield significantly as proven by the statistical
analysis (P = 1.0 %). However, the increasing effect was different at different lime
dosages. Without lime treatment, higher nitrogen doses resulted in a smaller yield
167

increment, than with higher calcium carbonate dosaging. It has to be noted also, that
lime also had a positive effect on yield and it was more significant (P = 0.1 %) than that
of nitrogen.
Table 4: Dry matter production of perennial ryegrass (g pot -1 )
Újfehértó shifting sand, 1st+2nd cuts
Factors
N=80
Unlimed Limed Average
Ni=0 4.73 5.18 4.95
Ni=50 3.38 4.90 4.14
Ni=100 1.65 4.90 3.28
Average
N=160
3.25 4.99 4.12
Ni=0 5.85 6.63 6.24
Ni=50 4.85 7.05 5.95
Ni=100 0.88 5.80 3.34
Average
N average
3.86 6.49 5.17
Ni=0 5.29 5.90 5.59
Ni=50 4.11 5.97 5.04
Ni=100 1.26 5.35 3.31
Average
Table of variance
3.55 5.74 4.65
Factor MQ DF F SD5%
N 13.335 1 11.77 ** 0.62
Ni 22.811 2 20.14 *** 0.76
N*Ni 3.226 2 2.85 + 1.08
CaCO3 57.422 1 50.67 *** 0.62
N*CaCO3 2.385 1 2.11 0.88
Ni*CaCO3 12.393 2 10.94 *** 1.08
N*Ni*CaCO3 0.491 2 0.43 1.53
*** Significant at P=0.1 % level
** Significant at P=1.0 % level
+ Significant at P=10 % level
The yield-increasing effect of nitrogen had a significant interaction with nickel dosaging
at P = 10 % level. In treatments without nickel, nitrogen had a yield-increasing effect
both in limed and unlimed treatments. In the case of nickel treatment however, the
yield-reducing and yield-increasing effect of nickel and nitrogen, respectively, were
168

more definite in the non limed treatments. Under calcium carbonate treatment, a
significant yield-increment could be observed in all treatment combinations.
Nickel contamination had a significant yield-reducing effect in the sum of the two cuts
at P = 0.1 %. In unlimed treatments, a dosage of 100 mg kg -1 Ni had a stronger reducing
effect than the smaller dosage. Data of the table also indicate that in the average of N
treatments, the harmful effect of nickel was strong in the unlimed treatments (yieldreduction
from 5.29 g pot -1 to 1.26 g pot -1 ), while in the limed treatments, the effect was
much weaker (yield-reduction from 5.90 g pot -1 to 5.35 g pot -1 ).
Considering the effect of N treatment, it can be observed that yield-reduction was
smaller both in limed and unlimed treatments when smaller N dosage was applied than
at higher dosages. This is probably due to the fact that plants used more nutrients and
consequently they took up more Ni for producing higher yields.
CaCO3 treatment had a significant effect on yield in all treatment combinations. It
can be stated that as a result of the favourable soil pH casued by liming, the dry matter
production per pot increases. The ratio of this increment is higher than that of the yieldincreasing
effect of nitrogen.
By adding lime, the yield-reducing effect of nickel was reduced or even prevented.
This conclusion is supported by the fact that the significance level of Ni*CaCO3
interaction on shifting sand was P = 0.1 %.
As a continuation of the study, we plan to determine the constitution of elements in
the dry matter of perennial ryegrass. This would enable us to determine how treatments
influenced the uptake of elements by plants. This knowledge can lead to further
conclusions.
ACKNOWLEDGEMENTS
The research work was carried out by the support of “Tormay Béla” Honour Society of
the University of Debrecen, Centre of Agricultural Sciences.
REFERENCES
Balláné Kovács, A. - Filep, T. (2000): A különböző SO4 2- /SO4 2- +Cl - arányú K-, Ca-,
Mg-só adagok hatása az angolperje tápelem összetételére. Agrokémia és Talajtan
49: 3-4 p.
Csathó, P. (1994): A környezet nehézfém-szennyezettsége és az agrártermelés. MTA
TAKI, Budapest.
Filep, Gy. (1988): Talajkémia. Akadémiai Kiadó, Budapest
Kincses, S.-né – Nagy, P. T. – Balláné Kovács, A. – Filep, T. (2005): Az NPKtrágyázás
hatása a búza (Triticum aestivum) és a kukorica (Zea mays) Znfelvételére.
Acta Agronomica Óváriensis, 47: 239-252 p.
Láng, F. (1998): Növényélettan. ELTE Eötvös Kiadó, Budapest
Nagy, P.T. (2000): Égetéses elven működő elemanalizátor alkalmazhatósága talaj- és
növényvizsgálatokhoz. Agrokémia és Talajtan, 49: 521-534 p.
Simon, L. (1999): Talajszennyeződés, talajtisztítás. Környezetgazdálkodási Intézet
Környezet – és Természetvédelmi Szakkönyvtár és Információs Központ,
Budapest
Sváb J. (1981): Biometriai módszerek a kutatásban. Mezőgazdasági Kiadó, Budapest
169

TOXIGENIC FUNGI AND MYCOTOXINS IN GRAINS AND BAKERY
PRODUCTS FROM ROMANIA
Carmen Puia, Viorel Florian, Alexandra Suciu, Augusta Lujerdean*, Dana Pusta*
Department of Plant Protection, *Department of Chemistry, University of Agricultural
Sciences and Veterinary Medicine, Cluj Napoca, Romania
ABSTRACT
Our researches want to establish the mycoflora and the natural occurrence of
mycotoxins in grains for industrialization and fodder, bread and bakery products in our
country. We intend to compare these levels with the international ones and to change
the legal level of mycotoxins admitted by our laws and to align these levels with the
European legislation. The authors reported the identification of mycoflora in 130
samples of grains, bread, biscuits and cereal flakes from different districts of Romania.
Moulds evaluation was determined using conventional methods as blotting test and
Ulster test. The predominant genera were those of toxigenic fungi: Penicillium,
Aspergillus and Fusarium.
Our studies were focused on finding rapid methods for screening of aflatoxins B1,
(AB1), aflatoxin B2 (AB2), aflatoxin G1 (AG1), aflatoxin G2 (AG2) and ochratoxin A.
Mycotoxins were extracted in chloroform, separated on silicagel thin-layer
chromatography plates and quantificated using densitometric analysis. There were
analyzed 39 cereal samples (wheat and maize) from different Romanian districts.
The contamination range was: aflatoxin B1: 1.7 - 5.7 µg/kg, aflatoxin B2: 0.02 -2.8
µg/kg, aflatoxin G1: 1.1 – 5.7 µg/kg, aflatoxin G2: 0.12 – 1.8 µg/kg, total aflatoxins: 1.2
– 10.8 µg/kg and ochratoxin A: 4.4 - 30.0 µg/kg. The higher number of contamination
rate was in the case of wheat samples. The mycotoxin analyses reveal to us a high and
dangerous level of ochratoxin A and total aflatoxins. Our food and fodder are dangerous
and our legislation is way too permissive. We have to align our legislation with the
European one to take care about our grains.
Key words: mycoflora, toxigenic fungi, grains, bakery products, mycotoxins
INTRODUCTION
Man has long benefited from the nutritional properties of certain microscopic fungi,
such as ferments and yeasts used in making cheese, beer, bread, vinegar and yogurt. But
some moulds of fungal origin are potent poisons, especially the mycotoxin family that
has been the subject of increased attention over the past 15 years.
Most fungi produce a class of chemical compounds called secondary metabolites.
These compounds have a wide range of biological activities including antibiotic
(antibacterial and antifungal), acute and chronic toxicities (plant, animal, and humans),
hormones and growth regulation (plants and animals). Several species of fungi that
colonize grains produce mycotoxins, however not all isolates of a toxigenic species
produce mycotoxins and isolates capable of producing a mycotoxin do not always
synthesize the toxin (Mills, 1990).
Our researches, want to establish the mycoflora and the natural occurrence of
mycotoxins in grains for industrialization and fodder in our country. We intend to
170

compare these levels with the European ones and to change the legal level of
mycotoxins admitted by our laws and to align these levels with the European
legislation.
MATERIALS AND METHODS
Different random samples of grains (wheat, corn, Triticale and rye), bread and bakery
products (biscuits, cereal flakes) were collected from 14 districts of Romania. Each
sample (1 kilo) was put in sterilized bag and in laboratory was divided into 2 parts. One
part was used for mycological analysis and the second one was used for mycotoxin
analysis. The mycological analysis was made in 5 replications, by using the blotting
paper method (Raicu Cristina and Doina Baciu, 1978) and the Ulster method on
agarized plates (Hulea Ana and all, 1982). We used 10 grains from each sample in 3 or
5 replications. In this study was employed a Czapek - Dox agar medium with
Streptomycin as bacteriostatic agent, added after sterilization (Hulea Ana, 1969). The
identification of fungi was according to Raicu Cristina and Doina Baciu (1978) and
Domsch and Gams (1972).
Our studies were focused on finding rapid methods for screening of aflatoxins B1,
(AB1), aflatoxin B2 (AB2), aflatoxin G1 (AG1), aflatoxin G2 (AG2) and ochratoxin A.
Mycotoxins were extracted in chloroform, separated on silicagel thin-layer
chromatography plates and were quantified using densitometric analysis (Braicu and all,
2005). We analyzed samples from different Romanian regions.
The limits of detection of this method on TLC plate were 0.25 μg/kg for AG1, AG2,
AB1, AB2, respectively 3�μg/kg for ochratoxin A and the limits of quantification from
samples were 0.83 μg/kg AG1, AG2, AB1, AB2, 10 μg/kg for ochratoxin A
RESULTS AND DISCUSSIONS
The degree of infestation with fungi as presented in Table 1 is very high, above 50%, in
all types of samples collected from the country.
Table 1. The degree of infestation with fungi in grains and bakery products
Product
Analised
samples
(nr)
Number of samples infected with fungi
Potential toxigenic fungi (%)
Total
no (%)
Penicillium Aspergillus Fusariu
sp.
sp. m sp.
Wheat 130 78 / 60.0% 38.46 38.46 76.92
Maize 60 35 / 58.3% 85.71 77.14 68.57
Rye 6 5 / 83.3% 40.0 60.0 60.0
Triticale 6 5 / 83.3% 80.0 100 100
Bread 6 6 / 100% 100 83.3 33.3
Biscuits 6 4 / 66.7% 25.0 25.0 50.0
Flakes 6 3 / 50.0% 100 33.3. 100
Total 220 135 / 61.4% 56.29 53.33 74.07
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In wheat 60% of samples are infected, in corn 58.3% of samples but the most alarming
issue is that the fungal infection is present in bread (100%), biscuits and cereal flakes.
The results indicates that in the analyzed samples the percent of potential toxigenic
fungi is high too, we notice Penicillium from 25% to 100% (bread) infected sample,
Aspergillus from 33.3% to 100% (Triticale) infected samples and Fusarium from 33.3%
(bread) to 100% (Triticale and flakes) infected samples.
Analyzing the wheat samples by blotting test and Ulster test, we’ve determined
fungi related to 12 genera (Table 2) which have developed on bread grains: Penicillium,
Aspergillus, Fusarium, Alternaria, Cladosporium, Helminthosporium, Trichotecium,
Stachybotrys, Septoria, Epicoccum, Acremoniella and Rhizopus. On grains we can
observe fungi from field, typical storage mycoflora and intermediate mycoflora (Hulea
Ana and all, 1982; Boariu Carmen, 1988). We can notice higher values especially for
toxigenic fungi as Penicillium, Aspergillus and Fusarium.
Table 2. The occurrence of fungi in the analyzed samples of bread grains
Frequency of fungi determined by B.p.t. and Ulster test
Fungi
0% under 20% 20-70% above 70%
B.p.t U.t. B.p.t. U.t. B.p.t. U.t. B.p.t U.t.
Penicillium sp. 40,0 15.38 33.3 34.62 26.7 38.46 - 11.5
4
Aspergillus sp. 43.3 15.38 26.7 19.24 26.7 50.0 3.3 15.3
8
A. ochraceus 96.7 84.62 - 7.69 3.3 7.69 - -
A. niger 76.7 53.84 6.7 19.23 13.3 23.08 3.3 3.85
A. nidulans 100 84.62 - 7.69 - 7.69 - -
A. flavus 90.0 61.54 - 19.23 10.0 15.38 - 3.85
A. glaucus 96.7 84.62 - 15.38 3.3 - - -
A. versicolor 100 76.93 - 15.38 - 7.69 - -
A. fumigatus 93.4 92.3 3.3 3.85 3.3 - - 3.85
Fusarium sp. 10.0 26.92 26.7 30.77 53.3 34.62 6.67 7.69
Alternaria sp. 20.0 26.92 13.3 19.23 43.3 38.46 23.3 15.3
9
Cladosporium sp 56.7 38.46 26.6 30.77 16.7 30.77 - -
Trichotecium roseum 93.3 88.77 - 7.69 6.7 11.54 - -
Stachybotrys sp. 96.7 96.15 3.3 - - 3.85 - -
Rhizopus sp. 76.7 61.54 6.7 23.8 13.3 15.38 3.3 -
Septoria sp. 90.0 100 10.0 - - - - -
Helminthosporium sp. 96.7 100 3.3 - - - - -
Acremoniella atra 96.7 96.15 3.3 3.85 - - - -
Epicoccum purpurasc. 96.7 100 3.3 - - - - -
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Table 3. Natural occurrence of mycotoxins in cereals analyzed by TLC coupled with
densitometry from different Romanian districts
Samples Dist.
No of No. of Ochrat.
examin. contamin. A
samples samples µg/kg
AB1
µg/kg
AB2
µg/kg
AG1
µg/kg
AG2
µg/kg
Total
Aflat.
µg/kg
Wheat BC 1 1 0 5.3 5.5 10.8
BN 2 2 0 4.3 1.4 0
5.7
6.5 5.0 1.5
6.5
BV 3 2 0
4.2 1.0 5.2
30.0
2.0 1.8 3.8
0
0 1.2 1.2
BH 1 -
HD 3 2 0 5.7 2.6
8.3
3.9 0.9
4.8
CJ 2 -
MM 4 1 0 0.1 0.5 0.6
GR 2 -
SB 1 1 0 5.6 2.7 8.3
SM 1 1 0 4.0 4.0
SJ 1 -
Maize BC 2 2 0 1.7 0
1.7
3.9 1.4
5.3
BV 1 -
MM 1 -
SB 1 1 0 3.3 3.3
HD 2 1 4.4 1.7 0.6 2.3
GR 2 1 0 - 0.02 0.12 0.14
SM 1 -
Triticale BV 3 3 0
0.5 1.1
1.6
5.7 0.8 6.5
5.7 0.8 6.5
Rye SB 1 1 25.6 4.6 2.8 7.4
HD 1 - -
Bread CJ 2 -
Biscuits CJ 1 1 0 4.4 1.8 - 6.2
Limits by Commission of Regulation
(EC), No 472/ 2002
3-5

In conclusion, the fungal charge of grains for food and fodder on all samples is enough
high to alarm us, indifferent of the method of analyze. We have to corroborate this data
with mycotoxins analyses to know if our food and fodder is dangerous or not.
In the next table (Table 3) we present the natural occurrence of mycotoxins in
grains analyzed by TLC coupled with densitometry.
As shown in Table 3, we analyzed 39 samples from different Romanian districts,
20 samples were found contaminated with mycotoxins. Four samples were
contaminated with ochratoxin A and the concentration was higher than legal European –
we notice that in rye and a sample of wheat for bakery, the values were very alarming,
over 20 µg/kg. In 11 samples was identified AB1, in 9 samples the amont of this was
higher than norms; concerning the total concentration of aflatoxins 10 sample this was
higher than legal European norms. The concentration of aflatoxins is very dangerous in
biscuits for kids.
The contamination range was: aflatoxin B1: 1.7 - 5.7 µg/kg, aflatoxin B2: 0.02 -2.8
µg/kg, aflatoxin G1: 1.1 – 5.7 µg/kg, aflatoxin G2: 0.12 – 1.8 µg/kg, total aflatoxins: 1.2
– 10.8 µg/kg and ochratoxin A: 4.4 - 30.0 µg/kg. The higher number of contamination
rate was in the case of wheat samples.
I n conclusion, we can notice that the fungal charge of grains for food and fodder
and in bakery products on all samples is enough high to alarm us, no matter of the
method of analyze. The mycotoxin analyses reveal to us a high and dangerous level of
ochratoxin A and total aflatoxins. Our food and fodder are dangerous and our legislation
is way too permissive. We have to align our legislation with the European one to take
care about our grains.
REFERENCES
1. Boariu Carmen, 1988, Cercetări privind modificările fizico – chimice şi biologice
care au loc în timpul păstrării seminţelor destinate consumului., în „Contribuţii ale
cercetării ştiinţifice la dezvoltarea agriculturii din zona centrală a Câmpiei de
Vest”, C.M.D.P.A., 291-318.
2. Braicu Cornelia, Carmen Puia, C. Bele C., E. Bodoki, Carmen Socaciu, 2005,
Optimisation of screening systems to evaluate relevant mycotoxins from cereals
and bread, 4 th <strong>International</strong> Symposium of USAMV Cluj-Napoca, 16:144-149
3. Domsch K.H., W. Gams, 1972, Fungi in agricultural soils, published by Longman.
4. Hulea Ana, 1969, Ghid pentru laboratoarele de micologie şi bacteriologie, Ed.
Agrosilvică, Bucureşti, 304 p.
5. Hulea Ana, Gh. Tasca, C. Beratlief, 1982, Bolile şi dăunătorii produselor agricole
şi hortiviticole după recoltare, Ed. Ceres, Buc., 193 – 210.
6. Mills J.T., 1990, Mycotoxins and toxigenic fungi on cereal grains in western
Canada., Can. J. Physiol. Pharmacol. 1990 Jul; 68 (7):982-6.
7. Raicu Cristina, Doina Baciu, 1978, Patologia semintei, Ed. Ceres, Bucuresti, 207 p.
174

CORRELATIONS OF SOCIAL AND ECONOMIC FUNCTIONS IN
COMMUNITIES AROUND HORTOBÁGY
Erzsébet Csengeri
Tessedik Sámuel College Faculty of Agricultural Water and Environmental
Management Environmental Management Institute
1-3 Szabadság Sreet 5540 Szarvas
ABSTRACT
The change of ecosystems is mainly influenced by the material handling activity of man.
This is particularly accentuated in an area sensitive in other respects as well, where a
couple of decades ago man still fought hardly for a farming as efficient as possible on
this extensive area with saline, bad quality soils. A characteristic feature of Hortobágy is
that it represents a well-delimited natural unit from both botanical and zoological and
hydrographic respects, which preserves several cultural features. I will present the
environment-modifying activity of man mainly after the industrial revolution on the
example of communities with large agricultural lands.
Due to the social and economic changes brought about by the political change the
communities with dominance of agriculture got into more and more unfavourable
conditions which resulted in depopulation and the discontinuation of land use forms,
and thus, a new change of environment. From a demographic point of view, Hortobágy
can be divided into two main parts, the core village centre where 65% of the population
lives and the farm centres lying in a radius of 3 to 10 km, which used to be centres of
the former Hortobágy State Farm. By now, these communities partly became
depopulated, partly became parts of Hortobágy. All these communities are characterised
by aging of the population due to the lack of economic income. In respect of farming,
the tendency of shifting from the earlier intensive acitivities to extensive ones is typical.
Plants are produced only in areas with good quality soils which fulfills only the need for
feeds. Mainly the breeding of indigenous breeds is preferred for rehabilitation of the
original state of vegetation on the ploughed areas.
Keywords: human activity, land use industrial revolution economic changes
INTRODUCTION
As the human society plays an ever more determining role in the ecosystem
metabolism, the research of the activities and material handling of man as a key species,
i.e. of the society metabolism necessarily comes to the front. In this, the demography,
economy, culture, sociology, political and power structure and operation of local
communities are determining.
My objective is to show the impact of man as a key species on ecosystems on the
example of the settlements around Hortobágy. In the rural settlements to be presented, I
analyse in the first place the change of the role of agriculture from the point of view of
the demographic changes of the communities (Oláh, 2006).
There have been differences in the economic activity of the chosen settlements, both in
the past and present. The studied settlements were Árkus, Hortobágy-halastó, Kónya,
Máta, Szásztelek and the following farmsteads: Borsós, Kungyörgy, Kuntelek,
175

Várostanya, Malomháza, Faluvégháza, Villongó I and Hortobágy that administratively
unites these settlements.
The listed settlements are all typical agricultural, rural ones. The lack of independence
of the settlements shows already by itself that they are not viable on their own, neither
from a demographic nor from an economic point of view. According to the
classification of the rural human organization levels, they correspond to small clusters
of communities that are more isolated and offer less services compared to the town of
Hortobágy.
MATERIALS AND METHODS
The demographic and economic data were collected personally in the settlements. In
addition to my own collection, necessary information was obtained from the Central
Statistical Office and the T-STAR database. The demographic indices were determined
using the parameters of the Clocke index indicating the directions of the possible future
development of the given settlement. The studied settlements, Árkus, Hortobágyhalastó,
Kónya, Máta, Szásztelek, Borsós, Kungyörgy, Kuntelek, Várostanya,
Malomháza, Faluvégháza, Villongó I and Hortobágy are typically agricultural
settlements, and thus, their study over more years is interesting in view of the ever
decreasing population holding capacity of the agriculture.
Changes of the farming structure were studied from the 1960s, broken down by
ten-year periods, giving median values. The study targeted the different agricultural
land uses, changes of the ratios of agriculturally used crop lands and livestock. I made
an in-depth interview with Mr. Csaba Göcz who was in charge of agricultural
production on this territory of over 30 000 ha from the late 1960s.
RESULTS AND DISCUSSIONS
The changes of the resident population closely follow those of the economic life. Earlier
in time, in the 1960s and 1970s, the peripheries had higher population. In the eighties,
the fiscal goods coming from agriculture, especially from animal husbandry, permitted
the increase of the demands of the peasantry. Because of the better infrastructural
conditions, the population moved from the farmsteads to Hortobágy, while retaining
their occupation. This is when the total population of Hortobágy and the neighbouring
farmsteads reached its maximum of nearly 2000 persons (Figure 1). Following this, a
slowly decreasing trend started in the 1990s, which has lasted to our days.
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1600
1400
1200
1000
800
600
400
200
0
1960 1970 1980 1990 2000 2004
Figure1: Run of the population in periphery and core areas
As a result of the depopulation process, the farmsteads have either become part of
Hortobágy (e.g., Borsós) or totally depopulated like e.g. Várostanya, Malomháza,
Faluvégháza or Kuntelek. In Kungyörgy and Villongó I, not even ten families live,
while in Máta, Kónya, Hortobágy-halastó, Árkus and Szásztelek, there are more than 10
families left (Egyedné, 1999).
The parameters of the Clocke index show the dynamics of the rural population.
The parameters study the ratios of the following age classes that, in case of Hortobágy,
show the pattern presented in Table 1 in the studied two years.
Table 1: Demographic structure of, village population
perifery
1985 1995 2000 2004
Aging Ratio 0,10 0,30 0,57 0,70
Generation’s rejuvenation Ratio 0,20 0,20 0,20 0,19
Fertility Ratio 2,0 1,3 1,03 1,15
Sex Ratio 95 97 99 101
Dependants Ratio 0,47 0,35 0,3 0,34
As a result, it can be stated that the aging of the population, which corresponds to the
nation-wide trend, is even more pronounced in agricultural-type rural areas like
Hortobágy and the neighbouring farmsteads. The age composition of the Hortobágy
settlements shows the following pattern. In 1995, 21% of the population was children,
67% was middle-aged (15-19 years old), while 12% were over 60. On the basis of the
2001 statistical data, it can be seen that a part of the children of the 1995 statistics
moved to the middle-aged group, but this reduction was not compensated by
177
core

eproduction, and therefore, children represented only 19%. In the middle-aged group,
the same trend can be observed, their share is 69%. The population over 60 years of age
makes 11%. The generation rejuvenation can be considered good, although it is
gradually approaching one. The fertility ratio also shows a decreasing trend.
The dependants’ ratio can be considered good in the settlement, as the share of the
actively occupied population is predominant and unemployment does not exceed 10%.
These results show that the previous emigration process is now complemented by a
slow aging process, which can have unpredictable consequences in this area, important
from many respects. This process is also well reflected by the changes in the total
population of the settlement (Table 2).
Table 2:Run of the population
Years Total population/head
1960 1508
1970 1791
1980 1848
1990 1741
2000 1757
2004 1681
Next, I study if the changes of the economic data corroborate the demographic changes.
Here, I also go back to the early years of the industrial revolution. After the Second
World War, there several ideas were proposed regarding the better exploitation of the
Hortobágy puszta. However, all proposals concurred in that they considered animal
husbandry as one of the possible main directions. It is the principal branch of agriculture
today as well. The required feeds were produced in the ploughed areas of the
Hortobágy.
In view of the soil characteristics of Hortobágy (grassland clay, sodic, chernozem
loam, sandy loam, alluvial and sandy soils) the plant cultivation consists mainly of the
cultivation of corn and sugar beet satisfying the winter feed demand of livestock,
especially sheep.
In the 1960s, farming started on an area of 30 351 ha, whereof, in correspondence with
the planned agricultural profile, the largest share was given to meadows and pastures
that occupied approximately twice as much area as the plough-lands at the time.
This area would seem big enough to entirely satisfy the feeding needs of the warmblooded
livestock during all the summer and winter period. The real situation is much
worse. In fact, great part of the meadows and pastures are barren from the beginning of
June, they can be used only as sheep pastures. Areas with such characteristics make
65% of the farmed area of Hortobágy.
The lucerne growing on 22% of the study area also serves the feeding needs of
animal husbandry. Plants as poppy, tobacco or soybean were grown on a further 5% of
the farmed area, which did not produce adequate results. Autumn wheat was generally
cultivated on the sodic or highly sodic soils with the lowest production capacity, using
intensive wheat sorts and significant quantities of fertilizers (Table 3).
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Table 3: Land use/ha
1960 1970 1980 1990 2000
Plough-land 5379 5610 5350 3215 2802
Garden 3 4 7 7 7
Vineyard - - - - 0,36
Orchard - - - - 3,61
Meadow
Pasture-land
16179 10221 10100 12821 13006
Reed 551 570 1345 1869 2183
Forest 875 825 820 814 796
Fish-pond 7150 6920 6431 5011 4070
Total bearing surface 30137 24150 24053 23737 22870
Another result of the sixties was the systematic forestation programme started on the
lowest-lying parts of the pastures.
The land use trends have lasted throughout the years, although the usable crop land
has shrunk, partly because of their exclusion from cultivation and partly because of land
appropriations by the Hortobágy National Park founded in 1973.
The village of Hortobágy has its own administrative area since 1 January 1966 (a
total of 28 458 ha), previously it belonged to Balmazújváros. Of the 28 458 ha, the
surface used by the core areas and excluded from cultivation has been continuously
increasing, i.e. the crop land under agricultural cultivation has been diminishing.
The ploughing land use was the most widespread in the 1960s and 1970s because
of rice growing on ploughed areas, which terminated in 1980. Since that time, the
ploughed lands have been continuously shrinking, while the grasslands and – in the
recultivated wetlands – the reedy areas have been growing. The use of the fishponds
established in the early 20 th century has been constantly decreasing. The Hortobágy
National Park, established in 1973, has been continuously increasing the protected areas
and the area in its property has also grown. By 2000, over 80% of the crop land under
the jurisdiction of the village of Hortobágy was transferred under the administration of
the National Park, these areas are mainly used by the Hortobágy Non-Profit Company,
while a smaller part of them is leased by agricultural entrepreneurs (Szeifert, 1969).
Of the animal husbandry of the 1960s and 1970s, sheep breeding reached
exceptional economic levels resulting in significant exports, and thus, incomes, but the
fisheries and poultry farming show similar characteristics, whereof the highest was the
share of the guinea-fowl and duck farming. The cattle and horse breeding dealt only
with issues related to the rehabilitation of the genetic pool at the time (Table 4).
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Table 4: Livestock/pieces,
1960 1970 1980 1990 2000
Cattle 2172 2615 2980 2955 2681
Pig 7489 7820 7905 5015 1870
Horse, 630 715 608 520 385
Sheep 31838 33200 29020 14835 6485
Poultry 59949 450300 690100 71500 20891
Bee 48 69 36 55 46
The largest share in the exports and thus, the highest incomes came from fish exports,
however, interestingly, the statistical evaluations do not provide numerical data for
these stocks.
During the decentralized privatization of the State Farm, the large-scale intensive
animal farms were constantly privatized or liquidated between 1988 and 1992, which
resulted mainly in the reduction of the cattle and sheep stocks. In this area, pig breeding
had traditions mainly in household farms, but it has decreased to a minimum by today.
The poultry stock consisted of waterfowl (geese, ducks) and guinea-fowl. Between 1960
and 1980, the State Farm raised large quantities of so-called meat geese, pond-reared
broiler ducks and broiler guinea-fowl according to the market demand of the time. By
2000, the poultry stock has reduced to a minimum. It is typical that in the village of
Hortobágy, 100% of the cattle is kept industrially, while 30% of the horses, 65% of the
sheep and 70% of the pigs can be found in small-scale, privately owned farms.
As a result of the survey of the animal husbandry sector of the agriculture, it can be
stated that, in accord with the Hortobágy soil conditions and the productivity of the
natural vegetation evolved here, rearing cattle that is grazed in the open air all the year
round and sheep is more dominant even to this day, as well as the fish culture, done
within the frame of partly natural pond culture.
REFERENCES
Clocke P. J. (1997): An index of rurality for England and Wales. Regional Studies 11:
31-46
Egyedné Kertész I. (1999): A helyi önkormányzatok hatáskörébe tartozó adók
elemzése és ellenőrzése a bevételek alakulása folyamatában Hortobágy Község
Önkormányzatának 1991-1998. évi gazdálkodásán és adóztatási gyakorlatán
keresztül. Államigazgatási Főiskola, Budapest 12-13
KSH (2004): Egyéni agrárgazdaságok és népsűrűségük. Észak-Alföld. KSH
Házinyomda. Debrecen, Szolnok, Nyíregyháza 31-45
Oláh J.(2006): Környezetgazdálkodás. TSF MVKFK, Szarvas
Szeifert I.(1969): Hortobágyi Állami Gazdaság. Hajdú-Bihar Megyei Lapkiadó
Vállalat, Debrecen
180

RIDING TOURISM IN THE NORTHERN GREAT PLAIN REGION
(HUNGARY): STATUS AND POTENTIAL
Réka Incze and Gábor Hevessy
University of Debrecen, Center of Agricultural Sciences
4032 Debrecen, Böszörményi Str. 138
ABSTRACT
Only a small part (

assessed the different conditions of the Great Plain region inserted into a Hungarian
regional comparison system.
Nowadays, tourism is one of the most developing industrial branches; additionally, it
results the highest incomes from export in the world (UNO, 2002). Tourism offers for
127 million people possibility of work, with its share of 6 % of the world’s GDP, i.e.
each fifteenth person works in the field of tourism. If we include also workplaces
connected indirectly to tourism, then we get 260 million workplaces, i.e. approx. 10 %
of GDP (World Travel and Tourism Council, 2004). Incomes originated from tourism
achieved the value of 622 million USD in 2004 exceeding by 18.7 % the incomes of
2003. The above mentioned data do not contain inland travels, the number of which
may be the sextuple of international travels.
Tourism also plays an important role in the economy of Hungary. The value of the
foreigners’ consumption visiting Hungary was 822 billion HUF in 2004, within which
the value of tourist consumption was 596 billion HUF. Moreover, the Hungarian
population spent 385 billion HUF on tourist services. Summing these, the value of
touristical incomes achieved 981 billion HUF. The total GDP of touristical branches is 5
% of the whole Hungarian GDP. Taking into account also multiplier impacts 8.5 % of
the Hungarian GDP originates from tourism. The number of employees employed
directly in the field of tourism was 398,000 persons in 2004, which is 8.9% of the total
employment. Taking into account multiplier impacts the mentioned proportion increases
to 12.5 %, which means that each 8 th workplace has been produced by tourism.
Table 1.: touristically qualified riding establishments and services in Hungary
Number of horseshoes
5 4 3 2 1 Other* Total
Qualified establishments (pcs) 15 29 32 29 27 26 158
Accommodation (pcs) 831 702 572 445 880 1022 4452
Roofed riding ground (pcs) 9 13 8 1 2 7 40
Own horse number (pcs) 1635 778 757 574 471 1997 6212
Accommodation for foreign horses
(pcs) 232 359 377 166 221 649 2004
* E.g. presentation sites, horse-breeding, riding education establishments and
establishments rendering services of nomadic character.
The most popular destinations are Budapest, lake Balaton and spa resorts. The average
capacity utilisation of commercial accommodations was 39.5% in 2005. The best
utilisation, i.e. 48.2 % average utilisation was achieved by hotels. The average
utilisation of guest-houses was 22.7 %. The demand for higher quality accommodations
has increased, while the demand has decreased for lower quality services. The income
of commercial accommodation was 112.3 billion HUF and the income of hospitality
was 53.6 billion HUF and the income originated from other sources was 48.5 billion
HUF in 2005. The total income was 214.4 billion HUF, from which the share of hotels
was the highest (187.1 billion HUF). Anyway, the share of other commercial
accommodations was only 27.3 billion HUF. Accommodations are used almost by
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visitors belonging to all types of tourism. Henceforth we mention only the status of the
riding tourism. The riding tourism includes all branches of „riding and coach driving
seasonably or systematically pursued by tourists in their spare time, for hobby, for
maintaining their own health on their own at their own costs” (Fehér and Kóródi 2007).
According to the publication prepared by Magyar Turizmus Zrt. and Hungarian
Tourists’ Association (2006a) the touristically qualified riding establishments and
services presented in Table 1. can be found in Hungary.
Riding services can be divided into 3 groups according to their frequency (Fehér and
Kóródi 2007):
• frequently, generally offered services (e.g.: riding, riding education, lease keeping,
riding tour, coach driving, camping, accommodation and catering)
• often, but not generally services offered (e.g.: cross-country riding, show jumping,
dressage, competition, presentations, hunting and riding)
• rarely offered services (e.g.: visiting horse-breeding establishments, military)
Within the riding tourism active and passive services can be distinguished. Active
services are e.g. riding, riding education, western riding, coach driving and other
equestrian sports. Passive services are mainly presentation, e.g. presentation of races,
equestrian archery, coach driving presentation, five-in-hand and equestrian acrobatics.
Hungarian riders’ characteristics, like horse-herders’ attractions and five-in-hand
mean a special attraction both for Hungarian and foreign tourists. Horse-herders used to
guard herds of horses in the open air all the year in the plain. Nowadays horses are
rarely kept in such a way. Horse-herders maintain the tradition concerning their tools
and clothing. They use special horse-accessories, like saddles without girths and shortstock
ornamental long whip. The „five-.in-hand”, „Koch-five-in-hand” or “Hungarian
post” are daring equestrian attractions: the horse-herder is driving five or more horses
standing on the back of the last two horses.
Several groups of interest, e.g. hobby riders and their companions, persons
pursuing and organising equestrian sports, horse-breeders, resp. participants of riders’
performances, riding tours and camping are touched by the riding tourism. Furthermore,
groups rendering services connected with equestrian sports, e.g. persons or enterprises
following trades connected with horses. People belonging to different groups of interest
can be basically squaded into two big groups (Incze and Hevessy 2007). The first
groups is formed by persons touched rarely or as complementary activity by riding
services. The second group includes persons being engaged with horses as main
activity.
Nowadays riding tourism information is easily accessible in Hungary. Actual
riding services are published by different types of media, such as periodicals, radio, TV,
advertisement newspapers and Internet. The Internet is probably the most tailor-made,
because it offers the most flexible searching possibilities.
Enterprises rendering different riding services are co-ordinated by the Hungarian
Non-Profit Association of Riding Tourism (MLTKSZ). Aptitude test of horses as well
as the management of the horseshoe qualifying system belongs to the tasks of the
MLTKSZ. The “horseshoe” indication promotes the orientation concerning the quality
of different services. The indication of the quality extends from one to five horseshoes
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depending on the character and level of the service (it is similar to the conventional star
qualifying system of hotels).
Hungary has generally good natural conditions for equestrian programmes (Incze
and Hevessy 2007). The riding tourism in the Northern Great Plain can not be easily
assessed on the basis of up-to-date data. There are 9 touristical regions in Hungary,
which are used by Magyar Turizmus Zrt in general in its inland communication. The
Regional Tourist Service Committees also operate in accordance with this system.
However, only 5 regions are used in the external (i.e. international) communication. As
a result of this cumulation, the Northern Great Plain, the Lake Tisza and the Southern
Great Plain are called commonly as Plain and Lake Tisza cumulated region (Fig. 1.). It
is a question, why some recent Hungarian publication has also only 5 regions?
Fig. 1.: Northern Great Plain and division of Hungary into 5 regions
(Source: www.itthon.hu).
The horse stock of the Northern Great Plain is nationally important. The number of
horses is 7 031 in county Hajdú-Bihar (Lóska 2001), 4 091 in county Jász- Nagykun-
Szolnok and 7 047 in county Szabolcs-Szatmár-Bereg. The total is 18 169 horses,
which is approximately 20 % of Hungarian horse stock.
Several riding tourism services are present in the Northern Great Plain: e.g. riding,
hunter riding, dressage, military, voltage, cross-country riding, coach driving,
competitions, riding education, visiting of horse-breeding establishments, equestrian
shows. Additionally, there is catering and accommodation. In some places holiday
check and credit card can be also used.
The riding tourism conditions of the Northern Great Plain are presented according
to the elements of SWOT analyses, i.e. Strengths, Weaknesses, Opportunities and
Threats (Lisztes 2007, Lóska 2001, Borsy 1998). The micro-relief conditions of the
Northern Great Plain are varied and so, both advantageous and disadvantageous
conditions for riding tourism can be found. Extending steppes, sandy terrain, landscape
having mosaic character and gently sloping terrain are advantageous; while the plain
character, the cracked structure soil are disadvantageous. The narrowing continuous
terrains and strictly protected areas may mean danger, i.e. decreasing facilities. Anyway,
the relief is suitable for a lot of equestrian programs and it makes possible to elaborate
further paths for riding and coach tours.
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Concerning the weather long autumn and spring, hard winters and very hot summers are
characteristic. The weather should be taken into account during the organization of the
programs, e.g.: in summer, equestrian programs can be organized in early hours of the
morning or late the evening. Regarding surface waters, the network of rivers is rich,
here should be mention especially the Tisza. The presence of water may also cause
problems, e.g. in the case of marshes and bogs, not to mention floods. The combination
of water and riding tourism or perhaps organisation of combined programmes may offer
very interesting possibilities. The region has a very rich fauna and flora, which may be
attractive for naturalist tourists. Near communities, cultivated lands are often
predominant; these, of course, do not attract visitors interested for natural curiosities,
but these places are easily accessible. The (riding)tourism may disturb the natural
environment but undesirable impacts can be avoided with an appropriate organization.
The Hortobágy National Park has a positive attitude towards riding tourism within
reasonable limits (Lisztes 2007, Varga 2007). The horse-breeding establishment of
Máta, being on the territory of the National Park at issue, plans to renew the stage and to
use more efficiently the roofed riding grounds. There are also weak points in the
collaboration of the national park and touristically qualified riding establishments, e.g.
defective information flow. The collaboration with the national parks could offer further
possibilities, e.g. riding system of nature protection guards.
Summarized, tourism plays an important role both in Hungarian and in world
economy. Concerning, riding tourism Hungary has advantageous conditions and within
the country the Northern Great Plain is especially suitable for this activity. Besides the
above mentioned favorable conditions and riding establishments, the riding tour path
“Puszta” is a special attraction, it is listed among the best of the world (Harpers
magazine). The horse stock, touristical riding establishments, as well as natural and
economic factors offer a good possibility for further developments.
METHODS
Secondary data are obtained from documentation, while primary data are obtained by
carrying out own estimations, analyses and a case-study. The subject of the case-study
is the Debrecen Riding Academy. In the case-study, the documentation was completed
by interviews and field research (from September 2006 until January 2007). We have
considered the case-study important because through the Debrecen Riding Academy we
could present notions, types and possibilities of services by means of actual and
concrete examples. Furthermore, our objective was also to present strengths and
weaknesses, opportunities and threats trough a concrete situation. The data concerning
the Riding Academy are the data from 2005 and 2006 (there was no significant
difference between these 2 years). Our analyses and comparisons have been based on
the data of Magyar Turizmus Zrt. and Hungarian Riding Tourism Non-Profit
Association, and they were carried out by means of the software Microsoft Excel. In
comparisons, the conditions of the Plain and Lake Tisza cumulated region was
compared with those of the Transdanubia, Balaton, Budapest and its surroundings, as
well as with Eger-Tokaj Mountains region. Furthermore, we have assessed the
relationship between the Northern and Southern Great Plain within the Plain and Lake
Tisza cumulated region.
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RESULTS AND CONCLUSIONS
There are no data in literature concerning the total income originating of riding tourism.
We have made an estimation about the incomes of riding establishments having also
accommodations. The total number of nights passed in Hungary by tourists was 19 334
750 in 2005. Taking into account the 4452 accommodations shown in Table 1., and the
fact that the majority of establishments having accommodations belongs to the category
of guest-houses (the average use of which was 22.7%), the number of nights passed by
tourists is 368 870, which is 1.9 % of the total number of nights passed by tourists. The
guest-houses achieved an income of 18.8 billion HUF in 2005 with their average use of
22.7 %. This means an income of 3.3 billion HUF, which is 1.54 % of the total national
income of 214.4 billion HUF (connected to commercial accommodations).
We have assessed the situation of the Plain and Lake Tisza cumulated region in
comparison with other Hungarian regions and then, within the Plain and Lake Tisza
cumulated region the role of the Northern Great Plain. Concerning the number of
touristically qualified riding establishments the Plain and Lake Tisza cumulated region
has the first position among the 5 regions (Table 2.). Among the 55 touristically
qualified riding establishments 20 are in the Northern Great Plain, which is 12.5 % of
the total number (158) of Hungarian establishments. In the Southern Great Plain there
much more, 34 establishments, which is 21.5% of the national total number. There is
also a difference between the Northern and Southern Great Plain concerning the
categories of higher quality and so there are also proportionally more 5- or 4-horseshoeestablishments
in the Southern Great Plain. At Lake Tisza there is only one touristically
qualified establishment.
Table 2.: Touristically qualified riding establishments
Number of
horseshoes
5 4 3 2 1 Other Total
Plain and Lake Tisza cumulated region 5 14 11 7 7 11 55
Transdanubia 7 7 8 8 8 5 43
Balaton 0 4 5 1 2 3 15
Budapest and its surroundings 2 3 8 9 3 5 30
Eger-Tokaj Mountains 1 1 0 4 7 2 15
Concerning the number of accommodations, the status of the Great Plain is shown in
Fig. 2. The Plain and Lake Tisza cumulated region has the most (1868)
accommodations. Within this the Northern Great Plain, there are 810, and in the
Southern Great Plain 1032 accommodations.
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2000
1800
1600
1400
1200
1000
800
600
400
200
0
1868
Plain and
Tisza lake
cumulated
region
Transdanubian
1434
Balaton
239
602
Budapest and
its
surroundings
Fig. 2.: Number of accommodations of establishments having accommodations
The most of roofed riding grounds are also in the Plain and Lake Tisza cumulated
region (Fig. 3.). 10 out of 17 roofed riding grounds are in the Northern Great Plain and
so this region even as an independent region has an important status in national
comparison concerning this aspect. The most of own horses are also in the Plain and
Lake Tisza cumulated region (Fig. 4.).
18
16
14
12
10
8
6
4
2
0
17
Plain and
Tisza lake
cumulated
region
Transdanubian
13
Balaton
1
6
Budapest and
its
surroundings
Eger-Tokaj
Mountains
Eger-Tokaj
Mountains
Fig. 3.: Number of roofed riding grounds in each region
187
309
3
7

2500
2000
1500
1000
500
0
2137
Plain and
Tisza lake
cumulated
region
Transdanubian
1908
Balaton
367
1288
Budapest and
its
surroundings
Fig. 4.: Number of own horses in touristically qualified riding establishments
Out of 2137 own horses 649 are in the area of the Northern Great Plain. In the Southern
Great Plain, there is about the double of this number (1743 horses). In terms of assessed
parameters, the most significant difference lies in the number of horses between the two
regions.
Concerning the number of services rendered by different establishments there is a
balanced competition among regions (Fig. 5.). The Plain and Lake Tisza cumulated
region is bigger (in terms of its territory) than the other regions. Besides good riding
conditions, the size can also influence the favorable position of the region in
comparisons with other regions.
13
12,5
12
11,5
11
10,5
10
12,7
Plain and
Tisza lake
cumulated
region
Transdanubian
12,5
Balaton
12,2
11,1
Budapest and
its
surroundings
Eger-Tokaj
Mountains
Eger-Tokaj
Mountains
Fig. 5.: Number of service types in 5 regions.
The data concerning the riding tourism in different types of literature and media mostly
correspond to the reality, i.e. essential elements are exacts, but details may contain
errors. For instance Lóska wrote the following things about the Debrecen Riding
Academy in 2001: „Qualification: 3-horseshoe. The establishment is operated with the
188
512
11,3
8

technical supervision of the Debrecen University, Center of Agricultural Sciences. The
academy is engaged in dressage and show jumping, there is also a roofed riding ground
and there are 25 horses, which are Nonius breeds and Hungarian Sport Horses having a
medium education.” There is a mistake in the description: in the institutions there have
never been and there are no Nonius breeds. Otherwise, the descriptions corresponds to
the reality of that time. The Riding Academy is still operated with the technical
supervision of the University of Debrecen. The “only” 3-horseshoe qualification can be
attributed to the only medium education of the horses and to the relatively new horse
stock in that time.
The horseshoe qualifying system assists the orientation concerning the quality of
riding establishments. Nowadays the Debrecen Riding Academy belongs already to the
4-horseshoe category, which is distinguished ranking (the horseshoe qualifying system
is from 1 to 5). Nowadays, the horse stock does not have a medium, but a good
education. The institution has not got the 5-horseshoe qualification, because it does not
have its own accommodation. To obtain the 4-horseshoe qualification the institution has
to correspond to lots of requirements, e.g.: environmental aspects, prescriptions of
horse-keeping, professional skill of horses, aspects related to riding services,
professional and personal factors. Furthermore, the qualification indicates that there are
many horses in the institution and the Riding Academy renders several types of service.
The number, breed, age and education of horses are very important concerning the
popularity of an institution. The institution is nowadays suitable for keeping 48 horses.
In winter there are 42 to 48 and in summer 38 to 40 horses. The difference can be
attributed to the attractive force of the roofed rising ground (20 x 60 m), i.e. this
establishment offers advantageous conditions for riding (independent from the weather).
In terms of breeds, there are Gidrans, Hungarian and German Sport Horses, Shagya and
Ponies. Ten horses are owned by the University of Debrecen, while the others are in
private ownership. Nowadays, the youngest horse is 4, the oldest horse is 15 years old.
The placement of the establishment, horse stock, service types/prices and last but
not least attendants are determining aspects concerning the status of the riding
establishment. The Riding Academy is in the town, but at the same time it is in a green
belt, this is a double advantage. The riding establishment has only 4 regular staff: 2
grooms and 2 instructors. Besides the regularly employed instructors further 1 to 6
instructors work periodically in the institution. The main 3 services of the institution are
education (2200 -2800 HUF per occasion), rental keeping (40-50000 HUF per month)
and establishment use (1200-2200 HUF per horse per occasion). The number of riding
lessons is 20 to 30 lessons a week, which relates to both horses and instructors; this
means full house. There is no precise calculation concerning the number of visitors; a
daily average can be estimated as follows: 36 riders, 30 attending persons and 20
visitors. On the basis of this estimation, can be said, that the Riding Academy has about
4000 to 5000 visitors per year; 2000 of them arrive from Debrecen (and its
surroundings), a further 2000 of them from the other parts of Hungary and about 300
come from other countries.
Nowadays, a riding establishment has several possibilities to advertise its services.
For instance, the Riding Academy advertises its services in newspapers, on the Internet,
in radio and TV. Since there is full house, no marketing development is planned.
189
9

However, material development is probable because in winter there is a higher demand
for rental keeping and establishment use than the institution could nowadays offer.
Besides the above mentioned development ideas, the elaboration of a “Riders’
path” offers a further, very interesting possibility in the Northern Great Plain. This path
would be based on the collaboration of riding establishments; during this path visitors
could get acquainted with the biological characteristics, different riding activities as
well as occupations connected with riding.
LITERATURE
Borsy Z. (editor.). 1998. Általános természet földrajz. (General natural geography)
Budapest: Nemzeti Tankönyvkiadó (National Education Publisher).
ENSZ (2002) World Economic Situation and Prospects 2002 (Sales No. E.02.II.C.2),
United Nations, New York 2002
I.Fehér and M Kóródi. 2007. A vidéki turizmus sajátosságai (Characteristics of the rural
tourism) . Agroinform. January 2007. (extraordinary issue).
R.Incze ands G.Hevessy 2007. Viszonyunk a lóhoz (Our relationship with the horse).
Agroinform. February 2007..
L. Lisztes 2007. Hortobágyi Nemzeti Park: az értéktől az élményig (Natural Park of
Hortobágy: from value to experience). Presentation: Debrecen, 22.01.2007.
J. Lóska 2001. Az Észak-magyarországi és Észak-alföldi régió lovasturizmusának
helyzete és kibontakozásának lehetőségei (Status and possibilities of development
concerning riding tourism of Northern Hungary and Northern Great Plain)
(Regionális lovas turisztikai termékfejlesztési tervtanulmány és programjavaslat
(Preliminary study and programme proposal for product development regarding
riding tourism). Vanyarc-Sarlóspuszta.
Magyar Turizmus Zrt. 2006a. Lóháton Magyarországon (On horseback in Hungary).
Magyar Turizmus Zrt. 2006b. Turizmus Magyarországon (Tourism in Hungary) 2005.
World Travel and Tourism Council (WTTC) (2004) Annual Report: Progress &
Priorities 2004/05
Online sources
www.itthon.hu
http://www.mth.gov.hu/
www.equi.hu
www.utikonyv.hu
http://lovas.lap.hu/
Interviews
Boros Sándor. Instructor, Debreceni Lovasakadémia. Személyes interjú (Riders’
instructor, Debrecen Riding Academy, personal interview): Debrecen, 2007.01.09.
László Lisztes. Hortobágyi Nemzeti Park. Személyes interjú (National Park of
Hortobágy, personal interview): Debrecen, 22.01.2007..
Varga Emese. Mátai Ménes. Telefonos interjú (Phone interview): 2007.01.23.
190
10

TISSUE CULTURE METHODS FOR SCREENING RESISTANCE TO
PECTOBACTERIUM CHRYSANTHEMI IN POTATO
Ildikó Hudák 1 , Mária Hevesi 2 , Judit Dobránszki 1 and Katalin Magyar-Tábori 1
1 Research Centre of University of Debrecen, Westsik V. u. 4-6, H-4400 Nyíregyháza,
2 Corvinus University of Budapest, Faculty of Horticultural Science, Villányi út 29-43.,
H-1118 Budapest, Hungary
ABSTRACT
Pectobacterium chrysanthemi (Erwinia chrysanthemi) cause vascular wilts, stunting,
soft rot, spotting of leaves or parenchymal necroses of a wide range of plant species and
cultivars like Allium cepa, Capsicum anuum, Nicotiana tabacum and Solanum
tuberosum. This bacterium affects potatoes in almost every region in the world. At the
moment the chemical protection is not solved (Dobránszki and Hevesi, 2002.). Altought
the sensitivity of produced cultivars are different, resistant cultivars are not available for
management of these diseases. The resistance in potato usually comes from wild
Solanum species. In our Research Centre we have bred potato for more decades. Our
breeding work based on using of the different wild Solanum species bearing resistance
or tolerance against Pectobacterium species (Hudák et al., 2002.). The aim of our
research is working out new in vitro methods for evaluation of the level of resistance or
tolerance against potato soft rot. We infected in vitro planlets. For evaluation of the
data, disease rating was used. By the help of disease rate we quantified the differences
among potato clones. On the basis of our results the in vitro methods may be suitable
for fast test of the breeding materials, clones or cultivars against Pectobacterium-strains.
Keywords: Pectobacterium chrysanthemi, soft rot, disease rate, in vitro methods
INTRODUCTION
Pectobacterium species causes plant diseases that include blights, cankers, die back,
leaf spots, wilts, discoloration of plant tissues and especially soft rots variously
described as stalk rot, crown rot, stem rot or fruit collapse (Brenner et al., 2005.).
Pectobacterium chrysanthemi (Erwinia chrysanthemi) is one of the phytopathogenic
enterobacteria, which causes soft rot disease in various plants. This bacterium affects
potatoes in almost every region in the temperate, semitropical and tropical zone of the
world. Its pathogenecity is due to its ability to secret several extracellular enzymes
including pectate lyases, cellulases and proteases. These extracellular enzymes attack
cell walls and membranes of plants leading to plant tissue maceration (Kelman, 1990.).
Because the field experiments would be dangerous and prolonged therefore an in vitro
method should be elaborated. We carried out with Pectobacterium-strain isolated in our
country. We adopted and compared different in vitro infection methods. We infected in
vitro planlets. We described the symptoms after in vitro infection.
MATERIALS AND METHODS
We used the bacterial strain isolated by Mária Hevesi in Hungary. In vitro cultured
shoots of potato Boró, Réka, Rachel, 136/92 and 34/85 were used as test plants. The
shoots were propagated on hormone-free MS medium. The cultures were illuminated by
191

white light of 105 μMol s -1 m -2 using 16/8 hour light/dark cycles. The temperature was
20-22 C 0 . Three-week-old in vitro potato plants were infected during the experiments.
Pectobacterium chrysanthemi was grown at 26 C 0 on Nutrient medium. The bacterial
suspension contained 10 8 CPU. The inoculation was made by a forceps immersed in the
suspension. On control treatment we wounded the shoots with a forceps immersed in
sterile distilled water.
The symptoms were evaluated after one week. For evaluation of the data disease
rating were used. The diseased plants were divided into five categories based on the
number of the wilted leaves. The disease categories were determined as follows:
� Symptom-free plantlets
� 1-25 % of leaves wilted
� 26-50 % of leaves wilted
� 51-75 % of leaves wilted
� 76-100 % of leaves wilted
With modification of this method a special disease rate (DR) was worked out:
Σ [(N1 x 1) + (N2 x 2) + (N3 x 3) + (N4 x 4) + (N5 x 5)]
DR = --------------------------------------------------------------------------
Σ N
N1-5: Number of plantlets in each disease category
N: All obtained plantlets
RESULTS AND DISCUSSION
The results of experiments are summarized:
During the experiments we chose the virulent strains of Pectobacterium chrysanthemy.
We worked out in vitro methods of infection. We described the symptoms after in
vitro infection. Disease rate was used for evaluation of the data. With the help of
disease rate we quantified the observable differences (symptoms). We separated three
sensitivity categories:
If 1 < DR < 2 non sensitive
If 2 < DR < 3 middle sensitive
If 3 < DR very sensitive
Symptoms: The symptoms were evaluated after in vitro infection on the seventh day. In
the control treatment the plantlets were healthy. No any leaf or stem symptoms could be
observable. The place of wounding could be seen but became dry (Figure 1). In the case
of clone 34/85 were not any leaf symptoms, at the worst case below or beyond
wounding stem symptoms could be observed. The place of wounding could be seen
(Figure 2). Wilt of the one-one leaf could be observable in the Réka and Rachel. The
place of the wounding could be seen clearly. Stem symptoms were typical. From the
wounding place, the stem rotted off and split in two (Figure 3). Both stem and leaf
192

symptoms could be observable in the Boró and clone 136/92. Seventy percent of leaves
of the plantlets rotted off and strong stem symptoms were observable on the seventh day
after in vitro infection (Figure 4).
Fig. 1. Symptoms of the control treatment after in vitro infection on the seventh day.
Fig. 2. Symptoms of the 34/85 clone after in vitro infection on the seventh day.
Fig. 3. Symptoms of the Rachel and Réka after in vitro infection on the seventh day.
193

Fig. 4. Symptoms of the Boró and 136/92 clone after in vitro infection
on the seventh day.
We carried out the infection of the in vitro plantlets and determined values of DR in
average of three, independent experiments in the case of five clones (Figure 5).
Values of disease rate against Pectobacterium chrysanthemi:
• 34/85 clone (DR = 1,2) non sensitive,
• Rachel (DR = 2,3) middle sensitive,
• Réka (DR = 2,5) middle sensitive,
• 136/92 clone (DR = 3,2) very sensitive,
• Boró (DR = 3,3) very sensitive.
3,5
3
2,5
2
1,5
1
0,5
0
1,2
34/85
Disease rate (DR)
2,3
Rachel
2,5
Réka
3,2
136/92
3,3
Boró
Fig. 5. Values of disease rate against Pectobacterium chrysanthemi
in 34/85, Rachel, Réka, Boró and 136/92 clones.
It could be concluded that the in vitro methods are suitable for testing sensitivity of the
clones and cultivars. In the future we have the intention to extend over these
194
DR

experiments the other clones and we want to compare our results with in vivo
experiments.
ACKNOWLEDGEMENTS
This work was supported by GVOP (project N 0 .: GVOP-3.1.1.-2004-05-0041/3.0).
REFERENCES
Brenner, D. J., Krieg, N. R. and Staley, J. T. 2005. Bergey’s manual of Systematic
Bacteriology, Part II. The protobacteria. P. 721-730.
Kelman, A. 1990. Blackleg, bacterial soft rot. In: W.J. Hooker (ed.): Compendium of
Potato Diseases, Part II. Disease in the Presence of Infectious Pathogens. p. 27-29.
Dobránszki, J. and Hevesi, M. 2002. A burgonya fekete szártőrothadása és
baktériumos lágyrothadása. Burgonyatermesztés. 2002. augusztus. p. 16-19.
Hudák, I., Dobránszki, J. and Hevesi, M. 2002. Hazánkban izolált, burgonyát
károsító Erwinia-fajok vizsgálata. "a Magyar Tudomány Napja 2002" alkalmából
rendezett Szabolcs-Szatmár-Bereg Megyei Tudományos Konferencia. 2002.
november 11.
195

EFFECT OF AUXIN ON CALLUS FORMING CAPACITY OF DIFFERENT
PEA GENOTYPES (PISUM SATIVUM L.) 1
ABSTRACT
Katalin Magyar-Tábori, Judit Iszály-Tóth, Judit Dobránszki,
Nóra Mendler-Drienyovszki
Research Centre of Debrecen University,
Nyíregyháza, Westsik V. street 4-6, H-4400 Hungary
Effect of different auxins (IBA, NAA and 2,4-D) were tested in two concentrations (2.0
and 4.0 mgl -1 ) on callus forming capacity of eight genotypes (Graphis, Baccara, 2107,
Hanka, Hunor, Janus, M49060 and Erbi). Leaflets and stem section without node were
used as explants and they were placed on G57 media supplemented with 3 % sucrose,
0.8 % agar-agar and 1.0 mgl -1 BA. Cultures were incubated in dark for 3 weeks at 26 ºC,
after then they were cultured on 16 h photoperiod for further 4 weeks. Callus forming
capacity was observed and scored weekly. Our results confirmed that there are
significant differences between genotypes: 2107 breeding line showed the greatest,
while cv. Janus had the least callus forming capacity. The best results were achieved by
IBA in both concentrations, although the effect of higher level of NAA was similar,
depending on genotypes. 2,4-D was not effective especially in high concentration.
Induction of callus was more effective on leaflets than on stem segments.
Keywords: Pisum sativum L., callus, auxin, in vitro, explant type
INTRODUCTION
The pea (Pisum sativum L.) is an important protein and vegetable crop. Improving of
drought tolerance of peas has been one of the major tasks in most breeding programmes
and in vitro methods can help the breeding work. Results of Ezhova et al. (1995)
suggest that mutations affecting developmental characters may be expressed in tissue
culture. In vitro callus culture can be used to model breeding for heterosis and to predict
productivity and heterosis in pea plants (Rybtsov et al., 1997). However, the application
of in vitro selection system can only be achieved by means of plant cell and tissue
culture where the induction of somaclonal variation frequently is an inherent feature
(Lehminger-Mertens & Jacobsen, 1993). Because of there is a very few information
about callus culture of peas and responses of genotypes to in vitro conditions can be
different (Dobránszki et al., 1999, Vandorne et al., 1995) it is necessary to work out the
callus induction method, which can be used for the genotypes included in our breeding
programme. Although in the case of tobacco auxin autotrophic callus could be obtained
(Csiszár et al, 2004) and there is hormone-autonomous callus for sugarbeet (Saunders &
Daub, 1984) in the case of pea the presence of auxin was necessary for callus induction
(Gantchev, 1997). In our experiments effect of auxins and explant type on callus
development were tested for eight pea genotypes to select the optimal callusing
treatment.
1 This work was supported by GVOP-3.1.1.-2004-05-0041/3.0) project.
196

MATERIALS AND METHODS
Shoot cultures of eight pea genotypes (Graphis, Baccara, 2107, Hanka, Hunor, Janus,
M49060 and Erbi) were maintained in vitro and explants for the experiments were
isolated from micropropagated plants. Two kinds of explants were used: fully
developed leaflets of the upper third of stem or stem segments without node. Isolation
was made in a sterile solution (0.15 mgl -1 citric acid and 0.1 mgl -1 ascorbic acid) to
prevent the withering of explants and the deleterious oxygenation process. Explants
were placed on G 57 media (salts and vitamins) (Gamborg et al., 1968) supplemented
with MS-Fe (Murashige & Skoog, 1962), 3.0 % sucrose, 0.8 % agar-agar and 1.0 mgl -1
benzyl-adenine (BA) as cytokinin. Naphthalane-acetic acid (NAA), indolebutyric acid
(IBA) and 2,4-dichlorophenoxyacetic acid (2,4-D) were applied as auxins in two
concentrations: 2.0 and 4.0 mgl -1 . Cultures were incubated in dark for 3 weeks at 26 ºC,
after then they were cultured on 16 h photoperiod for further 4 weeks. Callus forming
capacity was observed and scored weekly as follows:
1- there is no callus on the explant
2- some calluses on some end of explant
3- there are some calluses on both end of explant
4- strong callus forming but the surface of the explant can be seen
5- the callus overgrows the explant.
Statistical analyses were made by variance analyses followed by Tukey test, using by
SPSS 9.0 for Windows programme.
RESULTS AND DISCUSSION
In our experiments callus induction and development were obtained in all treatments
and for each genotype. The statistical analysis showed significant interactions between
genotypes, explant type and auxins. In the average of all treatments the IBA was the
best auxin, and the 2,4-D was the least efficient. Effect of NAA in lower concentration
(2.0 mgl -1 ) was similar to the effect of IBA, although by the end of fourth week the
callus forming capacity was significantly lower (table 1.).
Table 1. The main-effect of auxin on callus forming capacity (in the average of both
explants and genotypes).
Callus development on the end of
1 st 2 nd 3 rd 4 th
Auxin
treatments
mgl -1 week
NAA 2.0 1.69 d 2.52 d 3.40 cd 4.25 cd
NAA 4.0 1.59 c 2.43 c 3.34 c 4.21 c
2,4-D 2.0 1.51 b 2.22 b 2.81 b 3.39 b
2,4-D 4.0 1.34 a 2.01 a 2.63 a 3.21 a
IBA 2.0 1.64 d 2.52 d 3.47 d 4.36 e
IBA 4.0 1.64 d 2.54 d 3.47 d 4.32 de
Means within column signed by the same letter belong to the same homogeneous group.
197

The genotypes differentiated considering the degree of callus development already from
the end of first week. By the end of fourth week the 2107 breeding line showed the
strongest callus forming capacity, while cv. Janus was the weakest. Only cv. Hunor and
Erbi belongs to the same group, the others differ from each other significantly (table 2.).
Table 2. Effect of genotypes on callus induction and growth in the average of all
treatments.
Callus development on the end of
1 st 2 nd 3 rd 4 th
Genotypes
week
Graphis 2.33 g 3.21 f 4.08 g 4.42 f
Baccara 1.88 f 2.69 d 3.51 e 4.30 e
2107 1.85 f 2.84 e 3.76 f 4.65 g
Hanka 1.76 e 2.67 d 3.20 d 4.05 d
Hunor 1.00 a 1.72 a 2.58 a 3.60 b
Janus 1.32 d 1.85 b 2.51 a 2.97 a
M49060 1.23 c 2.12 c 2.87 c 3.77 c
Erbi 1.10 b 1.79 ab 2.78 b 3.63 b
Means within column signed by the same letter belong to the same homogeneous group.
Similarly to Gantotti and Kartha (1983), who could induce the callus cultures on each
part of plant, excluding the root tip, we also obtained callus culture on both type of
explants (table 3.). Because of significant interactions and since we found significant
differences between callus forming capacity of different type of explants, we continued
the statistical analysis as one-way ANOVA.
Table 3. Effect of explant type on callus forming capacity in the average of all
treatments.
Callus development on the end of
1 st 2 nd 3 rd 4 th
Explant type
week
Leaflets 1.55 a 2.49 a 3.35 b 4.21 b
Stem segments 1.58 a 2.25 a 3.00 a 3.67 a
Means within column signed by the same letter did not differ significantly
Callus development was better on the leaf explants so we analysed the effect of auxins
on callus development on leaf explants. Molnár (1993) found 2,4-D to be suitable for
callus induction, and we also could induce callus development by the use of 2,4-D,
although the degree of callus development was decreased by 2,4-D, especially in higher
concentrations, excluding cv. Hanka (table 4). Gantchev (1997) also tested different
concentrations of 2,4-D (2.0-32.0 mgl -1 ) on internodal stem segments of pea, and found
198

that callus induction and development were obtained in the presence of the lowest level
of 2,4-D.
Table 4. Effect of auxins on calli developed on leaf explants of different genotypes by
the end of fourth week.
Genotypes
Callus development on the media supplemented with
NAA 2,4-D IBA
2.0 4.0 2.0 4.0 2.0 4.0
mgl -1
Graphis 5.00 c 5.00 c 3.80 a 4.14 b 5.00 c 5.00 c
Baccara 5.00 c 5.00 d 4.00 b 3.73 a 4.88 d 4.38 c
2107 5.00 c 5.00 c 4.70 b 4.28 a 4.95 c 4.93 c
Hanka 5.00 c 5.00 c 4.07 b 4.00 b 3.73 a 3.75 a
Hunor 3.60 a 4.06 b 3.94 b 3.86 ab 4.37 c 4.97 d
Janus 4.14 c 3.83 c 2.74 b 1.80 a 4.06 c 4.04 c
M49060 3.50 a 4.00 b 3.37 a 3.51 a 4.00 b 4.03 b
Erbi 4.49 b 3.67 a 3.49 a 3.71 a 4.46 b 4.26 b
Means within row signed by the same letter belong to the same homogeneous group
Presence of NAA and IBA irrespectively of their concentrations was enough to induce
and maintain an intense callus development of leaf explants of cv. Graphis, 2107 and cv
Janus. In the case of cv. Baccara the best results were obtained on media containing IBA
at 2.0 mgl -1 concentration, or NAA in both hormone levels. IBA applied in 4.0 mgl -1
was the best treatment for cv. Hunor, while both levels of IBA were favourable for
M49060 and cv. Erbi. In the case of latter cultivars the effect of NAA was the same
advantageous in concentration of 2.0 mgl -1 or 4.0 mgl -1 for cv. Erbi and M49060,
respectively. Only the cv. Hanka was the genotype, which responded to IBA treatments
by the weakest callus development, while induction of callus development was the most
successful on media with NAA. Even if the 2,4-D is generally used for callus induction
in peas (Gantchev, 1997, Molnár, 1993, ) and in other species (Bayliss & Dunn, 1979,
Arora & Singh, 1978, Conger et al., 1982) we found the other auxins (NAA and IBA) to
be more effective to increase callus forming capacity of the majority of pea genotypes
tested in our experiments.
Our results suggested that callus formation can be induced on all genotypes and by use
of each auxins, although the genotypes required an optimal hormone content in medium
for intensive callus growth. Further research is underway in our laboratory to examine
the effects of different osmotic materials on the rate of callus growth.
REFERENCES
Arora, I.K., Singh, R.N. (1978): Growth hormones and in vitro callus formation of
papaya. Scientia Horticulturae, Vol. 8, Iss. 4, 357-361.
199

Bayliss, M. W., Dunn, S. D. M. (1979): Factors affecting callus formation from
embryos of barley (Hordeum vulgare). Plant Science Letters, Vol. 14, Iss. 4, 311-
316.
Conger, B.V., Hilenski, L.L., Lowe, K.W., Carabia, J.V. (1982): Influence of
different auxins at varying concentrations on callus inductions and growth from
embryo and leaf-explants in gramineae. Environmental and Experimental Botany,
Vol. 22., Iss. 1, 39-48.
Csiszár, J., Szabó, M., Erdei, L., Márton, L., Horváth, F., Tari, I. (2004): Auxin
autotrophic tobacco callus tissues resist oxidative stress: the importance of
glutathione S-transferase and glutathione peroxidase activities in auxin
heterotrophic and autotrophic calli. Journal of plant Physiology, Vol. 161, iss. 6,
691-699.
Dobránszki, J., Takács-Hudák, Á., Magyar-Tábori, K., Ferenczy, A. (1999): Effect
of medium on the callus-forming capacity of different potato genotypes. Acta
Agronomica Hungarica, 47 (1), pp. 59-61.
Ezhova, T. A., Bagrova A. M., Gostimski, S. A. (1995): Cell selection as a possible
reason for the specificity of somaclonal variation in pea. Plant Breeding 114 (6):
520-524, dec.
Gamborg, O. L., Miller, R. A., Ojima, K. (1968): Nutrient requirements of
suspension cultures of soybean root cells. Exp. Cell. Res. 50, 151-157.
Gantchev, L. S. (1997): Initial stages of callus development on internodal stem
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P.V., Yamada, Y. (eds): Handbook of Plant Cell Culture. Vol. 4. Techniques and
Applications. McMillan Publ. Co., New York, 370-418.
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protoplast of pea (Pisum sativum L.). Biotechnology in Agriculture and Forestry,
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Engineering III: Springer-Verlaug Berlin, Heidelberg.
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(Vicia faba L.) and peas (Pisum sativum L.) breeding. Acta Agronomica
Óváriensis Vol. 35, No. 1., 119-223.
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study heterosis in the pea. Genetika, 33 (11): 1517-1522, Nov.,
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pea (Pisum sativum L.) – effect of explant, genotype and culture conditions.
Annals of Applied Biology 126 (1): 169-179.
200

COMBINING ABILITY STUDIES IN A SEVEN- PARENTAL DIALLEL
CROSS OF DIFFERENT PEAS VARIETES
(PISUM SATIVUM L.)
Nóra Mendler-Drienyovszki, Mándi Lajosné, Katalin Magyar-Tábori,
Györgyi Csontos
Research Centre of University of Debrecen,
Westsik V. u. 4-6. H-4400. Nyíregyháza, Hungary
ABSTRACT
Results of a 7x7 diallel cross containing parents and F1 without reciprocals are
discussed after Model 1 and Method 2 of Griffing. Affecting three yielding capacity
traits were studied and analized: pod length, pod per plant, seed per pod.
The difference between genotypes in every traits were significant (P=0,01).
Predominance of additive effects in tested features were typical excluding pod pr plant.
The phenotypical performace is grater ofthe parents, which have higher GCA values.
In general the effects of SCA isn't correlate with performance of F1 generation.
INTRODUCTION
Peas have been grown as an important source of animal feed and human food for many
centuries. Over this time, pea has been selected for these uses. Several thousand
varieties exist throughout the world.
The majority part of our most successful pea varieties are more or less near
relationship with each other. The genetical background of high yield capacity can be the
same because of the crosses of the best pea cultivars seldom result in better new line
than the parents. However it can be supposed that the parents with good combining
ability can produced better lines than the average in a later generation. It's important in
the plan of different cross- combinations to choose the parents purposefully. In practice
the genotypes which hold a good phenotype values, not to be sure of inheritance their
desired traits, but another not so good varieties would be valuable like a parent in the
diallel cross.
The diallel analysis is a quantitative genetics method in plant breeding where we
can estimate the interaction between enviroment and genes in a population or the
offsprings from the selected paretnts (Mohay, 1986).
The diallel crosses are used in generally from mating modells, because we can get
the most completly genetic information about the genotypes.
The base methods are the analysis of Griffing (1956), Hayman and Jinks (1954), which
is based on diallel cross.
These diallel methods are popular all over the world, and it can be used
successfully in other species, for example maize (Daniel and Bajtai, 1975, Pepó et al.
1989), sunflower (Füredi and Frank, 1981), wheat (Matuz, 1976, Du et al. 1999), pea
(Csizmadia, 1985, Lőkös, 1985).
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MATHERIALS AND METHODS
Based on observations of several years we selected ten varieteies from our varieties
collection for diallel- parent. Seven varieteies got usable results (1:Léda, 2:Margó,
3:Foremos, 4:Presto, 5:Mariza, 6:Early sweet, 7:Debreceni korai velő) since the
crossing was not successful for all combination. We would like to produced a new early
pea varieteies with high yielding capacity parameters. We made the crosses in 2002, in
several repetitions depending the number of flowers. We sowed the parent and non-
reciprocal F1 generation beside net in 2003. We made the observations in more
qualitative and quantitative traits by instructions of UPOV. In this paper we analysed
the pod length, seed per pod, and pod number in one plan.
Statistical methods: The Model I. Method II of Griffing (1956) used for the combining
ability analysis of F1 data was:
xijk=μ+gi+gj+sij+rij+bk+eijk
where: μ= population means
gi= GCA effect for parent i,
gj= GCA effect for parent j,
sij= SCA effect for parents i and j,
rij= reciprocal effect
bk= block effect for block k,
eijk= error.
Diallel Analysis and Simulation Software by Burrow and Coors (1993) was used in this
study.
RESULTS
Phenotypic values
Seed per pod: the population average is 5,34 seed per pod. The parent number 3
(Foremos) has the most seed number (5,73), and the parent number 6 (Debreceni korai
velő) has the least (4,96). If we study these parent's combinations we can see that the
best combination is the 3x4. This combination has the highest seed number (6,45).
Combination of 4x6 has the least seed number (4,56).
Pod length: the population average is 5,35 cm. The Foremos has the longest pod, too
(5,70 cm) and the Margó ( parent number two) has the shortest pod length (4,96 cm).
We can get the best result for the 4x7 combination (6,25 cm) in our combination which
is better than the average. The worth combination is the 6x7 lines (4,20 cm).
The third trait which was analysed the pod per plant. The population average is 4,857
pods in one plant. Better then the average is the Léda (parent number 1) (5,50), worst
then the average is the Debreceni korai velő (4,27) (parent number 7).
To study the average of the combinations themselves have seen the most successfully
combination is the 1x6, the worst is the 2x4 combination (3 pods).
Effects of combining ability
The different btween genotypes in every traits were significant (P=0,01).
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Table 1: Mean squares of GCA and SCA in every traits
MS (GCA) MS (SCA)
Pod per plant 3,175 3,524
Pod length 1,817 0,852
Seed per pod 1,126 0,527
The GCA values are higher than one in every traits and larger than SCA values, expect
in the trait of pods per plant. The estimated GCA effects associated with each parent are
shown in Table 2.
Table 2: Generally combinig ability
Parents Trait
Pod per plant Pod length Seed per pod
1. 0,771 0,362 -0,001
2. -0,161 -0,489 0,021
3. -0,095 0,415 0,466
4. -0,228 0,027 0,099
5. 0,104 -0,148 -0,068
6. 0,304 -0,354 -0,464
7. -0,695 0,186 -0,052
se [g(i)] 0,260 0,124 0,157
se [g(i)-g(j)] 0,397 0,190 0,240
The values of GCA very similar to each other, because of selected parents were very
similar respecting of yielding- features (excluding Foremos). The values of pod length
of GCA are correlated with the higher valueses of seed per pod of GCA in Margó,
Foremos, Presto, Mariza. The estimated of SCA effects are given in Table 3.
Table 3: Specific combining ability effects
Combination Traits
Pod per plant Pod length Seed per pod
1x2 0,200 -0,700 -0,495
1x3 -0,533 -0,152 -0,546
1x4 0,600 0,439 0,319
1x5 -0,066 -0,068 0,151
1x6 1,066 0,714 0,423
1x7 -1,266 -0,232 0,148
2x3 1,400 0,546 -0,056
203

2x4 -1,466 -0,535 -0,249
2x5 0,200 0,003 -0,101
2x6 -1,000 0,393 0,270
2x7 0,666 0,292 0,632
3x4 -0,533 -0,514 0,535
3x5 -0,200 0,185 0,537
3x6 0,600 -0,255 -0,094
3x7 -0,733 0,190 -0,375
4x5 1,600 -0,183 -0,126
4x6 -0,600 0,109 -0,417
4x7 0,400 0,685 -0,062
5x6 -1,266 0,018 -0,149
5x7 -0,266 0,044 -0,310
6x7 1,200 -0,979 -0,032
se [s(i,j)] 0,512 0,246 0,311
se [s(i,j)-s(i,k)] 0,794 0,381 0,481
se [s(i,j)-s(k,l)] 0,687 0,330 0,417
In the case of SCA the results were the same: we could not find any very different
results of SCA in either combination. There are relatively higher values of SCA for pod
per plant in 2x3 and 4x5 combination, and for pod- length in 1x6 and 4x7, and for seed
number of pod in 2x7 and 3x5 combinations. The standard deviations are higher in
number of pod per plant.
DISCUSSION
The parents in the diallel were selected consciously. Our aim is to produce an earlier
variety with high yielding capacity thus these results just are the part of the whole
experiment, which can help us to breed the new varieteies.
Predominance of additive effects in tested features were typical excluding pod per
plant thus the features can be improved any further in the respective combinations.
These results are similar to other author's results. The reports on inheritance of yield-
related traits in garden pea are variable as apparent from gene action that was reported
to be additive (Rathore et al., 1995) and epistatic (Narsinghani et al., 1982) for pods per
plant, additive (Singh and Singh, 1991) and non additive (Srivastava and Singh, 1988)
for seeds per pod.
The studied traits, which has higher GCA effects-values has higher phenotypic
values. According to our results the parents of the best phenotypic values combination
are the parents with the best GCA values parents (Table 4.) The best variety was the
Foremos in the pod length and seed per pod traits, and these were accompained the
highest GCA values. In the combination with the best phenotypic values, the one parent
is the Foremos (3.). Since the Foremos an earlier variety with long pod length (8-10 cm)
and many seed per pod (8-9). These traits are just in a few combination have improving
effects. These improving effects aren't so large rate, that we can stabilize the trait in the
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combination. We could not obtained that result what was required from the variety.
Table 4: The best phenotypic combination
The combinations with the best The parents with the best
performance
GCA
Pod per plant 1x6 1
4x5 6
1x4 5
Pod length 4x7 3
1x4 1
3x7 7
Seed per pod 3x4 3
3x5 4
2x7 2
(only the three highest values are presented)
We can say same connection about the parents with worth values as mentioned above,
those parents and combinations got the least seed number and the shortest pod length.
To study the SCA values the results are various (Table 5.). In general the effects of
SCA isn't correlate with performance of F1 hybrid, except the seed per pod, where are
the three combination give the rank in the performance and SCA values. Singh et al.
(1991) also found connection between performances and values of SCA.
Table 5. The combinations with the best performace and SCA, only the three highest
values are presented in the table.
The combination with the
best performace
Pod per plant 1x6
4x5
1x4
Pod length 4x7
1x4
3x7
Seed per pod 3x4
3x5
2x7
The combination with the
best SCA
4x5
2x3
6x7
1x6
4x7
2x3
2x7
3x5
3x4
Further research works are underway to analise the inheritance of plant-heigh and 1000
seed-yield by the use of diallel programe.
In this year the F5 generation will be observed by instructions of UPOV. We attend to
205

compare the results of statistical analise to the field performance of breeding lines.
REFERENCES
Burrow, M. D.- J. G. Coors (1993): Diallel analysis and simulation. User's guide.
Dep. of Agron. , Univ. of Wisconsin, Madison.
C. G. Du- L. R. Nelson- M. E. McDaniel (1999): Diallel Analysis of Gene effects
Conditioning resistance to Stagnospora nodorum (Berk.) in Wheat. Crop. Science,
Vol. 39, May-June. 686-690.
Csizmadia L. (1989): A borsó egyes mennyiségi tulajdonságának öröklődése.
Zöldségtermesztési Kutató Intézet Bulletinje Kecskemét 22: 29-40.
Daniel L.- Bajtai I.: (1975): Néhány mennyiségi tulajdonság alakulása 11
csemegekukorica beltenyésztett törzs diallél keresztezésben. Növénytermelés 24:
285-294.
Füredi J.- Frank J. (1981): Napraforgó vonalak kombinálódó- képességének
vizsgálata és a kombinációk genetikai elemzése Griffing- módszerrel.
Növénytermelés 30: 289- 300.
Griffing, B. (1956.): Concept of general and specific combinig ability in relation to
diallel crossig systems. Australian Journal of biological Sciencis, Melbourne 9.
4:463-492.
Hayman, B. I. (1954): The theory and analysis of diallel crosses. Genetics 39: 789-
809.p.
Jinks, J. L. (1954): The analysis of continuous variatio in a diallel cross of Nicotiana
rustica varieties. Genetics 39.:767-788.p.
Matuz J. (1976): Heterózis és kombinálódóképesség vizsgálatok őszi búzafajtában.
Doktori értekezés. Gödöllő.
Mohay, J. (1986.): Diallélkeresztezési rendszer. Genetikai kislexikon. Natura Kiadó,
Budapest. 40.
Narsinghani, V. G.,- U.S.N. Rao- S.P. Singh. (1982): diallel cross analysis for
quantitative traits in mutant pea types. Indian J. Agric. Sci. 52:364:367.
Pepó P.- Pásztor K.- Palij A. F.- Pepó P. (1989): Kukorica mutánsvonalak genetikai
analízie. Növénytermelés 38: 193-199.
Rathore, P. K.- V. P. Gupta- P. Plaha (1995): Genetics of seed yield and some other
traits in pea using NCIII and triple cross analysis. Indian J. Pulse Res. 8: 119-122.
Singh, M.N.- Singh, R.B. (1991): Genetics of seed number and seed weight in pea.
Indian J. Pulse Res. 4: 165-167.
Srivastava, C.P.- R.B. Singh (1988): genetics analysis of seeds per pod in peas (Pisum
sativum L.) Veg. Sci. 15:38-48.
Tóthné Lőkös K.- Heszky L. (1994): Az általános kombinálódó- képesség, a
fenotípusos érték és az örökölhetőségi értékszám kapcsolata borsófajták diallél
kísérletében. Növénytermelés, 1994. Tom. 43. No.2: 101-107.
206

SUSTAINABLE LAND-USE BASED ON WESTSIK’S CROP ROTATION
EXPERIMENT
János Lazányi
University of Debrecen Centre for Agricultural Sciences
H-4032 Debrecen Böszörményi út 138.
SUMMARY
Sustainable agriculture and land use seeks solutions for environmental, economic and
agricultural problems at the same time. The aim is to develop new production methods
which provide protection for nature. In any sustainable agricultural system, the
management of organic matter and, more widely, management of natural resources are
based on the total self-sufficiency of the farm. The entire cycle of organic matter
production and decomposition takes place within the farm boundaries. The rate of
metabolism and the organic matter cycle are characteristic features of each farm unit
and define each unit’s activity for a long time.
The increased demand for food and the change in ecological requirements on
agricultural production mean that scientists must revise commonly used traditional
principles, in order to develop alternative crop production practices. In this respect, they
must strive to adopt technologies which serve to maintain soil fertility, and, at the same
time, meet environmental protection goals. Westsik’s crop rotation experiment is an
efficient tool which provides a thorough scientific basis for solving this problem.
Westsik’s experiment models various possible methods for managing nutrients (fallow,
straw, farmyard and green manure), and assists in seeking answers to questions of the
sustainability of agricultural production.
A study of sustainability of agricultural production and the connections between
agriculture, land use and environmental quality have been initiated in developed
countries, but the difficulties in less developed countries without adequate food supply
are even more serious. The aim of this study is to examine the potential for
environmentally-friendly land use in Hungary, and to draw attention to the importance
of long-term field experiments in the study of agronomic sustainability.
Key words: Sustainable agriculture, land use, crop rotation experiment
INTRODUCTION
Sustainable development is development that meets the needs of the present generation
without compromising the ability of future generations to meet their own needs.
Sustainable development wants to secure higher standards of living now and for future
generations. An important recommendation of the United Nations <strong>Conference</strong> on
Environment and Development held in Rio de Janeiro (1992), was that individual
countries should prepare strategies and action plans to implement their part of the
agreements. Agenda 21 also lays great emphasis on the need for all sectors of society to
participate in the formation of effective national strategies for sustainable development.
The problem of agriculture is as old as agriculture itself. The core of the problem has
always been soil erosion and loss of soil fertility. Today, when energy input into
agricultural production has increased, a new aspect has been added to this problem,
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further exacerbating the old one. Historical and recent experience with agriculture
suggests that human demand are not likely to be achieved without negative effects on
natural ecosystems, environmental quality and rural communities. Agriculture in
developing countries moves from traditional practices relying on natural fertility and the
regenerative power of the soil, to practices depending on high levels of inorganic
fertilisers, the intensive use of chemicals for pest control, and crops produced mainly
under monoculture.
The best known and most remarkable example of continuous production in
Hungary is the Westsik’s crop rotation experiment established in 1929, which is still in
use to study the effects of organic manure treatment, develop models and predict the
likely effects of different cropping systems on soil properties and crop yields. In this
respect, Westsik’s crop rotation experiment provides data of immediate value to farmers
concerning the applications of green, straw and farmyard manure. The experiment also
provides a resource of yield, plant and soil data sets for scientific research, whether into
plant and those soil processes which control soil fertility, or into the sustainability of
production. Maintenance of Westsik’s crop rotation experiment can also be used to
illustrate the value of long-term field experiments.
Sustainable agriculture is both a philosophy and a system of farming. It is rooted
in a set of values that reflect an awareness of social and ecological realities. It
emphasises management which works with natural processes to conserve all resources
and minimise waste, as well as environmental damage, while maintaining or improving
the profitability of production based on nutrient and water cycles, energy flows,
beneficial soil organisms and natural pest controls.
Sustainable systems also aim at ensuring the well being of rural communities, and
producing food which is nutritious and not contaminated with products which might be
harmful for the ecosystem. A sustainable farming system seeks to reduce or entirely
avoid the use of synthetic fertilisers, pesticides, growth regulators and other agricultural
chemicals. The system relies on crop rotation, crop residues, animal manure, legume
crops and green manure as organic matter, conservation tillage and non-chemical pest
control, to maintain the fertility of soil and control insects, weeds and diseases.
MATERIALS AND METHODS
The crop rotation experiment established by Vilmos Westsik in 1929 offers an excellent
possibility to study soil fertility management in many respects. The experiment,
consisting of 15 treatments, makes it possible to study sustainable land use under
different applications of green, straw and farmyard manure treatments, to study the
ecological impact as well as economic aspects of different production methods. One of
the main practical objects of the Westsik's crop rotation experiment was to measure the
long-term effects of different organic manure and inorganic fertilisers on rye and potato
production. In this way, the 15 treatments can logically be grouped as follows (i) control
treatments where no fertilisers were applied, (ii) straw manure treatments, (iii) farmyard
manure treatments, (iv) green manure treatments, (v) second crop green manure
treatments (Westsik 1951, 1965).
The F-1 block received no fertilisers and organic material treatment except the rye
and potato roots and straw incorporated into the soil. The fallow in this block was green,
and the plant material produced was ploughed into the soil. The F-2 block represents
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green manure treatment, where lupine was grown as a main crop and incorporated into
the soil 4-5 weeks after flowering. The phosphorus and potassium fertilisers in this
treatment were applied the previous autumn, before the lupine was sown. The F-3
represents lupine root manure treatment, where lupine was grown for grain and the total
organic material, except for the grain, was incorporated into the soil.
Blocks F-4 - F-7 represent straw manure treatments. In the F-4 block, rye straw
was applied as mulch. In blocks F-6 and F-7, straw manure was fermented without
nitrogen, and in block F-6, with nitrogen addition. The straw manure was incorporated
into the soil 4-6 weeks before the sowing of rye. F-8 is the only block with 4 main
crops, where lupine grew twice in 4 years; once as a main crop produced for grain and
once as a second crop produced after rye and before potato, for green manure. In the F-9
block, lupine was grown as a forage crop and harvested 2-3 weeks after flowering.
Blocks F-10 and F-11 represent farmyard manure treatments without and with
supplementary fertilisers, respectively. In block F-12, lupine is grown after a green
forage crop and sown in May. This block is also evaluated with farmyard manure
treatments to measure the comparative effects of the two treatments. Blocks F-13, F-14,
F-15 represent green manure treatments, where lupine is grown as a second crop after
rye and before potato. The F-15 block received no supplementary fertilisers. The
difference between blocks F-13 and F-14 can be found in the time of the incorporation
of green manure.
RESULTS AND DISCUSSION
Minimum and maximum yields have varied considerably during the 77 years of the
experiment. Analysing the average rye yields of different treatments, we can conclude
that the poorest results were achieved in 1963, when the average of the various crop
rotations was 1.23 ton/hectare. The highest yields were harvested in 1984, when the
average was 3.15 ton/hectare (Figure 1). The difference is 188 %. The average yield of
potato was the lowest in 1972, and the highest, 12 times as much, in 1991 (Figure 1).
From the two-plant species, rye proved to be the more balanced in productivity. Its yield
variations may be attributed primarily to climatic factors, while, in the case of potato,
the variations can be closely related to changes in varieties or in the quality of
propagation material. The first improvement in the average yields of potato occurred in
the early 1940's, when a growing demand arose for potatoes as an important foodstuff.
During and immediately after World War II, the average potato yields decreased in both
crop rotation experiments, as well as nationally. The next increase was achieved in the
second half of the 1950's, and was related to the development in potato research and in
seed potato production at the Research Institute in Nyíregyháza (Figure 2).
209

Rye Yield (t/ha)
Potato yield (t/ha)
5.5
5.0
4.5
4.0
3.5
3.0
2.5
2.0
1.5
1.0
0.5
0.0
50
45
40
35
30
25
20
15
10
5
0
1931
Maximum Minimum Mean
1935
1939
1943
1947
1951
1955
1959
1963
1967
1971
1975
1979
1983
1987
Figure 1: Rye yields in the crop rotation experiment
Maximum Minimum Mean
1991
1995
1999
2003
1931
1935
1939
1943
1947
1951
1955
1959
1963
1967
1971
1975
1979
1983
1987
1991
1995
1999
2003
Figure 2: Potato yield in the crop rotation experiment
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In Westsik's crop rotation experiments, the average yields of potato increased again in
the late 1980's and early 1990's. This success could be explained by the breeding of the
virus resistant Boro variety and its involvement in the experiments. Similar yield
increases could be noticed during the analysis of Broadbulk's experiments in
Rothamsted, where the yields of wheat were doubled within only a few years, by
changing the variety and applying pesticides (Johnston 1969, Johnston and Poulton,
1980, Garner and Dyke 1969). Comparing Figures 1 and 2, it is difficult to show any
correlation between the yield trends of rye and potato; however, if we express potato
yields as a function of the average rye yields, a very strong correlation can be revealed,
except for crop rotations F-2, F-3 and F-9. Average potato yields can be characterised
with the linear regression: y = 0.79 + 4.21x. The correlation coefficient is r 2 = 0.9198.
This means that there is no significant difference in the effect of various crop rotations
on the yields of rye and potato (Figure 3). Control treatments, where fertilisers were not
used in addition to the occasional organic matter management, are on the lower part of
the curve. The deviation of F-2, F-3 and F-9 treatments from the regression line means
that lupine main crop has different effects on the average yields of potato and rye,
independently of the fact whether lupine is grown for green fodder, green manure or
seed.
Potato yield (t/ha)
16
14
12
10
8
6
4
2
0
F-1
F-2
Potato Rye
F-3
F-4
F-5
F-6
F-7
F-8
F-9
F-10
F-11
F-12
F-13
F-14
F-15
Figure 3: Rye and potato yield in the crop rotation experiment
Vilmos Westsik, who explained it with the infection of Fusarium species, which are
able to decrease the yields of both lupine and potato considerably, described this
phenomenon earlier. Furthermore, we have to add that lupine was ploughed under
directly before rye, so it was this culture that benefited from the favourable effect of
211
3.0
2.5
2.0
1.5
1.0
0.5
0.0
Rye yield (t/ha)

organic matter mineralization. The effects of treatments on potato yield were also
expressed as a function of control treatments to reduce variation between years. In this
case, the mean value of F-1, F-7, F-10 and F-15 was calculated for each year and
studied as to how the increasing mean value effects the value of selected treatments.
Agriculture in developed countries has moved progressively from traditional
practices relying on natural fertility and the regenerative power of the soil, to practices
depending on high levels of inorganic fertilisers, the intensive use of chemicals for pest
control, and crops produced mainly under monoculture (Lockeretz 1988, Carter 1989).
According to Edwards (1987), alternative agriculture is a farming practice avoiding, or
at least minimising, the use of non-renewable production inputs, such as fertilisers and
pesticides. Lowrance at al. (1986) and Senanayake (1991) defined sustainable
agriculture as agriculture which does not deplete soil or people.
For many years, farmyard manure has been the key element in the maintenance of
soil fertility (Johnston 1987, Keeney 1989). Composted manure increased its value,
stabilised the nutrients and reduced the risk of environmental pollution. Composting
provides more stable humus, which improves soil aeration, water infiltration and water
holding capacity. The high temperature of the composting process destroys microorganisms,
the seeds of most weeds, and reduces water contents and total volume. In
sustainable agriculture, crop and livestock systems should maximise for mutual support.
Crop residue is an excellent feed for animals. Livestock manure can be composted and
returned to the fields as a source of plant nutrient. Various crop and livestock
components can be integrated to minimise the effects of weather-related adversities, as
they can be crucial for the survival of a farm.
In sustainable agriculture, where the use of synthetic chemicals is reduced or
eliminated, the action of soil micro-organisms becomes a major factor in nutrient
cycling and also plant growth (Lazányi 1996, 1997). Sustainable agricultural cropping
systems should reduce non-farm inputs to decrease environmental and health hazards
associated with the use of agricultural chemicals and, at the same time, offset rising
production costs and maintain soil fertility. The reduction of leguminous plants in the
cropping system results not only from the availability of fertilisers, but also from the use
of fossil energy as a source of power in agriculture. Because of these changes, farmers
are no longer compelled to use one part of their land for the production of forage and
grain for their draft animals. In Hungary many farms has eliminated all livestock from
their operations, moving to cash grain enterprises and making them entirely dependent
upon fertilisers and other chemicals. However, when nitrogen fertiliser is expensive or
not available, the producers depend on the nitrogen fixed by legumes to maintain the
nitrogen cycle and to improve the fertility of the soil. The quantity of nitrogen fixed
varies greatly, from zero to several hundred kilograms per hectare, according to the soil
type, structure, soil pH and nutrient content, temperature, water regimes and
management of the legumes.
There are other benefits from using legumes in the crop rotation, but they are often
disregarded because of the difficulties in quantifying them. The yields of cereals grown
after a leguminous crop or in crop rotation are greater than those grown in monoculture,
regardless of the amount of fertiliser applied. This response is often referred to as the
leguminous effect in crop rotation. As additional nitrogen does not eliminate this yield
difference, most of the response must be due to factors other than nitrogen availability.
212

Crop rotation breaks the weed and insect cycles that often predominate in continuous
cropping. Crop rotation also enhances soil structure and improves water regime. In this
way, leguminous crops have long-term benefits in crop rotation, resulting in enhanced
soil organic matter content, which not only improves nitrogen availability, but also
improves soil structure, by reducing soil erosion and cultivation costs.
High input agriculture is relatively new in Hungary. Crop rotation was introduced
in the last century, extensive use of chemical fertilisers and pesticides became important
only in the second half of the XX. century. Agricultural production has become efficient
per unit of human labour and per unit of land. However, when efficiency is measured
against other criteria - capital, fossil fuel, energy, economic or social equity - the results
are less clear. The crop rotation experiment established by Vilmos Westsik in 1929
offers an excellent possibility to study soil fertility management. The experiment,
consisting of 15 treatments, makes it possible to study sustainability of agricultural
production under different applications of green, straw and farmyard manure treatments,
to study the ecological impact as well as economic aspects of different production
methods.
REFERENCES
Carter, H. O. (1989): Agricultural sustainability: An overview and research
assessment. Calif. Agric. 16-17.
Edwards, C. A. (1987): The concept of integrated systems in lower input sustainable
agriculture. American Journal of Alternative Agriculture 2(4):148-152.
Garner, H. V., Dyke, G. V. (1969): The Broadbalk Wheat Experiment. Rothamsted
Exp. Station. Report for 1969, 2, 26-45.
Johnston, A. E. (1969): Plant nutrients in Broadbalk soils. Rothamsted Experimental
Station Report for 1968. Part 2, 93-112.
Johnston, A. E. (1987): Effects soil organic matter on yields of crops in long-term
experiments at Rothamsted and Woburn. INTECOL Bulletin 15, 9-16.
Johnston, A. E., Poulton, P. R. (1980): Effects of soil organic matter on cereal yields.
Rothamsted Experimental Station, Report for 1979, Part 1, 234.
Keeney, R. D. (1989): Towards a sustainable agriculture. Need for clarification of
concepts and terminology. American J. of Alt. Agr. 4: 101-106.
Lazányi, J. (1996): Results of Westsik's crop rotation experiment for sustainable
agriculture. Hungarian Agricultural Research. 1996. Vol. 5. No. 1. 4-8. p.
Lazányi, J. (1997): Linking conventional and sustainable agriculture. In Filep, Gy. (ed)
Land Use and Soil Management. Debrecen. 68-86 p.
Lockeretz, W. (1988): Open questions in sustainable agriculture, American Journal of
Alternative Agriculture. 3:174-181.
Lowrance, R., Hendrix, P. F., Odum, E. P. (1986): A hierarchical approach to
sustainable agriculture. American Journal of Alternative Agriculture 1. 169-173.
Senanayake, R. (1991): Sustainable Agriculture: Definitions and parameters for
measurement. Journal of Sustainable Agriculture. 1:7-28.
Westsik, V. (1951): Homoki vetésforgókkal végzett kísérletek eredményei.
Mezőgazdasági Kiadó, Budapest.
Westsik, V. (1965): Vetésforgó kísérletek homoktalajon. Akadémiai Kiadó, Budapest.
213

THE EFFECT OF LONG-TERM PHOSPHORUS FERTILIZATION ON THE
TRYPTOPHAN AND ZINC CONTENT OF FEED PEAS
Ágnes Elek- Zoltán Győri- Mária Borbély
University of Debrecen, Centre of Agricultural Sciences
Faculty of Agronomy Institute of Food Science, Quality Assurance and Microbiology
ABSTRACT
The quality of food is determined by the chemical composition. One of the most
important nutritional parameter is the protein, and also its amino-acid composition. Pea
samples from the control (0+0+0 kg/ha) and the 20 th treatment of the 1995 OTK research
were analysed from Putnok, Mosonmagyaróvár, Keszthely, Bicsérd, Hajdúböszörmény
and Iregszemcse research sites. We analysed the tryptophan content and its change as a
result of phosphorus and zinc fertilization of the pea samples. HPLC analysis was
performed at 110° C for 20 hours after alkaline hydrolysis. Our results show that high
rates of phosphorus fertilizers decreased the tryptophan content of peas.
Keywords: tryptophan, HPLC, zinc, amino acids, hydrolysis
INTRODUCTION
The quality of food is determined by the chemical composition. One of the most
important nutritional parameter is the protein, and also its amino-acid composition.
Amino-acid analysis is usually a good challenge for the analytical laboratories, even they
have the suitable instruments for the determination. Most of the amino-acids of proteins
in food and their raw materials can be analysed by acidic hydrolysis. Tryptophan has a
sensitive indult-group, therefore it degrades in acidic solution. For HPLC determination
we introduced a sample preparation with alkaline hydrolysis (Ba(OH)2). We analysed the
tryptophan content and change of samples from OTK long-term field trials as an effect of
phosphorus and zinc fertilization.
Correct determination of tryptophan in food and feed is very important, as this
amino acid is essential for monogastric animals. On the other hand, there are less data
about tryptophan content of feed, than for the other amino-acids. With full knowledge of
the recent research results, tryptohan seems to have even more important role in human
nutrition. Less tryptophan than it is necessary for human promotes the development of
nervous depression. Its role was proved also in migraine headache. It is well known that
consumption of feed with high protein content will decrease the tryptophan-level in the
brain, due to a competition among different amino acids, regarding their intake.
Triptrophan content determination requires special methods because of its sensitivity
in acidic conditions in the presence of oxygen. Measurement can be performed by ionexchange
chromatography (Moore and Stein, 1963; Dévényi 1971), photometry
(Matheson, 1974; Basha and Roberts, 1977; Votisky 1984).
The highest tryptophan yield was found in the case of short term hydrolysis (60 min),
performed with mercaptoethanesulfonicacid at high temperature. The most common
method is the HPLC determination following an alkaline hydrolysis of proteins with
Sodium- or Barium-hydroxide (Delhaye and Landry, 1992). This method was used in our
laboratory to measure the tryptophan content of some plant originated food raw materials.
214

The reliability of the method was checked with recovery experiments with L- Tryptophan
(Fluka, Germany), and by the analysis of reference sample with known tryptophan
content.
MATERIALS AND METHODS
Pea samples from the control (0+0+0 kg/ha) and the 20 th treatment of the 1995 OTK
research were analysed from Putnok, Mosonmagyaróvár, Keszthely, Bicsérd,
Hajdúböszörmény and Iregszemcse research sites. Crude protein content of the analysed
samples varied from 6 to 23 %, according to the research site.
Hydrolysis: For hydrolysis 8,40 g Ba(OH)2x8H2O and 10 ml distilled water was added to
the sample that contained some 10 mg Nitrogen in a pressure-resistant test tube with
Teflon pipe. The reaction was performed at 110° C for 20 hours. The mixture was
washed to a flask with 30 ml distilled water, and Barium was precipitated with 0,05 M
Phosphoric acid in the form of Barium phosphate. The pH of the filtrate was adjusted to
3,0 with 6N Hydrochloric-acid and diluted to 100 ml with distilled water : methanol
80:20 solvent mixture. The solution was filtered through 0,45 µm membrane filter
(Millipore, USA) before HPLC analysis.
Chromatography: High performance liquid chromatography was performed by a
Merck-Hitachi AS 4000A equipment. Gradient program was used for separation on C18
column (LiChorCart 125x4) with eluent A:0,01mol/l KH2PO4 (Reanal) /Metanol 95:5 and
B: Methanol (Merck). For detection Merck-Hitachi F1050 fluorescent detector was used
(ex: 280 nm, em: 356 nm).
RESULTS AND DISCUSSIONS
According to the literature, the protein content of feed peas is highly dependent on the
crop year and the cropping site (Figure 1). Data show that the protein content of the
control plots varied from 19 to 25 % (Mosonmagyaróvár and Iregszemcse). The
fertilization did not increase the protein content of pees in Putnok, at the same time, it
decreased in Keszthely. More (in Mosonmagyaróvár from 19.59 to 23.63%) or less (in
Hajdúböszörmény from 23.4 to 24.93%)increase was found in the other research sites.
215

P mg/kg
fehérje(%)
30
25
20
15
10
5
0
MO BI KE IR PU HB
kontroll
20.kezelés
Figure 1: The effect of fertilization on the protein content of peas (1995)
7000
6000
5000
4000
3000
2000
1000
0
MO KE PU IR
1 kezelés
20kezelés
Figure 2: The effect of fertilization on the phosphorus content of peas (1995)
216

The increased phosphorus rates increased the phosphorus content in peas in
Mosonmagyaróvár, Keszthely, Putnok, and decreased only in Putnok (Figure 2).
Zn mg/kg
70
60
50
40
30
20
10
0
MO KE PU IR
Figure 3: The effect of fertilization on the zinc content of peas (1995)
1 kezelés
20kezelés
As regards zinc content both the cropping sites and the effect of fertilization had
significant differences. The fertilization treatments decreased the zinc content. The
highest effect was found in Putnok, furthermore, the lowest decreasing effect of
fertilization on zinc was found in Mosonmagyaróvár (Figure 3).
We determined the tryptophan content of pea samples by hydrolysis, in alkaline
media for 20 hours at 110 ºC. We had to take into consideration the different crude
protein content of the samples during the destructuring in barium-hydroxide media, 8,40
g Ba(OH)2x8H2O and 10 ml water was added to 10 mg N content. The rate ranged
between 89-98%.
217

Trp (fehérje %-a)
1,2
1,0
0,8
0,6
0,4
0,2
0,0
Mosonmagyaróvár
Keszthely
Putnok
Iregszemcse
1. kezelés
20. kezelés
SzD 5%
SD5% between treatments MO: 0,310** SD5% between treatments KE: 0,186*
SD5% between treatments PU: 0,198** SD5% between treatments IR: 0,166**
Figure 4 The effect of fertilization on the tryptophan content of peas at different
research sites
NPK fertilization significantly decreased the tryptophan content of peas at the four
examined research sites. These results verify the need for testing the relationship between
phosphorus and zinc in new varieties/hybrids, on the one hand by analysing plant samples
from the early stage of the growing year, on the other hand by analysing the peas, thus we
can obtain information on the trypthophan content as well.
Our earlier results showed that increasing rates of NPK fertilizers decresed the
tryptophan content, as the P-Zn ion antagonism decreased the intake of Zn.
CONCLUSIONS
1. It was verified with experiments that the determination of tryptohan can be reliable
performed by alkaline hydrolysis.
2. Data from literature show that the tryptophan content of peas can be reliably
performed using the presented method.
3. Increasing phosphorus rates decreased the zinc intake due to the P-Zn antagonism
and led to decreased tryptophan content.
4. Our data was included in the database, and the introduced method can be applied to
analyse samples from field trials examining varieties and fertilization of leguminous
crops as well.
218

REFERENCES
Basha S.M, Roberts R.M (1977) :A simple colorimetric method for the determination of
tryptophan ,Anal. Biochem. 378-386.
Csapó,J. (2006) (szerk.) : Élelmiszer és takarmányfehérjék minősítése Mezőgazda
Kiadó, Budapest
Debreczeni, B., Debreczeni, B.-né (1994)(szerk): Trágyázási kutatások 1960-1990.
Akadémiai Kiadó, Budapest
Dévényi T., Gergely J.(1971) :Aminosavak, peptidek, fehérjék Budapest: Medicina
Könyvkiadó
Ágnes Elek, Mária Borbely, Zoltan Győri(2007): Tryptophan content of some food
raw materials Cereal Research Communications, in press.
Izsáki Zoltán: (2006): A N- és P-ellátottság hatása a kukoricaszem (Zea mays L.)
fehérjetartalmára és aminosav-összetételére - Növénytermelés Vol 55 Nos 3-4
Landry, J; Delhaye,S(1992) : Simplifed Procedure for the Determination of Tryptophan
in Food and Feedstuffs from Barytic Hydrolysis. J. Agric. Food Chem. 40, 776-779.
Matheson, N. A.(1974): The determination of tryptophan in purified proteins and in
feeding-stuffs. British Journal of Nutrition, 393-400.
Magyar Takarmánykódex FVM értesítő (2003).LIV évfolyam 11.sz. VIII. sz. melléklet
1711-1715.
MSZ ISO 6496:2001
MSZ 6830-4:1981
Stein W.H., Moore M(1963): Chromatographic determination of amino acids by the use
of automatic recording equipment. In: "Methods in Enzymology", Vol. 6., Ed.
Colowick S. P. and Kaplan N. O., Academic Press, New York, 1963.
Votisky,E. Colorimetria determination of tryptophan in corn-cob-mix(CCM) (In:
Progress in Tryptophan and Serotomin Research Ed. H.G. Schlossberger,
W.Kochen, B. Linzen and H. Steinhard,W. de Gruyter and Co. Berlin-New York,
115-117
219

THE IMPORTANCE OF CALCIUM AND MAGNESIUM SUPPLY
ON THE ACID BROWN FOREST SOILS IN THE NYIRSÉG REGION 1
Jakab Loch*, János Lazányi**, Péter Tamás Nagy* and Rita Kremper*
* Department of Agricultural Chemistry and Soil Science, University of Debrecen,
Centre of Agricultural Sciences, Faculty of Agronomy
** Department of Rural Development and land Use, University of Debrecen, Centre of
Agricultural Sciences, Faculty of Agroeconomics and Rural Development
ABSTRACT
According to soil tests made in long-term field experiments and to the yield data of nine
years it can be concluded that on a certain habitat triticale yield can be duplicate with
appropriate NPK and Ca, Mg supply. NPK fertilization and especially N treatments
acidify the soil, and increase the Ca and Mg leaching. For maximum yield it is
necessary to add both Ca and Mg systematically beyond the well balanced NPK
portions. Ca supplement can be well characterised by AL-Ca values.
Keywords: long-term field experiment, brown forest acid sandy sols, fertilization,
liming, AL-Ca and AL-Mg content of soils.
INTRODUCTION
Soils in Nyírség are extremely varied. One characteristic type of sandy soils is the
brown forest soil with thin clay layer, which is usually poor in calcium and magnesium.
The lack of calcium and magnesium depends on the extent of leaching. Calcium and
magnesium can be adsorbed relatively in a greater portion only in clay stripes lying in
deeper zones. Blownsands are poorer in calcium and magnesium even than this.
The fulfilment of ecological aspects and the realization of environmentally friendly
plant cultivation require the study of long-term effects of nutrition supply and the
foundation of sustainable production. A good possibility for this is provided by a longterm
experiment adjusted in 1962 by Láng István. The yield data of the long-term
experiment reported so far (Láng I. 1973, Kádár - Szemes 1994, Kádár – Németh –
Szemes 1999, Márton 2006) unambiguously prove that appropriate yield on brown
forest soil with thin clay layer which is strongly leached in the upper layer can be
achieved only with balancing the pH conditions and with a well balanced nutrient
supply.
Productivity of sandy soils is determined by the low humus content accompanied
with low nutrient content and by the weak water management characteristics. According
to the soil tests and yield analysis of MTA TAKI long-term experiments in Nyírlugos
we already drew attention to the importance of the certain nutrients addition (Loch et al,
2002, Loch 2003, Loch et al 2005), and to the necessity of calcium supplement. Soil
acidity hinders the effectiveness of nutrients, lack of calcium and magnesium formed as
a consequence of leaching can limit the yield.
1 Authors are grateful to co-workers of MTA-TAKI for making soil sampling possible
and for providing the yield data of the experiment.
220

MATERIALS AND METHODS
Sandy soils were sampled from the „Nyírlugos” long-term field experiment, which were
established in 1962. The “Nyírlugos” LTFE was set up on acidic brown forest soil with
alternating thin layers of clay substance „kovárvány” in the Nyírség region. The main
characteristics of the experimental soils are: pH(KCl) 4.5, humus 0.7%, CEC 5-10
mgeq/100 g in the ploughed layer. The topsoil was poor in all five macronutrients (N, P,
K, Ca, Mg) and the groundwater depth was 2-3 m (Kádár et al., 1999).
The “Nyírlugos” experiment is a long-term small-plot (50m 2 ) fertilization and
liming experiment. The experiment consists of 32 treatments, 4 replications altogether
128 plot experiments. The indicator plant is triticale since 1980.
Fertilization and liming treatments in the experiment were changed several times in
the last few decades but the levels are constant since 1980 (Table 1.).
Table 1: Treatments of LTFE in “Nyírlugos”, since 1980
Treatments (kg/ha/year)
Levels N P2O5 K2O CaCO3 MgCO3
Control - - - - -
1 50 60 60 250 140
2 100 120 120 500 280
3 150 180 180 1000 -
Soil samples were collected from 5 different places in each plot after harvest of triticale,
in August 1998 and 2001.
Collected soil samples are handed and prepared as follows: samples were dried and
drilled. Before grinding, samples were cleaned from plant remains and other possible
dirt, and pass a 2 -mm screen. Finally, samples were stored in plastic boxes in dry place
until the examination.
In our earlier studies, the effects of treatments were described by the 0.01M CaCl2
soluble nutrient contents measured from soils collected in 1998 (Nagy et al., 2002; Loch
et al 2005). In this study, the long term effect of treatments on the average yield (1996-
2004) of triticale (based on the data of Márton (2006)) was described by the pH and AL
soluble Ca and Mg values measured from soils collected in 1998 and 2001.
RESULTS AND DISCUSSIONS
1. Effects of treatments on the yield and soil characteristics:
Among N1, N2, N3 treatments only the N1=50kg/ha increased the yield significantly, N2
= 100kg/ha dose held on a level, while N3 =150kg/ha dose caused depression. We
referred (Nagy et al 2002) previously that in case of this treatment the amount of
inorganic N fraction increased. Ammonium-nitrate applied in excess acidifies the soil,
pH decreases, and Ca and Mg leaching into deeper level intensifies.
In N2P1, N2P2, N2P3 treatments the yield increases, phosphorous doses improve the
utilization of N, the acidifying effect of ammonium nitrate decreases, which is proved
by pH values also and leaching of Ca and Mg is also smaller.
221

Yields of N2K1, N2K2, N2K3 treatments are less than that of NP treatments, yield
increase presents only in case of the largest potassium dose. The N2P2K2 treatment
combinations verify that the supplement of the three elements is necessary, as the yield
increases significantly. Maximum yield can be achieved only by NPK treatments
completed with calcium carbonate. In these treatments soil pH, AL-Ca and AL-Mg
content increase (Figure 1.).
AL soluble Ca (mg/kg)
1200
1000
800
600
400
200
0
Kontroll
N1
AL-Ca
pH
Yield
N2
N3
N2P1
N2P2
N2P3
N2K1
N2K2
N2K3
N2P2K2
N2P2K2Ca2
N2P2K2Mg2
N2P2K2Ca2Mg2
Figure 1: Effect of treatments on triticale yield, the AL-Ca and pH values (0-20 cm)
2. Formation of triticale yield is in accordance with the soil parameters:
AL-Ca values in accordance with the pH values show that in treatments without liming
a great lack of calcium formed as a long-term effect of NPK treatments. Based on
Balogh (1988) investigation and our own experiences if AL-Ca value in soil is smaller
than 100-150mg/kg with lack of calcium has to be accounted, and pH values under 4.5
are also considered as very critical values. According to Iváncsics (1988) if AL-Ca
values in soil is smaller than 120mg/kg the Ca supply level is weak.
As the figures show, in the 0-20 cm soil layer AL-Ca level is under the critical
value in case of N treatments and in the NPK treatments combination.
222
t
7
6
5
4
3
2
1
0
Yield (t/ha)and pH (0.01M CaCl 2)

AL soluble Ca (mg/kg)
350
300
250
200
150
100
50
0
Kontroll
N1
AL-Ca
pH
Yield
N2
N3
N2P1
N2P2
N2P3
N2K1
N2K2
N2K3
N2P2K2
N2P2K2Ca2
N2P2K2Mg2
N2P2K2Ca2Mg2
Figure 2: Effect of treatments on triticale yield, the AL-Ca and pH values (20-40 cm)
AL soluble Ca (mg/kg)
200
180
160
140
120
100
80
60
40
20
0
AL-Ca
pH
Yield
Kontroll
N1
N2
N3
N2P1
N2P2
N2P3
N2K1
N2K2
N2K3
N2P2K2
N2P2K2Ca2
N2P2K2Mg2
N2P2K2Ca2Mg2
Figure3: Effect of treatments on triticale yield, the AL-Ca and pH values (40-60 cm)
223
6
5
Yield
4
(t/ha) and
3
pH
2
(0.01M
1CaCl
0
6
5
Yield
4
(t/ha) and
3
pH
2
(0.01M
1
CaCl
0
2)
2)

In 20-40 cm layer there is a definite differentiation (formed in the upper layer) caused
by the treatments, the difference in the upper layer decreases according to liming.
Finally in 40-60 cm soil layer beside the strong differentiation AL-Ca values of each
treatments are below 200mg/kg, and in case of certain treatments it is only 60 mg/kg.
The represented data unambiguously prove the necessity of calcium-carbonat addition.
AL-Mg values show similar differentiation to AL-Ca values. These indicate weak and
medium supply in treatments without Ca and Mg. Increase of magnesium content relate
to the fact that lime powder applied for liming contained magnesium also (Table 2).
Table 2: Effects of treatment on AL-Mg (mg/kg) values in different layers
Treatments
Depth (cm)
0-20 20-40 40-60 Átlag
Control 13.2 13.2 13.2 13.2
N1 7.6 8.3 9.4 8.4
N2 8.4 6.0 6.8 7.1
N3 5.3 8.8 10.1 8.1
N2P1 5.2 10.4 8.4 8.0
N2P2 6.9 10.6 10.4 9.3
N2P3 7.7 9.5 8.7 8.6
N2K1 6.4 5.6 13.0 8.3
N2K2 4.1 6.3 6.9 5.8
N2K3 4.5 7.7 7.1 6.5
N2P2K2 5.9 10.7 6.3 7.6
N2P2K2Ca2 8.8 9.1 9.2 9.0
N2P2K2Mg2 22.2 14.4 16.2 17.6
N2P2K2Ca2Mg2 13.9 15.4 24.6 18.0
CONCLUSION
Data of long-term experiment confirm that on one of the characteristic soil type in
Nyírség i.e. on acidic brown forest soil with thin clay layer triticale can be cultivated
effectively with an adequate nutrient management. The production can be duplicated
related to the untreated control. Nutrient management and adjusting of pH influence
fertility of soils and its maintenance to a great extent.
NPK fertilization using by itself do not make it possible to reach the maximum
yield. Acidic pH depending on the extent of leaching and the lack of calcium are
limiting factors for yield, therefore there is a need for ordinary liming and occasionally
magnesium addition.
One-sided N fertilization is especially to be avoided, while N dominance increase
soil acidifying and the leaching of Ca and Mg. Applying N, P, K fertilization together
adequately improve the utilization of N. Yields of NP and NPK treatments refer to the
importance of phosphorous and potassium fertilization on a given cropland.
AL-Ca values are good indicators of calcium lack. Data of the experiments confirm
the earlier experiences, values under 100mg/kg limit the yield, Ca and Mg are leached
224

gradually from soils containing 100-150 mg/kg AL-Ca therefore their addition is
necessary. The pH values under 4.5 can be considered as critical values.
Studies of deeper layers prove that the ordinary Ca and Mg supply has effect
primarily on surface layers.
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és magnézium visszapótlással. Kandidátusi értekezés. Karcag.
Iváncsics I. (1988): Kutatási eredmények a gyakorlatnak. Tápanyag-gazdálkodás.
Mezőgazdasági és Élelmezési Minisztérium, Agroinform Kiadó, Budapest 88-93.
Kádár I. – Szemes I. (1994): A nyírlugosi tartamkísérlet 30 éve. MTA – TAKI,
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Kádár I. – Németh T. – Szemes I. (1999): A nyírlugosi tartamkísérlet 1998 évi
eredményei. Kézirat, MTA-TAKI Budapest.
Láng I. (1973): Műtrágyázási tartamkísérletek homoktalajon. Akadémiai doktori
disszertáció. Budapest.
Loch J. (1999): A hazai talajok magnéziumellátottságának jellemzése és a
magnéziumtrágyázás. Akadémiai doktori értekezés. Debrecen.
Loch J. – Jászberényi I. – Nagy P.T. (2002): A kalcium- és magnéziumpótlás
jelentősége savanyú magnéziumhiányos nyírségi homoktalajon. In:
Tartamkísérletek, Tájtermesztés, Vidékfejlesztés Nemzetközi Konferencia (Szerk.:
Láng I. – Lazányi J. – Németh T.) I. kötet, 130-135. o.
Loch J. (2003): A magnéziumtrágyázás jelentősége a savanyú homoktalajok
termékenységének növelésében. In: Talajjavítás – talajvédelem. (Szerk.: Győri Z.
– Jávor A.) DE ATC 109-115.
Loch J. – Kiss Sz. – Vágó I. (2005): A 0,01 M kalcium-klorid talaj-kivonószer
alkalmazásának hazai tapasztalatai In: Fenntartható homoki gazdálkodás a
Nyírségben, (Szerk. Lazányi J.), Kiadó:Westsik V. Nyírségi Tájfejlesztési
Alapítvány. 137-155.
Márton L. (2006): a műtrágyázás, meszezés és a csapadék változékonyságának hatása
a tritikále termésére és terméskomponenseire. In: Loch J., Lazányi J. (ed.): A
tápanyag-gazdálkodást segítő talajvizsgálati módszerek alkalmazása a Nyírség
homoktalajain. Westsik Vilmos Nyírségi Tájfejlesztési Alapítvány. Nyíregyháza.
51-77.
Nagy P.T. – Jászberényi I. – Loch J. (2002): A trágyázás hatása a 0,01 M kalciumkloridban
oldható nitrogénformák mennyiségére a nyírlúgosi tartamkísérletben. In.:
Tartamkísérletek, Tájtermesztés, Vidékfejlesztés Nemzetközi Konferencia (Szerk.:
Láng I. – Lazányi J. – Németh T.) I. kötet, 143-148.
225

COMPARATIVE ANALYSIS OF CHEMICAL AND BIOLOGICAL SOIL
EXAMINATION TO DETERMINE THE PLANT AVAILABLE N CONTENT
OF SOIL IN THE NYÍRLUGOS LONG TERM FIELD EXPERIMENTS 1
Péter Tamás Nagy* – János Lazányi** – Jakab Loch*
* Department of Agricultural Chemistry and Soil Science, University of Debrecen,
Centre of Agricultural Sciences, Faculty of Agronomy
** Department of Rural Development and Land Use, University of Debrecen, Centre of
Agricultural Sciences, Faculty of Agroeconomics and Rural Development
ABSTRACT
Chemical and biological (incubation) analysis was carried out the acidic sandy soil of
“Nyírlugos” long term field experiment. For chemical analysis 0.01 M CaCl2 extractant
was used. From extractant mineral and organic nitrogen were determined.
For biological analyses a modified incubation technique was applied to determine the
easily soluble, above mentioned N fractions.
The chemical and biological analysis was carried out chosen samples from 1998
and 2001. From the results of chemical analysis it was evident that the amounts of
easily soluble N forms highly depend on the applied treatments. All treatments
increased the content of both mineral and organic N compared to the control. Moreover,
it was found that excess N doses (100-150 kg/ha) resulted significant yield-decrement.
Results of biological analysis were pointed out that logarithmic rate equation was
founded between the amount of mineralized N and time, during incubation. Our results
open the door to establish the potentially mineralizable N-contents (Npot) and the
constant of the mineralization (k) of soils of different treatments. It can be established
that the applied treatments has not remarkably effect on the values of Npot, but higher
constant rate of mineralization was achieved in the limed treatment group than the
treatments without lime. The lowest mineralization rate constant was established in the
control.
From the results it was evident that the used modified incubation method is a
reliable tool to plan and implement of professional N supplement.
Keywords: easily soluble N forms; incubation; long term field experiment;
INTRODUCTION
The balance between supplying sufficient nitrogen for optimal plant performance and
minimizing pollution from excessive nitrogen application is delicate and very important.
It is widely know that N misuse can lead to pollution of the environment but it effects
on soil productivity and yield is well-known. The availability of soil N to the crop is
therefore of great importance in agricultural systems.
The determination of actual N-supply are very important and obligate for sustainable
1 Authors are grateful to coo-workers of MTA-TAKI for making soil sampling possible
and for providing the yield data of the experiment.
226

agriculture in Hungary, especially sandy soils, which are very sensitive to
environmental effects and inefficient human treatment (Németh (1996)).
The formation and fate of soluble N-forms and its response to organic matter
mineralization is not obvious yet and often inconsistent despite the intensive research
(Lazányi et al. (2002)).
The aim of this paper is to present data on different easily mineralizable N-forms
of a long term field experiment (LTFE) by incubation and chemical method in the
Nyírség region, in Hungary.
MATERIALS AND METHODS
1. Soils and experimental designs
Sandy soils were sampled from the „Nyírlugos” long-term field experiment, which were
established in 1962. The “Nyírlugos” LTFE was set up on acidic brown forest soil with
alternating thin layers of clay substance „kovárvány” in the Nyírség region. The main
characteristics of the experimental soils are: pH(KCl) 4.5, humus 0.7%, CEC 5-10
mgeq/100 g in the ploughed layer. The topsoil was poor in all five macronutrients (N, P,
K, Ca, Mg) and the groundwater depth was 2-3 m (Kádár et al., 1999).
The “Nyírlugos” experiment is a long-term small-plot (50m 2 ) fertilisation and liming
experiment. The experiment consists of 32 treatments, 4 replications altogether 128 plot
experiments. The indicator plant is triticale since 1980.
Fertilisation and liming treatments in the experiment were changed several times in the
last few decades but the levels are constant since 1980 (Table 1.).
Table 1: Treatments of LTFE in “Nyírlugos”, since 1980
Treatments (kg/ha/year)
Levels N P2O5 K2O CaCO3 MgCO3
Control - - - - -
1 50 60 60 250 140
2 100 120 120 500 280
3 150 180 180 1000 -
Only ten (one-sided N- and N-combined) treatments are summarized in this paper.
Chosen treatments can be divided four groups according to applied nutrients and lime
(Table 2.) Soil samples were collected from 5 different places in each plot after harvest
of triticale, in August 1998 and 2001.
Table 2: Chosen treatments and their variations of “Nyírlugos”
Control group N group NPK group Limed group
N1 N2P2 N2P2 K2Ca2
Controls N2 N2K2 N2P2 K2 Mg2
N3 N2P2K2 N2P2 K2 Ca2 Mg2
227

2. Sample preparation
Collected soil samples are handed and prepared as follows: samples were dried and
drilled. Before grinding, samples were cleaned from plant remains and other possible
dirt, and pass a 2 -mm screen. Finally, samples were stored in plastic boxes in dry place
until the examination according to Hungarian guideline (MSZ 20135:1999).
3. Procedure of biological and chemical soil analysis
A modified periodical incubation experiment is carried out in the laboratory to
determine the actual rate of N-mineralization (Stanford and Smith, 1972; Filep and
Tóthné, 1980; Filep and Ferencz, 1999). We improved conditions of this modified
incubation apparatus.
The enlarged amount of soil (without quartz sand) is supported in a leaching glass tube.
A thin glass wool pad is placed above and below the soil sample to avoid soil dispersion
when the leaching solution is poured into the tube. It is leached free of mineral N and
then incubated at optimum temperature (35°C) and moisture (WC=75%) for 16 weeks.
100 ml of 0.01 M CaCl2 solution is used for each N removing operation. So this way the
mineralized and easily soluble organic N of soil was removed periodically. Vacuum is
applied only to remove excess solution.
The leachates of incubation and the chemical extractions are analysed for mineralized N
(sum of nitrate and ammonium nitrogen) and organic N according to the method
described by Houba et al., (1986). The different N forms were analysed by autoanalyser
(SKALAR Segment Flow Analyser - CFA system).
The main advantage of this method that both the 0.01 M CaCl2 soluble inorganic Nform
and the UV digestable organic-N form of soil are determined simultaneously
(Nagy, 2004).
We used N-free nutrient solution containing 0.007 M CaSO4∗2H2O; 0.002 M MgSO4;
0.01 M Ca(H2PO4)2 and 0.015 M K2SO4 for use in replacing nutrients. The leaching
process is repeated after 2., 3., 5., 7., 9., 12., and 16 weeks.
The incubation can be terminated when cumulative N mineralization approaches a
plateau.
RESULTS AND DISCUSSIONS
The incubation was applied for chosen samples 1998 and 2001 as mentioned above. The
Nmin and Norg content of soil was measured along the incubation. The pH, the cumulated
values of Nmin and Norg and the results of chemical extraction are shown in Table 3.
1. Results of chemical examination
The pH of soil of experiment was strongly acid as earlier results (Kádár et al., 1999) and
the result of control showed.
Mostly the one sided N and NP, NK, NPK treatments had decreased the pH, due to
the acidic effect of applied fertilizers and their long term using. Note, that only the
liming treatments could hinder the further acidification and increased the pH.
The amount of 0.01 M CaCl2 soluble mineral N was between 2.55 and 9.63 mg/kg
228

according to the treatments. Obtained values correspond to the soil type and ruling soil
conditions. Results were pointed out that in these circumstances the mineralization
processes were hindered.
The amount of 0.01 M CaCl2 soluble organic N was between 4.08 and 7.25 mg/kg
according to the treatments. This fraction has changed lesser than the above mentioned
mineral fraction according to the treatments.
The obtained results of 0.01 M CaCl2 soluble N forms can be explained the effects
of applied dosage of fertilizers. The easily soluble N content of soil (Nmin + Norg) was the
lowest in the control and the highest in the N2P2K2Ca2Mg2 treatment. Increasing dose of
N has a negative effect on pH, slightly changed the amount of Norg and significantly
increased the amount of Nmin, especially N3 treatment (Table 3).
Table 3: Results of chemical and biological analysis (“Nyírlugos” 1998-2001)
Chemical method Biological method
pH Nmin Norg Nmin + Norg Nmin Norg Nmin + Norg
Treatments (mg/kg)
Control 4.31 3,20 4,08 7,28 180.72 47.07 227.79
N1 4.22 4,18 4,15 8,33 175.84 54.81 230.65
N2 4.04 3,40 4,98 8,38 157.59 42.77 200.36
N3 3.74 9,63 4,55 14,18 176.37 55.60 231.97
N2P2 4.07 3,31 4,53 7,84 172.34 52.75 225.08
N2K2 3.93 8,05 6,43 14,48 156.91 48.23 205.14
N2P2K2 4.05 2,55 4,75 7,30 157.41 45.81 203.22
N2P2K2Ca2 4.40 6,15 4,83 10,98 190.67 49.24 239.91
N2P2K2Mg2 4.78 3,75 5,63 9,38 190.47 45.38 235.85
N2P2K2Ca2Mg2 5.09 9,10 7,25 16,35 211.29 39.02 250.30
Average 4.26 5.33 5.12 10.45 177.99 49.07 225.03
LSD5% 0.25 1.66 0.64 2.09 10.82 3.26 16.81
The treatments of NPK group resulted an adjusted balance between mineral and organic
N forms and slightly decreased the soil pH compared to the control.
The treatments of limed group increased the pH and the Nmin and Norg content of soil
compared to the control. The amount of sum of N forms was about approximately
twofold in these treatments than the control.
2. RESULTS OF INCUBATION
From the cumulated data of incubation the following results can be establish:
Only the treatments of limed group have increased the Nmin content of soil compared to
the control (Table 3). One-sided N-treatments decreased the content of mineralized N
forms of soil compared to the control during the incubation. It can be explained with the
acidification effects of one-sided N fertilization which has caused unfavourable
conditions for mineralization processes.
Poorer mineralization conditions were caused applying treatments of NPK group as
229

well. The effect of these treatments can be interpreted by the decreasing soil pH. While
improving soil pH (limed group) the conditions of mineralization would be better and
the amount of Nmin was higher than the control (Table 3).
The cumulated Norg was varied between 39.02 and 55.60 mg/kg according to
treatments. The lowest value was obtained in N2P2K2Ca2Mg2 treatment. It can be
explained by the better circumstances of mineralization. Results of applying one sided
N treatments pointed out that the rate of mineralization was lower compared to the
control and especially the limed treatments. Furthermore, it was found that the
treatments of NPK group had not effect on the amount of Norg.
The amount of sum of N forms was the highest in the case of treatments of limed group.
Treatments of NPK group have a negative effect on the amount of sum of N forms
compared to the control. It can be established that increasingly applied N not improved
the rate of mineralization remarkably.
Furthermore, the 0.01 M CaCl2 soluble N-fractions are in mainly inorganic forms
under incubation period but the content of organic forms also significant (Table 3.).
The relationship between the Nmin and time could be studied from the periodical data of
incubation (Table 4 and Figure 1).
According to results typical relationship was established N mineralization vs. time
(Figure 1). In spite of the different treatments the shapes of the obtained curves were
analogous. Logarithmic rate equation was founded in the case of all treatments.
Difference was established between coefficients of correlations and intercepts (Table
4.). Differences were explained the above mentioned effects of treatments. Liming
increased, one-sided N treatments decreased the amount of Nmin.
Cumulated Nmin (mg/kg
250
200
150
100
50
0
0 4 8 12 16
Time (weeks)
Figure 1: Relationship Nmin vs. Time
230
Control
N1
N2
N3
NPK
Limed

Table 4: Results of regression analysis
Treatments Regression equation R value
Control y = 11,83Ln(x) + 105,02 0.7652
N1 y = 14,097Ln(x) + 93,719 0.7971
N2 y = 12,989Ln(x) + 89,133 0.8198
N3 y = 11,83Ln(x) + 105,02 0.7652
NPK group y = 13,019Ln(x) + 88,636 0.8043
Limed group y = 15,419Ln(x) + 111,97 0.8153
In the 1970s Stanford and co-workers (Stanford and Smith (1972), Stanford (1973),
Stanford et al. (1974)) advanced the concept of potentially mineralizable N (Npot) and a
related mineralization rate constant (k) for use in characterizing soil-available N. Since
then this concept has been used, modified, and discussed in great detail by numerous
scientist, but they theory has been accepted generally.
This theory assumed that organic nitrogen mineralization at optimum temperature and
moisture followed first-order kinetics. Accept the above mentioned theory our results
have made possibility to estimate the potentially mineralizable N of the soil and its rate
constant. Our results have confirmed that the correlation between 1/Nt and 1/t is linear.
The potentially mineralizable N-contents (Npot) and the constant of the mineralization
(k) were calculated according to Filep and Tóthné (1980) (Table 5.).
It can be established that the applied treatments has not remarkably effect on the values
of Npot, but higher constant rate of mineralization was achieved in the limed treatments
than the treatments without lime. The lowest mineralization rate constant was
established in the control.
Table 5: Calculated Npot and k values
Treatments Npot (mg/kg) k (35ºC)
Control 277.799 0.056
N1 233.247 0.057
N2 233.451 0.058
N3 234.028 0.058
NPK group 233.551 0.058
Limed group 249.885 0.064
3. Interpretation of results of chemical and biological analysis to yield
Obtained results of biological analysis can be compared to the yield (Figure 2.).
The constant of the mineralization was the lowest in the control. Increasing dose of onesided
N slightly increased the k value and decreased the yield, especially N2 and N3.
This effect can be explained by the increasing acidization of soil and the increasing
content of easily soluble N fractions in these treatments. The higher k was obtained in
the case of the limed group. It can be explained the above mentioned more favourable
soil conditions. The result of NPK treatment-group was better than the results of N
treatments and control but worse compared to the limed group.
231

Figure 2: Relationship between the yield and the constant of the mineralization (k)
From obtained values of Npot it was evident that liming improves the conditions of
mineralization of soil. It can be explained by higher microbiological activity and higher
pH of the soil (Kátai et. al. (1999)). Furthermore the mineralization potential of soil is
depended not only the actual mineralized N content and pH of soil but the amount of
easily soluble, low molecular weighted and mobilizable organic N fraction as well.
From the results it was evident that the used modified incubation method is a reliable
tool plan and implement of professional N supplement.
Results of chemical analysis can be compared to the yield as well (Figure 3.).
N (mg/kg)
k value
12
10
8
6
4
2
0
0,066
0,064
0,062
0,06
0,058
0,056
0,054
0,052
Control N1 N2 N3 NPK Limed
Nmin
Norg
Yield
k
yield
Control N1 N2 N3 NPK Limed
Figure 3: Relationship between the yield and the 0.01 M CaCl2 soluble mineral and
organic N
232
7
6
5
4
3
2
1
0
7
6
5
4
3
2
1
0
Yield (t/ha)
Yield (t/ha)

From obtained data, it can be established that the content of 0.01 M CaCl2 soluble Nmin
and Norg fractions were the lowest in the control. One-sided N treatments mostly
increased the amount of mineral N forms, but excess doses of N (>50 kg/ha) resulted
significant yield-decrement. Moreover, 150 kg/ha N dose (N3) highly increased the Nmin
in the upper layer. This excess amount of mineral N (mostly nitrate) can be dangerous
for the quality of soil water. Liming treatments produced higher yields without excess
content of nitrate.
REFERENCES
Filep Gy. - Tóthné Bíró Á. (1980): Hazai talajok mineralizálható N-készletének és Nszolgáltatásának
mérése és számítása. Agrokémia és Talajtan. 29. 229-244. (in
Hungarian)
Filep Gy. - Ferencz G. (1999): A talaj N-szolgáltató képességének becslésére használt
néhány számítási módszer értékelése. DATE Tudományos Közleményei. Tom.
XXXIV. 73-82. (in Hungarian)
Houba, V. J. G. - Novozamsky, I. - Huijbregts, A. W. M. - Van Der Lee J. J.
(1986): Comparison of soil extrctions by 0,01 CaCl2 by EUF and by some
conventional extraction procedures. Plant and Soil 96. 433-437. p
Kádár I. - Németh T. - Szemes I. (1999): Triticale trágyareakciója a nyírlugosi
tartamkísérletben. Növénytermelés, Tom. 48. No. 6. 647-661. (in Hungarian)
Kátai J. – LazányI J. – Veres E. (1999): Talajmikrobiológiai vizsgálatok a Westsik
vetésforgó tartamkísérletben. DATE Tiszántúli mezőgazdasági tudományos napok.
Konferencia kiadvány (szerk.: Loch J. – Vágó I. – Jávor A.): 175-184. o. (in
Hungarian)
Lazányi J. – Loch J. – Jászberényi I. (2002): Analysis of 0.01 M CaCl2 soluble
organic nitrogen in the treatments of Westsik’s crop rotation experiment.
Agrokémia és Talajtan. 51:79-88. o.
MSZ 20135:1999: Determination of the soluble nutrient element content of the soil.
Hungarian Standards Institution. Budapest (in Hungarian)
Nagy P. T. (2004):
Németh T. (1996): Talajaink szervesanyag-tartalma és nitrogénforgalma. MTA
Talajtani és Agrokémiai kutató Intézete, Budapest (in Hungarian)
Stanford G. - Smith S. J. (1972): Nitrogen mineralization potentials of soils. Soil Sci.
Soc. Am. Proc. 36:465-472. In. Page A. L. - Miller R. H. - Keeney D. R. (1982):
Methods of soil analysis. Part 2, Agronomy No. 9 (2) p. 711-733. Madison,
Wincosin
Stanford G. (1973): Rationale for optimum nitrogen fertilization in corn production. J.
Environ. Qual. 2:159-166. In. Page A. L. - Miller R. H. - Keeney D. R. (1982):
Methods of soil analysis. Part 2, Agronomy No. 9 (2) p. 711-733. Madison,
Wincosin
Stanford G. - Carter J. N. - Smith S. J. (1974): Estimates of potentially mineralizable
soil nitrogen based on short-term incubations. Soil Sci. Soc. Am. Proc. 38:99-102.
In. Page A. L. - Miller R. H. - Keeney D. R. (1982): Methods of soil analysis. Part
2, Agronomy No. 9 (2) p. 711-733. Madison, Wincosin USA
233

EFFECT OF FERTILIZATION SYSTEMS AND LIMING ON MOBILE
PHOSPHORUS CONTENT IN DIFFERENT SOIL TYPES
ABSTRACT
Augusta Olivia Lujerdean
University of Agriculture Science and Veterinary Medicine Cluj-Napoca,
Faculty of Animal Breeding and Biotechnology
3-5 Calea Manastur Street 400732, Romania
Liming and application on acidic soils of inorganic fertilizers or organic manure had the
considerable influence on the mobile phosphorus content and pH of soil. P(AL)
determinations suggest that differences in the soil physical-chemical properties and the
forming which P is applied influences the effectiveness of fertilization and P balance.
The phosphorus transformation process was changed under the influence of liming
intensity, soil liming changed phosphate forms and the ratio between mineral fractions
of phosphates.
The aim of this study was to establish the effect of liming and fertilization on the
amount of available soil phosphorus in different classes of North-Western Romanian
soils: albic luvisoil, clay illuvial brown, clay illuvial chernozem.
Observations showed a maximum effect of pH increase and P mobile content fertilising
inorganic NPK together with manure on albic luvisoil.
Keywords: soil liming, mobile phosphorus, inorganic fertilizer, farmyard manure
INTRODUCTION
Different classes of soils in North-West Romania present low productivity inadequate
soil management, irregular supply of nutrients, the acidity being pointed out as the
cause for declined yield.
Phosphorus availability in most soils is optimal at the soil pH of 6.5. Below this value
as in more the soils of this part of Romania phosphorus will react with iron (Fe)/
aluminium (Al) oxides and start forming precipitates which prohibit the plant from
being able to fixate phosphorus. Liming acidic soils will generally increase availability
of phosphorus to plant. Ideally a liming program should be planned so that the pH can
be kept between 5.5 and 6.8 if maximum benefit is to be derived from applied
phosphorus (Havlin et al.2005). Over liming can depress phosphorus solubility due to
the formation of insoluble precipitates with calcium. A decrease in phosphorus
availability because of this formation of Ca-P minerals also happen in an alkaline soil
with a pH value higher than 6.5. Several other characteristics affect the process and
availability of phosphorus fixation. The texture and type of soil strongly influences the
level of mobile phosphorus.
MATERIALS AND METHODS
Soil samples were obtained from long time experiments established in 1961 at Livada,
Satu Mare County. This experiment includes treatments with different levels of mineral
fertilizers, organic manure and lime inputs.
234

Tops soil (0-30 cm) samples were taken from fields with an average size of 1.0 ha. They
were representative for the following classes of soil: luvisoil, clay illuvial brown, clay
illuvial chernozem. Each type of soil was fertilized as follows: N100, P 70, mineral NPK
(N100 P70 K60) mineral and organic NPK (N100 P70 K60 + 20t ha -1 farmyard) and lime
addition at various rates: 0.5 and 10 t ha -1
Detailed land use and agricultural management data were collected through
Romanian Statistic Annual (2004-2005). Each field was considered as a separate unit
because it is not managed in a coherent rotation.
In 2006 five representative points of each type of soil were selected for sampling. The
soils were air dried and ground ≤ 2mm sieve at the time of sampling.
Extractions with acetate-lactate solution (AL) were performed using soil to
solution ratio of 1:20 and the shaking time was 30 minutes. 5ml filtrate was treated with
(NH4)2 MoO4 0,15% and 5 ml fresh prepared 1,5% ascorbic acid standard curve was
prepared using suitable volumes of standard solution in place of sample. It has been
used a series of 6 standard solutions within the approximate range of 0.01 – 2.0 mg P/l.
After 10 minutes absorbance was measured at 880 nm using a reagent blank as a
reference solution.
Soil pH was measured in water using 1:2.5 ratio.
All analyses were carried out in 4 repetitions.
RESULTS AND DISCUSSIONS
1. Chemical analysis of soils showed that the pH and mobile phosphorus content was
exactly related to lime ratio and type of fertilizer. Impact of treatment (liming and
fertilizing) on different types of soil is shown in the below tables:
1. ALBIC LUVISOIL (initial pH = 4.90, P=15.4 ppm)
In this experiment the strongest effect on improving soil acidity and phosphorus content
of albic luvisoil was observed when fertilizing with mineral and organic manure
mixture (N100 P 70 K60 + 20t/ ha -1 FYM) and applying 10t ha -1 limestone.
On the background without organic fertilizer the greatest efficacy was recorded when
albic luvisoil had been applied with N100 P 70 K60 and 10t / ha -1
Table 1: Effect fertilizing and liming 5t/ ha -1 CaCO3 on albic luvisoil
Treatment ∆pH P (ppm)
Control 0.49 21.5
N100 0.21 10.1
P70 0.47 28.0
N100 P70 K60 0.96 40.1
N100 P 70 K60 + FYM 0.89 46.5
FYM 0.35 40.0
∆pH = pH variation
FYM = farmyard manure
235

Table 2: Effect of fertilizing and liming 10t/ ha -1 CaCO3 on albic luvisoil
Treatment ∆pH P (ppm)
Control 0.85 20.5
N100 0.24 15.4
P70 0.88 39.0
N100 P70 K60 1.09 85.4
N100 P 70 K60 + FYM 0.90 91.5
FYM 0.90 58.0
2. CLAY ILLUVIAL BROWN SOIL (initial pH = 5.10, P = 18 ppm)
Liming 10t/ ha -1 and fertilizing N100 P 70 K60 + FYM increased soil pH by 1.02 units and
mobile phosphorus raised to 70.1 ppm on clay illuvial brown soil.
Table 3: Impact fertilizing and liming 5t/ ha -1 CaCO3 on clay illuvial brown soil
Treatment ∆pH P (ppm)
Control 0.29 24.5
N100 -0.20 19.2
P70 0.29 28.4
N100 P70 K60 0.88 38.4
N100 P 70 K60 + FYM 0.80 45.3
FYM 0.32 33.0
Table 4: Impact fertilizing and liming 10t/ ha -1 CaCO3 on clay illuvial brown soil
Treatment ∆pH P (ppm)
Control 0.65 20.5
N100 -0.10 23.4
P70 0.50 38.2
N100 P70 K60 1.00 60.9
N100 P 70 K60 + FYM 1.02 70.1
FYM 0.70 39.0
3. CLAY-ILLUVIAL CHERNOZEM (initial pH = 5.20, P = 25.4 ppm)
Applying only mineral fertilizer or together with manure and liming = 10t/ ha -1 pH of
clay illuvial chernozem significantly increased: 1.06, respectively 1 pH units. Lime has
positive effect on P concentration, an increase to 48.9 ppm, respectively 50.4 ppm was
found.
236

Table 5: Impact fertilizing and liming 5t/ ha -1 CaCO3 on clay illuvial brown soil
Treatment ∆pH P (ppm)
Control 0.15 28.9
N100 0.10 21.4
P70 0.35 30.0
N100 P70 K60 0.90 42.3
N100 P 70 K60 + FYM 0.88 48.2
FYM 0.20 34.9
Table 6: Impact fertilizing and liming 5t/ ha -1 CaCO3 on clay illuvial brown soil
Treatment ∆pH P (ppm)
Control 0.55 14.3
N100 0.20 24.5
P70 0.40 30.4
N100 P70 K60 1.06 48.9
N100 P 70 K60 + FYM 1.00 50.4
FYM 0.35 40.9
CONCLUSIONS
1. The phosphorus content and pH value were found to be strongly dependent both of
treatment and soil type.
2. Soil supplementation with 10t/ha lime has the greatest effect on albic luvisoil pH
that increased with 0.85 units while P content of clay illuvial brown soil raised to
31.2 ppm.
3. Neutralization of soil acidity with limestone not only increases the soil pH, but also
changes the availability of P. Applying 10t ha -1 CaCO3 decreased to 20.5 ppm
(albic luvisoil), 14.3 ppm (clay illuvial chernozem) and 20.4 ppm (clay illuvial
brown soil).
4. This effect can be offset by fertilization. The application of NPK together with
manure increased the P level to 70.1 ppm (clay illuvial brown), 50.4 ppm (clay
illuvial chernozem) and 91.5 ppm (albic luvisoil).
REFERENCES
Havlin, J.L., James, D. Beaton, Samuel L. Tisdale and Werner L. Nelson, (2005):
Soil Fertility and Fertilizers: An Introduction to Nutrient Management 7 th Edition.
Economic Journal, 68: 302-316 p
Larsen, S. (1967): Soil Phosphorus. Advances in Agronomy 19: 105-207 p
Augusta Olivia Lujerdean (2003): Cercetari privind metodologia agrochimica de
determinare a formelor de fosfati din soluri. Teza de doctorat, Universitatea de
Stiinte Agricole si Medicina Veterinara, Cluj-Napoca
Pierysznski, G.M. (2000): Methods of Phosphorus Analyzis for Soils Sdiments
Residuals and Waters. Southern Coop. Series Bull No 396
237

Rusu, M., Boeriu, I., Kurtinec, P., Botha, P.V. (1981): Aspecte privind chimismul
fosforului in solurile acide din Nord Vestul tarii. Analele I.C.C.P. Fundulea, Vol 47
Irina Vintila, Borlan, Z., Rauta, C., Daniliuc, D., Tiganas, L. (1984): Situatia
agrochimica a solurilor din Romania, Ed. Ceres, Bucuresti.
238

COMPOSTED SLAUGHTERHOUSE BY-PRODUCTS EFFECT
ON CROP YIELD
Péter Ragályi, Imre Kádár
Research Institute for Soil Science and Agricultural Chemistry
H-1022, Budapest, Herman Ottó Str. 15, Hungary. E-mail: ragalyi@rissac.hu
ABSTRACT
Use of treated slaughterhouse waste as fertilizer can be a good way to maintain soil
productivity as well as reduce organic waste. Effect and residual effect of different
composts and meat powder were examined on crop development and yield on a
calcareous sandy soil. The soil contained 1-6% CaCO3 and 1-1.5% humus, humus layer
was 60-80 cm and pH(H2O) 7.0-7.4 as an average. The site had a poor N, P and K
supply. Water table was at 6-8 m depth. Trials were set in 2002 and 2003 with 5 doses
(0, 25, 50, 100, 200 t/ha fresh compost and 0; 2,5; 5; 10; 20 t/ha meat powder once
applied) and 4 replications giving a sum of 20 plots with 5x8=40 m 2 area each, arranged
in split-plot design. Sown crops were maize in 2002, mustard in 2003 and triticale
monoculture from 2004.
In the first two years of the trial the yield was negligible due to draught, though
tendencies showed 50t/ha dose mature compost and 25 t/ha immature compost as
optimal treatment.
All of the composts had significant residual effect on triticale in 2004 as the weather
was favourable and immature compost increased harvested straw and grain air-dried
yield from 1.6 to 5.3 t/ha. Maximal yields occurred at the maximal compost doses
without depression. In following years residual effects were moderated, but differences
were significant in the case of stronger composts even in 2006, in the 4 th and 5 th
experimental years.
Keywords: compost, fertilization, crop yield, slaughterhouse by-product
INTRODUCTION
Fodder use of animal origin wastes is strictly regulated by EU since BSE (bovine
spongiform encephalopathy) disease showed up, so alternative utilities have to be
found. In Hungary 100-120 million tons of wastes produced of which 5% is so called
“hazardous”. Hazardous wastes from animal bodies reach 300-400 thousand tons of
which 70-90 thousand are composted. After heat treatment these wastes become nonhazardous,
and other treatments like composting make land application possible. This
practice may improve soil parameters like organic matter, nutrient content, water
holding capacity and also reduces the yearly deposited 55 000 tons of slaughterhouse
wastes and sewage sludge (Vermes 1998, Kiss et al. 2001). Burying or incineration of
these materials is expensive and can result in environmental pollution (Izsáki 2000).
Debreczeni and Izsáki (1985) set up pot experiments to study the changes of field crops
grown on calcareous sandy soil and chernozem meadow clayey soil mixed with various
doses of sewage sludge from a tannery. On calcareous sandy soil 13 g sludge D.M. / kg
soil increased mustard above ground parts with 74% but higher doses caused
depression. Maize yields (above ground) were doubled in 33 g sludge D.M. / kg soil
239

treatment, while spring barely had 60 and 20% more yield due to 13 and 66 g sludge
D.M. / kg soil resp.
Effect of fertilization using various rates of tannery sewage sludge was studied in
field experiment on crop yields on calcareous humic sandy soil. The 60 t/ha (15-20 t/ha
D.M.) sludge dressing applied for four years completed with phosphorus and potassium
fertilizers was found to increase by 23 and 15% the grain yield of spring barely and rye
resp. as compared to the fertilized control in the first two years, however in the third and
fourth years yield dropped by 20%. 120 t/ha sludge applied for a four-year period was
found to cause yield depression in spring barley in first year, in further years rye and
winter wheat grain yields were roughly equal with those obtained in the fertilized
treatment. The 60 + 60 t/ha dose for two-two years did not cause depression and
resulted in 2-16% higher yields (Izsáki and Debreceni 1987). A slight residual effect
over 2-3 year of this experiment has been observed in 120 t/ha and 60 + 60 t/ha
treatments (Debreczeni and Izsáki 1989).
Simon and Szente (2000) set up field experiments with municipal sewage sludge
compost in maize on slightly acidic loamy-sand textured brown forest soil with 0, 10,
40 t/ha loads 2 weeks before sowing (in 1996) and 0, 10, 20, 40 t/ha loads 1 week
before sowing (in 1997). In the first year 9.6 t/ha yield of the control plot was increased
with 14% and 4% by 10 and 40 t/ha compost resp. In second year 20 t/ha dose gave
68% higher yield while 40 t/ha gave similar yield to 3.7 t/ha of control.
Application of sewage sludge is often limited by N content and not by relatively low
heavy metal content. Total N content release and utility parallel with decay take more
years. Unfortunately only a few data is available about N release rate of different
sewage sludge in different soil types, weather conditions and cultivation practice.
Kádár et al. (2002) set up a field experiment with dried communal sewage sludge
and slaughterhouse compost with 0, 25, 50, 100, 200 t/ha rate on clayey brown forest
soil applied 5-6 weeks before sowing of sugar beet. Despite of draught the crop could
develop well on treated plots while control plants were depressed. The highest 200 t/ha
load of slaughterhouse compost resulted the highest sugar yield of 8.7 t/ha compared to
6.5 t/ha of control. Optimal 25 t/ha dose of sewage sludge enhanced yield to 7.2 t/ha,
above this rate N overdose worsened quality parameters. Sludge and compost improved
structure, water management and fertility of soil.
MATERIALS AND METHODS
The long-term field experiment was set up at experimental site of the Research Institute
for Soil Science and Agricultural Chemistry at Őrbottyán on a calcareous sandy soil
with 1-6% CaCO3, 1-1.5% humus, 10-15% clay fraction, pH(H2O): 7.0-7.4 in average
in the ploughed layer. The water table was at 6-8 m, the site was prone to drought and
weakly supplied with N, P and K. Materials were applied at 5 different levels in 4
replications making 20 plots for each experiment (compost form). The plots had an area
of 5x8=40m 2 and arranged in split-plot design. In each experiment the applied rates
were 0, 25, 50, 100, 200 t/ha fresh compost. In case of meat powder the doses were 0,
2.5, 5.0, 10.0, 20.0 t/ha. As the soil and the composts were poor in potassium, 200 kg/ha
K2O fertilization was given in all the experiments in spring 2003. The composts were
processed by ATEVSZOLG Inc.
240

The mature compost became friable, inodorous, homogeneous material after 2-month
air-exposure and 10-month maturation. Immature compost was stinking, rough,
heterogeneous, after 6-week air-exposure and without maturation. Meat powder based
semi-mature compost was the material of immature compost maturing 6 months more,
but was still stinking and rough. Cooked meat based semi-mature compost received 2month
air-exposure and 8-month maturation.
Testing immature and semi-mature composts as well as extreme high application
doses served experimental purposes. Before application 2-2 composite samples (from
20-20 core samples) were taken from the materials. Analyses were made in the
Research Institute for Soil Science and Agricultural Chemistry. The average
compositions of composts are shown in Table 1.
Table 1: Composition of composts and meat powder in D.M.
with cc.HNO3+cc.H2O2 digestion
Measured Unit Mature Immature Semi-mature Semi-mature Meat
parameters compost compost compost mp* compost cm** powder
Dry Matter % 38.9 45.8 60.0 55.7 95.0
Org. Matter % 26.3 41.7 40.3 43.8 58.6
Organic C % 15.2 24.1 23.3 25.3 33.9
C/N ratio 7.5 7.7 7.1 8.7 5.3
Ca % 9.31 12.65 11.25 11.69 7.02
P % 2.22 5.56 4.26 5.26 4.06
N % 2.04 3.12 3.26 2.89 6.41
K % 0.76 0.76 0.83 0.50 0.41
Mg % 0.70 0.36 0.37 0.54 0.18
Na % 0.52 0.79 0.69 0.63 0.45
S % 0.50 0.70 0.62 0.75 0.60
Zn mg/kg 540 270 164 237 104
Cu mg/kg 109 46 19 42 13
NH4-N mg/kg 169 3006 941 882 167
NO3-N mg/kg 2480 1135 61 122 1
*meat powder based, **cooked meat based
In the first experimental year (2002) maize (Zea mays), in the second mustard (Sinapis
alba) and from the third triticale (X Triticosecale) monoculture were grown. The
amount of precipitation on sandy soil has strong effect on yields. The years 2002 and
2003 were drought. The maize received 237 mm and the mustard 52 mm of
precipitation during their growing season. Between 2004 and 2006 the triticale had
satisfactory amount of precipitation evenly distributed (Table 2).
241

Table 2: Precipitation of the experimental station, mm (Calcareous sandy soil,
Őrbottyán, 2002-2006)
2002 2003 2004 2005 2006 45-year
average
January 6 40 46 7 51 35
February 13 27 49 52 39 30
March 14 0 53 8 35 39
1 st quarter 33 67 148 67 125 95
April 30 12 39 96 23 46
May 46 32 42 41 82 59
June 41 8 68 48 83 67
2 nd quarter 117 52 149 185 188 173
July 52 57 35 85 30 61
August 98 13 67 124 118 51
September 59 17 13 74 29 46
3 rd quarter 209 87 115 283 177 168
October 52 79 48 14 14 44
November 32 45 55 33 15 55
December 40 7 36 76 4 45
4 th quarter 124 131 139 123 33 144
Total 483 337 551 658 523 569
RESULTS AND DISCUSSION
The average 20-30 t/ha rate of farmyard manure decays in 3-4 years in soil. About half
of its 2.0-3.0% D.M. nitrogen content can be utilized during this time of which 50% in
the first year, 30% in the second and 20% in the third (Sarkadi 1975).
In 2002 the 200 t/ha immature compost caused depression in yield, otherwise no
significant effect could develop. Trends showed that up to 50 t/ha mature compost and
25 t/ha immature compost application resulted in slightly better maize stand. Above
ground D.M. yield was 3 t/ha as an average. Drought season continued in 2003 when
mustard yields were economically negligible.
Effects of the composts were enhanced in 2004 as the crop received sufficient
precipitation during the 9 months growing season. The plantation could develop better
and reach higher yields even at lower doses of the applied materials, but maximal load
boosted the results. The 2 nd year residual effect of mature compost was moderate, but
significant. Immature compost could increase 3-fold the control yield, that is from 1.6
t/ha to 5.3 t/ha grain (table 3).
Highest dose of meat powder based semi-mature compost had also strong after
effects on triticale and doubled the yield of control, while the cooked meat based one
increased it with nearly 50% (table 4). Meat powder had also similar positive effects,
242

maximal yield could be reached at 5 t/ha load, but considerably decline did not occur
even at higher levels (table 5). The soil could have 320 kg/ha N through the applied 5
t/ha meat powder, which could cover the N demand of the higher biomass production
(Kádár and Ragályi 2004).
Table 3: Effect of slaughterhouse composts on air-dried triticale yield, t/ha
(Calcareous sandy soil, Őrbottyán)
Compost t/ha fresh material LSD5% Mean
0 25 50 100 200
Mature compost (applied 09 May 2002)
Triticale 2004
Grain 2.2 2.2 2.0 2.9 2.6 0.6 2.4
Straw 3.8 3.8 3.3 4.8 4.5 1.0 4.0
Total
Triticale 2005
6.0 6.0 5.3 7.6 7.1 1.4 6.4
Grain 1.7 1.9 1.7 2.2 2.3 0.6 1.9
Straw 2.8 2.9 2.6 3.6 3.5 1.1 3.1
Total 4.5 4.8 4.2 5.8 5.8 1.6 5.0
Immature compost (applied 09 May 2002)
Triticale 2004
Grain 1.6 2.8 3.2 4.5 5.3 1.8 3.5
Straw 2.8 4.5 5.0 6.7 8.0 2.8 5.4
Total
Triticale 2005
4.4 7.3 8.2 11.2 13.4 4.6 8.9
Grain 1.8 1.9 2.2 3.1 3.2 1.2 2.4
Straw 2.6 2.7 3.1 4.9 5.3 1.8 3.7
Total
Triticale 2006
4.4 4.6 5.2 8.0 8.5 3.0 6.1
Grain 0.8 0.8 1.1 1.1 1.5 0.4 1.1
Straw 1.5 1.8 2.1 2.2 3.1 1.2 2.1
Total 2.3 2.6 3.2 3.4 4.6 1.5 3.2
Comparing the composts the most effective was the meat powder based semi mature
one, but the other semi mature and the meat powder had also definitive effects. Mature
and immature composts were applied one year earlier so they had milder effects.
In 2005, in the 3 rd - 4 th years of the treatments mature compost residual effects
weakened and was not able to produce significant differences in yield so further
experiments were terminated. Meat powder based semi mature gave the highest yields
even in this year, which was 30% less, than in previous year in average. Maximal doses
were able to rise 5-6 t/ha control plot yield to 8-10 t/ha, but the effects are already
significant between 25 and 100 t/ha compost as well as 5 and 20 t/ha meat powder
treatment. Lower doses of the applied materials however caused no significant effect
any more.
243

Average yields in 2006 were only half of that in 2005 and treatment effects were just
above the significant limit except for meat powder in the case of grain yield.
Table 4: Effect of slaughterhouse composts on air-dried triticale yield, t/ha
(Calcareous sandy soil, Őrbottyán)
Compost t/ha fresh material LSD5% Mean
0 25 50 100 200
Meat powder based semi-mature compost (applied 18 Nov 2002)
Triticale 2004
Grain 2.4 3.8 4.3 4.4 5.4 1.7 4.1
Straw 3.9 5.9 6.3 6.5 8.1 2.1 6.2
Total 6.3 9.7 10.6 10.9 13.6 3.8 10.2
Triticale 2005
Grain 2.3 2.2 3.0 3.3 3.2 0.9 2.8
Straw 3.3 3.2 4.3 5.2 5.6 1.3 4.3
Total 5.6 5.4 7.3 8.5 8.8 2.2 7.1
Triticale 2006
Grain 1.2 1.2 1.4 1.5 1.6 0.4 1.4
Straw 2.3 2.4 2.9 3.0 3.4 1.1 2.8
Total 3.5 3.6 4.3 4.5 5.0 1.4 4.2
Cooked meat based semi-mature compost (applied 06 May 2003)
Triticale 2004
Grain 3.2 3.1 3.8 4.7 4.6 0.9 3.8
Straw 4.9 5.0 5.7 6.6 6.7 1.2 5.8
Total 8.1 8.0 9.4 11.3 11.4 2.0 9.6
Triticale 2005
Grain 2.5 2.4 2.3 3.0 3.2 0.5 2.7
Straw 3.9 3.7 3.3 4.5 5.1 0.9 4.1
Total 6.5 6.1 5.6 7.5 8.3 1.3 6.8
Triticale 2006
Grain 1.1 1.0 1.0 1.4 1.4 0.2 1.2
Straw 2.3 2.1 2.0 2.6 2.6 0.5 2.3
Total 3.4 3.2 3.0 4.0 4.0 0.7 3.5
The main results and conclusions can be summarised as follows:
1. Composts and meat powder are valuable fertilizers having much higher content of
N, P, Ca, Zn and Cu compared to the farmyard manure. These materials could be
successfully used in cultures having great fertilizer demand like sugar beet and also
fibre crops or cereals.
2. Composts and meat powder have considerable effect and even after 3-4 years
residual effect on yield.
3. According to results composts can be applied at 25-50 t/ha rate as a fertilizer
similarly to manure. On sandy soils, poorly supplied with K additional K
fertilization is recommended. Application above 50 t/ha can cause depression in
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unfavourable years. 100-200 t/ha doses can also cause environmental risk because
of the high N and P content.
Table 5: Effect of meat powder on air-dried triticale yield, t/ha. Applied 18 Nov
2002 (Calcareous sandy soil, Őrbottyán)
Meat powder t/ha LSD5% Mean
0 2.5 5 10 20
Triticale 2004
Grain 2.7 2.8 4.7 4.5 4.2 1.3 3.8
Straw 4.6 5.0 6.9 6.9 7.0 1.7 6.1
Total
Triticale 2005
7.3 7.8 11.6 11.4 11.2 3.0 9.9
Grain 1.9 2.0 1.9 3.0 3.9 1.3 2.6
Straw 3.0 3.2 2.7 4.4 6.7 2.5 4.0
Total
Triticale 2006
4.9 5.2 4.6 7.4 10.6 3.7 6.6
Grain 1.0 0.9 1.0 1.1 1.2 0.4 1.1
Straw 1.6 1.6 1.9 1.8 2.3 0.6 1.8
Total 2.6 2.5 3.0 2.9 3.5 1.0 2.9
REFERENCES
Debreczeni I., Izsáki Z. (1985): Tannery sewage sludge effect on crop element
composition. Agrokémia és Talajtan 34: (3-4) 421-432. (in Hungarian)
Debreczeni I., Izsáki Z. (1989): Tannery sewage sludge fertilization effect and residual
effect on cereals on a sandy soil. Növénytermelés 38:3 231-239. (in Hungarian)
Izsáki Z., (2000): Collecting, disposal and utility of agricultural wastes. Tessedik
Sámuel Főiskola, Szarvas. 94 pp. (in Hungarian)
Izsáki Z. – Debreczeni I. (1987): Tannery sewage sludge fertilization effect on a sandy
soil. Növénytermelés 36:6 481-489. (in Hungarian)
Thaer, A. (1809): In: Kádár I. (ed.) (1996): Albrecht Thaer: “Basics of rational
agriculture. Manuring.” MTA TAKI. Budapest. 99 pp. (in Hungarian)
Kádár I., (2003): Report of ATEV compost effect on soil fertility. MTA TAKI.
Manuscript. (in Hungarian)
Kádár I., Hámori V., Morvai B., Petróczki F. (2002): Soil load and pollution limit
values; sewage sludge and slaughterhouse waste compost effect on sugar beet. In:
Cukorrépa termesztési/termeltetési tanfolyam és tanácskozás. 37-40. Szerk.:
Várnainé J.A. Cukoripari Egyesülés. Budapest. (in Hungarian)
Kádár I., Ragályi P. (2004): Final report in 2004 about long-term field experiments set
up with slaughterhouse composts. In: MAE Talajtani Társaság Talajvédelmi Klub.
FVM Növény és Talajvédelmi Szolgálat. Budapest. 6 pp. (in Hungarian)
Kiss J., Simon M., Horváth Z., Kádár I., Kriszt B., Szoboszlay S., Morvai B., Csomor
L., Szántó G. (2001): Biological degradation of animal origin fatty wastes. In: XVI.
Országos Környezetvédelmi Konferencia. (Szerk: Elek Gy.– Vécsi B.) 351-360 pp.
Siófok. (in Hungarian)
245

Sarkadi J. (1975): Methods for estimating fertilizer demand. Mezőgazdasági Kiadó,
Budapest. 252 pp. (in Hungarian)
Simon L., Szente K. (2000): Sewage sludge compost effect on maize N content and
yield. Agrokémia és Talajtan. 49: 231-246. (in Hungarian)
Vermes L. (1998): Waste management, waste utility. Mezőgazda Kiadó. Budapest. 201
pp. (in Hungarian)
246

THE EFFECT OF BENTONITE ON SPECIFIC SOIL PARAMETERS AND
MICROBIAL CHARACTERISTICS OF THE CARBON CYCLE
János Kátai 1 , Magdolna Tállai 1 , János Lazányi 2 ,
Edina Veres Lukácsné 1 , Zsolt Sándor 1
1 University of Debrecen, Centre of Agricultural Sciences,
Department of Agrochemistry and Soil Science
2 University of Debrecen, Centre of Agricultural Sciences,
Department of Rural Development and Land Use
ABSTRACT
The effect of increasing dosages (5, 10, 15, 20 t/ha) of bentonite (Bentonite experiment)
and bentonite composted with manure (Bentonite + manure experiment) was studied in
a small plot experiment on acidic sandy soil at the Experimental Farm of the
Nyíregyháza Research Centre, UD CAS. The effect of treatments on the physical,
chemical and microbial soil parameters can be summarized as follows:
• Bentonite significantly increased the water-holding capacity of soils. In samples
treated with bentonite moisture content was 2-3 % higher than that in treatments
with bentonite and manure.
• The pH value of the soil was shifted towards neutral. In bentonite treatments, pH
was more increased than in treatments of bentonite composted with manure.
Accordingly, hydrolytic acidity was also reduced.
• When studying the nutrient content of soil, we found that bentonite increased,
though not always significantly, the readily available potassium content of soil. As
a result of the composted treatments, the readily available nitrate, phosphorus and
potassium content of the soil significantly increased.
• Regarding the studied microbial parameters, we found that the composted
treatments caused a higher (though not always significantly) increment in the total
number of bacteria, the number of microscopic fungi and cellulose-decomposing
bacteria and the biomass-C content of soils. In both series of treatments, the high
dosages caused a reduction in the number of microbes.
High bentonite dosages also reduced carbon dioxide formation.
Saccharase activity was significantly increased by small dosages of bentonite, while
composted treatments resulted in a lower, not significant increment in enzyme
activities.
• In the experiment, several relationships were found between the physical, chemical
and microbial soil parameters.
Keywords: bentonite, moisture content, pH, quantitative changes in microbes,
microbial activity
INTRODUCTION
Nowadays, sustainable agriculture is a worldwide known notion, a basis of which is the
protection of the environmental elements and reducing the environmental loading to the
minimum. These aspects should also be considered in soil melioration, therefore,
increasing attention is paid to the study and application of natural, environmentally
247

enign materials in agriculture. Numerous researchers studied products from mining
and industry for increasing the organic and inorganic colloid content of sandy soils and
for improving their water and nutrient management (Blaskó, 2005). Zeolites (Kazó,
1981; Köhler, 2000), ground rhyolite tuff, ground basaltic tuff, and bentonite (Köhler,
2000; Lazányi, 2003) have already been examined.
Carbon is an essential element of life, a composer of live and dead organic matter,
the carbon cycle is a basic process of the biogeochemical cycle (Jolánkai et al., 2005).
Microorganisms play an extremely important role in transforming organic matter into
inorganic matter and in the cycles of the different elements including carbon. For
describing the transforming processes, intensity of cellulose-decomposition (Kádár et
al., 2005), carbon-dioxide production of soils (Zsuposné Oláh, 2005; Varga et al. 2005),
and the amount of carbon in the microbial biomass are used (Szili-Kovács et al., 2005;
Szili-Kovács 2006; Lukácsné Veres et al., 2006).
With special regards to the requirements of sustainability, we joined an experiment
of sand melioration, in which we studied the effect of bentonite, a natural, clay-like
stone rich in colloids, and bentonite composted with manure on the moisture content,
pH, nutrient management and microbial activity of soil, which is demonstrated via
specific elements of the carbon cycle.
MATERIAL AND METHODS
The small-plot experiment was set up at the experimental farm of the Nyíregyháza
Research Centre of UD CAS in October 2002. Samples were collected from the top 2-
20cm layer of soil. The in vitro experiments were carried out at the soil microbial
laboratory of the Department of Soil Sciences, UD CAS FA between 2003 and 2005.
The soil type was brown forest soil with alternating layers of clay (”kovárvány”), with
sandy texture and acidic pH (pH(H2O) 5.28; pH(KCl) 4.33 ). The soil was deficient in
nitrogen, the level of phosphorus and potassium supply was medium and satisfactory,
respectively. Treatments were performed with increasing dosages of bentonite and
bentonite composted with manure (Table 1).
From the physical parameters, moisture content was determined after drying to
constant mass at 105 0 C (Klimes-Szmik, 1962). From chemical soil parameters, pH was
measured in suspensions with distilled water and KCl (Filep, 1995). The nitrate-N
content of soil was determined with the Na-salicylate method (Felföldy, 1987), while
the easily soluble phosphorus and potassium content of soil was measured with 0.1m
ammonium-lactate (AL) extractor (Gerei, 1970).
The amount of microbes in the soil was determined by the plate dilution method
from soil-water suspension as the total number of bacteria (on Bouillon agar) and the
number of microscopic fungi (on peptone glucose agar) (Szegi, 1979). The number of
cellulose-decomposing bacteria was determined by the method of the most probable
number of bacteria according to Pochon et al. (1962). Regarding soil microbial activity,
the carbon-dioxide production of soil was measured according to the method of
Witkamp (1966. cit. Szegi, 1979), while the soil biomass-C production and saccharase
enzyme activity were determined by the fumigation-incubation method (Jenkinson et
al.,1976) and the method of Frankenberger et al., respectively. The statistical analysis
was performed using the SPSS 9.0 program
248

RESULTS
The effect of treatments on the studied soil characteristics is presented as a mean of the
three years (2003-2005).
Soil moisture content (Figure 1) varied between 12.69 and 14.38 % (mass %) in the
average of three years. In treatments with bentonite, a significant increase was observed
in moisture content, 2-3% higher than in treatments with bentonite composted with
manure.
Moisture content
(mass %)
15
13
11
9
7
5
1 2 3 4 5 6 7 8 9 10
Treatments
Figure 1: The effect of bentonite on soil moisture content
The pH value of soils (Figure 2) increased significantly in both treatment series and
solutions (H2O; KCl). Bentonite treatments increased the pH value more than the
bentonite + manure treatments.
pH
7
6
5
4
3
1 2 3 4 5 6 7 8 9 10
pH (H2O)
Treatments
pH (KCl) y1
18
13
8
3
Hydrolytic
acidity
(y1)
Figure 2: The effect of bentonite on the pH and hydrolytic acidity of soil
A considerable reduction was observed in the hydrolytic acidity (y1) (Figure 2) with
increasing pH. In both treatment series, even small dosages (5 t/ha; 5+9 t/ha) caused a
significant reduction, while the change was smaller in the case of higher dosages.
The NO3-N content of soil (Table 1) reduced, though not always significantly, as a
result of bentonite treatments. This cannot be stated about the bentonite + manure
treatments where NO3-N content increased in proportion to the increase in dosages. As a
249

esult of higher dosages, no considerable changes were observed in nitrate content.
However, the nitrate content of treatments 8, 9 and 10 was significantly higher than that
of the control.
We have also measured the readily available phosphorus and potassium content.
The AL-soluble phosphorus content of soils significantly reduced as a result of
bentonite treatments, regardless of dosage. The phosphorus content of composted
treatments, however, increased significantly, and no considerable reduction was found
even in the high dosage treatments (treatments 9 and 10).
The readily available potassium content of soils increased slightly as a result of
bentonite treatments, but the values were not significantly different from those of the
control. However, the composted treatments significantly increased the potassium
content of soils.
Table 1. The effect of treatments on the readily available nutrient content of the soil
(average of 2003-2005)
Sample
Dosage
Treatment
Nitrate-N
(mg - kg -1 )
AL-P2O5
(mg kg -1 )
AL-K2O
(mg k g -1 )
Bentonite
1. - 0 (t/ha) 6.50 148.77 250.97
2. 1.× 5 (t/ha) 6.23 119.43 253.77
3. 2.× 10 (t/ha) 5.93 118.67 248.77
4. 3.× 15 (t/ha) 5.90 141.53 277.50
5. 4.× 20 (t/ha) 5.80 124.57 237.60
Bentonite+manure
6. - 0 6.70 145.90 252.20
7. 1.× 5 (t/ha)+9 (t/ha) 7.17 156.87 301.80
8. 2.× 10 (t/ha)+18 (t/ha) 8.20 153.53 312.93
9. 3.× 15 (t/ha)+27 (t/ha) 8.50 163.87 283.53
10. 4.× 20 (t/ha)+36 (t/ha) 8.40 154.87 307.20
Significant Difference (5%) 0.25 3.65 15.6
The impact of treatments on soil microbial parameters (Table 2) is demonstrated via
specific characteristics of the carbon cycle.
Bentonite treatments increased slightly the total number of bacteria and the
amount of microscopic fungi, the increase of the latter was not significant. Higher
dosages resulted in a decrease in the number of microbes. In both groups of microbes,
the medium dosages had an inducing effect. Composted treatments had a higher
increasing effect on the amount of microbes, though it was not always significant.
The number of cellulose-decomposing bacteria was increased considerably by
treatments of bentonite no. 3 and treatment no. 2 of the combined series. High bentonite
250

dosages significantly reduced the amount of cellulose-decomposing microbes. The high
dosage treatments with bentonite composted with manure had a weaker inhibiting
effect.
In treatments with the two high dosages of bentonite, CO2 production was lower
than that of the control. Medium and high dosages of bentonite + manure also
significantly reduced the formation of CO2 in the soil.
The soil biomass-C value reduced under that of the control already at applying medium
dosages of bentonite. The small dosages of composted treatments caused a significant
increment, while the two high dosages reduced its values and treatment 10 resulted in a
significant reduction.
Regarding saccharase enzyme activity, a higher value was measured in treatment
2 among the bentonite treatments. Further treatments reduced the enzyme activity,
though not significantly. Composted treatments resulted in a smaller, non-significant
increase in enzyme activity.
Sample
Table 2. The effect of treatments on the amount of microbe groups and their
microbial activity (average of 2003-2005)
Dosage
Total
bacteria
(*10 6 g -1 soil)
Total fungi
(*10 3 g -1 soil)
Cellulose
decomposing
bacteria
(*10 3 g -1 soil)
CO2production
(CO2 mg
100g -1 )
Biomass-C
(µg g -1 )
Saccharase
(glucose
mg 100g -1 )
1. - 3.20 82.01 11.59 8.47 99.14 7,96
2. 1.× 3.80 64.69 9.89 8.73 100.71 9,10
3. 2.× 5.35 82.92 12.79 8.45 85.35 8,75
4. 3.× 3.94 54.91 7.90 5.74 85.27 8,76
5. 4.× 4.14 55.18 4.01 7.98 83.27 7,55
6. - 5.33 82.73 9.03 8.23 117.72 8,01
7. 1.× 6.15 71.21 20.95 8.10 136.18 8,94
8. 2.× 6.07 95.47 9.49 6.89 127.27 8,24
9. 3.× 6.05 88.73 7.03 8.37 118.67 8,34
10. 4.× 6.60 91.37 8.10 7.16 83.05 8,07
(SD5%) 0.25 3.54 4.73 0.30 3.86 0.72
In correlation analysis, we searched for correlations between the physical, chemical and
microbial soil parameters for both treatment series (Table 3). In our analysis, we
emphasize only the strong correlations.
In bentonite treatments, a strong correlation was found between the nitrate-N
content and readily available phosphorus content of soil (r=0.71). A strong correlation
was observed between changes in soil nitrate-N content and biomass-C content
(r=0.91). Among the microbial parameters, a strong positive correlation was found
between the total number of bacteria and the number of cellulose-decomposing bacteria
251

(r=0.71), and between the total number of bacteria and CO2 production of the soil
(r=0.82).
Table 3. Correlation ( r ) between the examined physical, chemical, and
microbiological parameters of soil
* Correlation is significant at 0.05 level (2-tailed)-medium
** Correlation is significant at 0.01 level (2-tailed)-tight
In treatments of bentonite composted with manure, a strong correlation was found
between pH and readily available phosphorus content of the soil (r=0.71). Nitrate-N and
the total number of fungi also correlated closely (r=0.80). Among microbial parameters,
the total number of bacteria showed a strong positive correlation with CO2 production
(r=0.81), similarly to bentonite treatments, and a strong correlation was found between
the changes in the total number of bacteria and the biomass-C content of soil (r=0.79).
Composted treatments had a favourable effect on the amount of cellulose-decomposing
bacteria, but the number of fungi showed a strong negative correlation with their
increasing numbers (r=-0.71).
252

DISCUSSION
In our experiments, we found, similarly to the earlier observations of Lazányi (2005),
that bentonite has a favourable effect on the soil moisture content.
Szegi et al. (2003) studied the effect of bentonite on the biomass of pea and sweet
corn. According to them, the fertility of soil improved with improving physical and
chemical soil parameters. Our experiments have confirmed the earlier results (Szegi et
al. 2002, 2003), according to which bentonite reduced the acidic pH of the soil ( pH(H2O)
5.28 ) and simultaneously the values of hydrolytic acidity were also reduced. Bentonite
composted with manure increased the readily available phosphorus, potassium and
nitrate content of the soil.
No data were found in the literature about the changes in the amount and activity
of soil microbes. According to our experience, the amount of bacteria and fungi were
increased primarily by treatments of medium dosage. There were only slight changes in
CO2 production and saccharase enzyme activity, while the amount of microbial
biomass-C was considerably higher in treatments with bentonite composted with
manure.
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254

INVESTIGATION OF MINERAL CONTENT IN WINTER WHEAT GRAIN
SAMPLES FROM LONG-TERM FIELD EXPERIMENTS
Zoltán Győri – Árpád Tóth – Diana Ungai
University of Debrecen Centre of Agricultural Sciences Faculty of Agronomy
Institute of Food Science, Quality Assurance and Microbiology
ABSTRACT
Besides the technological quality of the wheat flour consumers are primarily interested
in the nutrition-biological value (for example minerals, vitamins, etc.) of the final
products.
The assay demonstrates the results of the researches in different soils in Hungary
within the framework of the National Long-term Field Trials. The winter wheat grain
samples are from three research sites: Karcag, Nagyhörcsök and Iregszemcse. The
analyzed winter wheat grain samples are from the corn – corn - winter wheat - winter
wheat crop rotation. The applied fertilizer doses were the following: 0 – 150 – 200
kg/ha N, 0 – 100 – 120 kg/ha P, 0 – 100 – 200 kg/ha K, the other two active substances
remained the same. The concentrations of calcium, magnesium and manganese of wheat
samples are shown in this article.
The results of the analysis conducted in our accredited laboratory demonstrate that
the effect of the K and P fertilizers on the Mg and Mn concentration depends rather on
the production site, while the effect of the N fertilizer depends on the crop year. The
application of N fertilizer increased the Ca content of the wheat grain samples in all
three research sites in every statistically proved case.
Keywords: Long-term field experiment, winter wheat grain, NPK fertilization, Ca-,
Mg-, Mn-content
INTRODUCTION
Besides the technological quality of the wheat flour consumers are primarily interested
in the quality of the final product, i.e. food and the attractiveness of its exterior,
nutrition-biological value and their healthiness and usefulness for the human organism.
Most of the attention is usually directed at micro-nutrients, such as vitamins and
minerals. In human nutrition in Europe, winter wheat constitutes one of the most
important sources of micro-, and macro-elements (Mahan et al., 1996., Lagua et al.,
1996).
Pais (1981) claims that wheat grains contain 6.8 mg kg -1 of copper, 62 mg kg -1 of
iron, 34 mg kg -1 of manganese and 31 mg kg -1 of zinc. Sugiura et al. (1998) obtained the
following average results for wheat flours: protein content 15.5%, ash content 1%,
potassium content 4800 mg kg -1 , calcium content 300 mg kg -1 , phosphorous content
3200 mg kg -1 , magnesium content 1400 mg kg -1 , copper content 2.5 mg kg -1 , iron
content 27 mg kg -1 , manganese content 21.6 mg kg -1 and zinc content 13 mg kg -1 .
The variability of mineral content of wheat flours can be attributed to
environmental and genetic factors and their interaction. Several mineral elements show
high positive correlation with protein content in both flours and bran (Posner and Hibbs,
1997).
255

Western Canadian hard red spring wheat crops were characterized for cadmium, copper,
iron, manganese, selenium and zinc by Gawalko et al. (2002). Samples were received
from crop districts in Manitoba, Saskatchewan and Alberta for the 1996, 1997 and 1998
crops. Year-to-year variations in grain chemistry are small for Cd, Mn, Se and Zn, but
Cu and Fe contents show 12% and 9% decreases respectively over the three years. The
overall variability for the plant-essential trace elements, Cu, Fe, Mn and Zn, is low in
comparison with Cd and Se. It is demonstrated that the spatial variation in crop
chemistry across the Canadian Prairie wheat producing region is greater than the year to
year variations.
To compare the effects of organic manuring and inorganic fertilization, the
contents of trace elements in wheat harvested from a field experiment were analysed.
Inorganic fertilization compared to organic manuring increased the content of
manganese in wheat. Kirkham (1983) studied the effect of the sewage sludge on the
non-essential elements (Cd, Cr, Ni, Pb), as well as nine essential elements (N, P, K, Ca,
Mg, Cu, Fe, Mn, Zn) content of wheat samples in Manhattan, Kansas. Soil and winter
wheat were sampled in 1980 after a total of 128 t ha −1 of sludge (dry-weight basis) had
been added to the soil (32 t ha −1 year −1 ) since 1976. Soil in sludge-fertilized fields had
more N, P, K, Mg, Fe, Cu, Mn, Zn, Cd, Cr and Ni than did soil from the control fields.
In general, concentrations of essential elements in plants grown on sludge were similar
to concentrations of essential elements in plants grown with inorganic fertilizer.
Based on the results of long-term field trials, Kádár (1991) found that after
fertilization the following proved changes occurred in the concentrations of the
individual elements: N fertilization increased the Al and Mn content; P fertilization
increased the following: Al, B, Ca, P, Sr and decreased Zn content. K fertilization
increased the Ba and K content and decreased Mg content.
The nitrogen fertilizers decrease the soil pH (Debreczeni and Debreczeniné, 1994)
and it is affect the uptake of mineral elements with special regard to the Mn-uptake.
MATERIALS AND METHODS
The assay demonstrates the results of the researches in different soils in Hungary within
the framework of the National Long-term Field Trials. Field trials started in 5 fields in
1966 and in 4 fields in 1967. The fundamental conception was to use 4-year crop
rotation: winter wheat – corn – corn – peas, and winter wheat – corn – corn – winter
wheat, out of which peas were excluded in 2000. The researches are done in split-splitspot
arrangement, in four repetitions.
The analyzed winter wheat grain samples are from the corn – corn - winter wheat -
winter wheat crop rotation. Out of the different fertilization doses we used the results of
the nutrients in the control and high-dose plots in our calculations. The applied fertilizer
doses were the following: 0 – 150 – 200 kg/ha N, 0 – 100 – 120 kg/ha P, 0 – 100 – 200
kg/ha K, the other two active substances remained the same. When applying the
fertilizer, we have to consider that besides active substances additional elements are
given as well. The following includes the quantity of the additional elements in
Ammonium-nitrate, Superphosphate and Potassium-chloride fertilizers that are
important in our analysis (mg/kg; %):
256

Element Ammonium-nitrate Superphosphate Potassium-chloride
Ca 6,0 % 17,4 % 3470
Mg 23,8 570 302
Mn 0,299 152 19
Source: Győri (1998).
The winter wheat grain samples in our analysis are from three research sites: Karcag,
Nagyhörcsök and Iregszemcse. The soil type is meadow chernozem in Karcag and
calcareous chernozem at the other two research sites. Table 1 shows the characteristics
of the cultivated layer of the soil at the different research sites. The dates of control
plots are calculated from the averages of results of the 10 th , the 16 th and the 20 th years.
Table 1: Nutrient supply in different research sites
Parameters Karcag Nagyhörcsök Iregszemcse
Humus content (%) 2,7 (medium) 2,7 (good) 2,4 (medium)
AL-P2O5 (mg/kg) 34 (very weak) 81 (weak) 103 (medium)
AL-K2O (mg/kg) 270 (good) 147 (medium) 150 (medium)
pHKCl 4,7 7,2 7,4
KA 47 38 37
Source: Debreczeni and Debreczeniné (1994)
The analyses were carried out by the accredited laboratory of the Institute of Food
Science, Quality Assurance and Microbiology of the Faculty of Agriculture of the
Centre for Agricultural Sciences of the University of Debrecen. The mineral element
contents of the samples were determined with OPTIMA 3300 DV ICP-OES (Perkin-
Elmer Ltd. Wellesley, USA) using No. BCR CRM 189 certified reference material
(Kovács et al., 1996, 1998). The concentrations of calcium, magnesium and manganese
of wheat samples are shown in this article.
The data were evaluated by using one-way ANOVA, and there were carried out average
and standard deviation calculations run on SPSS 11.5 for Windows and Excel 6.0 for
Windows.
RESULTS
The effect of N fertilization on the mineral content of winter wheat samples
The applied N fertilization significantly increased the Ca content of wheat samples in
every studied crop year in Karcag (Figure 1). At the Iregszemcse research site, the
effect of N fertilization on the Ca content was proved by variance analysis only in some
cases, however, this time the 150 and 200 kg/ha N doses– similarly to the results in
Karcag– increased the Ca content in each case. In the Nagyhörcsök research site,
similarly to the effects in Karcag, the effect of the N fertilization was clear: the various
doses of N fertilizers had a positive effect on the Ca content of wheat samples.
According to Győri’s (1998) measurements the Ammonium-nitrate has 6% Ca as
additional element in the fertilizer. The amount of Ca was 0 kg/ha in the control
257

treatments and it was 12 kg/ha in the case of 200 kg/ha applied N-fertilizer. Take this
fact into account is important to evaluate accurately the results.
Ca conent (mg/kg)
800
700
600
500
400
300
200
100
0
2001 2004 2005 2001 2004 2005 2001 2004 2005
Karcag Iregszemcse Nagyhörcsök
Control 200 kg/ha
Figure 1: Ca content of winter wheat samples as an effect of N fertilization
The effect of N fertilization on the Mg content was inconsistent in all the three research
sites, depending on the crop year; it both decreased and increased the Mg concentration.
For example in 2002, applying 200 kg/ha N in Nagyhörcsök after winter wheat forecrop
significantly increased while in 2005 after corn forecrop decreased the Mg content of
the wheat grain samples.
In Karcag in 2001, after corn forecrop it significantly decreased while in 2005 after
winter wheat forecrop increased the Mg content of wheat grain samples. In the
Iregszemcse research site we found the opposite effect: N fertilization after corn
forecrop increased, while after winter wheat decreased the Ca content of wheat. In the
Nagyhörcsök research site in 2001 and 2002, the application of 200 kg/ha N doses
significantly increased the Mn content of winter wheat grain samples, while significant
effect was not fund in 2003-2005.
The effect of phosphorus fertilization on the mineral content of winter wheat
samples
The effect of phosphorus fertilization on the Mg and Mn content of winter wheat grain
samples were not significant either at 120 kg/ha dose, or in dry or humid crop years in
Karcag. The effect of phosphorus on the Ca content was statistically proved in one case:
in 2003, after corn forecrop the application of 120 kg/ha P2O5 active substance
decreased the Ca content of the wheat grain samples from 602,3±23,5 mg/kg to
544,0±24,0 mg/kg compared to the control plots. As regards samples from Iregszemcse
and Nagyhörcsök, the effect of phosphorus was significant in one crop year,
respectively. Applying 100 kg/ha P2O5 fertilizer significantly increased the Ca content
of wheat grain samples after winter wheat forecrop in Iregszemcse in 2002, and after
corn forecrop in Nagyhörcsökön in 2001.
258

According to our results, the effect of phosphorus fertilization on the Mg and Mn
content of the samples was not significant in Karcag.
In the Iregszemcse research site, the effect of phosphorus fertilizer on the Mg content of
the samples was only significant in 2002, while on the Mn content of samples was
significant in 2001, 2002 and 2004. In these cases the phosphorus doses increased the
concentration of these elements.
We found that in Nagyhörcsök the application of 100 or 120 kg/ha P2O5 active
substance– irrespectively of the forecrop – increased the Mg and Mn content of the
wheat grain samples in almost every crop year. (Table 2).
Table 2: The effect of phosphorus fertilization on the Magnesium and Manganese
content of winter wheat samples from Nagyhörcsök
Mg content (mg/kg) Mn content (mg/kg)
2001 2004 2005 2001 2004 2005
control 775,8±25,8 1013,3±29,8 959,0±29,8 24,8±1,4 29,2±2,0 10,7±0,3
100 kg/ha
933,3±24,5 1219,5±19,5 1086,3±48,7 38,3±1,5 45,1±1,0 12,5±0,1
P2O 5
p value 0,000 0,000 0,011 0,000 0,000 0,000
The effect of K fertilization on the mineral content of winter wheat samples
In 2004 in Karcag, the application of 200 kg/ha K fertilizer significantly increased the
Ca, Mg and Mn content of wheat grain samples after corn forecrop, while in the same
year this effect was not experienced after winter wheat forecrop.
In the Iregszemcse research site, the effect of K on the nutrient content of the samples
was statistically proved in one crop rotation in one year (2004): the average manganese
content of the control wheat samples was 12,2±0,1 mg/kg, and the application of 100
kg/ha K2O increased this value to 12,9±0,1 mg/kg. The effect was proved at P=1%
significance level.
Contrary to the results from the Karcag and Iregszemcse research sites, (Figure 2)
the application of K fertilizer decreased the Ca and Mg content of the wheat grain
samples in each case in Nagyhörcsök, the results are statistically proved. The
application of K fertilizer decreased the Ca content of wheat grain samples from
525,5±32,6 mg/kg to 442,3±39,8 mg/kg in 2001, and from 682,3±47,4 mg/kg to
321,3±12,8 mg/kg in 2005. In Nagyhörcsök, the changes in the Mg content proved to be
significant only in 2001, when the Mg content of the control wheat samples decreased
from 1022,8±20,2 mg/kg to 890,0±32,7 mg/kg.
259

Ca and Mg content (mg/kg)
1400
1200
1000
800
600
400
200
0
2001 2004 2005 2001 2004 2005
Ca content Mg content
Control 200 kg/ha
Figure 2: The effect of K fertilization on the Ca and Mg content of winter wheat
samples
DISCUSSIONS
In the calcerous chernozem soils in Iregszemcse and Nagyhörcsök the applied N
increased the Mg and Mn content of the wheat grain samples in dry years and decreased
in humid years. We cannot draw the same conclusion from the results of the meadow
chernozem soil with salt in the deeper layers in Karcag. The increase of the Mnconcentration
is probably related to the decrease of soil pH in consequence of nitrogenapplying.
The application of N fertilizer increased the Ca content of the wheat grain
samples in all three research sites in every statistically proved case. The unique Caincreasing
effect might be the result of the considerable amount of Ca as additional
element in the fertilizer.
As regards phosphorus fertilization, we found that its effect on the nutrient content
depends on the production site and the soil type. In Nagyhörcsök and Iregszemcse, the
application of phosphorus increased the Mg and Mn content of the wheat grain samples
in every statistically proved cases, while in Karcag it had no effect on the concentration
of these elements.
The K content of the soil was sufficient in Karcag, while in Nagyhörcsök and
Iregszemcse it was medium, therefore we expected that the effect of K fertilization
would have higher effect at these cropping sites. Contrary, the nutrient-increasing effect
of K fertilization was proved only in Nagyhörcsök. At the Iregszemcse research site,
although the K supply was only medium, the K fertilization had no effect on the nutrient
content probably due to other soil-chemical factors.
The results of the analysis conducted in our accredited laboratory demonstrate that
the effect of the K and P fertilizers on the Mg and Mn concentration depends rather on
the production site, while the effect of the N fertilizer depends on the crop year.
260

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Cereal Chemists, Inc., St. Paul, Minnesota, U. S. A.
Srikumar, T.S., Öckerman, P.A. (1991): The effects of organic and inorganic
fertilization on the content of trace elements in cereal grains. Food Chemistry, 42:
225-230 p.
Sugiura, S.H., Dong, F.M., Rathbone, C.K., Hardy, R.W. (1998): Apparent protein
digestibility and mineral availabilities in various feed in gredients for salmonid
feeds. Aquaculture, 159: 177-202 p.
261

COMPARISON OF SOIL-TEST EXTRACTANTS FOR ZINK AND COPPER
Rita Kremper - Sándor Berényi - Jakab Loch – Andrea Balla Kovács
Debrecen University, Centre for Agricultural Sciences
Department of Agrochemistry and Soil Science
ABSTRACT
KCl-EDTA, Lakanen Erviö and CaCl2- DTPA solutions were compared as soil Cu and
Zn extractant. The former two methods are the Hungarian official methods, while
CaCl2- DTPA is a frequently studied method word-wide. 27 agricultural soil samples
were collected for the study from the Soil Information and Monitoring System (SIMS)
database. LE and KCl-EDTA extracted similar amounts of Zn and Cu. CaCl2-DTPA
extracted much smaller amounts both for Zn and Cu (approximately half-third as much)
than the other two extractant. It can be attributed to the pH of the extractants. We
suppose that KCl-EDTA and LE solved Zn and Cu oxides-hydrates, Zn and Cu
carbonates at pH 4.36 and pH 4.6 respectively which can not be solved by CaCl2-
DTPA at pH 7.2. We found good correlation between the three methods despite of the
small sample number.
Keywords: extractrant, DTPA, KCl-EDTA, Lakanen-Erviö, microelements
INTRODUCTION
The three sequential steps generally followed to develop micronutrient cation soil tests
are extractant selection, greenhouse evaluation and field calibration (Martens and
Lindsay, 1990). The first step is to select an extractant that will solubilize a
proportionate part of labile forms of micronutrient cations from different soils. The
second step is to evaluate if amounts of cations extracted are related to quantities
absorbed by plants from different soils. This step commonly is completed int he
greenhouse rather than in the field. The third step field calibration of a soil extractant is
conducted if a suitable relationship is established during the greenhouse evaluation.
During field test scientist separate soils into responsive and non-responsive categories,
or determine the insufficient, sufficient and transitional zone of the available
micronutrient cations in the soil.
Most of the Zn and Cu in soils is unavailable to plants and may be divided by chemical
extraction into five chemical pools (McLaren and Crawford, 1973)
1. as free and complexed ions in soil solution
2. as non-specifically and specifically adsorbed cations
3. as ions occluded mainly in soil carbonates and hydrous oxides
4. in biological residues and living organisms
5. in the lattice structure of primarily and secondary minerals.
In soils Zn and Cu cations would be specifically adsorbed by carbonates, hydrous
oxides of Al, Fe and Mn, soil organic matter and phyllosilicates (Udo et al., 1970)
Soil extractants for plant available Cu and Zn forms can be divided into water and
neutral salts, weak and strong acids and chelating agents. Neutral salts by itself (as
CaCl2) proved to extract too small amounts for Zn and Cu which are not measurable
accurately. Dilute acids as (0.025-0.1M HCl) has been used on acidic soils as
262

micronutrient soil tests for many years. The disadvantage of this method is to solve
such Zn and Cu forms which are not available for plants, and unsuitable for calcareous
soils.
One of the major advances in micronutrient soil testing has been the development
of extracting solutions that contain DTPA and EDTA. Chelates reduce the activity of
free metal ionsin solution through the formation of soluble metal-chelate complexes.
Free ion concentration in the soil solution is replenished from solid phases in response
to this micronutrient chelation. The quantity of micronutrient extracted by a chelate
reflects both the initial concentration in the soil solution (intensity factor) and the ability
of the soil to maintain this concentration (capacity factor), (Viets and Lindsay, 1973).
Therefore, chelating agents may simulate nutrient removal by plant roots and the
replenishment of the micronutrient from labile phases in the soil.
In this study we focus on the first step i.e. laboratory test of the extractant
selection. We chose KCl-EDTA, Lakanen Erviö extractants as official Hungarian
methods and DTPA-CaCl2 as a frequently studied extractant word-wide. Molnáros et al.
have already made similar investigations on west Hungarian soils, but they have not
involved DTPA- CaCl2 in their studies.
Our aim was to examine the relationship among Cu and Zn extracted by the three
extractants on 27 soil samples with different soil properties.
MATERIALS AND METHODS
Charactaristics of the applied extractants are represented in Table 1. 27 soil samples
from Soil Information and Monitoring System (SIMS) of Hungary were used. The soil
samples were collected from Hajdú-Bihar and Szabolcs-Szatmár-Bereg counties. The
soils varied greatly in their physical and chemical properties and included sand, loam
and clay as well. Table 2 summarizes the most important soil properties.
Extactant
KCl-EDTA
DTPA-CaCL2
Lakanen
Erviö (LE)
Table 1. Some physical and chemical characteristics of soils
Extractant
composition
0.05 M EDTA
(as Komplexon III)
0.1 M KCl
0.005 M DTPA
0.1 M CaCl2
0.1 M TEA
0.02 M EDTA
(as Komplexon II)
0.5 M NH4OAc
0.5 M CH3COOH
Soil
extractant
ratio
Shaking
time
(min)
pH Reference
1:2 120 4.36 MSZ 20135
1:2 120 7.2
Lindsay et al.
1978
1:10 0 4.65 MSZ 20135
263

Table 2. Some physical and chemical characteristics of soils
Soil
number
pHKCl KA CaCO3% Hu% Texture
1 6.37 31

LE and KCl-EDTA extracted similar amounts of Zn and Cu and CaCl2- DTPA extracted
much smaller amounts both for Zn and Cu (approximately half-third as much) than the
other two extractant. The thirst two extractant were acidic (pH 4.6 and pH 4.35
respectively), thus KCl-EDTA and LE solved such micronutrient forms e. g. Zn and Cu
oxides-hydrates, Zn and Cu carbonates which can not be solved by CaCl2- DTPA at pH
7.2.
As we expected for each extractants the extracted Cu concentrations were the
largest in case of clay, followed by loam and the smallest for sand. Zn concentration
followed the same tendency for each extractant, but here we got greater values of
averages for sand than for loam.
By he investigated methods the different soil solution ratio and pH influence the
accurate measurement of low concentrations. Extracts of KCl-EDTA with 1:2 soil
solution ratio are better measurable than LE extracts with 1:10 soil solution ratio.
DTPA-CaCl2 solution was buffered to pH=7.2, thus the extracted amounts were smaller
than that of KCl-EDTA solution.
Results of the correlation analysis show (Table 4, Table 5) that in spite of the small
number of samples the extractant concentration of the three methods vary
proportionately. Former similar results were experienced by Balla Kovács et al. Though
the extracted amounts of Cu are similar for KCl-EDTA and LE, the correlation between
these two extractants is not greater than the correlation between these and DTPA-CaCl2
according to the summarised data. When we sorted the soil samples by texture in some
cases the correlation data were better than the summarized data, but in case of Zn we
got worse correlation for loam, and in case of Cu the correlation coefficients were
smaller for sand and loam.
Classifying the soils by lime content, for calcareous soils in case of Cu correlation
coefficient between LE and KCl-EDTA was greater than between DTPA-CaCl2 and the
other two extractant. Contrasted with the other two extractants, buffered DTPA-CaCl2
do not solve Cu forms attached to carbonates, which causes the differences in the
correlation coefficients.
265

Soil
Number
Table 3. Copper and Zn extracted by three extractants for various soils
KCl-
EDTA
Zn (mg/kg) Cu (mg/kg)
LE
CaCl2-
DTPA
KCl-
EDTA
LE
CaCl2-
DTPA
Sand
1 2.02 1.52 0.51 0.62 7.45 2.11
2 0.50 0.32 0.13 2.20 0.37 0.15
3 0.58 0.51 0.20 0.84 1.12 0.37
4 1.04 1.71 0.46 2.46 3.35 1.04
5 1.86 2.66 1.17 1.64 1.86 1.07
6 2.46 2.86 1.09 4.52 5.59 1.89
7 2.42 2.92 1.37 3.82 4.72 2.31
8 6.80 13.71 2.16 4.40 9.07 2.24
Average 2.21 3.28 0.89 2.35 3.50 1.28
Scattering 2.01 4.33 0.69 1.45 2.57 0.85
CV% 90.95 132.25 78.06 61.99 73.52 66.46
Loam
9 2.02 2.92 0.46 6.90 8.45 2.06
10 1.42 1.65 0.33 4.46 5.59 1.07
11 0.84 1.84 0.34 1.34 3.23 0.77
12 0.64 2.03 0.13 4.28 6.21 2.73
13 1.58 2.73 0.90 2.40 4.84 1.54
14 1.42 2.35 0.44 2.88 5.71 1.09
15 1.18 3.93 1.37 6.16 7.70 2.01
16 2.52 2.47 0.98 6.02 6.71 2.46
17 0.70 0.70 0.47 3.92 4.47 1.89
18 2.52 2.54 0.84 7.78 9.44 2.46
19 2.82 1.90 0.63 5.14 5.22 1.91
Average 1.61 2.28 0.63 4.59 6.49 1.85
Scattering 0.77 0.82 0.36 1.97 1.99 0.65
CV% 79.86 52.59 90.65 315.95 217.56 252.62
Clay
20 2.72 3.43 1.74 4.54 5.96 2.98
21 3.12 2.22 0.47 10.56 11.18 4.62
22 1.34 1.65 0.39 5.74 9.19 2.34
23 0.92 2.16 0.27 3.46 6.83 2.21
24 5.44 6.09 2.70 19.36 11.44 6.58
25 4.44 3.81 2.25 12.9 14.04 5.47
26 8.28 7.23 3.42 12.9 13.17 4.72
27 3.44 3.55 1.71 9.68 9.44 3.48
Average 3.71 3.77 1.62 9.33 9.63 3.85
Scattering 2.37 1.97 1.16 5.55 3.35 1.73
CV% 47.88 36.06 57.58 42.92 30.64 35.03
Summarised
Average 2.41 3.02 1.00 5.60 6.75 2.35
Scattering 1.90 2.60 0.85 4.30 3.43 1.51
CV% 79.11 86.34 85.36 76.70 50.81 64.32
266

Table 4. Correlation coefficient between extracting solutions sorted by texture
Aspects of sort Extractant Lakanen-Erviö CaCl2-DTPA Summarized
n=27
KCl-EDTA
CaCl2-DTPA
0.82
0.73
0.89
Sand KCl-EDTA 0.98
0.91
Zn
n=8
Loam
CaCl2-DTPA
KCl-EDTA
0.87
0.26
0.38
n = 11 CaCl2-DTPA 0.69
Clay KCl-EDTA 0.94
0.92
n=8 CaCl2-DTPA 0.95
Summarized KCl-EDTA 0.81
0.91
n=27 CaCl2-DTPA 0.87
Sand KCl-EDTA 0.17
0.36
Cu
n=8
Loam
CaCl2-DTPA
KCl-EDTA
0.91
0.83
0.69
n = 11 CaCl2-DTPA 0.66
Clay KCl-EDTA 0.79
0.96
n=8 CaCl2-DTPA 0.82
Table 5. Correlation coefficient between extracting solutions sorted by lime content
Aspects of sort Extractant Lakanen-Erviö CaCl2-DTPA Summarized
n=27
KCl-EDTA
CaCl2-DTPA
0.82
0.73
0.89
Zn
Non-calcareous
soils n = 18
KCl-EDTA
CaCl2-DTPA 0.88
0.93
0.87
Calcareous soils
n=9
KCl-EDTA
CaCl2-DTPA 0.98
0.95
0.96
Cu
Summarized
n=27
Non-calcareous
soils n = 18
KCl-EDTA
CaCl2-DTPA
KCl-EDTA
CaCl2-DTPA 0.81
0.87
0.83
0.89
0.91
0.92
Calcareous soils
n=9
KCl-EDTA 0.86 0.69
267

SUMMARY
27 soil samples with various soil properties were extracted by three different methods,
LE and KCL-EDTA extracted nearly the same Zn and Cu amounts, while the buffered
DTPA-CaCl2 extracted only half - third of that. We suppose that LE and KCl-EDTA
solve such micronutrients forms that are not available for plants. The extracted Zn and
Cu amounts showed good correlation in most of the cases. To determine the exact
relationship between the extractants further studies are required.
REFERENCES
Andrea Balla Kovács és Sándor Berényi- Imre Vágó 2006 Comparison of 0.01M
CaCl2-0.005M DTPA and 0,1M KCl-0,01M EDTA extractants for determination
of Cu and Zn in soils in greenhouse experiment Acta Agronomica Óváriensis
Vol.47. No.1
Brennan R. F. and Best, E. 1993: Copper in Soil Analysis an Interpretation Manual
commissioned by Australian Soil and Plant Analysis Council Inc 1993 CSIRO
publishing
Lidsay, W.L., and W.AQ. Norvell, 1978. Development of DTPA soil test for zinc,
iron, manganese and copper. Soil Sci. Soc. Am. J. 42:421-428
Maftoun, M., V. Mohaselli, N. Karimian, and A. M. Ronaghi 2003: Laboratory and
Greenhouse Evaluation of Five Chemical Extractants for Estimating Available
Copper in Selected Calcareous Soils of Iran Communication in Soil Science and
Plant Analysis Vol. 34. Nos. 9-10, 1451-1463, 2003
Martens, D.C., Lindsay, W.L., 1990 Testing soils for copper, iron, manganese and
zinc in Soil testing and plant analysis , 3 rd ed. SSSA Book Series, no.3 edited by
Westerman, R. L.
McLaren, R.G., and D.V. Crawford, 1973. Studies on soil copper. I. The fractionation
of copper in soils. J. Soil Sci. 24:172-181
Molnáros, I., Gráczol, Cs., 2000 A talajok réz-, cink- és mangántartalmának
összehasonlítása KCl-EDTA, Lakanen-Erviö és töménysavas feltárással a
Talajvédelmi Információs és Monitoring Rendszer vizsgálatai alapján. Agrokémiai
és Talajtan Tom. 49. No.1-2.
Udo, E.J., H.L. Bohn, and T. C. Tuckerm, 1970 Zinc adsorption by calcareous soils.
Soil Sci. Soc. Am. J. 34:405-407
Viets, F.G. and Lindsay, W. L., 1973 Testing soils for Zn, Cu, Mn, and Fe In Soil
Testing and Plant Analysis (eds L.M. Walsh and J. D. Beaton.) pp. 162-8 (Soil
Science Society of America Inc.: Wiscosin, USA)
268

YIELD OF RYE AS AFFECTED BY THE CROPYEAR IN THE WESTSIK`S
CROP ROTATION LONG-TERM FIELD EXPERIMENT
István Henzsel – Gyulané Györgyi
University of Debrecen, Centre of Agricultural Sciences,
Research Centre Nyíregyháza
Rye is grown on different types of soils, including sandy soils low in humus, brown
forest soils and on soils with shallow fertile layer, often eroded. Early growth and
tillering is favored by a mild, rainy and long autumn, and a cool, long spring. A critical
period for rye is the time when late frosts occur in May during flowering, accompanied
by cool, rainy days (Kruppa-Szabó, 2005). Cool and rainy weather at flowering may
result in reduced seed set. In case June is too hot, grains will not develop well, which
also results in reduced yield. The best yields can be expected when rainfall is abundant
in May and the weather is cool in June (Szabó, 1992).
The objective of the present study was to determine the effects of the yearly
rainfall and temperature on the yield component of rye under different regimes of
organic manure supply.
MATERIALS AND METHODS
The study was conducted in the Westsik’s crop rotation long-term field experiment
located at the Nyíregyháza Research Center of the University of Debrecen. The crop
rotation experiment was established by Vilmos Westsik in 1929 with the overall
objective to improve the fertility of sandy soils. The crop rotation experiment offers
possibility to study different regimes of organic matter supply. The experiment consists
of 14 rotation-treatments of three-year-cycle and one treatment of four-year-cycle.
The soil of the experiment is a loose sandy soil, slightly acid, low in O.M. This
study was conducted in four rotation regimes including one with no nutrient supply, one
with straw manure treatment, one with farmyard manure treatment, and one with a
second crop green manure treatment.
The design of the selected treatments is as follows:
Sand-improvement with the periodical use of fallow (I):
Crop 1: fallow (weeds are ploughed under before flowering)
Crop 2: rye without fertilization
Crop 3: potato without fertilization
Sand-improvement with fermented straw manure (V):
Crop 1: rye with N, P and K fertilizers after the application of straw manure
fermented with calcium-ammonium-nitrate
Crop 2: potato with N, P and K fertilization
Crop 3: rye without fertilization
Sand-improvement with farmyard manure (XI):
Crop 1: fodder crop with farmyard manure, P and K fertilization
Crop 2: rye with P and K fertilization
Crop 3: potato with N fertilization
269

Sand-improvement with lupin green manure as second crop ploughed under in autumn
(XVI):
Crop 1: rye with N, P and K fertilization, followed by lupin green manure
Crop 2: potato with P and K fertilization
Crop 3: rye with N, P and K fertilization
Rye yields from the above regimes were analyzed in the years 2002, 2003, 2004, 2005
and 2006.
Sampling areas were designated in each plot before harvesting the crop.
Measurements were performed on the height of rye plants, length of the spikes, number
of spikes, thousand grain weight, and rye yield. We were seeking for possible
correlations between yield and yield components of rye, and data of rainfall and
temperature at Nyíregyháza.
RESULTS
The yearly and monthly rainfall totals at Nyíregyháza are presented in Table 1. On the
basis of the yearly rainfall, the year 2003 can be characterized as deficient in rainfall
(432 mm), while in the year 2004 there was an abundance of rainfall (704 mm). Rainfall
in the rest of the years was around the average.
The yearly and monthly mean temperature values measured at Nyíregyháza are
presented in Table 2. On the basis of the yearly mean temperature, the coolest year was
2005 (9.6 °C). Warmer than 2005 and close to each other were 2003 (9.9 °C), 2004 and
2006 (10.1 and 10.2 °C). The hottest year was 2002 (11.1 °C).
Table 1: Rainfall at Nyíregyháza (mm)
Year 2001 2002 2003 2004 2005 2006
January 8.5 32.1 43.2 21.7 12.5
February 15.0 26.0 42.0 38.5 56.4
March 35.5 4.3 79.4 8.0 59.5
April 24.8 18.0 22.9 76.8 60.4
May 62.0 35.7 51.7 60.3 91.6
June 55.5 41.2 115.9 29.9 118.1
July 76.0 54.7 99.0 84.2 10.0
August 79.0 19.8 80.5 120.7 110.9
September 64.5 66.0 53.1 43.8 44.6 5.0
October 8.0 60.5 107.1 34.8 16.5 21.0
November 32.5 29.0 22.5 55.8 29.6 18.7
December 11.0 40.0 17.5 35.5 37.0 12.0
Total (mm) 552 432 704 568 576
270

Table 2: Mean temperature at Nyíregyháza (°C)
Year 2001 2002 2003 2004 2005 2006
January -1,3 -2,8 -3,0 -0,9 -4,5
February 4,0 -5,3 0,1 -3,3 -1,9
March 7,0 3,3 5,7 2,9 3,2
April 11,2 10,3 11,6 11,3 12,0
May 20,4 19,4 14,6 16,1 15,6
June 20,1 21,1 18,9 18,6 19,3
July 23,1 21,8 21,0 21,2 22,5
August 20,6 22,4 20,3 19,8 19,4
September 14,0 14,8 15,0 15,0 16,2 16,9
October 12,4 9,1 7,9 11,2 10,2 11,1
November 1,9 5,9 6,1 4,8 2,7 5,9
December -5,2 -2,4 -0,3 0,8 0,0 2,5
Mean (°C) 11,1 9,9 10,1 9,6 10,2
Rye yield data taken in the selected crop rotation regimes are presented in Figure 1.
Crop rotation regime with no nutrient supply:
The lowest rye yield was obtained in 2002 and 2006 (1.26 t/ha and 1.27 t/ha
respectively), years with an average amount of rainfall. Higher yield was obtained in
2003, a drier year (1.56 t/ha) and in 2005, a year with an average amount of rainfall
(1.73 t/ha). The highest yield was obtained in 2004, a year with high amount of rainfall
(2.03 t/ha).
Crop rotation regime with straw manure treatment:
Also in this regime, the lowest yield was obtained in 2002 (1.74 t/ha) and in 2006 (1.74
t/ha), followed by the years 2003 (2.56 t/ha) 2005 (2.68 t/ha). The highest yield was
obtained again in 2004 (2.87 t/ha).
Crop rotation regime with farmyard manure:
The lowest rye yield was obtained in 2002 (1.93 t/ha), followed by the year 2003 (2.52
t/ha). Close to each other were 2005 and 2006 with yields of 3.13 and 3.02 t/ha,
respectively. The highest yield was obtained in 2004, with a value of 3.82 t/ha, which is
1.9 t/ha more than that obtained in 2002.
Crop rotation regime with lupin green manure as second crop ploughed under in
autumn:
The lowest yield was obtained again in 2002 (1.44 t/ha), followed by 2006 (1.96 t/ha),
2003 (2.26 t/ha) and 2005 (2.60 t/ha) similarly to regimes with no nutrient supply and
with straw manure. The highest yield was obtained in 2004 (3.93 t/ha). Also in this
regime, there was a substantial difference between the lowest and the highest yield
obtained. The yield difference between 2004 and 2004 was greater than 100 %.
271

Yield (t/ha) .
4,5
4,0
3,5
3,0
2,5
2,0
1,5
1,0
0,5
0,0
I V XI XIV
Crop rotation regime
Figure 1: Yield of rye in the Westsik’s crop rotation long-term experiment
2002 was the least favorable year for rye. The weather was unfavourable for tillering,
which resulted in a low number of spikes/m 2 . There was sufficient rain in the critical
period for flowering, but during grain development the water requirement of rye was
not met, and in addition, both periods were hotter than usual. The spikes were shorter
and the thousand grain weight was less than the average. As a result, rye yield was the
lowest of the years studied. It is also unfavourable for rye, if rainfall is in excess during
flowering and grain development, as it was the case in 2006. Even if the number of
spikes, spike length, and the thousand grain weight is around the average, yield tends to
be lower under such weather conditions.
If the weather is dry, but conditions are favourable for tillering, resulting in an
above-average number of spikes per unit area, then an average level of yield can be
expected even with shorter spikes and lower thousand grain weight, as it was the case in
2003. The highest rye yield can be expected in cases, when the weather is favourable for
tillering. When rain is abundant in autumn, there will be sufficient amount of water in
the soil after sowing, and if the weather is mild, the soil will not get frozen too early. In
this case, an above average number of spikes per unit area can be expected. It is
favourable if rain is abundant in spring and early summer, and also, if weather is cool
during flowering and grain development. This will result in longer spikes with higher
number of grains and higher thousand grain weight, which all will produce an
outstanding yield.
SUMMARY
The objective of the study was to determine the effects of the yearly rainfall and
temperature on the yield and yield components of rye under different regimes of organic
manure supply.
272
2002
2003
2004
2005
2006

It was found that either way of applying nutrients affected the height of rye. When May
is cool, the amount of rainfall has less influence on the height of rye with farmyard
manure than with other ways of nutrient supply.
The number of rye spikes varies as a function of the ways of nutrient supply. There
was a smaller variation in the number of spikes in treatments where farmyard manure
was applied as compared with the application of green manure or manuring with straw.
However, if weather conditions are unfavorable during the tillering of rye, the number
of spikes will be lower, without respect to the way of nutrient supply.
If the weather is warm and dry in spring and in early-summer there is a great
variation in the length of spikes between rotation regimes with and without nutrient
supply. In case the spring is rainy, there is no difference in the length of spikes between
rotation regimes.
If the weather is warm and dry in spring and in early-summer there is a negative
correlation between spike length and spike number per unit area irrespective to nutrient
supply. In rainy and cool weather this negative correlation may not be evident.
The thousand seed weight was not significantly affected by the method of nutrient
supply, but the effect of the cropyear was evident. Warm weather during flowering and
graining may result in a lower value of thousand seed weight as compared with values
obtained in cool weather. When the weather is cooler in May and June, with sufficient
rainfall at flowering and graining, the thousand seed weight will be high, irrespective to
the way of nutrient supply. Even when the weather is cooler in late spring and early
summer with less-than-average amount of rainfall at graining, a higher value of
thousand seed weight can be expected than in spring with average rainfall and warm
weather. When May and June are cool and rainfall is in excess of the amount that rye
requires, thousand grain weight may be depressed.
Comparing the rotation regimes it turns out that each way of nutrient supply
increased the yield of rye over the rotation regime with no nutrient. In most of the cases,
the use of farmyard manure produced higher yield than manuring with straw or using
green manure. In treatments where straw manure was used, the effect of the cropyear on
the variation of rye yield was smaller than in treatments where farmyard manure or
green manure was used. In case weather conditions are favorable for rye, an outstanding
level of yield can be attained, similarly to the use of farmyard manure.
REFERENCES
Kruppa J. – Szabó M.: Rozs és évelő rozs. In: Növénytermesztéstan 1. Gabonafélék.
2005. Szerk. Jolánkai M., Mezőgazda Kiadó, Budapest, 228-237. p.
Szabó M.: Rozs. In: Szántóföldi növénytermesztés. 1992. Szerk. Bocz E. Mezőgazda
Kiadó, Budapest. 283-292. p.
273

1
RESULT OF ALFALFA CROPPING BY THE BASIC OF TIME- AND
VILLAGE TYPE DATA FOR COUNTY OF SZABOLCS SZATMÁR BEREG
László Nagy
Nyiregyháza Research Centre of DUASC
4-6. str. Westsik V. Nyíregyháza
ABSTRACT
The volume of alfalfa (Medicago sativa L.) cropping in Hungary depends on the district
of production but mostly on the possibilities of utilizations, because this species can
growth almost all over the country , (Késmárki 2005).
In Hungary on the first place by the production area volume stands the next
counties – Jász-Nagykun-Szolnok, Békés and Hajdú independently by the type of
utilization. The importance of the counties has altered in the different periods observed,
1st. table. By the average- and the safety of grain yield excellent districs are in Tolna,
Pest, Komárom-Esztergom counties for many years. To see on the hay production, the
excellent counties are in Györ-Sopron, Vas and Baranya if we observ the intensiv
period of alfalfa production in Hungary (Nagy 1989, 1996).
The purposes of my presentation firstly are, to show the formation of the districts
and villages of lucerne production, in the second instance - the connection of the first
one with the soil background.
The task was completed with data of three different courses: the basic one is 1936-
1962; the second one is 1961-1985; the third one is 2004-2006 period.
The main results are the next: the production increased with forthy percent in the
period of 1961-1960 and 1971-1980. In the course of 1981-1985 there was no change in
the production area relative to the basic period, but decreased relative to both previous
ones (Nagy 1989, 1996). Present (last) period can also characterized by the decreasing.
The value of this is only sixthy percent relative to the basic one.
By the data of basic period at some villages there are considerable percent alfalfa
production area relative to the whole (counties) one. These are: Nyíregyháza,
Tiszavasvári, Újfehértó independently the year observed. In the instances of some
villages the percent of lucerne relative to whole villages production area is very high,
more than 7,0%. These are: Milota, Tiszadada, Szamossályi, Hermánszeg
In the instances of the so called statistical districts show hudge values by two
cases. One of them is Tiszavasvári-, the another one is Fehérgyarmat dictrict. In both
cases the value of accumulated cropping area are more, than ten percent of the countie’s
one. Observing the area concentration relative to the soil characteristics of the district
given can see, there is 0,8 correlation coefficient betwen acid soil and the value of
production area.
Key words. Alfalfa, production, long term data, soil properties
INTRODUCTION
The volume of alfalfa (Medicago sativa L.) cropping in Hungary depends on the district
of production and the possibilities of utilizations purposes.
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2
The main modification properties are until now the animal husbandary and the seed
production for abroad, marketing line.
The most important production areas by the volume for hay and seed production are the
districts of the next counties – Jász-Nagykun-Szolnok, Békés, Hajdú, and recently Pest,
1st. table.
By the level of average- and the safety of grain (seed) yield excellent districts are
in Tolna, Pest, Komárom-Esztergom counties for many years. Also excellent districts
for good level of the hay production, are Györ-Sopron, Vas and Baranya counties if we
observ the intensiv (1961-1985) period of alfalfa production in, Hungary,(Nagy 1989,
1996). It was very good effect on to the level of seed- and hay production to develop
hungarian varieties which are much better in some indxes, than foreign ones nowdays
too.
The purposes of my presentation are: to show the formation of the districts and
villages of lucerne production and the importance of the soil background.
MATERIAL AND METHODS
The basic of the paper are statistical data have been collected from the Central
Statistical Bureau proceedings, from three different periods. The first (basic) is origins
from 1936-1962 years with the data of 1936,1937, 1938, 1948 1962 years; the second
one comes from 1961-1985 (25 years long period), the third one origins from 2004-
2006 (3 years long period). The (soil) data of the statistical district belong to the
Szabolcs-Szatmár Bereg counties origins from the book of Geczy published in 1968.
The villages belong to the different districts origins from the inter net. The correlation
coefficient was accounted by the programme of excell.
RESULT AND DISCUSSION
Until now the production area of alfalfa in Hungary has changed relative to the period
of 1936-1962. In the meantime, in the period of 1961-1985, area covered by alfalfa
increased with 64%. From this time the area, until nowdays has decreased with 41,5%
relative to the basic data. Relative to 1961-1985’s data the decreasing is 64,3%.
The position of alfalfa a bit altered by the excellent plantig activity of some county,
eg. Szabocs Szatmár Bereg, betwen 2004-2006. The percent proportion of some county
in the view point of alfalfa production also has changed in this period, because, nine
county has increased the production area. In ten county has been decreasedt it, 1.
táblázat
Among the statistical district of Szabolcs - Szatmár – Bereg county there are two
with very high value in the basic period, one of them is the Tiszavasvári, the another
one is the Fehérgyarmati.
The alfalfa production value both of them is more than 10 percent. To the first one
belong nine villages, to the second one belong forthy nine.
Studying the soil properties of the statistical districts, can establish, in Szabolcs
Szatmár Bereg, the position of the acid soils relates to the proportion of lucerne
utilization. The coefficient of correlation betwen them is 0,8, 2nd table.
There are high variability in the village range data. In the basic period some village has
considerable proportion relative to the total utilzation area. More than fithy percent of
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3
the total alfalafa area belong to the forthy four villages of different statitistical districts.
The list and their proportion from the whole area of the important villages can see in the
3rd and 4th tables.
Table 1: Proportion of alfalfa production area of the counties in Hungary in different
period (by the data of CSB)
1936-1962 1961-1985 2004-2006
County %
Jász-Nagykun.-Sz. 12,3 11,9 11,2
Békés 11,3 10,7 8,1
Hajdú 8,0 8,1 9,7
Pest 6,5 7,1 9,8
Baranya 5,9 4,8 2,8
Borsod 5,8 5,8 9,1
Csongrád 5,8 6,4 6,9
Fejér 5,1 5,3 3,4
Heves 5,1 4,0 2,3
Somogy 4,9 3,8 2,4
Győr 4,8 4,0 5,6
Szabolcs-Sz.-B. 4,5 3,8 3,2
Bács-Kiskun 4,1 7,8 8,5
Tolna 3,9 5,0 3,4
Veszprém 3,6 3,0 3,9
Nográd 2,2 2,1 1,5
Vas 2,2 2,0 2,6
Komárom 2,0 2,9 3,9
Zala 1,8 1,6 1,6
Total % 100,0 100,0 100,0
Total ha 249 942 409566 146534
North Great Plain % 24,86 23,76 24,12
r 1,0 0,92 (a/b) 0,73 (a/c)
Note a b c
The most important ones belong to this group of are the next: Nyíregyháza,
Tiszavasvári, Nagyhalász, Tyukod, Ökörítófülpös, Fehérgyarmat, Dombrád,
Vásárosnamény, Nagykálló, Nyírbogát, Nyírtass.
An another group with villages Milota, Tiszadada, Szamossályi, Hermánszeg is
important because here is very high, 7-8% the proportion of the lucerne in their total
sowing area. It can be stated, 29,1 percent of the villages have 3-5% lucerne proportion,
52,6 percent have 1-3% proportion and 9,1 percent have less than 1% lucerne area
proportion in 1936-1962 period. Recent statistical data isn’t convenient for this purpose
because of the missing of wide scale data collection. Because of this fact this data base
is very importan for the people dealing with this theme in any time.
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4
Table 2: Percent of alfalfa and- the soils types in the district of Szabolcs Szatmár
Bereg county (by the data of Géczy 1968)
Soil pH properties
alfalfa Strongly alkaline saturated acid Strongly
District
alkaline
acid
%
Tiszavasvári 15,2 0,3 21,9 1,5 75,9 0,1
Fehérgyarmati 12,6 0,1 0,8 0,1 64,9 34,3
Mátészalkai 9,9 2,2 13,5 0,9 70,1 13,3
Vásárosnaményi 8,6 0,1 0,1 14,4 61,4 23,8
Nyíregyházi 8,6 2,1 14,5 28,8 54,7 0,1
Kisvárdai 8,1 3,0 12,3 14,3 57,6 12,8
Nagykállói 5,9 3,4 6,8 33,0 52,0 1,8
Csengeri 5,6 0,1 0,1 1,4 58,9 39,7
Nyírbátori 5,0 0,1 20,6 16,1 57,8 5,4
Baktalórántházi 4,8 7,5 11,6 11,1
-
58,2 11,6
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)
Note a b c d e f
Sz.Sz.B. county 100,0 1,8 10,8 13,7 60,9 12,5
Table 3: The most important villages for alfalfa production in Szabolcs Szatmár
Bereg county (by the basic of CSB 1976)
District
Villages name with highest production area on
1st 2nd 3rd 4th
place
Total
number
of
villages
Tiszavasvári Tiszavasvári Tiszadada Tiszalök Tiszaeszlár 9
Nyíregyházi Nyíregyháza Nyírpazony Kótaj Nyírtelek 9
Ibrány –
Nagyhalászi Nagyhalász Gávavencsellő Ibrány Kemecse 17
Csengeri Tyukod Porcsalma Csenger Ura 11
Mátészalkai Ököritófülpös Szamosszeg Vállaj Mérk 26
Fehérgyarmati Fehérgyarmat Tunyogmatolcs Szatmárcseke Milota 49
Kisvárdai Dombrád Fényeslitke Kisvárda Zsurk 30
Vásárosnaményi Vásárosnamény Tarpa Aranyosapáti Gulács 27
Nagykállói Nagykálló Balkány Szakoly Kállósemjén 8
Nyírbátori Nyírbogát Nyírgyulaj Nyírbátor Nyírbéltek 20
Baktalórántházi Nyírtass Apagy Nyírmada Nyíribrony 19
Total observed 11 11 11 11 225
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5
Table 4: Of the most important villages in Szabolcs Szatmár Bereg county
by the alfalfa production areas (by the basic of CSB 1976)
Production area (ha)
Sum of
District 1st 2nd 3rd 4th the area
placed villages
ha
Tiszavasvári 597,6 278,8 258,4 224,4 1359,2
Nyíregyházi
Ibrány -
623,0 66,6 66,2 65,2 821,0
Nagyhalászi 168,2 164,4 113,0 91,6 537,2
Csengeri 231,2 129,6 74,2 72,5 507,6
Mátészalkai 228,2 104,4 71,8 62,3 466,7
Fehérgyarmati 131,7 99,8 96,6 94,6 422,6
Kisvárdai 143,3 114,6 74,0 68,8 400,7
Vásárosnaményi 177,4 59,0 55,0 50,9 342,3
Nagykállói 117,2 84,6 63,0 60,6 325,4
Nyírbátori 72,0 61,2 57,7 53,8 244,8
Baktalórántházi 72,4 67,4 40,8 33,2 213,8
Total 2562,1 1230,4 970,8 878,0 5641,3
Total of the county 11239
REFERENCES:
Géczy G. 1968: Magyarország mezőgazdasági területe. Akadémiai Kiadó, Budapest.
284. p
Késmárki I. 2005. Lucerna. p:357-384. In Antal J. (szerk). Növénytermesztéstan 2.
Mezőgazda kiadó. Budapest.
KSH. 1976: Növénytermelés. 2. kötet. Községsoros adatok 1936-1962. Történeti
Statisztikai Kötetek. Budapest. KSH Könyvtár és dokumentációs Szolgálat.
Nagy L. 1989: A nagyüzemi szarvaskerep magtermesztés eredményei, 25 éves
tapasztalatok a fejlesztés lehetőségei. Vetőmag folyóírat, Budapest. November, 11-
13.
Nagy L. 1996: Szarvakerep vetőmag és takarmány termesztés fejlesztése. Kandidátusi
értekezés.129 p.
278

THE IMPORTANCE OF LONG-TERM FIELD EXPERIMENTS IN THE
QUALITY ASSURANCE OF CROP PRODUCTION
ABSTRACT
Éva Széles, Zsuzsanna Szathmáry, Péter Sipos, Dóra Hovánszki,
Csilla Uri, Zoltán Győri
University of Debrecen, Centre for Agricultural Sciences
Institute of Food Science, Quality Assurance and Microbiology
The quality assurance and traceability of food products has become a major issue of
food safety during the previous years. Long-term filed experiments can give well
applicable data for food safety and food processing.
The objectives of long-term field trials are to measure the effectiveness of different
fertilizers and types of farmyard manure in different cropping environments, and to
determine the accumulation of various elements in the soil or the plant. This is the only
way to obtain statistically reliable data about the interaction between the effect of
fertilizers and other management techniques (soil cultivation, crop density, etc.), as well
as to set the limit values for the chemical soil examination.
In this work, applicability and necessity of long-term field experiments to the food
safety through a National Long-Term Field Trials was studied.
Keywords: National Long-Term Field Trials, element correlations
INTRODUCTION
As food production is getting more and more global, the multiple sources, suppliers and
process steps along production pose a threat to the safety of the product. Therefore, the
quality assurance and traceability of food products has become a major issue of food
safety during the previous years. Participants in the food chain are expected to know the
origin of the raw materials they use in order to comply with new regulations and with
consumer expectations. Regulation EC/178/2002, known commonly as the food law of
the EU has taken up a new integrated approach, stating that the regulations for food and
feed cannot be handled separately. The “farm to fork” or “stable to table” principle
covers all sectors of the food chain, including feed production, primary production, food
processing, storage, transportation and retail sale. Hungary, as an export-oriented and
EU member country also has to meet the new EU requirements in order to be able to
produce goods that meet the requirements of other countries.
Since the requirements in the crop production sector are less strict than in animal
production and the issue of the quality assurance of feed grain crops and forages is even
more neglected, efforts are continuous in the EU to involve primary production in the
scope of food regulation. Primary production (the agricultural sector) has a critical role
in ensuring safe raw materials, since deteriorations in the raw materials cannot be
eliminated later during the processing stage. Therefore, the quality assurance of food
products should start with the certification of safe seed and cultivation techniques.
This requirement can be met by using management techniques during which the
level of the possible contaminating factors in the soil and their intake by the crop is
measured and controlled. Long-term field trials are a system of researches conducted at
279

the same plots for several years or decades. The most famous field trials are in
Rothamsted (UK), Halle (Germany) since 1878, Askov (Denmark) since 1894, Poltava
(Ukraine) since 1885 (Sárdi, 2003). The objectives of long-term field trials are to
measure the effectiveness of different fertilizers and types of farmyard manure in
different cropping environments, and to determine the accumulation of various elements
in the soil or the plant. This is the only way to obtain statistically reliable data about the
interaction between the effect of fertilizers and other management techniques (soil
cultivation, crop density, etc.), as well as to set the limit values for the chemical soil
examination (Loch-Nosticzius, 2004).
MATERIAL AND METHODS
Examined samples are from the NLTFT (National Long-Term Field Trials) established
in 1966 and the main objective was to create a uniform trial network that gives points of
reference for the national planning of nutrient demand by involving the different micro-
and macro ecological features of different cropping sites. On the other hand, it provides
a good opportunity to examine the long term effect of mineral fertilizer levels on the
quality characteristics of yield, such as its nutrient status.
The samples used are from Bicsérd, Karcag, Nagyhörcsök, Iregszemcse,
Keszthely, Hajdúböszörmény, Kompolt, Putnok and Mosonmagyaróvár research sites.
The characteristic of these sites according to their soil, agrochemical and climatic
properties was presented by Debreczeni and Debreczeniné (1994) in details. Analysed
winter wheat, maize and pea samples were produced from 1999 to 2003. Different NPK
fertilizer doses and ratios were applied in the experiment. 3 levels of nitrogen (150, 200,
250 kg/ha), 4 levels of phosphorus (200, 100 150 and 200 kg/ha) and 2 levels of
potassium (100 and 200 kg/ha) fertilizer doses were used for winter wheat and maize in
different ratios. For peas, 2 levels of nitrogen (40 and 120 kg/ha), 2 levels of
phosphorus (100 and 150 kg/ha) and 1 level of potassium (100 kg/ha) were applied in
different ratios.
The analysis of the grain samples was performed in the Institute of Food Science,
Quality Assurance and Microbiology of the University of Debrecen, Centre of
Agricultural Sciences. Determination of element contents was performed by Perkin-
Elmer Optima 3300 DV inductively coupled plasma optical emission spectrometer
(ICP-OES) after HNO3 and H2O2 digestion (Kovács et al., 1996; Kovács et al., 1998).
In plant samples undergone wet destruction by HNO3 and H2O2 we determined B,
Ca; Cr; Cu; Fe; K; Mg; Mn; Ni; P; S; Zn content in the dry matter by Perkin Elmer,
Optima 3300 DV, ICP-OES (USA).
Samples were determined by Perkin-Elmer, Optima 3300 DV inductively coupled
plasma emission spectrometer.
RESULTS AND DISCUSSIONS
In our experiment, more than 3000 pea, wheat and corn samples from the NLTFT
(National Long-Term Field Trials) from 1999 to 2003 have been analysed. The database
with over 35000 data allowed for drawing several important conclusions.
The values of the elements represent the value of the plant as raw material for food
or feed when considering the supplementation of the plant with a given element, and
280

essential to the determination of the supplementation of the plant with specific nutrients.
Furthermore, they have an important role in the extension services and quality
assurance. Table 1. shows the values with the corresponding deviations.
Table 1: Basic data for the nutrient content of peas, corn and wheat
Pea Corn Wheat
Mean Dev. Mean Dev. Mean Dev.
B mg kg -1 6.48± 0.7 2.79± 0.6 0.748± 0.3
Ca mg kg -1 1015± 193 119± 47 510± 69
Cu mg kg -1 7.56± 1.9 1.57± 0.3 5.05± 0.8
Fe mg kg -1 63.4± 20 27.8± 7 59.4± 36
K mg kg -1 9820± 852 3158± 276 3695± 354
Mg mg kg -1 1195± 107 1112± 118 1146± 118
Mn mg kg -1 12.1± 2.8 7.18± 1.4 37.1± 4.1
Ni mg kg -1 3.50± 2.8 0.88± 0.3 0.59± 0.3
P mg kg -1 3897± 774 3057± 454 3319± 529
S mg kg -1 2120± 135 1407± 179 1725± 193
Sr mg kg -1 5.09± 1.7 0.659± 0.2 2.73± 1.0
Zn mg kg -1 36.4± 14.0 15.2± 4.2 23.7± 8.2
Protein % % 27.0± 2.2 8.56± 1.1 14.6± 2.1
Figure 1: Deviation in % of the specific elements at pea, corn and wheat samples
The average and standard deviation values are resulted from the different fertilizations
and from the average of the 50-100 samples from several cropping sites. The extent of
281

the standard deviation, the % deviation represents the extreme nutrient concentrations of
a given plant, as a result of the different soil types and different nutrient supply. The
high % deviation indicates that the intake of the specific nutrient is primarily influenced
by the environmental conditions, at the same time, the small standard deviation
indicates that the intake of the element is independent of the environmental factors and
the plant is supplied with the sufficient nutrients, but the amount built-in is significantly
controlled by genetics. Figure 1 demonstrates the % deviation.
Apparently, the standard deviation of some elements, which have an important role
in the build-up of the grain is small and are hardly determined by environmental factors.
As determined by genetics, the concentration of these elements in the grain have to be
small, they might have an impact on the yield. One of them may be the parameter that
can limit the yield, according to Liebig’s minimum theory. These elements include
sulphur, potassium, magnesium, manganese, nitrogen. Calcium, boron (especially at
wheat), iron, nickel, strontium and zinc are not expected to influence the yield of the
examined plants in the soils in Hungary. However, these elements can improve the
quality of the yield, as supplied in higher amounts, their concentration in the grain will
increase. Consequently, zinc has no influence on the quantity of the yield, but supplied
in higher amount, the plant can build more zinc into the grain, thus the raw material
produced will meet food and feed requirements. If these elements are not supplied, the
consumers of the products made from this raw material will develop malnutrition
symptoms of the specific microelement.
It is important to compare the microelement content of the specific plants in the
dietetics point of view. To compare the different data, we have normalized the nutrient
content of corn and peas to that of the wheat, as shown in Figure 3.
Figure 3: Comparing the nutrient content of the crops normalized to the element
content of the wheat
282

The comparison of the nutrient content of the crops shows that the sulphur, phosphorus
and magnesium content is similar, however, several nutrients show significant
deviations. The boron and nickel content of pea, being 6-8 times higher than that of
wheat is noticeable. At the same time, the manganese content of wheat is 5 times higher
than that of pea or corn.
To determine the protein content of the grain, we used the nitrogen content. The
sulphur content of the grain is determined by amount of the amino acids containing
sulphur (cystein, cystine). The amount and quality of protein in the grain significantly
influences the quality of the yield and the nutritional and feeding value.
It is noticeable that only at wheat was the correlation between the nitrogen and sulphur
content close, as shown in Figure 4.
Figure 4: Correlation of the nitrogen and sulphur content of the crops
The amino acids containing sulphur have an important role in the build-up of the protein
structure, therefore, the analysis of the nitrogen-phosphorus ratio as regards their buildin
the grain is of great importance in our further research. The samples from the NLTFT
(National Long-Term Field Trials) are an important basis for this work.
CONCLUSIONS
Summarizing, we can conclude that the results of the analysis of the samples from the
NLTFT (National Long-Term Field Trials) helped us to identify several new relations
and supports the scientific work in the fields of plant nutrition, sustainable development,
maintenance of soil fertility and the production of healthy raw materials. Consequently,
long term field trials are essential in quality assurance, since it takes long years to
identify the optimal plants and nutrient doses in a given crop rotation and their
concentration in the grain, in order to obtain the maximum quality for a given
production purpose.
283

MORPHOLOGICAL AND AFLP VARIATION IN SOME GENOTYPES OF
POA ANGUSTIFOLIA L. AND POA HUMILIS EHRH. EX HOFFM.
Zsuzsa Lisztes-Szabó 1 , Ákos A. Zubor 2 , Béla Tóthmérész 3 ,
Mária Papp 4 , József Prokisch 2
1 Institute of Plant Sciences, University of Debrecen, H-4015, Debrecen, P. O. Box 36
2 Department of Food Science and Quality Assurance, University of Debrecen,
3 Department of Ecology, University of Debrecen, H-4015, Debrecen, P. O. Box 36
4 Department of Botany, University of Debrecen, H-4015, Debrecen, P. O. Box 36
ABSTRACT
Quantitative morphological characteristics of P. humilis and P. angustifolia individuals
collected from different habitats in Eastern Hungary were compared after growing at the
same site for three years. The individuals were also investigated using AFLP technique.
The study showed that the morphological plasticity of these species is considerable and
the quantitative morphological characteristics measured were of only limited usefulness
for taxonomic separation. There was no difference in utility between the vegetative and
reproductive features. AFLP analysis of genomic DNA showed a high degree of
polymorphism within the species. Eastern Hungarian populations of P. humilis and P.
angustifolia could not be distinguished on the basis of molecular characters using 3
AFLP primer combinations. The Poa pratensis L. species group demonstrates that in
cases of polymorphic taxa the genetic variability allows effective adaptation to slight
environmental changes.
Keywords: morphological characters, AFLP, transplantation, multivariate analysis
INTRODUCTION
Poa angustifolia L. and Poa humilis Ehrh. ex Hoffm. are among the most widespread
native grass species in Hungary and are widely used in cool season turfs and forage
grasses, with many known cultivars. P. angustifolia (or P. pratensis subsp. angustifolia)
and P. humilis belong to Poa pratensis L. aggregation. The species of this aggregation
differ in their leaf width and a few other – in some cases not obvious – morphological
features. Many individuals with intermediate morphological characters can be observed.
The morphological variability of P. pratensis genotypes is demonstrated by several
authors (Bonos et al., 2000; Frenot et al., 1999; Rua, 1996; Reader and Bonser, 1993).
The identification of the species within the P. pratensis aggregation is difficult and
accurate and reliable methods are necessary for distinguishing and characterising them.
Since the influence of environmental factors can be eliminated using molecular
approaches, the use of DNA based molecular markers is an effective method for the
separation of species. Recently, a PCR-based assay for plant DNA fingerprinting,
AFLP, has been developed which reveals significant levels of DNA polymorphism (Vos
et al., 1995).
The aim of this study is to contrast the morphological and genetic variability of P.
angustifolia and P. humilis populations collected from East Hungary to provide
additional information on the extent of genetic divergence. Individuals of P. humilis and
P. angustifolia collected from different habitats were cultivated in the same
284

environment for three years, then the morphological features of the control individuals
(which remained in the original habitat) were compared with the transplanted ones. The
study looks for answers to the following questions: (1) whether there are differences in
morphological features between the two species, excluding those used for identification
and collection, (2) whether the transplanted individuals differ from the controls in their
morphological features (3) whether reproductive features more readily reveal taxonomic
differences than do vegetative features (4) whether there is any obvious separation
between the two species - and among other Poa species - based on AFLP markers.
MATERIALS AND METHODS
Plant material: The shoot clusters for transplantation were collected in May 2003 from
sandy, sandy loam and alkaline soil pastures in Hajdú-Bihar County, Eastern Hungary
(Table 1). Two individuals of P. pratensis and an individual of P. compressa were also
included in the molecular part of this study because the genetic distance among P.
pratensis, P. angustifolia and P. humilis may give more information about their
taxonomic status.
Table 1: Origins of studied P. humilis, P. angustifolia, P. pratensis and
P. compressa populations.
Population Origin Lat. (N) Long. (E) Association
P. humilis (1) Józsa 47°34’ 21°36' Agrosti-Alopecuretum
P. humilis (2) Pallag 47°36’ 21°40' Cynodonti-Poetum
P. humilis (3) Martinka 47°35’ 21°46' Astragalo-Festucetum
P. humilis (4) Mikepércs 47°28’ 21°39' Trifolio-Lolietum
P. humilis (5) Hajdúbagos 47°25’ 21°41' Agropyro-Convolvuletum
P. angustifolia (1) Pallag 47°36’ 21°40' Cynodonti-Poetum
P. angustifolia (2) Martinka 47°35’ 21°48' Astragalo-Festucetum
P. angustifolia (3) Martinka 47°36’ 21°50' Cynodonti-Festucetum
P. angustifolia (4) Zsáka 47°09’ 21°26' Agrosti-Alopecuretum
P. pratensis (1) Bagamér 47°28’ 22°00' Caricetum acutiformis-ripariae
P. pratensis (2) Martinka 47°35’ 21°51' Caricetum vulpinae
P. compressa Mikepércs 47°29’ 21°39' Trifolio-Lolietum
Additionally, to estimate the genetic relation within the aggregation it is informative to
compare the three species with P. compressa as a distinct species outside the
aggregation.
The shoots of the 12 populations were separated to small tiller units and planted in
the Botanical Garden of University of Debrecen in the same plot. There were 10 rows in
the plot, one row for each population. A row contained 15 tillers. The distance between
tillers was about 100 mm, the width of a row is 500 mm. Plastic dividing walls of 200
285

mm depth were sinked into the soil between the rows to prevented spreading of stolons
to the neighbouring rows.
Morphometrical analysis: In May 2006, shoots were collected from the labelled shoot
clusters of the original habitats (controls) and others were collected from the
experimental plot. The following characteristics of the vegetative organs were
measured: (1) height at flowering, (2) lengths of the longest leaf-blade of lateral tillers,
(3) maximum widths of these leaves, (4) length of the flag leaf-blade, (5) and the flag
leaf sheath, (6) width of the flag leaf-blade, (7) length of the ligule of the flag leaves.
The following characters of the reproductive organs were measured: (8) length of the
panicle, (9) length of the longest panicle branch, (10) length of the spikelets from the
end of the lowest panicle branches, (11) length of the lower and (12) upper glume, (13)
length of the palea, (14) length of the lemma. We measured 15 individuals from each
population. The average of 10 data per individual was used for the following features: 2,
11, 12, 13 and 14. A slide-gauge was used for measuring.
AFLP method: For AFLP, each sample consisted of about one leaf per plant for ten
plants per population. ZenoGene kit (ZenonBio Ltd, Szeged, Hungary) was used for
DNA extraction following manufacturer’s instructions. Extracted DNA samples were
stored at –20 °C until used. After the AFLP procedure (K. Szabó et al., 2006) the final
amplified and selected PCR fragments were appeared by capillary electrophoresis with
ABI Prism 3100 Genetic Analyzer in fragment analysis mode at the Agricultural
Biotechnology Centre (Gödöllő, Hungary) using 360 mm length capillaries and POP-4
gel. GeneScan HD-400 Rox Size was applied as standard marker. AFLP fragments were
identified by GeneScan 3.7 computer program.
Data analysis: Morphological data were analysed by principal component analysis
using the R statistical program (R Development Core Team, 2005). For cluster analysis
of AFLP data each detected band above 80 threshold limit was scored as present (1) or
absent (0) in all samples. The unweighted pair group method using arithmetic means
(UPGMA) was used and a dendrogram was constructed based on Jaccard’s similarity
coefficient (Jaccard, 1908) by the statistical software package SPSS 11.0 for Windows
(SPSS Inc., USA). Genetic diversity and significant differences in the populations were
estimated by the proportion of polymorphic loci. The allele frequencies and
heterozygosity were estimated based on square root of recessive genotypes. These
estimates and Nei-genetical distances (1978) were analysed by TFPGA (Miller and
Mark, 1997).
RESULTS
Morphometrical analysis: We did not observe a sharp separation on the PCA biplot
diagram based on all features of the two species (Fig. 1). Clusters of the individuals of
the two species show complete overlap. However, the variables show unambiguously
two different tendencies, depending on whether they are vegetative (v1-v8, v14), or
reproductive (v9-v13) characteristics.
286

PC 2
5
4
3
2
1
0
-1
-2
-3
-4
V2
V5
V4
V10
V11
V14
V9
V1
V6
V12 V13
V7
-6 -4 -2 0 2 4 6
PC 1
Figure 1: PCA biplot of the two species based on all variables.
(1-8, 14 vegetative, 9-13 reproductive variables.) □: transplanted P. angustifolia, ○:
transplanted P. humilis, �: control P. angustifolia, �: control P. humilis. v1: height,
v2: panicle length, v3: panicle width, v4: flag leaf sheath length, v5: flag leaf blade
length, v6: ligule length of the flag leaves, v7: the widest width of leaves, v8: flag leaf
blade width, v9: spikelet length, v10: upper glume length, v11: lower glume length,
v12: palea length, v13: lemma length, v14: longest leaf blade lengths of lateral tillers.
There was no separation based on the features of reproductive organs (Fig. 1). However,
the individuals were concentrated into two, partly overlapping groups based on the
characteristics of their vegetative organs. The first is the group of transplanted
individuals which have larger vegetative organs (Fig. 1). The second is the group of
individuals with smaller organs which were collected from the original turf clones. An
explanation for this is that in the experimental environment the interspecific competition
was suppressed as seedlings of other species were eliminated and the transplanted plants
were able to spread and occupy all of the available area.
The PCA results separated the transplanted and control populations of P.
angustifolia into two groups but the transplanted and control populations of P. humilis
did not show such separation (Fig. 1). The quantitative morphological characteristics of
P. angustifolia responded more sensitively to the transplantation.
Neither species nor transplanted populations were separated based on the
reproductive characters. The two species did not show differences considering the
vegetative characteristics, but the transplanted and control individuals of P. angustifolia
separated slightly.
In the AFLP analysis three primer combinations were used and generated clear banding
patterns. A total of 338 DNA fragments were produced across 12 genotypes of four Poa
287

species (plus P. pratensis and P. compressa) with 333 fragments being polymorphic
(98.52% polymorphism) (Table 2). A representative example of the amplification
products obtained at P. angustifolia genotypes using the primer combination EcoRI-
ACC+Tru1I-CAG is shown in Fig. 5. The number of bands from each primer
combinations ranged from 84 (EcoRI-ACC+Tru1I-CAA) to 159 (EcoRI-ACC+Tru1I-
CAG). In the present study, a wide variation in the number of polymorphic bands (83-
157) and the percentages of polymorphic bands within a primer combination (98.80-
98.74 %) were observed. The mean polymorphic rate was 98.47 %.
Table 2: The number of bands and degree of polymorphism revealed
by AFLP primer combinations.
Primer combinations Total bands Polymorphic bands Polymorphism rate (%)
EcoRI-ACC+Tru1I-CAG 159 157 98.74
EcoRI-ACC+Tru1I-CAC 95 93 97.89
EcoRI-ACC+Tru1I-CAA 84 83 98.80
Total 338 333 98.52
A presence (1) or absence (0) binary data matrix containing 333 polymorphic AFLP
fragments was used to generate the genetic similarity estimates. The Nei-genetical
similarity, between 0.87 and 0.95, shows remarkable genetic variation among the
studied Poa populations. At the same time, the species of the P. pratensis group are
close to each other, only P. compressa shows some separation. Inside the group, the
similarity is the biggest between P. pratensis and P. humilis, and the smallest between
P. pratenis and P. angustifolia.
UPGMA clustering analysis was carried out in order to demonstrate graphically the
genetic similarities among the Poa genotypes (Fig. 2), to present a dendrogram of 338
AFLP fragments obtained from three primer combinations at 12 Poa populations. P.
humilis (2) is separated at a high level from other populations. P. humilis (1) and P.
humilis (3) are close to each other. P. angustifolia (2) and P. angustifolia (3) is similar,
as are P. angustifolia (1) and P. angustifolia (4). But the populations of different species
are not separated from each other as would be expected. The two populations of P.
pratensis are in different clusters.
288

Figure 2: Genetic similarity among Poa genotypes revealed by UPGMA cluster
analysis based on AFLP data.
The heterozygosity is bigger (25.6 %) in P. humilis than in P. angustifolia (23.4 %).
The heterozygosity and polymorphism in P. humilis populations are bigger than P.
angustifolia populations (Table 3).
Table 3: Mean heterozygosity and polymorphism rate in populations of P.
angustifolia and P. humilis.
Heterozygosity (%) Polymorphism (%)
P. humilis 25.60 94.40
P. angustifolia 23.40 83.80
The mean theta value of 0.2581 (0.2307 lower limit and 0.2827 upper limit) does not
mean significant differences among the populations of studied species with 95%
confidence interval and 1000 bootstrap repeat.
Populations were found to be genetically similar and there was a wide range of genetic
variability among germplasm sources. The different leaf blade width of the tillers of the
two species is apparent, but sharp separation cannot be seen between the two species in
the clusters on the basis of AFLP markers. This questions the value of the leaf blade
width as a taxonomic character. The species level difference is ambiguous between the
four species.
289

DISCUSSION
The species level segregation of the two observed grass species is not supported by the
morphological features measured. Rua (1996) reported similar results, as the
quantitative morphological features did not reveal sharp statistical segregation between
some Poa species. In our study the basis of the identification was the width of the leaf
blades whilst other features, such as the length values, also used in identification keys,
did not prove to be useful.
Our previous observations suggested that the habitat adaptation, due to the habitat
variations, destroyed the sharp border in morphological features between the two
species. The results of the present study also supported that there was no phenotypic gap
between these species, based on the measured characteristics. After three years growing
in the same plot, tillers showed differences in morphological characteristics compared
to individuals of original habitats. In some cases it seems there are not correspondences
between morphological characters and genotypes (McElroy, et al. 2002; Gillespie and
Boles, 2001). These observations are consistent with the results of our study.
The present study confirms the previous results that behind the habitat-sensitive
features of grass shoots there is high within-species genetic variability. This could be
due to a relatively high proportion of potentially sexually produced off-types (Larson et
al., 2001; Curley and Jung, 2004). AFLP variation of population in Poa pratensis L.
aggregation does not show species level separation between the studied species based
on the generated AFLP markers. The Poa pratensis L. group serves as an example of
the fact that in cases of polymorphic taxa the genetic variability results in adaptation to
small environmental changes and that separable ecotypes exist. Thus, this study
provides further evidence for the utility of AFLP for turfgrass genetics as well as
important genetic information for turf breeders and managers.
Our study showed that the morphological plasticity of our species was
considerable, which was related to the small scale variability of the habitats. Thus, the
measured quantitative morphological characters were not useful for taxonomic
separation. There was no difference in the usefulness of the vegetative and generative
features considering for the separation of the species.
The study of AFLP variation of populations of the two species suggests complex
population structures with high levels of introgression. Such results are consistent with
the morphological variation seen in genera Poa generally. The high degree of
polymorphism in Eastern Hungarian populations resulted in continuous forms from
apparently pure P. humilis to apparently pure P. angustifolia. The transplantation
experiment showed that, despite considerable phenotypic plasticity, the morphology is
determined by genotype (Howland et al., 1995). Poa taxa have high genetic potential
for microevolutionary development.
REFERENCES
Bonos, S. A., Meyer, W. A., Murphy, J. A. (2000): Classification of Kentucky
bluegrass genotypes grown as spaces-plants. Hortscience 35 (5): 910-913.
Curley, J., Jung, G. (2004): RAPD-based genetic relationships in Kentucky bluegrass.
Crop Sciences 44: 1299-1306.
290

Frenot, Y., Aubry, M., Misset, M. T., Gloaguen, J. C., Gourret, J. P., Lebouvier,
M. (1999): Phenotypic plasticity and genetic diversity in Poa annua L. (Poaceae)
at Crozet and Kerguelen Islands (subantarctic). Polar Biology 22 (5): 302-310.
Gillespie, L. J., Boles, R. (2001): Phylogenetic relationships and infraspecific variation
in Canadian Arctic Poa based on chloroplast DNA restriction site data. Canadian
Journal of Botany 79: 679-701.
Howland, D. E:, Oliver R. P., Davy, A. J. (1995): Morphological and molecular
variation in natural populations of Betula. New Phytol. 130: 117-124.
K. Szabó, Zs.; B. Tóthmérész, Á. A. Zubor, M. Papp, Z. Győri, J. Prokisch (2006):
AFLP variation in Poa pratensis L. agg. in Eastern Hungary. In: Proceedings of
41st Croatian and 1st <strong>International</strong> Symposium on Agriculture, ISBN 953-6331-
39-X, p. 225-226.
Larson, S. R., Waldron, B. L., Monsen, S. B., St. John, L., Palazzo, A. J.,
Mccracken, C. L., Harrison, R. D. (2001): AFLP variation in agamospermous
and dioecious bluegrasses of Westers North America. Crop Sciences 41: 1300-
1305.
McElroy, J. S., Walker, R. H., van Santen, E. (2002): Patterns of variation in Poa
annua populations as revealed by canonical discriminant analysis of life history
traits. Crop Science 42 (2): 513-517.
Miller, Mark P. (1997): Tools for population genetic analysis (TFPGA) 1.3: A
Windows program for the analysis of allozyme and molecular population genetic
data. Computer software distributed by author.
R Development Core Team (2005): R: A language and environment for statistical
computing. R Foundation for Statistical Computing, Vienna, Austria. ISBN 3-
900051-07-0, URL http://www.R-project.org.http://her.bio.nau.edu/miller/tfpga.htm.
Reader, R. J., Bonser, S. P. (1993): Control of plant frequency on an environmental
gradient – effects of abiotic variables, neighbors, and predators on Poa pratensis
and Poa compressa (Gramineae). Canadian Journal of Botany - Revue Canadienne
de Botanique 71 (4): 592-597.
Rua, G. H. (1996): The genus Poa in Patagonia: a phenetic analysis of species of Poa
subgenus Poa. Botanical Journal of the Linnean Society 121 (3) 229-241.
Vos, P., Hogers, R., Bleeker, M., Reijans, M., van de Lee, T., Hornes, M., Frijters,
A., Pot, L., Peleman, J., Kuiper, M., Zabeau, M. (1995): AFLP: A new
technique for DNA fingerprinting. Nucleic Acids Res. 23: 4407-4414.
291

THE EFFECT OF SOME FOLIAR COMPOSITIONS ON THE PRODUCTION, NUMBER OF
INFLORESCENCES AND FRUIT IN THE TECHNOLOGY OF CULTIVATING
TOMATO IN THE FIELD
Popa Alina-Grigoriţa
University of Oradea, Faculty of Enviromental Protection
ABSTRACT
The experiments proved the influence certain foliar compositions have on the production of
tomato fruit, the number of inflorescences and fruit on the plant in the technology of
cultivating tomato in the field, with an organic-mineral fertilization applied on the soil
Key words: Foliar compositions, nutritive elements, production, number of inflorescences,
number of tomato fruit.
INTRODUCTION
Compared to other horticultural soils (especially vine and tree growing), the soils cultivated
with vegetables in the field are usually more fertile. This initial fertility must be maintained
and developed in order to sustain the intensive character of the technology and nutrition of
tomato culture.
The real needs of the field tomatoes towards the main nutritive elements (Cs= specific
consumption N, P, K kg/ t product) are very important in the fertilization system of field
tomatoes, the relations between the determining nutrients and the total quantity of nutritive
elements “exported” from the soil together with the crops, at the area unit (Cg= global
consumption N, P, K kg/ ha).
As regards the specific and global consumptions, the tomatoes take part in the group of
species with an average consumption. The fertilization systems must ensure a management
of the nutrients, which, by sorts, doses and methods, must determine the accomplishment of
great quality productions (Rusu M. et. al, 2005).
Taking into account the intensive character of the technology of tomato culture,
according to the fertilization on soil, it was studied the effect of some foliar compositions in
order to finish some systems of fertilizing tomato, based on the positive interaction of the
fertilization on soil to the foliar one; the latter having an integrated character.
MATERIAL AND RESEARCH METHOD
The field culture was initiated on argic faeoziom soil.
The experiments were carried out in the year 2003, in locality Oradea, using Unirea soil.
The experiment was done with the blocks method, in 3 repetitions, the area of a variant
being 6, 4 mp.
The effect of 4 foliar fertilizers was studied, and the witness was showered with water.
292

Three foliar fertilizations were done: the first treatment at the first inflorescence and the
following ones at an interval of 14 days.
The effect of the foliar fertilizers on the production of tomato fruit and on the number of
inflorescence and fruit on the plant was carried out.
The foliar compositions sorts were applied on an agrochemical background resulted from
the interaction of the organic fertilization (50 t/ ha semi fermented manure) to the complex
application of the mineral manure (N120P120K120).
RESULTS AND DISCUSSIONS
It can be noticed in table 1 that significant results were obtained at all foliar fertilized
variants, particularly at the variant fertilized with Bionutrifag F (table no.1, figure no.1).
Table 1 The effect of the foliar fertilization upon the production in the crop of tomatoes
(UNIREA) from the field (Oradea, 2003)
No.
var.
Foliar assortment
Production
Concentration
of the
solution t/ha Difference
Significance of
difference
1 Martor stropit cu apă - 35,82 - -
2 Nutrifag 1% 41,50 5,68 ***
3 Bionutrifag F 1% 44,60 8,78 ***
4 Ferticare 24-8-16 1% 42,10 6,28 ***
5 Polyfeed 19-19-19 1% 42,16 6,34 ***
DL (5%) 1,74 DL (1%)
2,53 DL (0,1%) 3,79
293

Production t/ha
50
45
40
35
30
25
20
15
10
5
0
Control sprinkled
with water
Nutrifag Bionutrifag F Ferticare 24-8-16 Polyfeed 19-19-19
Foliar assortment
Producţia t/ha
Figure 1 The effect of the foliar fertilization upon the production in the crop of tomatoes
(UNIREA) from the field (Oradea, 2003)
The significant results of the application of foliar manure on the tomato field culture are due
to an organic mineral under layer and to its recovery by an extra-radicular application of
manure as phase/additional fertilizations (three treatments at an interval of 14 days).
In addition, the foliar fertilizations have determined a greater number of inflorescences and
fruit on the plant, fact that prolonged the period of fructification and cropping (table no. 2).
Tabel 2 The influence of foliar fertilization upon the number of inflorescences and plant
fruits
No.
var.
Foliar assortment
Number of
inflorescences
Număr fructe
number of fruit
1 Martor stropit cu apă 12,1 36,3
2 Nutrifag 13,0 39
3 Bionutrifag F 16,0 48
4 Ferticare 24-8-16 14,7 44,1
5 Polyfeed 19-19-19 15,2 45,6
From the efficient foliar sorts, in the first place Bionutrifag F and Polyfeed 19-19-19 are
focused at the field tomato culture, because they contain a balanced representation of
nutrition elements and a superior recovery of the fertilization on soil.
294

CONCLUSIONS
The rigorous experimentation of the foliar manure, with an intensive technology for
tomatoes, shows that these compositions with extra radicular application can be framed in
the context of fertilizing on soil, as a measure of an “integrated” character. In the intensiveconventional
fertilizing technologies, the foliar fertilization contributes more intensely at the
productive recovery, of the quantity and quality of fruit, due to the nutrients’ contribution on
the soil.
The field cultivated tomatoes, on an agrochemical organic-mineral background (50t/ha
manure + N120P120K120), present significant production increases at the applied foliar
sorts and in the first place at the complex product Bionutrifag F (with macro- micro
elements and biological active substances). These effects are due especially to the
accomplishment of some productivity elements (number of inflorescences, number of fruit),
and also to prolonging the ripeness and cropping.
REFERENCES
1. Alina Grigoriţa Popa, 2007, Optimizarea agrochimică a sistemului sol-plantă în
tehnologia de cultivare în spaţii protejate a tomatelor, Teză de doctorat, USAMV Cluj-
Napoca, 168-170;
2. Mihai Rusu, Marilena Mărghitaş, Ioan Oroian, Tania Mihăiescu, Adelina Dumitraş,
2005, Tratat de Agrochimie, Ed. Ceres, Bucureşti, 80-82, 569-572.
295

METHODS AND MEASURES FOR AN AGROCHEMICAL OPTIMIZATION OF THE GREENHOUSE
SOIL AT THE TOMATOES CULTURE
Popa Alina Grigoriţa
University of Oradea, Faculty of Enviromental Protection
ABSTRACT
The suggested subject represents a synthesis of the methods for agrochemical optimization
of greenhouse soil cultivated with tomatoes, included in the doctorate thesis called: Agro-
Chemical Optimization of the Soil-Plant System in the Protected Space Cultivation
Technologies of the Tomatoes
The optimization of the content of greenhouse soil cultivated with tomatoes in organic
matter constitutes an essential objective, because an optimized agrochemical support
prevents and reduces some disturbing factors as: salinity-salinization, acidity-acidification,
toxicity of other ions, polluting effects.
Key words: Agrochemical optimization, organic matter, soil reaction, soluble salts,
nutritive elements.
1./ The optimization of the organic matter content from the greenhouse soil at
tomatoes culture
The agrochemical optimization of soils implies a concomitant solving of problems related to
correcting the extreme reaction states and the requests of fertilization
The system of vegetables nutrition in protected areas differs fundamentally from the
vegetables nutrition and fertilization on the field because of certain particularities such as:
� The intensive growth of the vegetal mass, influenced by the environment factors
(temperature, humidity);
� The relation between the aerial part and roots is in great favor of the aerial part;
� Using the soil mixtures with an increased content of organic matter and nutritive
elements, with very diverse physical-chemical characteristics;
� The immobilization of some ions by the organic matter (bore, cuprum, manganese);
� The intense microbiological activity in the soil that favors the development of some
pathogen agents, imposes the taking of some prevention and control measures;
� The possibility to control the environment factors and soil;
� Difficulties in ensuring a nutritive balance in the soil, according to the plant’s age and
metabolism.
Because of the intensive character of the system of greenhouse culture, the application of
organic manure on the basic fertilization is necessary, for obtaining an optimum content of
organic matter in soil, which in fact is the main nutritive support of the culture (Tisdale S.,
1993).
For the greenhouse culture of tomatoes, decomposed manure is used and applied on the
basic fertilization, with about 2 weeks before the seed planting. Because of the presence of
296

some pathogen agents, a chemical and thermic disinfection is necessary, with hot vapors
(150 o C), at 90 o -95 o C, for 6-7 hours, at 30cm deep.
The manure doses are 80-100t/ha once a year in greenhouses with older cultures and
twice a year (cycle I and cycle II) in new greenhouses (Davidescu D. and Velicica
Davidescu, 1992; Ciofu et al, 2003).
Composted birds manure can be also applied with 3 months in advance in doses of 15-
25t/ha and forestry composts from resins bark or beech (Rusu, 1993).
In greenhouse conditions, the mobility of nutritive elements from the organic manure is
more intense than at the same culture in the field, thus considering that for each 100t manure
in a vegetation cycle in the soil it is liberated in average: 250-300 kg N, 120-170 kg P2O5
and 350-450 kg K2O (after Davidescu D., 1992, cited by Avarvarei, 1997).
The technological steps for optimizing the content of organic matter (MO) is a
compulsory request for this technology, so that every interpretation and practical measure
takes into account this value of the organic component. An optimized agrochemical support
solves the nutrition effects and prevents or diminishes some disturbing factors (salinitysalinization,
acidity-acidification, toxicity of other ions, polluting effects) (Rusu, 1968,
1988).
2./ The agrochemical optimization of the reaction state and of the content of soluble
salts in the soil from the greenhouse at tomato culture
As regards the optimization of the content of soil in nutritive elements, the concentrations of
the nutritive ions have to be guided towards the levels of optimum sufficiency in soil, after
fertilizations.
The greenhouse soils are frequently prone to salinization and alkalization, following
the accumulation of soluble salts obtained from the fertilization with organic and chemical
manure, irrigation water, water from the phreatic layer and the process of chemical
alteration of the soil mineral fraction.
The soil disinfection using vapors in greenhouses has the effect of increasing
salinization due to the acceleration of the transforming process of the ammoniac azoth
(NH +
4 ), which inhibits the activity of nitrification bacteria. These two processes lead to a
temporary accumulation of NO2 (Davidescu D. and Velicica Davidescu, 1992).
Heavy irrigation water, as well as the superficial phreatic layer, constitutes a source of
enriching the surface soil layer with mineral salts (bicarbonate, carbonate, chlor, sodium
sulphate, magnesium).
Sweet irrigation waters (rain water) diminish the salinization effect with the dilution of
the soluble salts.
For this reason, shortly after being in use, the reaction of the greenhouse soil tends to
become neuter-alkaline or even alkaline, fact that has negative influences on the cultures
development.
The application of the amendments is imposed when the value of pH exceeds 8.5.
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The total content of soluble salts has to be adjusted and a correct system of soil fertilization
has to be established, for the greenhouse soil to correspond from the agrochemical point of
view (Neaţă Gabriela, 2002).
As regards the salinity tolerance it is well known that tomatoes do not support
concentrations over 0.17% in light soils, over 0.20% in medium soils and over 0.25% in
compact soils (Maria Apahideanu, 1998).
The salinity tolerance of the main greenhouse cultures is appreciated on the limits of the
concentrations (table 1):
Culture
Table 1 Salinity tolerance of the main greenhouse cultures
(mineral residue, g/100g sol) (after Bernstein, 1987 cited by Rusu, 1993)
The limit where the culture starts to
be affected
Fine
texture
Thick
texture
Middle
texture
The limit where the production
decreases with 50 %
Thick
texture
Middle
texture
Fine
texture
1. Cucumbers 0,15 0,20 0,23 0,25 0,30 0,35
2. Green
peppers
0,15 0,18 0,20 0,23 0,30 0,43
3. Lettuce 0,15 0,18 0,20 0,23 0,30 0,43
4. Cabbage 0,25 0,20 0,23 0,26 0,40 0,55
5. Tomatoes 0,27 0,20 0,25 0,30 0,45 0,60
6. Spinach 0,20 0,25 0,30 0,30 0,45 0,60
The prevention of the exaggerated increase of the salts content is solved by taking some
preventive and curative measures:
� Applying fertilizers in best economic doses, especially those with azoth;
� Using some fertilizers with well balanced relations between the nutritive elements
depending on the requests of the species with the needed reports;
� Application of extra-radicular fertilization during the vegetation period as a measure of
diminishing the dose applied on the soil;
298

� The pH decrease in the greenhouse soil can be obtained by using oligotrophic peat
(highly acid, with pH 3-4,5 and Ah of 90m.e./100g peat) or mesotrophic peat (with pH
5-5,5 and Ah of 50-60m.e./100g peat)(Rusu, 1982);
� Washing the soluble salts by irrigations, when the soil is permeable and the phreatic
layer is under the critical level. In the greenhouse soils with difficult draining where
prophylactic washings cannot be done, the low mineralized oligotrophic peat, which
contains 90-95% organic matter, is applied;
� Correcting the pH with gyps, when it exceeds the value of 8,2-8,4 and the PSA value is
greater than 10% (Rusu, 1993);
� On the soils with a low content of mineral azoth it is necessary to apply 0,25-0,50 kg
azoth for every ton of applied peat, in order to prevent a temporary immobilization of
the soluble azoth.
Depending on the applied doses and knowing the contribution of the peat’s nutritive
elements, it can be calculated the quantities of mineral fertilizers that have to be applied to
ensure the plants’ needs regarding the adaptation to different concentrations of salts.
REFERENCE
1. AVARVAREI I., VELICICA DAVIDESCU, R. MOCANU, M.GOIAN, C. CARAMETE, M. RUSU,
1997, Agrochimie, Ed. Sitech, Craiova;
2. DAVIDESCU D., VELICICA DAVIDESCU, 1992, Agrochimie horticolă, Ed.Academiei
Române, Bucureşti, 91-497;
3. MARIA APAHIDEAN, 1998, Contribuţii la stabilirea unei metode noi de cultură, “fără
sol”, a tomatelor în seră, Teză de doctorat, Cluj-Napoca;
4. R. CIOFU, NISTOR STAN, VICTOR POPESCU, PELAGHIA CHILOM, SILVIU APAHIDEAN,
ARSENIE HORGOŞ, VIOREL BERAR, KARL FRITZ LAUER, NICOLAE ATANASIU, 2004,
Tratat de legumicultură, Ed. CERES, Bucureşti, 129-136, 617-643;
5. RUSU M, V. MUNTEANU, N. METEŞ, J. JANCU, 1988, Probleme ale optimizării
agrochimice a solurilor sub influenţa măsurilor agrochimice, Analele ICCPT, vol.LVI,
261-267;
6. RUSU M., I. BOERIU, I. VLĂDUŢU, 1968, Potenţialele activităţii ale unor ioni în solurile
acide calcarizate, Stiinţa Solului, nr.4, vol.VI, 26-32;
7. RUSU M., V. MUNTEANU, I. IANCU, DANIELA DAVID, ALEXANDRA IONESCU, MARIA
FLOREA, ANA STANCIU, 1982, Cercetări privind regimul microelementelor din sol în
interacţiune cu îngrăşămintele şi amendamentele, Analele ICCPT, vol.XLIX, 121, 130;
8. RUSU M., V. MUNTEANU, I. IANCU, MARILENA MĂRGHITAŞ, S. ALB, 1993, Efectul
magneziului pe solurile modificate agrochimic, Bul. USAMV, A-H, Cluj-Napoca, 47/2,
117-124;
9. TISDALE S.L., 1993, Soil fertility and fertilizers, Macmillan Publ.Co. New-York, USA.
299

THE INFLUENCE OF THE SEED’ BIOLOGICAL LINKS ON THE
WHEAT PRODUCTION QUALITY
Mariana Popovici
University of Oradea, Faculty for Environmental Protection
ABSTRACT
The quality of wheat production is a complex characteristic, depending on both quantitative
elements (content of useful substances: proteins, fats, carbohydrates etc.) and on their
chemical composition (essential aminoacids, saturated and non-saturated fat acids, etc.) but
also on quantitative elements (physical, biochemical, mechanical and technological
characteristics of the final products).
INTRODUCTION
The quality characteristic of the wheat is much more complex than the production capacity
and it is influenced by several factors having a specific contribution in its completion; if one
adds the influence of the environmental factors and the modifications that occur in the
technological process one may complete this overall picture. Thus, the wheat quality cannot
be expressed in terms depending on a single trait but it depends on numerous characteristics,
each having its precise importance in assessing the value of the final product.
The main way through which one can take action on the quality and substance of the
wheat proteins consists in their amelioration, all these characteristics being genetically
controlled. The agricultural researches and practice showed that even the technological
factors are making their contribution to largely impact on the content and the amount of
protein production per hectare. Among these factors, one can place first the chemical
fertilisers, especially those based on azote (nitrogen) which condition the aminoacids and
proteins biosynthesis and, implicitly, the harvest’ quality. An important role is detained by
the crops’ rotation, irrigations, soil workings, which all compete to increase the
efficaciousness of the fertilizers appliance.
Within the performed experiments, the indices for quality of the autumns wheat are
been showed on the basis of the analysis regarding the protein content in gluten.
MATERIAL AND METHOD
Among the wheat quality indices, there have been analysed the following: the protein
content of the B,SC1,SC2 biological links and other “replications” during the 2004-2005 and
2005-2006 periods and the amount of gluten present in the biological links of “Dropia”
autumn wheat, employed in the 2004-2006 two-year research period within the premises of
the Leş production farm, Bihor County.
300

RESULTS AND DISSCUSSIONS
1. The protein content influence of the biological links.
The protein content of the wheat grains is a hereditary trait, strongly influenced both by the
environmental conditions and by the crop ones. The research carried-out clearly shows the
strong reaction of the protein content function of the ecological circumstances, the variation
amplitude from one location to another and, especially, from one year to another is reaching
5 up to 7%.
Within the framework of the carried-out experiences, there have were carried out
analysis on the protein content of the B,SC1,SC2 biological links and other “replications”
and the results achieved are presented in the table 1 bellow.
Table1: The impact of the biological links of wheat seed on the protein content of grains
Biological 2004-2006 2005-2006 2004-2006 Average
category/class Protein % Protein % Protein %
Base (B) mt 12.3 100.0 11.8 100.0 12.1 100.0
SC1 12.0 97.6 11.8 100.0 11.9 98.3
SC2
Other
11.2 91.1 11.3 95.7 11.3 93.3
replications I 8.4 70.7 000
7.3 61.8 000
Other
7.9 65.3
replications II 7.1 57.7 000
7.3 61.8 000
7.2 59.5
DL 5% 5.3 10.4
DL 1% 14.4 21.5
DL0.1% 25.4 30.4
According with the obtained data, two main aspects are mainly emphasised, namely:
� The biological link of the seed puts its mark on the protein content. The differences
between the biological links obtained and maintained through a conservative
amelioration process (B,SC1,SC2) are not significant ones, as they are maintained
within some high-levelled parameters during the two-year research period. An evident
depreciation can be observed in the case of the “other replications” in which case, the
protein percentage diminishes to the statistically ensured quotas, but which are negative
in comparison with the Base (Mt) biological link. The striking differences are due to
the lack of selection pressure exercised on this biological material, the biological nonpurified
amount thus negatively marks the wheat grains’ quality.
� The differences, although less relevant, show the influence of the climatic year on the
protein percentage, soil factors making their impact on the wheat harvest’ quality.
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2. The influence of the biological links of the autumn wheat’ seeds on the gluten
content.
Among the component traits of the bread-making quality, a key-role is detained by the
protein quantity present in the grain, especially of those proteins components of the gluten.
The efficaciousness (force) of dough, the water absorption’ capacity and the bread tolerance
to obsolescence, all largely rely on these latest proteins component.
The bread volume depends in an essential manner on the gluten quality which, unlike
the quality, is on the first hand genetically established, being very little influenced by the
environmental conditions. At last, the quality is influenced by the bio-chemical properties of
the glutenins, main components of the gluten.
The gluten has some properties which confer to it a unique position among the vegetal
proteins, namely: cohesion, elasticity and extensibility in hydrated state. As a consequence,
the (wheat) flour characteristics are, on the first hand, a function of gluten and of its
behaviour in aqueous environment. The gluten analysis supposes an assessment of its
content and its quality.
The data on gluten amount contained in the biological links of the
(“Dropia”) autumn type of wheat for the 2004-2006 two-year period are
being presented in the table 2 bellow.
Table 2: The impact of the biological links of wheat seed on the gluten content of the
grains
Biological 2004-2006 2005-2006 2004-2006 Average
category/class Gluten % Gluten % Gluten %
Base (B) mt 28.0 100.0 26.8 100.0 27.4 100.0
SC1 28.0 100.0 26.0 97.0 27.0 98.5
SC2 27.5 98.2 26.0 97.0 26.8 97.8
Other
replications I 21.3 76.1 000
19.9
74.2 000
20.6 75.2
Other
replications II 20.4 72.8 000
19.2
71.6 000
19.8 72.3
DL 5% 12.3 11.4
DL 1% 20.1 21.4
DL0,1% 25.8 28.5
The content of humid gluten has a similar range with the protein content. Thus, at the level
of both years of research, the issue of the positive influence of the conservative amelioration
process positively occurs. The gluten content, given the conditions of a controlled
replication of seed’ production, varies almost imperceptibly; this small variation determines
us to take into consideration that, in order to reach at a good quality bread production, one
302

may employ (wheat) flour produced from the SC2 biological link, having the same value of
the humid gluten as in the case of the B and SC1 (biological) links.
At the level of “other replications” (biological) links the discrepancies are striking. The
lack of selection pressure at the level of these replications, leads inevitably to the qualitative
decrease of the flour, which, as above signalised, is associated with a production. From all
above mentioned, it is occurs the fact that the proper seed for obtaining some top-quality
bread-making products must be placed at the level of the SC2 seed type.
CONCLUSIONS
1. The gluten content of the wheat is strongly influenced by the biological category (class)
of the seed used, high differences concerning the content occur between the biological
links obtained through amelioration (B,SC1,SC2) and “other replications”.
2. The gluten content, in a similar manner with the protein content, has increased values in
the case of B,SC1,SC2 biological categories (classes) while the “other replications”
category has quite reduced values, from which it results the fact that the seed used to
obtain bakery products must be placed at the level of the SC2 seed.
3. The protein content and the gluten one show less significant differences in the two-year
research period; these are due to the soil and climatic factors.
REFERENCES
1. Haş, Ivan (2006)-Producerea seminţelor la plantele agricole, Editura Academic Pres,
Cluj-Napoca
2. Munteanu, S.L., I.Borceanu, M.Axinte, G.V.Roman(2001) - Fitotehnie, Editura Ion
Ionescu de la Brad, Iaşi
3. Savatti,M., G.Nedelea, M.Ardelean,(2004)-Tratat de ameliorare a plantelor,Editura
Marineasa,Timişoara
303

THE INFLUENCE OF SOIL OIL POLLUTION ON AGRICULTURAL CROPS
Maria Şandor, Nicu Cornel Sabău, Cornel Domuţa, Cristian Domuţa, Radu Brejea
Agricultural Research Station Oradea
The Faculty for Environmental Protection, University of Oradea
Calea Aradului St. No. 5. email: scdaoradea@yahoo.com.
Gen Magheru Blvd., No. 26.email: nsabau@uoradea .ro.
ABSTRACT
The paper presents the results of researches reguarding oil polluted soils, that took place
at the Agricultural Research station in Oradea, from 1993 to 2002.
The experimental device was made out of 1 m² microparcels, spread out in a random
order in a Latin square; these parcels were polluted under control with petroleum from
Suplacu de Barcău, Bihor County, with the following concentrations : 0, 1, 3, 5 and 10
% petroleum on the ploughed layer, with 4 repetitions.
The experience was set out on a luvosoil , and the soil was cultivated with millet, a
plant which is considered to be tolerant to soil pollution, in the first 3 years, and with
spring wheat in the last 7 yeras of research.
The results of the research have shown that the yield losses are proportional with
the petroleum concentration, and had a descending evolution. For instance, in the case
of the 1% pollution, losses are insignificant after 7 years of crops.
Keywords: pollution, petroleum, biodegradation.
INTRODUCTION
Soil pollution with oil residue is a very complex phenomenon which involves knowing
the chemical nature and concentration of the pollutive agent and the soil conditions.
Pollution with oil residue is manifested especially in the upper layer of the soil, but in
more seriuos cases of pollution, effects were encountered at 80 cm depth on the soil
profile, the depth at which the pollutive agent got into the soil was influenced by
quantity, time of action on the soil, local microenvironment, phisycal and chemical soil
properties.
Oil extraction, processing and transport in Bihor took place at the sites in Suplacu
de Barcău, Marghita and Oradea, which have become nowadays stations for OMV and
Petrolsub SA Suplacu de Barcău Refinery, today in conservation. Following these
activities, the soil is affected by histortical pollution on a surface of 200 ha, and is in
need of measurements of ecological rehabilitation. [3.]
The researches carried out in Romania by Toti Mh (2003) concerning the pollution
effects on agricultural land from the Southern part of Romania , have prooved that the
plant`s average life expectation diminished after a pollution of 1kg waste / m² (0,3%) in
the ploughed layer. [4.] The authors consider that a pollution of 1,5 – 3,0 kg waste / m²
is a moderate one, between 3 kg – 15 kg waste / m² the pollution becomes strong, and
between 15 – 30 kg waste / m² it is extremely strong, and thus the plants seeds no longer
germinate, and over 30 kg waste / m² it is excessive.
304

For the conditions in from Western Romania, Colibaş I publishes in 1995 the first
partial results of researches regarding yield losses in millet, in the first year of
controlled pollution with different doses of petroleum. [2.]
MATERIALS AND METHOD
The researches carried out in Oradea wanted to establish the effects controlled pollution
with petroleum from Suplacu de Barcău had on agricultural yield and on the
biodegradation period, without any ameliorative measures.
The oil reserve in Suplacu de Barcău is located at relatively small depth, in a layer
of Pontian sands, and has a high content of asphaltines.
Almost half the soils in Romania (49,397%) which are affected by pollution with
oil are luvosoils, and that the soil from Suplacu de Barcău is also a luvosoil, the
experience carried out was placed also on a luvosoil. (Table 1.)
Table 1: Some physical and chemical properties of the luvosoil from Oradea
Horizont Depth Textural DA
(cm) Class (g/cm 3 pH V Humus N C/N
) (H2O) (%) (%) (%)
Ap 0-25 LP 1,35 5,51 64,3 2,40 0,116 13,0
El 25-44 LP 1,48 5,78 67,3 2,23 0,111 13,5
Bt1w 44-60 TT 1,56 6,24 77,9 1,91 0,096 13,5
Bt2 60-93 TT 1,58 6,46 83,4 1,73 0,087 13,4
BC 93-110 TT 1,62 6,51 86,6 0,70 0,036 13,3
C 110-152 TT 1,61 6,60 92,4 0,50 0,026 13,2
The soil is dusty and with a content of clay, in the worked layer and in the eluvial one,
and with medium clay in the layers underneath these ones. The values of the bulk
density are medium in the eluvial layer and becomes large in the layer where clay
gathers, and at the base of the profile, which indicates that the soil is poorly ploughed at
the surface and poorly settled from a 25 – 44 cm depth , and moderately settled on the
rest of the profile.[1.] The soil reaction is moderately acid in the first two horizons and
weak acid deeper. The total humus and N content is small in the ploughed layer, and the
values of the C/N index are high, which comes to prove a poor N supply in the soil.
The experimental field set out in 1993 is made out of parcels of 1m² set out in a
latin square, randomised, in four repetitions, which were willingly polluted with
petroleum from Suplacu de Barcău with 0, 3, 9, 15, and 30 l/m², thus resulting
concentrations of 0 in the ploughed layer (unpolluted witness) and 1, 3, 5, 10 %.
The field was cultivated in the first three years with millet (1993 – 1995) a plant
with a high tolerance to pollution, and afterwards for the next seven years with spring
wheat, Speranţa breed.
RESULTS AND DISCUSSIONS
1. The climate conditions, characterised by rainfall and by annual temperatures,
show that the period of the ten years observed had more rainfall than the average multi
annual with 26,2 m and was warmer with 0,5 ºC.
305

The annual rainfall was between 367 mm in 2000 and 886 mm in 1996, the deviations
from the normal standard values ranged from – 268mm to +251 mm. Temperatures
varied between 9,6 – 12 ºC, these values were registered in 1996 and 2000, and in
comparison with the multi annual average they had a -0,9 – +1,5 ºC variation.
2. The annual millet production of the unpolluted parcels, vary one from another,
because of the different climate conditions in the three years of research, starting at a
very small yield (1993 – 17,6 q/ha hay) and grew to 44,2 q/ha. The productions from
1995 are smaller than the ones in the previous year, but the difference of 3,5 q/ha is
insignificant.
The various petroleum concentration in the ploughed layer determined millet hay
productions, correlated linearly. The inverse linearly correlations are very significant
statistically, for every one of the years researched, the correlation coefficients are : R =
0,9045 in the first year of controlled pollution, R = 0,8456 for the 2 nd year of controlled
pollution and R = 0,8615 for the 3 rd year. (Figure 1.)
Millet yields (q/ha)
60
50
40
30
20
10
0
1993 y = -1.026x + 15.994
1994 y = -2.6898x + 37.401
1995 y = -1.8137x + 38.192
0 2 4 6 8 10
Petroleum concentration in ploughed layer (%)
1993 1994 1995
Figure 2: The influence of petroleum pollution on millet hay production (1993-1995)
The slope of the regression lines for the years points out that the average yield losses,
registered with the increase in the petroleum concentration with 1 % , are of 1,026 q/ha
millet hay, in the first year of controlled pollution, 2,6898 q/ha millet hay in 1994 and
1,8137 q/ha in the 3 rd year of research. In order to compare the yields from the polluted
and the unpolluted witness parcel, seeing that the yields in the witness were very
different in the 3 years of research, the differences between the polluted and unpolluted
parcels were used, expressed in percentages.
The relative yield losses of the polluted parcels, from the three years of study have
had an evolution described best by a polinominal curve of the II degree. The
correlations that were established like this show correlation coefficients with no
statistical insurance for the 1% concentration R= 0,06790, statistically significant for the
concentration 3 %, R=8890 and R=0,9520, distinctively significant, for the
concentrations of 5 % and 10 %. (Figure 2.).
306

Yield losses millet hay (%)
80
70
60
50
40
30
20
10
0
3% y = -14.385x 2 + 57364x - 6E+07
R = 0.6790 *
10 % y = -10.806x 2 + 43086x - 4E+07
R = 0.9520**
5 % y = -22.876x 2 + 91227x - 9E+07
R = 0.8890**
1993 1994 1995
3% 5% 10%
Figure 2. The evolution of relative yield losses (%) 1993 – 1995
If we analyse the shape of evolution curves for the yield losses we notice that these
curves have grown in the second year of experiments, for all the variants of petroleum
concentration, and that year also marked significant yield losses : 23,08 % for the 1 % ,
42,31 % for the 3 %, 61,54 % for 5 % and 65,61 % for the maximum concentration of
10 %. In the 3 rd year of controlled pollution, the yield losses diminish indicating, the
possibility of the soil naturally recovering from the pollution.
The average yield for an experience cultivated with millet (1993 – 1995) was of
34,2 q/ha in the unpolluted witness variant, and ranged between 28,3 – 14,5 q/ha in the
variants with controlled pollution, bigger where the concentration was 1% and smaller
at a concentration of 10 %. (Table 2).
The statistical analysis of the losses from the medium yields (5,9 – 19,7 q/ha) show
that they are all significant, in all the concentrations studied. The annual yield
differences for 1 % pollution in the ploughed layer are significant in the first year (-34
q/ha), significantly different in the second year (-10,2 q/ha) and do not have any
statistical importance in the 3 rd year (-5,9 q/ha). The variant polluted with 3 % shows
significant differences only in the first two years (-5,5 and – 18,7 q/ha), just to become
only significant from a statistical point of view in the 3 rd year (-10 q/ha).
The differences from the 3 rd year of observation, insignificant for the 1%
concentration, suggest that most of the oil residue have been biodegraded and the soil is
recovering ecologically. The fact that the 3 % shows also only some small differences
can suggest that the biological recovery has started.
These speculations are also sustained by the statistical analysis of the interaction.
Variants (concentrations) x years indicates that the average yield losses are insignificant
when talking about the pollution of the ploughed land with 1 % and distinctively
significant when it comes to 3 and 5 % concentration of petroleum.
307

Table 2: Yield losses millet hay due to petroleum polution. (q/ha) 1993-1995
Variant
Years (Y) Average Semnification
(V) 1993 1994 1995 1993-1995
(%) (q/ha) (q/ha) (q/ha) q/ha Diference V VxY
0 - - - 34,2 - - -
1 -3,4 -10,2 -4,0 28,3 -5,9 ooo -
3 -5,5 -18,7 -10,0 22,2 -11,4 ooo oo
5 -7,4 -27,2 -14,3 17,9 -16,3 ooo oo
10 -11,2 -29,2 -18,7 14,5 -19,7 ooo ooo
1993 1994 1995 Variant (V) Variant x Years (VxY)
DL 5 % 2,46 5,25 5,56 2,49 7,83
DL 1 % 3,46 7,37 7,80 3,33 11,39
DL 0,1 % 4,88 10,41 11,02 4,39 17,09
3. Average annual yield in spring wheat from the unpolluted witness parcels, were
between 21,8 q/ha , in the 4 th year of observation (1996) and 4 q/ha in 2000 (which was
considered to be one of the years with most drought ever) (Figure 3.)
Yields (q/ha)
25
20
15
10
5
0
2002
2001
2000
1999
1998
1997
0 1 3 5 10 1996
Petroleum concentration (%)
Figure 3. Annual spring wheat yield (q/ha)
The yields from the variant`s repetition had a linearly evolution in each and every one
of the seven years with spring wheat crop (1996-2002), inverse proportional with the
petroleum concentration from the ploughed layer. The linearly correlations thus
established are significant statistically, in the first five years of the interval (R = 0,8096
308

– 0,6093) and extremely significant in the last two years (R = 0,4539 – 0,4058) (Table
3.)
Table 3. Regression equations between spring wheat yield and petroleum
concentration (1996-2002)
Nr. Anul Ecuaţia Coeficient de Semnificaţie
crt.
corelaţie
1 1996 Y = - 1,4818 X + 18,271 R = 0,8723 oo
2 1997 Y = - 1,2357 X + 19,696 R = 0,8906 oo
3 1998 Y = - 0,4377 X + 7,9734 R = 0,7833 oo
4 1999 Y = - 0,4169 X + 7,7041 R = 0,7531 oo
5 2000 Y = - 0,1430 X + 3,9634 R = 0,6093 oo
6 2001 Y = - 0,0885 X + 5,6811 R = 0,4539 o
7 2002 Y = - 0,3566 X + 10,0100 R = 0,4058 o
The relative production losses for spring wheat range between 24,3 % for 1 %
concentration to 74,7 % for 10 % concentration. These losses are bigger than the ones
registered during the previous year in millet. This is because wheat is less tolerant to
pollution than millet.
The evolution of relative yield losses in spring wheat from the last seven years of
observation has a rising tendency, described in polynomial equations of the 2 nd degree,
extremely significant for high concentrations (5 and 10 %), significant for 3 % and with
no importance for a 1 % concentration of petroleum. (Figure 4.)
The average values of relative yield losses decrease annually and reach about 10 –
15 % for the small 1-3 % concentrations, 25 % for the 5 % concentration and 37 % for
the maximum 10 % concentration. The average yield differences in spring wheat from
the last seven years of research in spring wheat pollution are of 1,1 q/ha for a 1 %
pollution – significant statistically, 3,3 q/ha for 3 %, 5,0 q/ha at 5 % and 6,3 % for 10 %,
the last two are very significant. (Table 4.)
The interaction (petroleum concentration) x years does not have any statistical
value for the variant polluted under control with 1 %, and is significant for the
concentration of 3 % and extremely significant for the 5 % and 10 % concentration in
the first 20 cm of the soil profile.
If we analyse the annual yield losses evolution from the period in which spring
wheat was cultivated we notice that in the variant polluted with 1 % petroleum, the yield
losses are extremely significant statistically in 1996, significant in 1997 – 1999, only be
become insignificant in the last three years of the research.
In the variant with a 10 % pollution, the yield in the last year is not significant
statistically, is significant in the ninth year, distinctively significant in the eighth year,
and very significant in the first seven years.
309

Yields losses, spring wheat (%)
90
80
70
60
50
40
30
20
10
0
-10
-20
-30
-40
-50
-60
3% y = 0.4101x 2 - 1645.2x + 2E+06
R = 0.4791 *
10 % y = 1.0655x 2 - 4267.5x + 4E+06
R = 0.7760 **
1996 1997 1998 1999 2000 2001 2002
5% y = 0.8161x 2 - 3269.5x + 3E+06
R = 0.7144 **
3% 5% 10%
Figure 4. The evolution of relative yield losses (1996 – 2002)
Tabel 4 Yield losses due pollution with petroleum in spring wheat (q/ha)
1996 - 2002
Years
No. Variant
Yield
Semnification
Crt. (V)
1996-2002
(%) q/ha Difference V V x Y
1 0 11,5 - - -
2 1 9,6 -1,1 o -
3 3 8,2 -3,3 ooo o
4 5 6,5 -5,0 ooo ooo
5 10 5,2 -6,3 ooo ooo
D.L. Variant Variant x Years
(V)
(VxY)
5 % 1,0 2,7
1 % 1,5 3,6
0,1 % 2,1 4,8
If we sum up all this data we see that the annual yield losses loose their statistical value
after the 7 th year of controlled pollution, for a 1 % concentration, after the 8 th year in 3
% and 5 % petroleum concentration, and in the 9 th year for a 10 % concentration.
310

CONCLUSIONS
The experimental field with microparcels, set out in a randomised latin square, from the
luvosoil at the Agricultural Research Station Oradea, was polluted in a controlled
manner with petroleum brought from Suplacu de Barcău, in different concentrations of
0, 1 ,3, 5 and 10 % in four repetitions. Between 1993 – 1995 the field was cultivated
with millet, a plant which is considered to be tolerant to pollution (first 3 years) and
with spring wheat Speranţa for the next 7 years.
The annual yields in the variants with different concentrations of pollution are
linearly correlated both for millet and wheat, the correlations are distinctively
significant for the first three years of millet crop and for the next 5 years of wheat crop.
The evolution of relative yield losses from the research period are described by a
polynomial regression of the second degree, both for wheat and millet. They are
significant for the variant polluted with 3 %, distinctively significant for the variants
polluted with 5 and 10 % and with no statistical significance for the 1 % concentration.
The biggest yield losses in millet crop were registered in the 2 nd year of research, only
to decrease in the 3 rd .
The average yield losses in three years, in millet were very significant, and ranged
between 5,9 q/ha hay for the variant of 1% petroleum and 19,7 q/ha hay for the 10 %
petroleum. The influence of the interaction variant x years of growing show that these
differences are insignificant for the 1 %, distinctively significant for the 3 and 5 % and
very significant for the 10 % concentration.
In wheat, the average yield losses for seven years are significant for the small 1 %
concentration and very significant for the other variants. The statistical significance of
the yield losses from the 4 th year of experience come to prove that wheat is less tolerant
to petroleum pollution than millet.
The evolution of annual yield losses show that the oil residue in the soil are
biodegraded, the periods of time needed to do that are direct proportional with the
initial concentrations. Annual yield losses loose their statistical significance after 7
years in crop at a 1 % concentration, 8 years in average concentration of 3-5 % and 9
years in big concentrations of 10 %. The yields from the 10 th year of experiments are
still smaller than in the unpolluted variant with 12 – 13 % at the variant polluted with 1
– 3 %, 25,6 % at the 5 % and 36,9 % for the 10 %.
REFERENCES
1. Colibaş I, Colibaş Maria, Tirpe Gh (2000): Solurile brune luvice, caracterizare şi
ameliorare – Ed. Mirton Timişoara.
2. Colibaş I., Colobaş Maria, Şandor Maria (1995): Măsuri de ameliorare a solurilor
poluate cu rezidii petroliere. – Cum să cultivăm pământul în zona centrală din
vestul ţării – Staţiunea de Cercetări Agrozootehnice Oradea.
3. Sabău N.C., Domuţa C., Berchez O. (2002): Geneza degradarea şi poluarea solului,
Partea a II-a Degradarea şi poluarea solului – Editura Uniersităţii din Oradea;
4. Toti Mh., Dumitru Mh., Rovena Voiculescu Anca, Mihalache Mh., Mihalache
Gabi, Constantinescu Carolina (2003): Metodologia de biodegradare a solurilor
poluate cu ţiţei, cu ajutorul microorganismelor specifice selecţionate din microflora
autohtonă – Edituta GNP Minischool;
311

EFFECT OF BIOFERTILIZATION ON PARSLEY YIELD AND N CONTENT
ON TWO SOILS
Ida Kincses 1 - Tibor Filep 2 - Andrea B. Kovács 1 - Péter T. Nagy 1 – Imre Vágó 1
1 University of Debrecen, Department of Agricultural Chemistry, Debrecen
2 Research Institute for Soil Science and Agricultural Chemistry of the Hungarian
Academy of Sciences, Budapest
ABSTRACT
A pot experiment was carried out to investigate effect of N fertilizer and biofertilizer on
parsley (Petroselinum tuberosum) yield and N content of plant, as well as the 0.01 M
CaCl2 soluble N concentrations in soils. Two soils were used, one of them is a
chernozem with slightly acidic pH and with high content of organic matter, and the
other one is an acidic sandy soil with low organic matter content.
On chernozem, the N fertilization significantly increased the yield of plant, while
there was no significant biofertilizer effect. On sandy soil, the dry matter of plants
increased with increasing N doses comparing to control, but no correlation has been
found between biofertilizer and dry matter weight.
Investigating N content of plants on chernozem soil, it was found that N
fertilization enhanced the N concentration in plant, but there was no relationship
between N content and biofertilizer rates at all. For sandy soil, at high rates of N the
biofertilization caused an increase in N concentration of plant, while at lower doses just
the opposite was noticed. So between two soils there was a remarkable difference: in
case of acidic sandy soil, the adding of biofertilizer caused an increase in both plant
biomass and in N content of leaf. These effects are not appeared for chernozem. Due to
the very low N supply ability of sandy soil, the increment of microbes due to
biofertilization caused N immobilization, because the enhanced microbial biomass take
up soluble N.
Based on data from experiment, it can be concluded that positive effect of
biofertilization on yield, N content of plant and soil displayed only on chernozem soil
during that short period of investigation. On the acidic soil with low organic matter
content, these effects are no appeared because of N immobilization by microbes.
Keywords: N fertilization, biofertilization, parsley, pot experiment, 0,01M CaCl2
soluble N-forms
INTRODUCTION
In horticulture cropping, the main goal is to reach appropriate quality and quantity of
crops, which mainly determined by genetics parameters of plants, ecological
circumstances and the nutrient status of soil. The nutrient supply is the one of the most
important factor to get maximum yields. In our country, the nutrient supply is based on
adding to soils chemical fertilizers.
Biofertilizers are an alternative way to substitute mineral fertilizers for increasing
soil fertility and crop production. Biofertilizers are products containing living cells of
different types of microorganisms, which have an ability to convert important elements
from unavailable to available form through biological processes (HEGDE et al., 1999).
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Evaluation of effect of fertilization on plants needs an examination on plant-soil system.
Amount, biodiversity and activity of living beings in soils depend on physical and
chemical parameters, while soil microorganism impact soil processes (SZABÓ, 1986).
Application of chemical fertilizers may cause acidification in soils due to its
chemical attributes (CHANDER – ABROL, 1972; FELIZARDO et al., 1972; POWER,
1972). To decrease the harmful effects of fertilization, farmers are trying to decrease the
amount of chemicals, and to substitute that to manure or biofertilizers, which are
permitted in biofarming. In biofarming, the sustainability and improving of soil fertility
plays the greatest role (AUBERT, 1981; NAGY et al.2001).
Our experimental data can be used, in the main objective of crop production, in
improving of environmental protection, and the site-specific fertilization.
MATERIALS AND METHODS
An experiment was carried out with parsley in spring of 2006 with 12 treatments and 4
replications at the Experimental Site of Debrecen University. Soils used were from
Latókép (calcouorus chernozem) and Újfehértó (acid arenosol). Important parameters of
soils are the followings:
Látókép: pH (H2O): 6.7; pH(KCl): 5.9; pH(CaCl2): 6.23; Humus (%): 2.5; KA: 42;
Újfehértó: pH (H2O): 4.1; pH(KCl): 3.7; pH(CaCl2): 6.23; Humus (%): 1.3; KA: 27;
10 kg of Látókép and 12 kg of Újfehértó soil was dispensed to pots. The N, P and K
treatments were added as NH4NO3, KH2PO4, KCl solution. The amount of applied
biofertilizer was in accordance with the prescribed volume and two times of that.. The
Phylazonit MC biofertilizer contained carboxi-methyl-cellulose (CMC), microelements,
Azotobacter croococcum and Bacillus megaterium soil bacteria, heteroauxin, gibberelin,
vitamin B. Treatmnets applied were in Table 1.
Table 1: Combination of treatments
Biofertilizer ml/pot
Treatment N g/pot Látókép Újfehértó P g/pot K g/pot
1 0.2 70 80 0.1 0.3 N1*B1
2 0.2 35 40 0.1 0.3 N1*B2
3 0.2 - - 0.1 0.3 N1*B3
4 0.1 70 80 0.1 0.3 N2*B1
5 0.1 35 40 0.1 0.3 N2*B2
6 0.1 - - 0.1 0.3 N2*B3
7 0.05 70 80 0.1 0.3 N3*B1
8 0.05 35 40 0.1 0.3 N3*B2
9 0.05 - - 0.1 0.3 N3*B3
10 - 70 80 0.1 0.3 N4*B1
11 - 35 40 0.1 0.3 N4*B2
12 - - - 0.1 0.3 N4*B3
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Seeds were sown on 5th may 2006. Pots were irrigated daily with deionized water
during the experiment, according to 60 % of maximum water capacity of soils. Plants
were harvested 20 th of July and soil samples were taken.
Beside plant biomass, the C, N and S content of plant also were measured by a dry
combustion method. The 0.01 M CaCl2 soluble N and P content of soil (HOUBA et al.,
1994) was determined by a continues flow equipment.
RESULTS AND DISCUSSIONS
We would like to get results with this experiment that how the N and biofertilization
affect dry matter of parsley, and N content of plant. Treatment effects on 0.01 M CaCl2soluble
total N and NO3-N concentration at the end of growing period were also
examined. The experiment was set up on a chernozem soil with good N supply, high
organic carbon content, around neutral pH, which gives good circumstances for
microbes; and on an another one, which have opposite properties, with poor N supply,
low content of organic C, and with very acidic character. The experimental design
opens the door to compare plant parameters on two soils.
As you can see from data of Table 2, in which dry matter weight of parsley was
presented, the N fertilization significantly increased the yield of plant. Although there
was no significant biofertilizer effect, but it can be concluded that bio-treatments
enhanced the dry matter.
Table 2: Dry matter content of parsley on chernozem
Leaf dry matter (g pot -1 )
Phylazonit doses (cm 3 pot -1 N (g pot
)
-1 )
70 35 0
Mean LSD5% (N)
0.2 17.7 17.5 17.5 17.6
0,1 15.6 15.1 15.1 15.3
0,05 14.6 14.4 14.3 14.4 1.4
0 15.6 15.6 14.7 15.3
Mean 15.9 15.7 15.4 15.7
LSD5% (phylazonit): 1.2
According to data presented in Table 3, the dry matter of plants increased with
increasing N doses comparing to control on sandy soil. However, there were no
significant differences among N treatments.
No correlation has been found between biofertilizer and dry matter weight on sandy
soil. It is also true that biofertilization increased dry matter of parsley at high N doses,
while at lower ones or at control the application of biofertilization decreased.
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Table 3: Dry matter content of parsley on sandy soil
Leaf dry matter (g pot -1 )
Phylazonit doses (cm 3 pot -1 N (g pot
)
-1 )
80 40 0
Mean LSD5% (N)
0.2 5.4 5.6 4.4 5.1
0.1 5.8 4.5 4.9 5.1
0.05 4.7 5.0 5.0 4.9 0.8
0 4.0 4.1 4.3 4.1
Mean 5.0 4.8 4.6 4.8
LSD5% (phylazonit): 0.7
Investigating N content of plants on chernozem soil, it was found that N fertilization
enhanced the N concentration in plant, but there was no relationship between N content
and biofertilizer rates at all (Table 4).
Table 4: Nitrogen content of parsley on chernozem soil
Leaf N content (%)
Phylazonit doses (cm 3 pot -1 N (g pot
)
-1 )
70 35 0
Mean LSD5% (N)
0.2 3.4 2.6 2.7 2.9
0.1 2.9 2.5 2.9 2.8
0.05 2.9 3.0 2.3 2.8 0.3
0 2.2 2.0 2.5 2.2
Mean 2.9 2.5 2.6 2.7
LSD5% (phylazonit): 0.3
Similarly to chernozem, with increasing of N fertilizer rates the plant N concentration
increased, and this treatment effect was confirmed statistically (Table 5).
Relationship between biofertilizer rates and N content has been shown a very similar
trend like in case of dry matter content of parsley. At high rates of N, the biofertilization
gave an increase in N concentration of plant, while at lower doses just the opposite was
noticed.
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Table 5: N content in plant on sandy soil
Leaf N content (%)
Phylazonit doses (cm 3 pot -1 N (g pot
)
-1 )
80 40 0
Mean LSD5% (N)
0.2 3.2 2.8 2.8 2.9
0.1 2.5 2.5 3.2 2.7
0.05 2.1 2.4 2.0 2.2 0.4
0 1.9 1.9 2.6 2.1
Mean 2.4 2.4 2.6 2.5
LSD5% (phylazonit):0.3
Between two soils there was a remarkable difference: in case of acidic sandy soil, the
adding of biofertilizer caused an increase in both plant biomass and in N content of leaf.
These effects are not appeared for chernozem. Due to the very low N supply ability of
sandy soil, the increment of microbes amount due to biofertilization may cause N
immobilization, because the enhanced microbial biomass take up soluble N.
To take further investigation on this problem, 0.01 M CaCl2 soluble total N and nitrate
was measured from soil samples taken at the end of growing period. Both N fertilization
and biofertilization significantly increased the soluble total nitrogen concentration on
the chernozem soil (Figure 1).
25
20
15
10
5
0
B1 B2 B3
N1 N2 N3 N4
Figure 1: Effect of N and biofertilization on 0.01 M CaCl2 soluble
total N content of chernozem (LSD5% (N): 2.90 LSD5% (B): 2.51)
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On sandy soil, the total N content increased with increasing N doses, while adding of
biofertilizer to soil caused no significant increase in soil CaCl2-soluble total nitrogen
concentration (Figure 2).
15
10
5
0
Figure 2 Effect of N doses and biofertilization on 0.01 M CaCl2
soluble total N content on sandy soil (LSD5% (N): 3.9 LSD5% (B):
3.37)
Moreover, biofertilizer have a negative influence during the investigated period on
0.01M CaCl2-soluble total N content of sandy soil with acidic character and low content
of organic matter.
The nitrogen fertilization, beside the total N, enhanced the nitrate concentrations in
0.01 M CaCl2 extractions (Figure 3). Biofertilizer also have an increasing effect on
nitrate, but only at high doses of N.
12
10
8
6
4
2
0
B1 B2 B3
N1 N2 N3 N4
B1 B2 B3
N1 N2 N3 N4
Figure 3: Effect of nitrogen rates and biofertilization on nitrate
concentration on chernozem (LSD5% (N): 3.78 LSD5% (B): 3.28)
317

There was a significant increase in nitrate-N comparing to control after N fertilizer
adding, while biofertilizer doses decreased (not significantly) the nitrate concentrations
on the sandy soil (Figure 4).
1
0,8
0,6
0,4
0,2
0
B1 B2 B3
N1 N2 N3 N4
Figure 4: Effect of nitrogen and bio-fertilization on nitrate-N on
sandy soil (LSD5% (N): 0.36 LSD5% (B): 0.31)
Based on data from experiment, it can be concluded that positive effect of
biofertilization on yield, N content of plant and soil displayed only on chernozem soil
during that short period of investigation. On the acidic soil with low organic matter
content, these effects are no appeared because of N immobilization by microbes.
REFERENCES
Aubert, C. (1981): Organischer Landbau. Verlag Eugen Ulmer, Stuttgart
Chander,-H. - Abrol,-I. P. (1972): Effect of three nitrogenous fertilizers on the
solution composition of a saline sodic soil. Commun. Soil Sci. Pl. Anal., 3. 1. 51-56. p.
Felizardo,-B. C. - Benson,-N. R. - Cheng,-H. H. (1972): Nitrogen, salinity, and acidity
distribution in an irrigated orchard soil as affected by placement of nitrogen
fertilizers. Soil Sci. Soc. Amer. Proc., 36. 803-808. p.
Hegde, D. M. – Dwived, B. S. – Sudhakara, S. N.: 1999. Biofertilizers for cereals
production in India – a review – Indian J. Agric. Sci. vol. 69 73-83 pp.
Houba, V. J. G. - Novozamsky, I. - Temminghoff, E. (1994): Soil analysis procedures
Extraction with 0,01 M CaCl2. (syllabus) Dept. Of Soil Sci. And Plant Nutrition
Wageningen Agricultural University – The Nederlands
Nagy J. -Német T. -Szabó J. -Pásztor L. -Dobos A. (2001): Tájgazdasági körzetek
kialakítása a Kreybig-féle „Átnézetes Talajismereti Térképsorozat” alapján.
Agrártudományi Közlemények 1. Acta Agraria Debreceniensis, 20-25.
Nagy P.T. (2000): Égetéses elven működő elemanalizátor alkalmazhatósága talaj- és
növényizsgálatokban. Agrokémia és Talajtan. Tom. 49. No. 3-4. 521-534.
Power,-J. F. et al. (1972): Effect of nitrogen source on corn and bromegrass
production, soil pH, and inorganic soil nitrogen. Agron. J., 64. 341-344. p.
Szabó I. M. (1986): Az általános talajtan biológiai alapjai. Mezőgazdasági Kiadó,
Budapest.
318

THE INFLUENCE OF CHEMICAL AND BIOFERTILIZERS ON THE YIELD
AND NITROGEN CONTENT OF LETTUCE (LACTUCA SATIVA L.)
Andrea Balla Kovács, Ida Kincses, Imre Vágó, Rita Kremper
University of Debrecen Faculty of Agricultural Science
Department of Agricultural Chemistry and Soil Science
ABSTRACT
The greenhouse pot experiment on calcareous chernozem soil from Debrecen-Látókép
and on sandy soil from Újfehértó was performed to study the effects of biofertilizer in
the presence of reduced doses of chemical fertilizers on the yield and nitrogen content
of lettuce and the water soluble nitrogen forms (NO3 - and total-N) measured in 0.01 M
CaCl2 extracts. The experimental plant selected was cabbage head lettuce (Lactuca
sativa L.). The bi-factorial trials were arranged in a randomized complete block design
with four replications, applying four levels (0, 0.05, 0.1, 0.2 g pot -1 ) of N as NH4NO3
with or without application of biofertilizer as Phylazonit MC. P and K doses applied
were identical in all pots.
The results obtained suggested: Nitrogen doses markedly increased growth (fresh
and dry weight) of lettuce, had significant positive influence on N content of plant and
on values of NO3 - , total N measured in 0.01 M CaCl2 soil extract . Its increasing effect
was the same on two types of soils, only significance of variance varied.
The application of biofertilizer was not as effective as N doses and its effect was
dependent on soil caracteristics. The Phylazonit MC had an increasing influence, but
not significant effect on the fresh and dry weight of lettuce leaves and slightly changed
N content of plant. On chernozem soil, we have found an increasing effect in values of
N content of plant, NO3 - and total N measured in 0.01 M CaCl2 extracts. On the
contrary, on sandy soil a slight decreasing effect of biofertilizer was found in N content
of plant and total N measured in 0.01 M CaCl2. It seems that in our experiment, (over
the entire growing season) application of phylazonit on sandy soil might cause N
immobilization, in consequence decreased the mineral N fraction available for plant and
so decreased the N content of plant. To gain more, reliable applicability of Philazonit
MC, further studies are needed.
Keywords: lettuce, mineral fertilizers, nitrogen, biofertilizers
INTRODUCTION
Nitrogen as a major constituent of all plants is one of the most important nutrients. It
has got a unique position because relatively high amounts are required by most
agricultural crops for optimal yields (Black 1968, Stevenson and Cole 1999). Most
plants take up N as nitrate (NO3 - ) and ammonium (NH4 + ) from the soil solution
(Blackmer 2000). The use of nitrogen fertilizers is standard practice in vegetable
production systems. Increasing concern over nitrate contamination of soils, nitrate
leaching in groundwater and nitrate content in vegetable may require a reduction in N
application in crop production, while maintaining optimal productivity (Gutezeit and
Fink 1999). Lettuce is an important horticultural crop. High nitrate concentration in
leafy vegetables constitutes a health hazard and is therefore undesirable. Applications of
319

iofertilizers may help to avoid environmental hazards for plants, animals and human
beings.
The present work was planned to study the effect of biofertilizer, Philazonit MC in
the presence of reduced doses of chemical fertilizers on the yield and nitrogen content
of lettuce and the water soluble nitrogen forms (NO3 - and total N) measured in 0.01 M
CaCl2 extracts.
MATERIALS AND METHODS
The greenhouse pot experiment was performed using soil samples from two regions,
calcareous chernozem soil from Debrecen-Látókép (1) and sandy soil from Újfehértó
(2). The important properties of soils used are included in Table 1.
Table 1. Properties of experimental soils
Soil no. pHKCl pHH2O pHCaCl2 Hu% KA
1 5.9 6.7 6.2 2.5 42
2 3.7 4.1 4.8 1.3 27
KA: Plasticity index according to Arany
10 kg and 12 kg soil (chernozem soil from Debrecen-Látókép and sandy soil from
Újfehértó, respectively) were weighed into Mitscherlich type pots. The experimental
plant selected was cabbage head lettuce (Lactuca sativa L.). Four seed of lettuce were
sown in the soil per pot. Ion exchanged water was added to all pots to keep the soil at
constant moisture (60% of the water-holdig capacity) using daily weighing.
The bi-factorial trials were arranged in a randomized complete block design with four
replications, applying four levels of N with or without application of biofertilizer
(Table 2.). P and K doses applied were identical in all pots (0.2 g P2O5 pot -1 and 0.2 g
K2O pot -1 ).
Treatment N doses
g pot -1
Table 2. Scheme of treatments applied
Phylazonit doses in
sandy soil
cm 3 pot -1
Phylazonit doses
in chernosem soil
cm 3 pot -1
code
1. 0 0 0 control
2. 0 40 35 N0+phyl
3. 0.05 0 0 N1
4. 0.05 40 35 N1+phyl
5. 0.1 0 0 N2
6. 0.1 40 35 N2+phyl
7. 0.2 0 0 N3
8. 0.2 40 35 N3+phyl
The nitrogen, phosphorus and potassium were added in solution of NH4NO3, KH2PO4
and KCl, respectively. The applied biofertilizer was Phylazonit MC (phyl), which
contains carboxi-methyl-cellulose (CMC), microelements, Azotobacter croococcum,
320

Bacillus megatherium soil bacteria, heteroauxin, gibberelin and vitamin B. The soil in
each pot was stirred well to ensure uniform mixing of soil and nutrients.
At the end of vegetation period soil samples were taken, air dried and sieved
(

The fresh and dry yields of lettuce grown on calcareous chernozem soil were much
higher in all pots than in the case of plants cultivated on sandy soil. The fresh and dry
weight on chernozem soil ranged 210.6 - 248.4 g pot -1 and 11.3 - 13.6 g pot -1 , on sandy
soil ranged 55.5 - 77.8 g pot -1 and 4.3 – 6.2 respectively.
Table 5. Mean fresh weight of lettuce cultivated on calcareous chernozem soil
(Látókép) as influenced by N and phylazonit doses
fresh weight (g pot -1 )
phylazonit doses (cm 3 pot -1 N
)
g pot -1 0 40
mean
0 210.6 217.4 213.9
0.05 239.8 242.3 241.1
0.1 244.1 245.9 245.0
0.2 245.3 248.4 246.8
mean 234.9 238.5
LSD5% (phylazonit): n.s.
236.7
LSD5% (N)
15.02
Table 6. Mean dry weight of lettuce cultivated on calcareous chernozem soil
(Látókép) as influenced by N and phylazonit doses
dry weight (g pot -1 )
phylazonit doses (cm 3 pot -1 N
)
g pot -1 0 40
mean
0 11.3 11.4 11.4
0.05 13.3 13.6 13.5
0.1 13.2 11.3 12.3
0.2 13.5 13.6 13.6
mean 12.8 12.5
LSD5% (phylazonit): n.s.
12.7
Source of
variation
LSD5% (N)
0.73
Table 7. Summary of ANOVA (F-test) for different source of variance
fresh weight dry weight fresh weight fresh weight
on sandy soil on chernozem soil
N fertilization *** * *** ***
Phylazonit doses n. s. n. s. n. s. n. s.
n.s.:non-significant; *:significant at P

N content of plant: Studying the nitrogen content of lettuce leaves we found that there
was significant relationship between N treatments and plant N content. Increasing
application of N from 0.05 g pot -1 up to 0.2 g pot -1 had significant positive influence on
N content of plant on both types of soils (Figure 1, Table 8). Biofertilizer did not cause
marked effect on N content of plant on chernozem soil. Inoculation of this type of soil
with Philazonit slightly increased N content with the exception of treatments with no
mineral fertilizer.
On sandy soil application of Phylazonit significantly (P

CaCl2- NO3 - -N (mg kg -1 )
9
8
7
6
5
4
3
2
1
0
5.7
6.1
5.8
7.0
5.5
0.4 0.4 0.6 0.6 0.5 0.8
Control
N0+phyl
Újftó
Látókép
N1
N1+phyl
N2
N2+phyl
8.1
N3
2.0
7.2
2.2
N3+phyl
Figure 2. Mean NO3 - - N (mg kg -1 ) measured in CaCl2 soil extracts from Látókép
and Újfehértó as influenced by N and phylazonit application rates. For interpretation
of treatments see Table 2.
totalN(mg kg -1 )
16
14
12
10
8
6
4
2
0
4.1
Control
11.3
3.7
N0+phyl
12.7
N1
Újftó
Látókép
4.0
11.7
3.8
N1+phyl
12.7
N2
4.5
14.3 14.7 14.5 14.8
4.2
N2+phyl
N3
8.5
6.0
N3+phyl
Figure 3. Mean total N (mg kg -1 ) measured in CaCl2 soil extracts from Látókép and
Újfehértó as influenced by N and phylazonit application rates. For
interpretation of treatments see Table 2.
By the end of the growing season nitrate content of sandy soil becomes very low (0.4-
0.8 mg kg -1 ). Only with the application of highest N doses (0.2 g pot -1 ) caused increased
CaCl2-NO3 - values (2.0-2.2 mg kg -1 ). This increasing effect of N doses on NO3 - content
was significant. We measured higher concentrations of NO3 - by turns on chernosem soil.
This might be due to higher content of organic matter which could mineralize during the
growing season. Both N doses and biofertilization significantly increased the CaCl2-
NO3 - on chernozem soil. The effect of chemical fertilizer was higher. Measured data
show that similarly to CaCl2-NO3 - , the values of CaCl2-total N were higher in
324
8.2

chernozem soil in all pots either. The increased N fertilizer doses caused significant
improvement in CaCl2-total N concentration measured on both types of soils. Effect of
Phylazonit slightly increased values of CaCl2-total N on chernozem soil, but
significantly decreased these values on sandy soil.
Table 9. Summary of ANOVA (F-test) for different source of variance for NO3 - - N
measured in CaCl2 soil extracts
Source of
variation
NO3 - - N
on sandy soil on chernozem soil
significance LSD5% significance LSD5%
N fertilization *** 0.41 * 1.04
Phylazonit doses n. s. - ** 0.73
n.s.:non-significant; **:significant at P

Jászberényi, I. – Loch, J. – Sarkadi, J. (1994): Experiences with 0.01 M CaCl2 as an
extraction reagent for use as a soil testing procedure in Hungary – Comm. Soil Sci.
vol. 25 1771-1777 pp.
Nagy P.T. (2000): Égetéses elven működő elemanalizátor alkalmazhatósága talaj- és
növényvizsgálatokban. Agrokémia és Talajtan. Tom. 49. No. 3-4. 521-534.
326

THE INFLUENCE OF USING BIOLOGICAL ADDITIVES ON THE
NUTRITIONAL QUALITIES OF CORN SILAGE AND ON THE FATTENING
PERFORMANCES OF BULL CALVES
Daniel Mierlita; I. Chereji; Cristina Maerescu
University of Oradea, Romania, 26, Magheru Street, Oradea;
ABSTRACT
Ensiling corn by using a biological additive based on homofermentative lactic acid
bacteria (Lactobacillus plantarum + Pediococcus pentosaceus) ensures, by comparison
with natural fermentation, a forage with higher nutritional qualities, with a lower pH
(acid), a lower content of ammoniacal N and raw cellulose and more undegraded
protein. The use of corn silage treated with inoculant in the food of young bull calves
intended for fattening, has ensured a significant growth in weight gain, DM
consumption and degree of the food’s capitalization, as opposed to the lot which
received naturally fermented forage in the rations. Thus by using inoculant to ensile
corn, the period of fattening was decreased by 26 days and 82 kg of concentrates and
120 kg of corn silage were saved. The biological additive used for the ensiling has not
influenced the output of slaughtering, but it has led to the increase of the muscle area,
which is positively correlated with the proportion of superior quality meat in the
carcass.
Keywords: corn silage, inoculants, fattening up bull calves.
INTRODUCTION
For fattening up young bulls, using fermented forage (especially corn) represents the
most efficient form of feeding, taken into account the technical and economical
advantages that it provides (Halga, 2005). The process of ensiling the forage depends on
the dominance of the lactic acid bacteria over the rest of the microflora, which, in
anaerobic conditions, increase the acidity of the environment, thus reducing the
competition between microorganisms. The efficiency of the ensiling process is given by
the way in which sugars are fermented in acid and by how the altering processes
determined by the clostridium bacteria (butyric acid bacteria) are avoided (Woolford-
2004). It is important that the decrease of pH in the ensiled forage be done as quickly as
possible (during the first 2-3 days) in order to avoid the degradation and loss of the
protein (Webb, 1998). During the uncontrolled processes of fermenting the silage, 45%
of the protein is decomposed up to NH3N (Jones, 1995; 1996).
The control of the silage’s fermenting was accomplished by using inorganic acids
for a long time (sulfuric, phosphoric and hydrochloric acids) or external organic acids
(formic and propionic), which had a series of negative effects in the short run (ruminal
and metabolic acidoses, ruminal parakeratosis) but especially in the long run,
materialized usually in the reduction of fertility as an effect of the acids collecting the
microelements (Suttle, 1998). At the present time, because of modern biotechnologies,
the guiding of silage fermentation is performed by using biological additives that
contain selected strains of lactic acid bacteria, especially lactobacillus, or lactic acid
bacteria and enzymes (cellulase, hemicellulase, amylase). In the USA only the
327

iological additives (the inoculants) have been approved by the FDA and can be used in
ensiling forages (Anon, 1997).
The research performed in this domain (Wilkins, 1996; Steen, 1997 ; Keady, 1998,
2000 ; Mierlita 2002) has shown that using inoculants for corn ensiling has led to a
significant reduction of the ammoniac level (a criterion for evaluating proteolysis), to a
greater consumption (3.8 – 5.2%) and a better digestibility of DM (1.7-4.0%), to a
greater weight gain in bulls intended for fattening (3.8-12.2%), to a better quality and
quantity of cow milk (1.7 -2.2 l/day and an increase of the protein level by 0,05%) as
well as to the improvement in the degree of food capitalization (by 6%).
In our research we have analyzed the effect of using an inoculant with mixed types
of homofermentative lactic acid bacteria with high tolerance to acidity, which would be
able to control the fermentations throughout the ensilaging period (Lactobacillus
plantarum + Pediococcus pentosaceus – 10 6 units forming colonies/g), on the
nutritional qualities of corn silage as well as on the fattening performances of bull
calves.
MATERIALS AND METHODS
The corn used in the ensiling process was harvested in an advanced state of vegetation
(kernel: wax-glass-like) with a content of 33-34% DM. Two silages were created : one
by natural fermentation and the other by using an inoculant with mixed types of
homofermentative lactic acid bacteria with high tolerance to acidity, composed of
Lactobacillus plantarum + Pediococcus pentosaceus (10 6 units forming colonies/g).The
research concerning the productive effect was performed on two lots of weaned bull
calves of the Romanian Baltata breed (12 heads / lot), aged about 3 months give or take
14 days. The fattening period lasted 426 days (14 months), respectively from the age of
3 months (after the weaning of the calves) and up to the age of 17 months. Of the two
lots, one was considered a control group (GC) and received naturally fermented corn
silage in its food, and the other was considered an experimental group (GE) and
received corn silage obtained by adding an inoculant (treated). The corn silage ensured
50% of the ration’s DM in both groups; the ration was completed by hashed alfalfa hay
(10% of DM) and a mix of concentrates (40% of DM) (table1). The food was
distributed in abundance in the form of unique forage mixtures.
Table 1: Experimental scheme
Associated activities
Objective Control lot (GC) Experimental lot
(GE)
1. The influence of
using inoculants with
mixed types of lactic
acid bacteria on the
nutritional qualities of
corn silage.
Corn ensiled by
means of natural
fermentation.
328
Corn ensiled by
adding inoculant :
Lactobacillus
plantarum +
Pediococcus
pentosaceus (10 6
units forming
colonies/g).
Surveyed aspects
- The chemical
composition of the
fermented forage
(DM, raw protein,
raw cellulose) ;
- pH ;
- ammoniacal N;
- lactic acid (g/kg
DM);

2. The influence of
using inoculants in corn
ensiling on the fattening
performances of bull
calves.
Structure of the ration (% of DM)
Corn silage - 50
Alfalfa hay – 10
Concentrates* - 40
Corn silage - 50
Alfalfa hay – 10
Concentrates* - 40
- evolution of
weight gain;
- consumption of
DM;
- degree of food
capitalization;
- metabolic
indices in the blood;
- ruminal
parametres (pH , no.
of bacteria/ml,
structure of AGV);
-the quality of the
carcass and meat.
* Mixture of concentrates composed of : corn, barley, soy bean groats, sunflower groats and
cereal-vitamin premix. It ensured 19.8% Pb in the 3-6 months period, 17.5% Pb. in the 6-12
months period and 15.4% in the 12-17 months period.
At the end of the experiment, control slaughterings were performed (n=3), establishing
the main butchery indices. The obtained data was processed and interpreted statistically
by using the ‘Student’ test.
RESULTS AND DISCUSSIONS
The use of the homofermentative lactic acid bacteria based inoculant for ensiling corn,
resulted in a forage having nutritional qualities superior to the one obtained through
natural fermentation (table 2), because the inoculant determined a rapid reduction of the
pH in the forage, which led to the quick reduction of the level of Coliform bacteria, thus
reducing the proteolysis process. This aspect is emphasized by the lower level of
ammoniacal N and the higher level of whole protein in the inoculant treated forage.
Moreover, the inoculant caused a significant decrease in the level of raw cellulose and a
rise in the quantity of lactic acid (g/kg DM).
Table 2: The effect of the inoculant (Lactobacillus plantarum + Pediococcus
pentosaceus) on the quality of the corn silage
Parameters
Control lot
(GC)
Experimental
lot (GE)
Changes due to
the inoculant
treatment (%)
DM (%) 33.75 34.19 + 1.30
pH 4.4 3.8* + 13.64
Raw protein (g/kg DM) 79.2 87.5* + 10.47
Ammoniacal N (%of total N) 6.2 4.1** - 33.88
Raw cellulose (% of DM) 26.9 23.8* - 11.53
Lactic acid (g/kg DM) 128.4 135.2* + 5.29
* p< 0.05; ** p

The positive effects of using the inoculant in the production of fermented forage on the
fattening performances of bull calves were materialized in the weight gain increase of
about 6.38%, the DM consumption improved 5.23% and the degree of food
capitalization increased 4.94% (fig. 1 and table 3). Therefore, the period of fattening
was reduced by 26 days (6.1%) when inoculated silage was administered, by
comparison with the naturally fermented silage, thus saving 82 kg of concentrates and
120 kg of corn silage.
12
11
10
9
8
7
6
5
4
3
2
488.1 515.0
940 1000
Final weight (kg) Weight gain
(g/day)
Group C Group Exp.
9.36
9.85
Consumption
DM (kg/day)
7.91 7.52
Specific
consumption
(UNC/kg gain
Figure 1: The influence of the biological additive used for ensiling corn on the
fattening performances of bull calves
330

Table 3: The evolution of weight gain, DM consumption and degree of food
capitalization
Control lot Experimental
Control Experim
Specification (GC) lot (GE) Specification lot ental lot
(GC) (GE)
Evolution of
Evolution of
body mass
average daily
(kg/head):
gain (g/day):
- 3 months 84.5±1.31 85.3±1.13 - 3-6 months 773 833*
- 6 months 154.8±3.15 161.1±2.97 - 6-9 months 916 970
- 9 months 239.1±4.73 250.3±5.45* - 9-12 months 976 1013
- 12 months 326.9±6.17 341.5±7.02* - 12-15 months 1016 1100*
- 15 months 420.4±5.39 442.7±6.17* - 15-17 months 1110 1186*
- 17 months 488.1±7.34 515.0±5.98** Total
period:
average
Total gain 403.6±5.75 429.7±4.41* 3-17 months 940 1000*
Average daily
Specific
consumption of
consumption
DM (kg/head):
(UNC/kg gain):
- 3-6 months 3.65 3.79 - 3-6 months 4.47 4.30
- 6-9 months 6.46 6.59 - 6-9 months 6.66 6.42
- 9-12 months 8.94 8.76 - 9-12 months 8.65 8.17
- 12-15 months 10.81 11.47 - 12-15 months 10.04 9.85
- 15-17 months 13.18 12.99 - 15-17 months 11.21 10.34
Total average
Total average
period:
period:
3-17 months 9.36 9.85 3-17 months
UNC = 6.2 MJ EN (1481 kcal EN) * p

The positive effect of the silage’s biological additive can be observed in the rumen,
where an improvement of pH and of the relation between the acetic and propionic acids
in favor of the latter, which is beneficial to the production of meat, can be noticed (table
4). This aspect is due to the fact that lactic acid bacteria in the inoculant at the level of
the rumen present probiotic effects (Weinberg, 2003).
There were no noticed differences between the two lots regarding the output at
slaughter, but the surface of the muscle content, which is positively correlated with the
quantity of superior quality meat in the carcass (McDonald, 2002) and the chemical
composition of the meat (leg) were better in the lot that received inoculated silage in the
food (table 5).
Tabel 5: The effect of the biological additive used in the ensiling of corn on the
quality of the carcass and meat
Specification Control lot Experimental lot
(GC)
(GE)
Output at slaughter, % 56.2 56.4
Weight of renal suet, g
Dimension of muscle area, cm
3210 3740
2
- rib section 8-9
- rib section 11-12
Chemical composition of the meat:
- DM, %
- Raw protein, %
- Raw fat, %
61.29
80.62
26.66
21.81
1.88
72.16
81.26
27.81
23.23
1.68
CONCLUSIONS
1. The ensiling of corn by using a biological additive based on homofermentative
lactic acid bacteria (Lactobacillus plantarum + Pediococcus pentosaceus) ensures,
by comparison with natural fermentation, a forage of higher nutritional quality,
having a lower pH (acid), a lower content of ammoniacal N and raw cellulose and
more undegraded protein.
2. The usage of inoculated corn silage in the food of bull calves intended for fattening
ensured a significant improvement in the weight gain, DM consumption and degree
of food capitalization, as opposed to the lot which received naturally fermented
silage in the food. Thus by using inoculant for the corn silage, the period of
fattening decreased by 26 days and 82 kg of concentrates as well as 120 kg of corn
silage were saved.
3. The biological additive used for ensiling did not influence the output at
slaughtering, but it has led to an increase in the dimension of the muscle area,
which is positively correlated with the proportion of superior quality meat in the
carcass.
332

REFERENCES
4. Anon D.L. (1997): – Silage additives: making the best choice. London: Milk
Development Council, 5-17 pp.
5. Halga P.; I.M. Pop; Teona Avarvarei; Viorica Popa (2005): – Nutritie si
alimentatie animala. Ed. ALFA, Iasi.
6. Jones R. (1995): – Forage additive supplement. Farmers Weekly, 26 th November,
24-43 pp.
7. Jones R.; Winters A.; Cockburn J. (1996): – Changes in amino acid content of
inoculated grass silage and its effect on animal production. In: Biotechnology in the
Feed Industry, Proceedings of the 12 th Annual Symposium. Nottingham University
Press, UK, 249-264 pp.
8. Keady T.W.J. (1998): – The production of high feed value grass silage and the
choice of compound feed type to maximize animal performance. In: Biotechnology
in the Feed Industry, Proceedings of the 14 th Annual Symposium. Nottingham
University Press, UK, 439-452 pp.
9. Keady T.W.J. (2000): – Beyond the Science: what the farmer looks for in the
production of silage. In: Biotechnology in the Feed Industry, Proceedings of the
16 th Annual Symposium. Nottingham University Press, UK, 157-174 pp.
10. McDonald P.; Edwards R.A.; Greenhalgh JED; Morgan CA. (2002): – Animal
nutrition. Pearson, Prentice Hall.
11. Mierlita D.; B Nagy; Gh. Salajan, Antonia Odagiu; Mariana Dinea (2000): –
Bioefficiency of bacteria cultures used for corn ensiling. Buletinul USAMV, Cluj-
Napoca, seria Zootehnie, Biotehnologii si MV, vol. 54; 91-102 pp.
12. Steen R.W. (1997): – Factors affecting the utilisation of grass silage for beef
production. In: Efficient beef production from grass. Symposium of the British
Grassland Society, No. 24, 129-137 pp.
13. Suttle N.F. (1998): – Sulphur in forages. Proceedings of Symposium An Foras
Taluntais, Dablin, 197-206 pp.
14. Webb K.E.; Matthews J.K. (1998): – Peptide absorption and its significance in
ruminant protein metabolism. In: Peptides in Mammalian Protein Metabolism.
Portland, London; 104-142 pp.
15. Weinberg ZG.; Muck ME.; Weimer RJ. (2003): – The survival of silage
inoculant lactic acid bacteria in rumen fluid. Journal of Applied Microbiology, 94:
1066-1078 pp.
16. Wilkins RJ (1996): – Silage production and utilization-Perspectives from the
Silage Research <strong>Conference</strong>s. Proceedings of the 11 th <strong>International</strong> Silage
<strong>Conference</strong>, Aberystwyth 37-48 pp.
17. Woolford M.K. (2004): – Inoculantii pentru silozuri: aspecte moderne. Alltech
Technical Publications, 73-108 pp.
333

PHYTOPHTORA INFESTANS - ECOLOGICAL CONTROL WITHIN
ROMANIAN (TRANSYLVANIAN PLANE) POTATO CROP
Ioan Oroian, Liviu Holonec, Viorel Florian, Laura Paulette, Antonia Odagiu
University of Agricultural Sciences and Veterinary Medicine, Faculty of Agriculture,
3-5 Mănăştur St., 400372 Cluj-Napoca, Romania, e-mail: neluoroian@gmail.com
ABSTRACT
The research aimed the following objectives: recording blight appearance, evolution and
action within a technology of organic culture; testing new products and/or products used
in other cultures for potato pathogens; the identification of other natural products used
in disease control; recording the behavior of different potato varieties against mycotic
pathogens in organic agriculture and establishing the sorts of varieties in areas of
interest. Treatments with nine organic products, used for blight control, were performed
in experimental field: nutritive clay 1%, 1/1whey, copper sulfate in association with
lime 1%, bentonite 0.5%, copper oxychloride 4kg/ha, degreased milk 1/1, copper
hydroxide 4kg/ha, cosmetic clay 1% and potassium permanganate 0.3%. The
experimental results and statistical interpretation reveal the validity of the well known
classical products in potato blight control. Those based on copper (4 kg/ha copper
oxychloride, 4 kg/ha copper hydroxide, 1% copper sulfide in association with lime), and
other products tested within this trial and susceptible for favorable effects due to their
pH, pellicular protection, or other properties recoded no significant efficacy, except
whey1/1.
Key words: Phytophora infestans, potato, treatment
INTRODUCTION
The organic agriculture appeared as alternative to intensive, conventional
(industrialized) agricultural practice, based on production maximization using inputs,
and high quantities of the energo-intensive stimulators of production, with the aim of
continuous increase of agricultural production for a preponderantly urban population in
full process of development (Morar et al., 2003). Within organic agriculture, an
assembly of balanced culture techniques (technological chains) in concordance with
natural environmental rules describes the potato culture, with the aim of obtaining crops
of high nutritional and health quality. Because in our country, only the first stages of the
organic agriculture are developing and elaborated studies on potato culture within
organic system were not yet performed, the study of the suitability of potato culture
within this agricultural system is imposed. The control of the main mycotic pathogenic
agent, which produces blight in this culture, will be focused (Donescu et al., 1996;
Orian, 2003; Oroian et al., 2003; Plămădeală, 1999).
MATERIAL AND METHOD
The experiments were performed in pedo-climatic conditions of the Transylvanian
Plane, with a transition temperate clime and specific traits determined by the general
orientation of the summits and slopes of Jucu village. This area is part of the wet and
colder specific spreading area of the Transylvanian Plane, Someş Plane. The vertic clay
334

chernozem represents the dominant soil, which exhibits the succession of the Am-Bty-
Cca horizons characteristic for the forest steppe areas, with marls rich in CaCO3. They
confer the soil two essential traits: heavy texture and high degree of basic saturation.
The determined agro-chemical indices are characterized by the neutral soil reaction,
with values between 6.9 - 7.1 (in water), 3.56 - 3.92% humus content in arable layer,
0.183 - 0.196% total nitrogen supplies, very high phosphorus content (15 ppm) and
mobile potassium (240 ppm).
The potato varieties used within this trial were planted at 70 cm distance between
rows using 4SaBP equipment. Two rows by variety were planted. The following
varieties were used: Tentant, Frumoasa, Robusta, Rozal, Redsec, Roclas, Dacia, Rodas,
Amelia, Agatha, Sante and Cristian.
In experimental filed, treatments with nine blight control organic products were
performed: nutritive clay 1%, whey 1/1, copper sulfate in association with lime 1%,
bentonite 0,5%, copper oxychloride 4kg/ha, degreased milk 1/1, copper hydroxide
4kg/ha, cosmetic clay 1% and potassium permanganate 0,3%.
The experimental data were statistically processed and interpreted using the dispersion
analyzes (ANOVA) and Duncan test.
RESULTS
The results concerning the blight attack degree (AD%) on leafs in studied varieties are
presented in figures 1 - 11. The observation was performed after two weeks from
treatment application.
Figure 1 represents the control diagram, and it is the reference point for the results
obtained after treatment with unconventional products used in fight against potato
blight. Within control variant, Robusta, Rozal, Dacia, Sante and Cristian varieties
recorded the lowest blight attack degree, while Roclas variety recorded the highest.
AD%
5
4,5
4
3,5
3
2,5
2
1,5
1
0,5
0
Tentant
Frumoasa
Robusta
Rozal
Redsec
Roclas
Figure 1: The behavior against mushroom Phytophthora infestans attack in control
Positive results after treatment with nutritive clay were recorded in Tentant, Roclas and
Robusta varieties (figure 2). The attack degree recorded in Robusta variety was 0 (zero),
while Agatha and Rivera varieties had negative reaction.
335
Dacia
Amelia
Agatha
Rivera
Sante
Cristian

AD%
4
3,5
3
2,5
2
1,5
1
0,5
0
Tentant
Frumoasa
Robusta
Rozal
Redsec
Roclas
Dacia
Amelia
Agatha
Rivera
Sante
Cristian
Figure 2: The influence of the nutritive clay treatment on the attack degree
One positive reaction was recorded when whey treatment was applied in Roclas variety
(figure 3). 10% decrease was obtained compared to control. Neutral reaction was
obtained in Tentant variety.
AD%
5
4,5
4
3,5
3
2,5
2
1,5
1
0,5
0
Tentant
Frumoasa
Robusta
Rozal
Redsec
Roclas
Dacia
Amelia
Agatha
Rivera
Sante
Cristian
Figure 3: The evolution of the blight attack when whey was used
After treatment with copper sulfate in association with lime, the Roclas variety recorded
a positive reaction, with 20% decrease of blight attack degree compared to control. In
Tentant, Agatha and Rivera varieties a neutral reaction was recorded (figure 4).
The bentonite treatment determined positive reaction in two varieties, meaning
Roclas, where the attack degree decreased by 20% compared to control and Agatha
where the attack degree decreased by 10% compared to control. In Tentant, Dacia and
Rivera varieties a neutral reaction was recorded (figure 5).
336

AD%
AD%
4
3,5
3
2,5
2
1,5
1
0,5
0
Tentant
Frumoasa
Robusta
Rozal
Redsec
Roclas
Figure 4: The effect of the copper sulfate in association with lime on the attack degree
4
3,5
3
2,5
2
1,5
1
0,5
0
Tentant
Frumoasa
Robusta
Rozal
Redsec
Roclas
Figure 5: The effect of the treatment with copper bentonite on the attack degree
Only in two varieties a significant positive reaction was recorded when the treatment
with copper oxychloride was applied: in Roclas variety the attack degree decreased by
20% compared to control and in Tentant the attack degree was 10% lower compared to
control. In the other variants, a neutral (Agatha and Rivera) or negative reaction was
recorded (figure 6).
337
Dacia
Dacia
Amelia
Amelia
Agatha
Agatha
Rivera
Rivera
Sante
Sante
Cristian
Cristian

AD%
4
3,5
3
2,5
2
1,5
1
0,5
0
Tentant
Frumoasa
Robusta
Rozal
Redsec
Roclas
Figure 6: The effect of the treatment with copper oxychloride on the attack degree
The degreased milk treatment determined a positive reaction in Roclas variety where the
blight attack degree decreased by 20% compared to control. In Tentant, Agatha and
Rivera varieties, a neutral reaction was determined. In Robusta, Rozal, Redsec and
Cristian varieties, de attack degree increased by 20% compared to control (figure 7).
AD%
4
3,5
3
2,5
2
1,5
1
0,5
0
Tentant
Frumoasa
Robusta
Rozal
Redsec
Roclas
Figure 7: The influence of the treatment wioth degreased milk on the attack degree
When copper hydroxide treatment was applied, a positive reaction was recorded in
Tentant and Roclas varieties, with a 10% decrease of the attack compared to control in
each variety. In Rivera variety a neutral reaction was recorded (figure 8).
A positive reaction was recorded in Roclas and Tentant varieties when cosmetic
clay treatment was applied. The blight attack degree decreased by 40% compared to
control in Roclas variety and by 10% in Tentant variety (figure 9).
338
Dacia
Dacia
Amelia
Amelia
Agatha
Rivera
Agatha
Rivera
Sante
Cristian
Sante
Cristian

Negative reaction was recorded in the following varieties: Cristian, where the blight
attack degree increased by 30% compared to control, and Frumoasa, Robusta, Rozal,
Redsec, Dacia, Amelia and Sante varieties, where the blight attack degree increased by
10% compared to control.
AD%
4
3,5
3
2,5
2
1,5
1
0,5
0
Tentant
Frumoasa
Robusta
Rozal
Redsec
Roclas
Figure 8: The effect of the treatment with copper hydroxide on the attack degree
AD%
4
3,5
3
2,5
2
1,5
1
0,5
0
Tentant
Frumoasa
Robusta
Rozal
Redsec
Roclas
Figure 9: The effect of the treatment with cosmetic clay on the attack degree
The potassium permanganate treatment determined a positive reaction in Tentant and
Roclas varieties, where the blight attack degree decreased by 10% compared to control.
The negative reaction was recorded in: Cristian variety, where the attack degree
increased by 30% compared to control, and Rozal, Dacia and Sante varieties, where the
blight attack degree increased by 20% compared to control (figure 10).
339
Dacia
Dacia
Amelia
Amelia
Agatha
Agatha
Rivera
Rivera
Sante
Cristian
Sante
Cristian

AD%
4
3,5
3
2,5
2
1,5
1
0,5
0
Tentant
Frumoasa
Robusta
Rozal
Redsec
Roclas
Figure 10: The effect of the treatment with potassium permanganate on the attack degree
The treatments applied with the aim of blight control determined a different reaction in
studied varieties, except Roclas variety, which was the only variety that positively
reacted to all applied treatments (figure 11).
Control
Nutritive clay
Whey
Copper sulphate in association with lime
Bentonite
Copper oxychloride
Degreased milk
Copper hydroxide
Cosmetic clay
Potassium permanganate
Figure 11: The evolution of the blight atack within the interaction between
variety and treatment
340
Dacia
Tentant
Amelia
Frumoasa
Robusta
Agatha
Rozal
Redsec
Rivera
Roclas
Sante
Dacia
Amelia
Cristian
Agatha
Rivera
Sante
Cristian
5
4
3
2
1
0

The Tentant variety positively reacted to five treatments, and had a neutral reaction to
the other applied treatments. The Robusta variety positively reacted to the treatment
with nutritive clay, where recorded the attack degree was 0 (zero), to other two
products, and a negative reaction to the other applied treatments. The Agatha variety
positively reacted only to one product, negatively to one product, too, and neutral to the
others. The Rivera variety positively reacted to one product, negatively to one product,
and neutral for the rest of them. In the other varieties a negative reaction was recorded.
DISCUSSIONS
The well-known copper based classical products (copper oxychloride 4kg/ha, copper
hydroxide 4kg/ha, copper sulfate in association with lime 1%) are efficient in potato
blight control. The other tested products susceptible of favorable effects due to their pH,
pellicle protection or other properties were not efficient, except the variant with whey
1/1.
Within experimental conditions, the lowest potato blight attack was emphasized in
Amelia and Robusta variety, and the lowest alternariosis attack in Sante, Robusta and
Rozal varieties, which recommend them as suitable for organic agriculture.
Among the tested varieties, five are suitable to be used in organic agriculture: Robusta,
Rozal, Redsec, Amelia and Sante. High production and degree of blight and alteriosis
resistance characterize these varieties during testing conditions. Further research is
needed in order to identify other varieties suitable to be used within organic agriculture
system.
The use of unconventional products susceptible to have properties against potato
pathogenic agents or capacity of their attack limitation represents an alternative which
can be developed within the context of implementation and extension of the sustainable
and organic agriculture system.
REFERENCES
Donescu D., Enoiu, M., Frâncu G., Plămădeala, B., Popescu, A. (1996): Ghid practic
de protecţie a cartofului. Ed. Ceres, Bucureşti
Morar, G., Fiţiu, A., Cernea, S., Vâtcă, S. D., Oltean, M. I., Sârbu Camelia (2003):
Tehnologii în agricultura ecologică, Ed. Risoprint, Cluj-Napoca
Oroian, I. (2003): Protecţia şi nutriţia plantelor, Ed. AcademicPress, Cluj-Napoca
Oroian, I., Fiţiu, A., Florian, V., Carmen Puia, Dumitraş Adelina, Roiban, G.,
Paulette Laura (2003): Controlul patogenilor în agricultura ecologică,
Ed. AcademicPress, Cluj-Napoca
Plămădeală, B. (1999): Mana cartofului. Biologie şi control, Ed. Ceres, Bucureşti
341

THE DYNAMICS OF THE POPULATIONS OF CEREAL PESTS AND
PATHOGENS IN THE ROMANIAN WESTERN PLAIN
Elena Bucurean, Nicolae Csép
University of Oradea, Faculty for Environmental Protection Oradea
SUMMARY
In a given agroecosystem, the exact determination of pest species, meaning primary,
occasional, potential and migrating pests, the identification of harmful plant pathogens,
requires particular attention: these species are different for each crop or zone, an aspect
that imparts certain characteristics to the fighting schemes.
Over the years, it has been noticed that the damages made by pests and diseases in
the western part of the country are not always of equal magnitude: these damages
greatly depend on the environmental and biotic factors’ influence on the species present
in the area (Bucurean, 2004; Csep, 2006). Consequently, there are some species that are
present and cause damages every year, species of both cyclical and seasonal economic
importance and there are other species that are simply part of the area’s fauna.
The spectrum of plant-specific pathogens also undergoes changes, especially under
the influence of the regime of temperature and humidity during the crop’s vegetation
period. Performing the meteorological observations and establishing the inoculum
reserve holds special importance in the application of environment-friendly
technologies.
Key words: wheat, pests, diseases, damages.
INTRODUCTION
Taking into account the fact that winter wheat is cultivated in over 45 countries, feeding
35–40% of the world’s population, as well as the fact that it ensures about 40% of the
total need of calories in human diet, it is absolutely imperative to be aware of the
entomofauna and pathogens present in cereal crops every year, in order to foresee and
then prevent the appearance of pest invasions (Ghizdavu et al.,1998; Perju et al.,1993)
and epiphytotic infections due the harmful pathogens. This aspect also stands with
regard to other local crops, with the addition that it is necessary to be aware of the
populations’ tendencies and the reserve of inoculum, in relation to the evolution of the
abiotic (temperature, humidity, soil, air currents etc.) and traffic factors (food and the
relations between predators, parasites and host pests).
RESULTS
a.) Pest populations: In cereal crops, the tendency of the existing pest populations in
the area is of slight increase in cereal bugs (Eurygaster spp, Aelia spp), wheat thrips
(Haplotrips tritici), wheat chafer (Anisoplia spp), cereal leaf beetle (Oulema
melanopus), cereal green aphid (Schyzaphis graminum) and various species of
phytophagous diptera (Oscinella sp, Agronyza sp, Phytomiza sp), wire worms (Agriotes
spp) as well as Continental field vole (Microtus arvalis), whose attack however has a
cyclical and seasonal character.
342

Table 1. The density of harmful pest species in wheat crops from Oradea in the 2005
– 2006 agricultural year
Species Date of control Density
Zabrus tenebrioides October 8 2.7 larvae/ m²
Eurygaster maura May 11 6.2 adults / m²
Eurygaster integriceps May 11 3.1 adults / m²
Aelia rostrata May 11 4.1 adults / m²
Oulema melanopus April 9 12 adults / m²
Oulema melanopus May 13 220 larvae / m²
Schyzaphis graminum June 24 56 aphids / plant
Haplotrips tritici June 16 27 adults and larvae / ear
Anisoplia spp June 29 9 adults / m²
Oscinella frit October 22 11.3% attacked plants
Cephus pygmaeus June 29 3.4% attacked plants
Macrosteles laevis October 28 11 adults / m²
Agriotes spp April 5 6 larvae / m²
Naturally, some of these species hold a minor economic importance in the area, partly
as a result of the fact that plants rebuild their vegetative mass, thus no production losses
due exclusively to pests are registered.
Of the total number of pests collected in cereal ecosystems, the dominant species were
the ones whose frequency is presented in table 2. The numbers represent the average
values of mowings at different dates.
Table 2. The frequency of pest fauna (%) in cereal crops
Crop Wheat Barley Oats Triticale
Species % % % %
Phyllotreta spp 29.7 22.7 26.7 42.9
Eurygaster maura 4.5 2.8 0.7 0.2
Aelia acuminata 1.9 1.4 0.7 0.5
Oulema melanopus 58.9 11.8 77.8 10.7
Cephus pygmaeus 12.4 6.7 2.9 16.5
Haplotrips tritici 71.0 41.6 - 0.2
Schyzaphis graminum 46.2 44.4 45.7 2.9
Anisoplia spp 4.9 2.6 - 1.5
Agromyza nigra 18.0 2.3 - -
As to maize crops, taking into account the fact that the majority of soils in the area have
an acid or weak acid reaction, particular difficulties are caused by the larvae of Agriotes
spp, which populate these soils. Their density fluctuates from one year to another,
depending on the quantity of rainfall during autumn – spring and on the humidity of the
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soil during the sprouting period, this density varying between 12 – 16 larvae of different
ages / m². Moreover, important damages are caused by the larvae of the turnip moth
(Agrotis segetum), which, due to their accentuated polyphagism, bring about damages
not only in maize crops, but also in beet, potato and leguminous plants. The density of
the butterflies caught by the pheromonal traps varied from one month to another,
reaching a peak during the flight period.
b.) Plant pathogens: Regarding the culture of winter wheat, the structure of the
complex of pathogens underwent modifications as to the cultivated species, as well as to
the climatic changes. The last 5 years, having had a rather droughty character, have
been less favorable to pathogen infections, favoring the proliferation of insects instead.
Table 3. The main winter wheat diseases in the Romanian Western Plain
Symptomps Disease
Cause agent Economical
expressed on: common name
importance
Leaves Barley Yellow
Dwarf
BYDV medium
Leaves Powdery Erysiphe graminis f.sp. low
mildew tritici
Leaves leaf spot H. tritici-repentis medium
Leaves leaf rust Puccinia recondita low
Leaves stripe rust P. striiformis variable
Seed / head head blight Fusarium spp. medium
Head common bunt T. foetida, T. Caries variable
Lower stem eyespot Pseudocercosporella
herpotrichoides
variable
Lower stem sharp eyespot Rhizoctonia sp. low
The symptoms of barley yellow dwarf virus were frequently identified between the
years 2000-2002, when the installation of vectors (Aphidae) in autumn was noticed in
strongly developed crops. Because no control is available for many viral diseases, such
as barley yellow dwarf, we must supervise the installation (and, when needed, the
fighting) of vectors.
Of the foliar diseases, the importance of the powdery mildew (Erysiphe graminis)
decreased, while that of the yellow leaf spot (Helminthosporium tritici-repentis Died.)
increased, the symptoms of the latter on the upper leaves being easily mistaken for the
effects of the drought. The pathogen is causing, in the current conditions, the greatest
yield losses (up to 250-350 kg/ha), associated with the reduction of seminal quality.
Fungicidal treatment with systemic fungicides, according to regional advice at heading,
balanced fertilisation can contribute effectively to the limitation of losses.
The brown leaf rust (Puccinia recondita) has been present every year, without
affecting more than 5 % of the leaves’ area of assimilation. The stripe rust (Puccinia
striiformis) appeared on some very susceptible wheat species, at the end of the
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vegetation period, without affecting more than 3% of the area of upper leaves. However,
the species represents a potential danger, due to possible damages.
The evolution of droughty climate during the past few years has significantly
reduced head blight due to infections with Fusarium spp. The infections are tightly
connected to the evolution of the climatic factor in the post – heading period.
In the years following the re-establishment of former properties, as a result of not
performing or inadequately performing seed dressing, the losses in production caused
by the common bunt have resurfaced. Of the species of Tilletia sp., the most frequent in
this ecological area are T. caries and T. foetida. During those years, frequencies of
attack on the ears of up to 15-20% were recorded. The correct application of fungicides
has reduced this indicator below the frequency of 0.2%. Although it is present in many
Transylvanian counties, T. nanifica has not been recorded in this area.
Lower stem diseases (eyespot - Pseudocercosporella herpotrichoides; sharp
eyespot -Rhizoctonia sp.; fusarium rot – Fusarium sp.) are present only occasionally,
drawing attention on some technological causes that have favored their appearance:
monocultures, unbalanced fertilization, and very susceptible wheat species. Crop
rotation (avoid planting wheat and barley after take-all infested wheats, balanced
fertilisation) are the most efficient preventive measures.
Table 4. The main winter barley diseases in the Romanian Western Plain
Symptoms Disease common Cause agent Economical
expressed on: name
importance
Leaves Barley yellow
dwarf virus
BYDV medium
Leaves Powdery mildew Erysiphe graminis
f.sp.hordei
low
Leaves / head Leaf stripe Pyrenophora
graminea
medium
Leaves Rhynchosporium Rhynchosporium medium
leaf-spot
secalis
Head True loose smut Ustilago nuda variable
Head Covered smut Ustilago hordei low
As to the cultures of winter barley that have profited from adequate seed dressing, the
presence of seed-born pathogens, true loose smut (Ustilago nuda), covered smut (U.
hordei), barley leaf stripe (Pyrenophora graminea), was below the limit of provoking
some important yield losses.
CONCLUSIONS
1. The pests of cereal crops represent limitative factors for the production and
supervising them in the area is a necessity. Therefore, it is imperative to monitor
their numerical evolution in time, on groups of cultures. The pest fauna is widely
345

epresented in this area, and annual damages are recorded as a result of the attacks
of the cereal leaf beetle, cereal bugs and wire worms.
2. The largest number of harmful pest species was registered in wheat crops, in
comparison with the other cereal crops (barley, oats, triticale). The cereal leaf
beetle’s presence corresponded to percentages between 10.7 and 77.8%, which
proves that this species represents a real problem for the culture of cereals.
3. The specific evolution of climatic factors has caused a temporary reduction of some
pathogens considered dangerous (head blight, eyespot, fusarium rot), but which,
due to their high infectious potential, must be under constant surveillance in order
to avoid possible damages.
4. Seed dressing , avoiding early seedling, balanced fertilisation, crop rotation,
fungicidal treatments especially after heading, ensure the most effective and safe
protection of cereals against pathogens.
REFERENCES
1. Bucurean Elena, 2004: Prognoza si avertizare in combaterea daunatorilor la
culturile agricole, Ed. Universităţii din Oradea.
2. Bucurean Elena et al., 2006: Protecţia eficientă şi sigură a plantelor faţă de
dăunători. Ed. Universităţii din Oradea.
3. Csep N.,2006: Protecţia eficientă şi sigură a plantelor faţă de boli.Ed.Universităţii
din Oradea.
4. Ghizdavu I. et al., 1998: Entomologie agricola, E.D.P. Bucuresti
5. Perju T. et al., 1993: Protectia integrata a culturilor de leguminoase, Ed. Ceres
Bucuresti
346

THE IMPORTANCE OF CONTROLLING FORMALDEHYDE FUMIGATION
Lucian Bara, Camelia Bara
University of Oradea, Faculty of Environmental Protection
26, General Magheru Street, Oradea
ABSTRACT
A comparative study of formaldehyde (HCHO) fumigation was carried out by controlled
vaporization, using an electric vapor generator, and by the Formalinpermanganate.method.
Determination of vapor levels as well as bactericidal action showed the generator to be more
effective. Maximum achievable fumigant levels were temperature dependent and related to
the equilibrium vapor concentration of HCHO. At a room temperature of 21°C, vaporization
of more than 2,000 ytg of HCHO per liter resulted in conversion of HCHO to
paraformaldehyde, which condensed on surfaces and contributed to prolonged residual
vapor levels.
An electronic monitor is described which is capable of detecting HCHO levels as low
as 10 Ag/liter and can be used to monitor the complete fumigation process.
INTRODUCTION
Formaldehyde fumigation has long been an accepted method of sterilization for areas where
microbiological cleanliness is required. Fumigation is usually carried out by mixing
potassium permanganate and an excess of Formalin (40% formaldehyde) solution in a
suitable container; sufficient heat is generated by oxidation of part of the formaldehyde
(HCHO) with permanganate to vaporize the remaining formaldehyde and water.
This method of fumigation is violent, messy, and potentially explosive. In this paper,
an alternative procedure is described for fumigation by controlled vaporization, in an
electrical vapor generator, of either Formalin or paraformaldehyde, a polymer of HCHO
which is converted to HCHO vapor upon heating. Formaldehyde levels were compared in
fumigations of several work areas by the controlled vaporization and Formalinpermanganate
methods, and the effectiveness of each procedure was determined from the
survival of resistant Bacillus subtilis spores.
MATERIALS AND METHODS
Reagents. A commercial Formalin solution consisting of 40% HCHO containing 10%
methanol as a depolymerizing agent was used throughout. Paraformaldehyde powder was a
white polymer consisting of 95% paraformaldehyde.
Areas fumigated. A low-security area and a highsecurity area were monitored throughout
the study. The low-security area was a large processing laboratory; during fumigation the air
conditioning was turned off, but no attempt was made to seal air supply or exhaust ducts.
The high-security area was a large sealed laboratory whose air supply was carefully
controlled; exhaust air was passed through a gas-fired furnace to ensure microbiological
347

security. The air conditioning for this area was turned off during fumigation, and the air
supply and exhaust outlet valves were closed.
Assay of HCHO. Initially, formaldehyde vapor concentrations were measured by taking air
samples through rubber tubes protruding into the room at various sample points. Four
samples were taken per sampling time, using an evacuated liter flask. Forty milliliters of 0.5
M (NH4)2SO4 was added to each flask, which was then cooled to 4'C to dissolve the
formaldehyde. One milliliter of a suitable dilution of the resulting solution was then mixed
with 1 ml of a freshly prepared chromotropic acid reagent (1% chromotropic acid in 18 M
H2SO4), and 8 ml of 18 M H2SO4 was added. After standing for 10 min, the mixture was
diluted to a final volume of 25 ml, and the optical density was determined at 540 nm.
HCHO was estimated from a standard curve in the range 5 to 40 yg/ ml, and the mean
concentration from the four samples was determined.
Biological test systems. The spore suspension used throughout was prepared from B.
subtilis NCTC 8233 prepared by culture for 7 days on sporulation agar and suspension of
the resultant growth in water. The suspension was subjected to two cycles of heating to
65°C for 30 min followed by centrifugation and resuspension in distilled water. A sample
was then stained and examined microscopically for spores. Glass rods 5 mm in diameter and
150 mm long, with a loop at one end and a graduation mark 50 mm from the other, were
dipped into the spore suspension to the mark. They were then suspended in a laminar flow
cabinet to dry and stored in a sterile jar. Before fumigation, rods were placed in different
parts of the areas to be tested and transferred to a sterile screwcapped test tube at the end of
each procedure, immediately after evacuation of the fumigant. Twenty milliliters of saline
was added to each tube to remove the spores from the rods, and an estimate of viable spores
was obtained by plating dilutions of the suspension onto nutrient agar. This was carried out
within 2 h of removal of rods from the area. Plates were incubated at 37°C, and then
colonies were counted after 3 days and checked after 7 days.
Electric vapor generator. Initially, HCHO was vaporized on a 500-W heating plate in a
stainless-steel beaker. The apparatus used subsequently, consists of a stainless-steel 5-liter
vessel with a 1,200-W heating element. Sufficient capacity was present to allow the addition
of water for humidification and to contain the resulting foam. A copper plate was interposed
between the element and the bottom of the vessel to ensure a uniform distribution of heat at
a thermostatically controlled temperature of approximately 170°C.
Electronic formaldehyde monitor. The apparatus used was based on a Figaro 812 N-type
semiconductor device, the conductivity of which changes in the presence of adsorbed
molecules. No significant interference was observed from substances other than HCHO in
the areas fumigated or by temperature changes normally encountered. The monitor was
calibrated in an enclosed system against HCHO released by paraformaldehyde vaporization.
RESULTS
HCHO levels by the Formalin-permanganate and hot plate vaporization methods. For
Formalin-permanganate fumigations, quantities of Formalin recommended have ranged
from 12 to 59 ml/m 3 of air space , and ratios for Formalin-permanganate have ranged from
348

3:1 to 5:3 . In the following experiments, fumigations were carried out by using a mixture of
17.8 ml of Formalin and 8.9 g of potassium permanganate per m 3 of air space in a lowsecurity
area at room temperatures of 16, 24, and 300C. In preliminary experiments, room
air samples were taken from each of four positions, including the ceiling and the floor, and
HCHO levels were determined by chemical assay. Because no differences in concentration
were observed between any positions, all results from the sampling points were averaged.
HCHO levels were also measured after vaporization of 12.5 ml of Formalin (70% of that
used in the Formalin-permanganate fumigations to allow for losses due to permanganate
oxidation) on a hot plate at a room temperature of 27°C.
Survival of B. subtilis after fumigation by the Formalin-permanganate method. B.
subtilis spores survived on some rods located at various places in a low-security area during
a 24- h Formnalin-permanganate fumigation at an initial temperature of 1800. Viable counts
of 2 x 106 were obtained from control rods, but only two of nine rods exposed in the treated
area were sterile; viable counts of 20 to 600 were obtained from the others. It is clear that,
although this procedure was fairly effective, sterilization was not achieved.
Two further experiments were carried out in a sealed jar at 20°C in controlled HCHO vapor
levels of 150 and 300 Ag/liter of air, with relative humidity maintained at approximately
100%. Exposure to 150, ug of HCHO per liter of air did not result in sterilization even after
exposure for 6 h. Exposure to 300, Ag/liter was much more effective, and no viable spores
were detected at 3 h.
Use of electric vapor generator. The results from several fumigations are presented, using
the electric vapor generator and different amounts of Formalin in a low-security area at
different initial temperatures. In procedure 1, persistent HCHO levels similar to those
obtained by the hot plate vaporization method were achieved. No viable B. subtilis spores
were detected after 24 h of exposure on the 18 spore rods located in the main laboratory
area, but a 0.5% survival was detected on a rod located in a closed cupboard. The fumigant
level in the cupboard was 160, ug/liter after 6 h. Procedure 2 was carried out with the same
volume of Formalin and an initial room temperature of 15°C. No surviving spores could be
detected after 7 h of exposure on 12 spore rods located around the room, but two located in
cupboards showed a survival of 1%. Procedures 3 through 5, which were carried out for
different times and temperatures and with varying amounts of Formalin, are also described.
In procedure 6, a mixture of paraformaldehyde and water equivalent to 8.9 ml of Formalin
per m 3 of air was used. Again, no surviving spores could be detected from rods distributed
throughout the main laboratory area in any of the procedures. All subsequent fumigations
were carried out with 3.7 g of paraformaldehyde per m3 of room volume, which is
converted to HCHO equivalent to that present in 8.9 ml of Formalin.
Humidity in each area was increased to 80%, a level which has been shown to increase
the effectiveness of fumigation by causing HCHO to dissolve in a film of moisture around
microorganisms, where it is more bactericidal than in vapor form .Use of Formalin at a
room temperature of 200C would have contributed approximately 30% to the relative
humidity. More reliable humidity control could be achieved with paraformaldehyde, which
was also more convenient to use. Use of electronic HCHO monitor. The electronic monitor
was developed because of the time involved in chemical assays of air samples. When the
349

output of the monitor was plotted against HCHO vapor levels obtained by chemical
analysis, a power-law relationship was obtained. Such a relationship is particularly useful,
since good sensitivity was achieved at the lower vapor levels and the total fumigation range
was covered on the same scale.
Relationship between temperature and equilibrium vapor concentration. The
relationship between temperature and the equilibrium vapor concentration is shown in. For
normal air-conditioned laboratories at 20 o C, the equilibrium vapor concentration is
approximately 2000 ,ug of HCHO per liter. HCHO vapor in excess of the equilibrium vapor
concentration will recondense on the coldest surfaces, producing a paraformaldehyde film
over surfaces within the room. This film slowly evaporates, producing an unpleasant odor.
HCHO levels after fumigation. The persistence of HCHO vapor after completion of the
fumigation and restoration of ventilation is a significant problem, especially in high-security
laboratories. The extent of the problem can be seen from chart recordings from the
electronic monitor during fumigation of a high- and a low-security area.
DISCUSSION
The data presented above demonstrate several advantages of the HCHO generator over the
Formalin-permanganate method in HCHO fumigation. Rapidly falling vapor levels in the
permanganate method are due, in part, to the formation of water condensate on cold surfaces
which, because of the high partition of coefficient of HCHO in water , rapidly takes up the
HCHO vapor. High-localized HCHO levels generated by rapid vaporization in this method
also lead to excessive condensation of paraformaldehyde.
From the standpoints of safety, convenience, and effectiveness, the HCHO generator
method was more successful, particularly where large areas were to be fumigated. It is not
necessary to rapidly leave the area after addition of Formalin. Instead, it is only necessary to
adjust a timer well in advance of the fumigation, and any necessary sealing of doors and
other outlets can then proceed at leisure and without the discomfort of leaking vapor. The
importance of adequate sealing is illustrated by the rapid loss of fumigant from a lowsecurity
area. Slow diffusion of vapor into closed cupboards was demonstrated in procedure
1. Cupboards, drawers, and other such fittings in the room must be opened to allow
adequate penetration of fumigant.
The value of maintenance of a uniform temperature was particularly apparent at a
fumigant concentration of 1.500, ug of HCHO per liter. At this level, the restoration of
ventilation has rapidly removed all traces of the fumigant down to the level of human
detection, commonly accepted as 1 ug/liter. This level complies with most of the
international standards currently in use.
350

REFERENCES
1. Bartzokas, C. A., K. McCarthy, W. B. Shackleton, and B. F. Baker. 1978, Observations
of the effects of formaldehyde on cockroaches and their flora. 1. Survival
of vaccinia virus infected cockroaches during fumigation with formaldehyde, J. Hyg.
80:125-129.
2. BraswelI, J. R., D. R. Spiner, and R. K. Hoffman. 1970, Adsorption of formaldehyde by
various surfaces during gaseous decontamination. Appl. Microbiol. 20: 765-769.
3. Tucker, J. F., E. G. Harry, and H. E. Wainman. 1975, The effect of fumigation with methyl
bromide or formaldehyde on the infectivity of poultry house litter naturally
contaminated with Salmonella virchow. Br. Vet. J. 131:474-485.
4. Wade, A. 1977. Disinfectants and antiseptics, p. 520. In W. Martindale (ed.), The extra
pharmacopoeia, 27th ed. The Phannaceutical Press, London.
351

THE CA AND MG SUPPLY OF THE SOIL EXTRACTED WITH DIFFERENT
SOLVENT SOLUTIONS
István Kocsis 1 Lajos Szabó 1 Simándi Péter 1 László Pásztor 2
1 Tessedik Sámuel College, Faculty of Agricultural Water- and Environment Management
H-5540 Szarvas, Szabadság str. 1-3., Hungary
2 Plant and Soil Protection Service of the Jász-Nagykun-Szolnok County, Szolnok
INTRODUCTION
Information upon Ca content of the soil used to be crucial
• if in the arable layer, calcium carbonate is lacking, and
• if calcium content of the soil is excessive in relation to the colloidal capacity of the
respective layer (Filep, 1988, Németh, 1971, Filep, 2002.).
There are alternative methods for characterising the Ca content of soils. The soils are well
supplied with calcium:
• if the content of CaCO3 is between 1 and 5 % measured with the method of Scheibler
• if the measured AL-Ca values are about 50-100 mg/100 mg in sandy soils, 100-200
mg/100 g in loam soils, and 500-700 mg/100 g in clayey soils.
• if the Ca content is exchangeable at a 70-80 % rate of the S value
• if the Ca values obtained with the EUF method are within the interval 35.0-60.0 mg/100
(Filep 2002)
Results of the method according to Scheibler are not informative for soils containing little
Ca, whereas extraction with AL gives erroneous results for calcareous soils. Determination
of exchangeable cations is performed with different solvents than with other nutritive
elements. The EUF method measures Ca together with other elements, however, it requires
a special device.
Excessive or insufficient Ca disturbs the uptake of other elements in the soil. The rule
of calcium-potassium influencing the absorption of phosphorus by Ca as well as the
antagonisms of Ca:Mg and Ca:K are widely recognised. The role of Ca and Mg influencing
the uptake of meso- and micro-elements is also known (e.g. abundance of Ca may cause
chlorosis due to iron deficiency). The Ca-EUF Mn ratio indicates that two agricultural
elements, the liming and the loosening of the soil could be applied.
The ratio between the contents of lime and colloids is considered as an important parameter
of the soil. The Ca status should reasonably reflect the availability of both macro- and
microelements. Information related to the Mg status is necessary mainly in sandy and in
alkaline soils. Relevant methods are largely similar to those applied in the case of Ca
determination. The solvent solution however should not contain the element in question:
(e.g. Ca salt is suitable for assessing Mg e.g. 0.01 M CaCl2). For Ca and Mg extraction KCl
solution is recommended. In assessing the Ca content available for the plants, mild solvents
give more reliable information than the more concentrated ones. For the determination of Ca
content, we used mild solutions, which facilitated the measurement of other elements too.
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For micro-elements, EDTA+KCl 15 times diluted was applied. Instead of EDTA,
EDTE+NH4Cl at the same concentration proved to be useful to measure Na and KCl
content. For the determination of Ca content, mild solvents completed with a complexforming
substance have been employed.
MATERIAL AND METHOD
The macerate with HNO3+H2O2 was prepared according to the standard MSZ 21470-
50:1998 (For all elements). Many elements of the soil are indifferent from the point of view
of plant nutrition; however, their registration fixes the “upper point” or the maximum value
for comparing it with the data obtained with other extraction methods.
Ethylene-diamine-tetra-acetate + KCl, or EDTA according to MSZ 20135:1999, serves for
the measurement of micro-elements but it is less used in the international practice. The KCl
is an active solvent that carries a complex forming effect and keeps the micro-elements in a
dissolved status.
Ammonium-lactate, MSZ 20135:1999 or AL. The acid buffering system (pH = 3.75)
overestimates substantially the available P and Mg content of alkaline (carbonatecontaining)
soils, whereas underestimates them in acid soils.
Ammonium-chloride with EDTE (EDTE+NH4Cl) solution.
It means 0.0033 M ethylene-diamine-tetra-acetic-acid, 0.0067 M ammonia-solution and
0.0067 M ammonium chloride per 1 dm 3 .
Preparation: a 1000 cm 3 test-tube is filled with 0.96 g ethylene-diamine-tetra-acetic-acid
(selection-B) and 40 cm 3 distilled water added. Then 0.9 cm 3 25 m/m % ammonia-solution
is mixed to the selection-B until being completely dissolved (3-4 minutes).
Further 0.33 g ammonium-chloride is dissolved and filled to about 950 cm 3 with distilled
water. The pH is adjusted to 7.00 ± 0.05 with 0.01 M hydrochloric acid or ammonia
solution, if necessary, and completed until the caesura with water.
RESULTS AND DISCUSSION
For the characterisation of soil profiles, the important moment is the assessment of the Ca
and Mg profile. From the point of view of fertility, a deeply penetrating humus layer and a
thin, upper, lime-less layer are advantageous.
The Ca and Mg supply of the arable layer signalises a favourable nutritional status and
granular structure. Ca content of the soil is not only considered in the upper layer but also in
the deeper strata. Especially calciphilous plants with a large root system are affected, e.g.
alfalfa. Subsequently, 5 soil profiles are presented from the point of view of Ca content
using different solvent solutions (Table 1).
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Table 1. Ca supply of 5 soil profiles measured by using different solutions
for extraction
Samples pH(KCl) CaCO3(%) AL-Ca EDTE-Ca KCl-Ca Exch. Ca H2O-Ca
Cs1 7.27 3.2 16284 698 2992 515.0 25.30
Cs2 7.31 15.0 58494 768 2636 497.2 25.70
Cs3 7.55 26.0 98023 712 2030 392.2 20.90
Cs4 7.54 22.0 90226 648 1661 350.2 20.80
Cs5 7.84 12.0 47998 575 1076 334.0 20.10
KR1 7.05 1.1 29539 852 3676 594.6 32.40
KR2 7.20 6.2 51742 689 3136 570.6 16.10
KR3 7.32 13.0 65102 587 2820 459.8 19.10
KR4 7.30 12.0 7588 557 2400 379.8 23.00
KMR1 4.92 0.0 4439 374 3408 436.8 8.57
KMR2 5.03 0.0 4528 417 4037 496.8 5.20
KMR3 5.59 0.0 4841 559 4175 524.0 2.74
KMR4 6.38 0.1 4791 571 3495 361.4 1.68
KMR5 6.91 4.9 19683 610 3590 389.0 3.35
KH1 7.17 3.8 18902 726 2499 411.8 24.80
KH2 8.10 18.0 72984 454 688 437.8 13.30
KH3 8.39 12.0 43129 355 253 114.2 8.70
KH4 8.37 10.1 31476 347 168 100.4 9.31
KMH1 6.94 0.0 1436 389 483 60.4 7.03
KMH2 7.11 0.0 588 329 302 50.8 3.30
KMH3 7.07 0.0 591 324 185 53.4 3.23
KMH4 7.49 0.0 464 280 197 35.2 2.58
The measurements prove that the solutions used for extraction produced highly variable data
of Ca content. Most quantities appeared in concentrated acid extracts, intermediate in
extracts with neutral solutions, the less was extracted with watery solutions forming
complexes. The results obtained could be also converted from one method to the other.
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Ca CO3(%) = AL − Cax2,5
10000
EDTE − CaxKA
Exch.-Ca (mg/100 g) =
75
The data obtained with mild complex forming solvents approach the quantities of
exchangeable Ca, moreover, they are informative in a mathematical sense. That means,
results obtained with EDTE-Ca signify the values of exchangeable Ca content.
With Mg, the same test has been performed with different solvents. Results are shown in
Table 2.
Table 2. Mg supply of 5 soil profiles measured by using different solutions for extraction
Samples pH(KCl) AL-Mg EDTE-Mg KCl-Mg Exch.-Mg H2O-Mg
Cs1 7.27 485 34.20 174 28.56 2.43
Cs2 7.31 747 26.00 105 23.84 1.54
Cs3 7.55 1194 37.20 138 25.53 1.84
Cs4 7.54 2123 108.00 335 60.50 5.51
Cs5 7.84 2867 226.00 572 86.88 12.40
KR1 7.05 645 70.30 349 45.62 4.25
KR2 7.20 931 57.10 332 55.66 2.25
KR3 7.32 1335 59.10 410 66.67 3.08
KR4 7.30 1920 80.00 526 80.83 4.16
KMR1 4.92 809 93.30 747 78.65 3.18
KMR2 5.03 934 94.50 938 99.83 3.20
KMR3 5.59 1068 104.00 1019 115.07 2.14
KMR4 6.38 837 92.60 758 80.22 1.84
KMR5 6.91 847 71.00 581 77.44 1.21
KH1 7.17 588 38.60 186 24.68 2.91
KH2 8.10 1793 111.00 252 54.21 5.42
KH3 8.39 1080 54.30 88 18.15 3.17
KH4 8.37 1020 63.60 79 19.36 3.70
KMH1 6.94 109 25.40 54 8.59 2.13
KMH2 7.11 61 18.20 39 8.59 1.63
KMH3 7.07 85 24.70 36 10.77 2.07
KMH4 7.49 53 14.10 33 7.62 2.38
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Measurements prove that most Mg was extracted with AL, then with KCl. Values of
exchangeable Mg are largely equal with the results obtained with EDTE as Mg content.
Result is convertible according to the formula:
EDTE − Mg
Exch.-Mg (mg/100 g) =
9−K CONCLUSIONS
Different solutions are used for extraction to assess the Ca and Mg content of soil samples.
Measurements prove that most quantities were obtained with AL-solution, lowest quantities
with water as solvent.
Values of exchangeable Ca and Mg content largely coincide with the results of EDTE-Ca
and EDTE-Mg. The results obtained with the different methods are convertible to each
other.
REFERENCES
Filep T. (2002): Characterisation of the nutrient buffer capacity of K, Ca, Mg in two
Hungarian soils. Agrokémia és Talajtan 2002. 73-78 p.
Filep Gy. (1988): Talajkémia. Akadémiai Kiadó. Budapest.
Németh K. (1971): Talajvizsgálatok elektromos ultraszűrővel. Agrokémia és Talajtan. 20:
515 p.
356
A

ABSTRACT
BIOLOGICAL INVESTIGATION ON AN EROSION CATENA
B. Szeder 1 , B. Simon 1 , M. Dombos 2 , T. Szegi 1
1 Szent István University, Department of Soil Science and Agrochemistry,
2103 Gödöllő, Páter K. u. 1. Hungary
2 Research Institute of Soil Science and Agrochemistry,
Budapest, Herman O. U. 15. Hungary,
Biological investigations were carried out on an erosion catena on Szent István
University Experimental Farm, north central Hungary. Four soil profiles (Chernozems
and Calcisols) were examined in detail for the general physical, chemical parameters.
The aim was to find out quantitative relationship between the physical and the
biological (and biochemical) degradation processes. The level of physical degradation
was expressed by the moisture retention capacity and the depth of humus rich horizon.
The degradation of soil fauna was examined by the number and the species composition
of collembolans along the transect in eleven sampling points. The physical degradation,
biochemical and biological parameters showed strong relationship: with increasing
erosion and decreasing depth of the humus layers most investigated parameters showed
degradation. The number and the abundance of Collembola had significant decreasing
tendency with increasing level of degradation. The results suggest that soil degradation,
decreasing moisture holding conditions affect strongly the collembolan community. The
physical, chemical parameters seemed to be more resistant to changes in soils compared
to the biological aspects, thus, it is assumed that the biological parameters can be used
as early indicators.
Keywords: degradation, biological indicators, Collembola, erosion catena
INTRODUCTION
Loss of soil organic matter is a global problem related to soils and the environment.
Most of the fertile Chernozems of agricultural soils in Hungary developed on calcareous
loess. Chernozems have been cultivated for several centuries in Hungary (Stefanovits,
1999). However, most of the Chernozem soils in the country has experienced some kind
of soil degradation, including decrease in organic matter, compaction, erosion and/or
structural degradation, mainly due to inappropriate land use practices. These
degradation processes are generally accompained by biological degradation, i.e. loss of
species richness. Soil fauna is essential in the decomposition processes of biomass,
synthesis of humus, mixing soil particles, development of soil structure and it influences
several other soil characteristics that determine normal soil functions. Soil degradation
processes influence the composition, population size and activity of the soil fauna
(Larsen, 2004; Bíró, 2005; Veisz, 2004).
Soil quality can be evaluated by using several soil parameters, depending on the
scale and objective of the evaluation. These parameters can be, for instance, the soil
organic matter and soil aggregate stability (Gardi et al., 2002). In case of an erosion
risk, soil quality can be characterized by using different parameters, such as organic
matter content, aggregate stability, and enzyme activities. However, these parameters
357

tend not to give enough information about the biological state or activity and diversity
of soils.
Collembola (springtails), as a representative group of soil fauna, has relatively low
biomass, but it is extremely important in influencing the structure of some soils. For
example, “alpine pitch rendzinas” on limestone have a black humus layer that is formed
almost entirely of Collembola feces. However, most soils have millions of collembolan
fecal pellets on each square meter (Hopkin, 2002). Springtails play important role in
decomposition and soil respiration through feeding on fungal hyphae. These effects are
beneficial in the aspect of soil structure, moisture regime, soil organic content, cation
exchange capacity, nutrient availability, that determine healthy soil functions (Cole,
2005). It is believed that soil degradation processes lead changes in the activity and
composition of the soil fauna (Larsen, 2004). Soil scientists have been considering a
highly diverse community of soil organisms to indicate good soil quality (Brady and
Weil, 1999).
The objective of this presented study was to find quantitative relationship between
the physical and the biological (and biochemical) degradation processes along an
erosion catena. The degradation of soil fauna was examined by the number and the
species composition of collembolans. It was expected that not only the abiotic factors
(soil-moisture, aeration, compaction) but also biotic factors, such as microbiological
activity are important ecological factors for soil animal communities. The appropriate
measurements and scaling of these effects can serve as a basis for soil biological
degradation assessments.
MATERIALS AND METHODS
1. Study area: The study was conducted on Szent István University Experimental Farm
in Józsefmajor, north central Hungary. Four soil profiles were examined in detail on a
topographic catena, representing different levels of erosion (no, low, high erosion and
accumulation profiles). The first two profiles are Chernozem soils, the third one, which
is the most eroded profile, is a Calcisol and the fourth one at the bottom of the valley is
a Cumulic Chernozem.
2. Laboratory analyses: The following general laboratory analyses were carried out on
the four soil profiles: soil organic matter (SOM %) determined by Walkley–Black
method (Walkley, 1947); cation exchange capacity (CEC) by Mehlich method;
pH(H2O) and pH(KCl) determined electrometrically (Buzás, 1988); texture; CaCO3
content by Scheibler method; and bulk density (BD) determination (Buzás, 1988). The
simplified moisture retention was also determined on the soil samples to find out the
level of physical degradation.
Besides the general laboratory analyses the following biochemical properties were
determined: water soluble carbon (WSC) and carbohydrates (WSChy) (Garcia, 1997),
enzimatic characterization (Veisz, 2004), urease, and ß-glucosidase activity (Garcia,
1997).
3. Experimental design for biological aspects: Collembola was sampled by regular
sampling, along a transect in eleven points with 20 m distances from each other. The
first four points belonged to the non eroded region, whereas the 5-8th points were low
eroded and the 9th, 10th points represented the most eroded parts of the transect. Soil
358

profiles were typical for catena positions, i.e. the plateau position was the non-eroded
region, the middle slope position was low eroded, the lower slope position was the most
eroded and at the valley the accumulated soil profile was found.
Four soil core samples were taken in each points along the transect from the upper 5 cm
by using 5 cm diameter sampling pots.
RESULTS AND DISCUSSIONS
The results of the laboratory analyses are given in Table 1. The depth of soil organic
matter was 90 cm in the non eroded profile, whereas it was only 10 cm at the highly
eroded site showing high physical degradation. The soil organic carbon (SOC) content
and the cation exchange capacity (CEC) of the soils decreased with increasing erosion
rate. The water soluble carbon (WSC) showed similar tendency. As for the bulk density
(BD), higher values were found in the upper slope positions. Concerning the enzymatic
characterization, less activity of urease (involved in N cycle) and
ß-glucosidase (involved in the C cycle) were experienced in the highly degraded slope
positions, suggesting less biological activity related to degradation.
Table 1: General laboratory parameters of the top horizons
of the four soil profiles along the catena
Profile DH SOC WSC WSCh CaCO3 CEC pH Ure B-Gl Texture BD
(upper h.) cm % ppm ppm % cmol·kg -1
μmols μmols FAO g·cm -3
NE 90 2,2 106,2 2,1 0 30 6,1 0,75 21,14 CL 1,4
LE 40 1,5 103,6 1,4 0 28 6,1 0,66 17,20 CL 1,5
HE 10 0,5 95,5 7,6 12 24 6,1 0,49 12,44 CL 1,3
AP 80 2,5 119,2 6,4 2 40 7,6 1,28 35,61 CL 1,2
Profile designations: NE – Non Eroded, LE – Low Eroded, HE – High Eroded,
AP – Accumulated Profile; DH - depth of humus layer (cm); SOC – Soil organic carbon
(%); WSC - Water soluble carbon (ppm); WSCh - Water soluble carbohydrates (ppm);
CEC - Cation exchange capacity (cmol·kg -1 ); Ure – Urease activity (μmols N-
NH4 + /g.h); B-Gl – B-Glucosidase activity (μmols PNF/g.h); CL- clay loam; BD - Bulk
density (g·cm -3 ).
The degradation of soil fauna was examined by the number and the species compostion
of collembolans along the erosion gradient. Eleven sampling points (Figure 1) were
chosen along the slope along the four soil profiles that were examined in detail. Except
for the highest point (number 0 in Figure 1) in the transect, which might have been a
sampling error, abundances of springtails decreased highly. Concerning the erosion
grades, the soil living springtails showed significant differences in their abundances
(F(2,41)= 9.2, p

The average values of abundances, derived from the four subsamples, showed strong
correlation with the moisture retention of soil (r=0.89) (Figure 4.).
Figure 1: Number of Springtails
(Collembola) along the erosion gradient
Figure 3: Simplified moisture retention at
the erosion levels
Figure 2: Number of Springtails
(Collembola) at the erosion levels
Figure 4: Correlation between water
retention and springtail abundance
Figure 5 shows that most springtails preferred the pH 6-7. Less individuals were found
above pH 7. As far as the Collembola species composition is concerned, we found much
less species on the high eroded area than the reference and low eroded areas (Table 2).
360

Table 2: Collembola species composition among the erosion gradient
Reference, non
eroded profile
Low eroded
profile
High
eroded
Collembola species
ref 1 2 3 4 5 6 7 8 9 10
Entomobria handschini 2 1
Entomobria multifasciata 27 10 7 3 2
Heteromurus nitidus 35 12 11 1 2 4 15
Folsomia cf. penicula 5 2 1 29 1 1 2 1 4
Lepidicyrtus cyaneus 8 7 22 1 4 1
Lepidicyrtus paradoxus 1 1
Ceratophysella cf. armata 3 1 1 1
Orchesella cincta 5 9 8 7 6
Orchesella sp. 1 1
Poduromorpha sp. 1 2
Sminthurus elegans 8 5 1 8 7 27 1 3
Sminthurinus aureus 2 1 1
Tomocerus sp. 1
abundance
30,00
20,00
10,00
0,00
The pH and the abundance of springtails
A
A
6,50 7,00 7,50
A
pH
A A
A
A
A
A
A
Linear Regression
abundance = 110,69 + -13,88 * pH
R-Square = 0,64
Figure 5: Correlation between pH and abundance of springtails
361

CONCLUSIONS
The physical degradation, biochemical and biological parameters showed strong
relationships on the studied erosion catena. With erosion and decreasing depth of the
humus layers most investigated other parameters showed degradation.
The number and the abundance of Collembola were investigated in this study along a
transect representing different levels of erosion and soil degradation (organic matter
decrease, soil compaction, etc.). We found that along a transect the abundance of
springtails has significant decreasing tendency with increasing level of degradation. The
results suggest that soil degradation affects strongly the collembolan community. In
case of the eroded area we experienced higher pH (above 7) because of the distance of
the calcareous parent material from the soil surface were less than the other sites. This
result suggests that due to degradation processes soil moisture conditions strongly affect
the belowground animal communities, such as springtails.
The examined enzyme activities are important information to characterize the measure
of soil degradation. The land degradation seems to be a key factor for variation in the
behaviour of the examined enzyme.
The physical, chemical parameters compared to the biological parameters showed the
following correlations: the physical features are more resistant to changes in soils
compared to the biological aspects, thus, it is assumed that the biological parameters can
be used as early indicators.
ACKNOWLEDGEMENTS
INDEX Nr. GOCE-CT-2003-505450
Hungarian Scientific Research Fund (OTKA – Nr. D 048592)
REFERENCES
Bíró, B., Villányi, I., Füzy, A., Naár, Z., (2005): Baktériumok és gombák
kolonizációja génmódosított (Bt-) és izogénes kontroll kukorica rizoszférájában,
Agrokémia és Talajtan, 2005, vol. 54, N o s 1-2, 189-202 pp.
Brady N. C., Weil R. R. (1999): The Nature and Properties of Soils. Twelfth Edition.
Prentice Hall. N. J. 404-405, 760-761.
Buzás, I. (Szerk.) (1988): Talaj- és agrokémiai talajvizsgálati módszerkönyv 2.
A talajok fizikai-kémiai és kémiai vizsgálati módszerei. (Methods of soil and
agricultural chemistry analyses 2. Physical-chemical and chemical methods of soil
analyses.) Mezőgazdasági Kiadó. Budapest. Hungary.
Cole L., Buckland S. M., Bardgett R. D. (2005): Relating microarthropod community
structure and diversity to soil fertility manipulations in temperate grassland. Soil
biology and biochemistry 37:99, 1707-1717.
Garcia, C., T. Hernandez, A. Roldan, J. Albaladejo (1997): Biological and
biochemical quality of a semiarid soil after induced devegetation. Journal of
Environmental Quality 26, 1116-1122.
Gardi, C., M. Tomaselli, V. Parisi, A. Petraglia, C. Santini. (2002): Soil quality
indicators and biodiversity in northern Italian permanent grasslands. European
Journal of Soil Biology. 38, 103-110.
Hopkin, S.P. (2002): Collembola. Encyclopedia of Soil Science, p. 207-210.
362

Larsen, T., P. Schjonning, J. Axelsen. (2004): The impact of soil compaction on
euedaphic Collembola. Applied Soil Ecology. 26, 273-281.
Stefanovits, P., Gy. Filep, Gy. Füleky (1999): Soil Science. (In Hungarian.)
Mezőgazda Kiadó. Budapest. 92-123.
Veisz, O, Bencze, S, Janda, T, (2004): Changes in the activity of antioxidant enzymes
in cereal species during the winter CEREAL RES COMMUN 32 (4): 493-500.
Walkley, A. (1947): A critical examination of a rapid method for determining organic
carbon in soils: Effect of variations in digestion conditions and of inorganic soil
constituents. Soil Sci. 63. 251–263.
363

THE PRESENT STATE OF THE ROMANIAN AGRO-ALIMENTARY
CONSUMPTION
Anca Monica Brata
University of Oradea, Faculty of Environmental Protection
ABSTRACT
The alimentary consumption is not and cannot be identical in the countries and areas of the
entire world, no matter how attractive the international consumption patterns would be. The
specialists in alimentation and the international organisms with responsibilities in this field
recommend the remodelling of the alimentary consumption and encourage having a healthy
alimentation, based on each nation’s accumulations and showing their value as being a
cultural nutritional thesaurus. It is considered that the alimentary education of the population
and the improvement of the health insurance systems can contribute to the limitation of
hunger.
Amongst the well developed countries and the developing ones, there are great gaps
regarding the average consumption per inhabitant, but the deficiencies are especially
structural and qualitative.
Keywords: alimentation, annual and monthly consumption.
INTRODUCTION
The humanity develops continuously. This is a fact that cannot be ignored, which modifies
the humans’ way of thinking and behaviour. This evolutional process takes place in every
field of life, including the alimentary one.
The evolution of the human species implied permanent changes in the alimentary
behaviour, in the alimentation manner, but also in the process of obtaining alimentary
products. The increase of people’s culture and civilization, the technical and scientific
discoveries, the research in the alimentation and alimentary field, all these have contributed
in a great extent to these changes. The diversity of the alimentary products, the
quantification of their metabolic effects and relating the aliments to the population health
state, have led to a need of a careful design of alimentary products, that takes into account
the nutritious value of aliments and the expected effect on alimentation.
The problem of insuring a sufficient level of the population’s alimentation from the
quantitative, qualitative and structural point of view has become for most countries and for
the governmental and non-governmental organisms – national, regional and international – a
more powerful preoccupation in the last half of the century. This preoccupation is the
consequence of the conscious knowing of the connection between health and the access to
various sources of food. Because the expansion and the limits of the access to the sources of
alimentation differ from an area to another, from a country to another and even within the
same country from an area to another, the taking of some measures regarding the
amelioration of the alimentation matter becomes essential [Bulgaru, 2001].
364

The alimentary problem has constituted and still constitutes a major preoccupation,
nationally and internationally, mainly focusing on the most efficient solving of the relation
between resources and consumption necessities.
At every people, along its existence and development, alimentary traditions, habits and
skills have installed more or less durable, that continue and will continue to play a
predominant role in the nutrition of the respective population, even under the impact of the
modern alimentation principles, based on their scientific and rational character. The main
factors of this phenomenon are the ambient geographical environment, family and external
influences [Gavrilescu, Giurca, 2000].
At present, the evolution of alimentation generates a rational alimentary behaviour,
implies the knowing of the body’s needs and the efficient ways to satisfy them. The
supporting of a rational alimentation represents a preoccupation of both the specialists of
nutrition and the governments of the states and of the national and international organisms
founded and organized for this purpose.
Thus, the reconsidering of the aliments’ consumption has become necessary, its
relation with the needs of the body, its adaptation to the climate and working conditions.
RESULTS AND DISCUSSIONS
In Romania, in the late years, the population’s real incomes were continuously decreasing.
Under these conditions, the housekeepers’ incomes were almost entirely intended for
covering the daily needs – especially the alimentary needs – and the compulsory payments
(taxes, fees, etc.), without being able to save significant amounts of money, except for the
patrons’ families.
In order to focus on the evolution of the goods’ consumption after 1989 we will
observe table 23 where we can find the structure of the consumption expenses on groups of
families as employers, peasants, pensioners.
Table 1. The structure of the consumption expenses on groups of families, after the
monthly income per person, in the year 1990 comparatively to the year 2000
Expenses for food Expenses for non- Expenses for payment
alimentary goods
of services
1990 2000 1990 2000 1990 2000
Employers 46,8% 50,6% 39,2% 27,8% 14,0% 21,6%
Peasants 63,7% 57,4% 28,3% 34,8% 8,0% 7,8%
Pensioners 59,2% 54,3% 22,7% 29,1% 18,1% 16,6%
Source: Anuarul Statistic al Romaniei, (The Statistic Annual of Romania), 2001
The following aspects can be noticed [Anuarul Statistic, 1991-2001]:
In the expenses for alimentary products there can be noticed an increase with 3.8% at
employers, and at peasants and pensioners a decrease with 6.3%, respectively 4.9%.
365

In the expenses for the non-alimentary goods there can be noticed a decrease with 11.4% at
employers, and at peasants and pensioners a decrease with 6.5%, respectively 6.4%.
The expenses for duties increase with 7.6% at employers and at peasants and
pensioners they decrease with 0.2%, respectively with 1.5%.
On resistance media, the alimentary expenses represent, in the total of the consumption
expenses, 56% in urban environment and 58% in rural environment. The above mentioned
data indicate the fact that, in our country, not even the families with the highest income have
a structure of the consumption expenses comparable with that of the western-European
countries, where the alimentary expenses do not exceed 20% of the total expenses for
housekeeping.
As regards the consumption on population categories in the post-December period, it
has been achieved in different conditions, following the decline line of the national
economy, the significant decrease of the agricultural and industrial production, the
agriculture system, as well as the new system of distribution under the conditions of private
actions. One thing that can be said is that the average annual level of the peasants’
consumption showed a tendency of increase, overtaking the consumption in the employers’
families that recorded significant decrease. This fact can be explained by the fact that the
main source of forming the alimentary consumption is the own production.
The alimentary consumption in Romania during the analysed period is the following:
Table 2. The average annual consumption per inhabitant, for the main alimentary
products
1990 1995 2000 2005
Cereals 213,6 215,8 219,7 219,8
Potatoes 59,4 71 86,5 97,9
Vegetables 127 140,4 134,3 183,3
Fruit 59,5 45,8 44,5 77,4
Sugar 27,3 23,5 23 25,6
Milk 140,1 188,6 193 238,9
Eggs 246 197 206 289
Meat 61 51,2 48,3 65,5
Source: Anuarul Statistic al Romaniei, (The Statistic Annual of Romania), 1990-2006
There can be seen an increase of the potatoes, vegetables, cereals and milk consumption.
The meat consumption is low enough as compared with other countries [Anuarul statistic,
1991-2006].
The presentation of the average annual consumption of the main alimentary products
on countries is also very interesting.
366

Table 3. The average annual consumption per inhabitant, for the main alimentary
products, on countries, kg/inhabitant in 2002
Country Vege- Potato Fruit Meat Alcoholic Sugar Milk Cereals
ables
drinks
Romania 137,32 89,75 46,07 42,27 53,27 22,73 123,17 218,33
Poland 124,48 134,77 62,4 67,63 78,5 35,27 181,65 154,18
Slovakia 80,58 77,53 62,93 69,57 79,25 36,07 188,87 131,68
Hungary 106,67 67,62 63,01 72,28 83,8 37,77 190,12 125,32
Check
Republic
76,15 79,6 86,08 90,31 110,39 42,25 211,54 105,56
Bulgaria _ 32,16 126,58 106,22 171,92 48,48 239,77 104,41
Source: Anuarul Statistic al Romaniei, (The Statistic Annual of Romania), 2003
As regards the vegetables consumption we are situated on advanced positions, the average
annual consumption is 137.3 kg/inhabitant, over the average of the European Union (122.57
kg), or of the US (125.8 kg/inhabitant), while in other developing countries, vegetables are
consumed in an annual average of 98.8 kg/inhabitant [Anuarul statistic 2003].
In the developing countries, the potatoes consumption is over the average of the
developed countries. The average of the potatoes consumption is 78.65 kg/inhabitant
annually, and in the US there is an average of about 65.27 kg/inhabitant. A Romanian
consumes annually an average of 89.75 kg of potatoes (in 2002).
Romania is much under the average of the developed countries and developing
countries at the fruit meat, milk and sugar consumption. The cereals consumption in our
country has the highest consumption in comparison with the countries analysed in the above
table.
A basic aliment in the human diet is milk. From milk we can prepare various products,
which determined the occurrence of a real alimentary industry with influence on human’s
feeding manner. By introducing the technology of transforming the liquid milk in powder
milk, it can be transported at long distances and can be stored for long periods of time. This
thing allowed that in the periods when the production is higher than consumption, to create
stocks that can be used in the moment when the production is lower than the consumption.
Table 4. The average milk consumption on countries, 2003 (l/inhabitant)
Country Annual average milk consumption (l/inhabitant)
Bulgaria 163,8
Hungary 176,71
Romania 188,87
Poland 190,12
Check Republic 203,57
Source: Anuarul Statistic al Romaniei, (The Statistic Annual of Romania), 2004
367

Among the developing countries, Romania is on an average position as regards the annual
average consumption of milk, but in comparison to other countries in the European Union or
USA, Romania is on an inferior position than these countries [Anuarul statistic, 2004].
For an extended presentation of the alimentary consumption in Romania, the
presentation of the daily average calories consumption is necessary.
Table 5. Alimentary consumption expressed in calories and nutritious factors, in a daily
average, per inhabitant
Calories Protides Lipids Fats
Year animals total animals total animals total
1990 711 3038 42,8 96,7 49,8 93,4 433,3
1992 671 2758 38,3 88,3 46,2 80,2 403,2
1994 690 2872 39 92,9 47,1 78 431,7
Source: Anuarul Statistic al Romaniei, (The Statistic Annual of Romania), 1991-1995
As one can notice, the calories consumption has decreased starting from 1990. In 1994 it
was ameliorated. It is considered that in Romania it would be necessary a daily average
consumption of 3400-3500 calories per inhabitant with a structure of 1/3 calories of animal
origin and a consumption of 100 grams proteins, 60% of animal origin. It would mean a
decent average consumption, close to the balanced one. At present we have a deficit of
about 20-25% at the calories number and of 35-40% at the proteins of animal origin. There
is, as it can be explained, a great difference of the alimentary consumption according to the
income, fact that leads to more decreased levels, that is at surviving limits, of absolute
hunger but of specific hunger too, due to malnutrition, to the lack of some important
nutritious elements in the population’s consumption ratio as the protides, vitamins and
mineral salts [Anuarul statistic,1991-1995].
Examining the low level of the alimentary consumption in Romania, it has to be taken
into account the important and well-known fact that it has a significant impact on the
population’s general behaviour, of the labour’s productivity, of health state, of demographic
indices, the level of general and infantile mortality. We have to note the fact that the
infantile mortality in our country is 2-3 times higher than in other western-European
countries, and the average life time is situated with 6 years in minus than in most European
countries, and the mortality affects more and more the poor population, and the less fed
people.
REFERENCES
1. Mircea Bulgaru – Dreptul de a minca, Editura Economica, 2001
2, Dinu Gavrilescu, Daniela Giurca – Economie Agroalimentara, Editura Expert, Bucuresti,
2000
3. Anuarele statistice ale României -1991-2006
368

FROM FARM TO FORK – EVALUATION OF CROPPING FIELDS WITH
REGARD TO FOOD AND FEED SAFETY AFTER LONG-STANDING
CULTIVATION
Péter Sipos, Zsuzsanna Szathmáry, Árpád Tóth, Diána Ungai,
Dóra Hovánszki, Zoltán Győri
University of Debrecen, Centre of Agricultural Sciences, Institute of Food Science,
Quality Assurance and Microbiology
138 Böszörményi Street, Debrecen, H-4032, Hungary
ABSTRACT
From the aspects of quality control system in accordance to food safety, the pesticide
residues and toxic or potentially toxic elements are important parameters to examine not
only from the products and its raw materials point of view, but from the external
resources (as soil and applied matters, e.g. manure) as well. The sensitivity and
accumulation properties of the different plants and cultures is different according to the
above mentioned factors, and the aim of production also determines the upper level of
limit values. On the other hand, in regulations of several countries like Hungary there is
no differentation as regards utilization and accumulation properties.
In a case study we have examined the soil parameters of food safety of differently
utilised (intensive cereal production, extensive meadow and bio production) cropping
fields which were cultivated for a long time, so the presence of contaminants in the soil
profile was assumed. We found that the examined fields are suitable for producing safe
foods, but in the case of intensive cultivation some accumulation of pesticides and
heavy metals is observable.
Keywords: food safety, pesticide remains, toxic elements, different levels of land use
INTRODUCTION
In latest decades there has been an increasing demand to extend the more or less
developed quality control system of food product processing to the raw material
production and supply. This would lead to getting closer to the fully introduced food
and feed safety in the whole plant-animal processing line. Beside this, it was obligatory
to apply the HACCP system by food processors and traders. The picture of applied
quality control systems is very diverse nowadays; strict application and monitoring is
used in the processing of horticultural crops, but in the case of field crops, especially
cereals and forage crops it is far from being effective.
To examine the possibilities of the development of a quality control system in the
whole plant-feed-animal-food chain, i.e. “from farm to fork” we have started a research
program with the contribution of industrial partners. Our aim is to create a base
specification that helps to apply the ISO 22000 standard and to develop a quality control
and tracing system specified for Hungarian circumstances. To examine the process we
made a case study for each contributor. To determine whether the fields taken into these
examinations are suitable for safe production, we analysed their pesticide remains and
heavy metal content. On these sites the crop production activity has been lasting for a
long time, so these examinations seemed necessary, because of the applied
369

agrochemicals as fertilizers and pesticides, the probability of contamination or
accumulation was considerable.
The group of organochlorines are widely used pesticides and are important in the
aspects of food safety (Kan and Meijer, 2007). The peak of their application was in the
’50ies and ‘60ies. Many of them were banned later in the most countries in Europe and
North America, but in developing countries large amounts have been applied in the
‘90ies (Kannan et al., 1997) and are still in use nowadays in several countries. The
information about organochlorine contamination is mostly limited to DDTs and lindane,
but it is easy to accept that other dangerous pesticide remains are still in our soils
(Shegunova, 2007). It may be also proved by the fact that both Hungarian and European
Union soil and water quality standards contain target and intervention values for DDTs,
drins as aldrin and endrin, and HCH and HCB (Oldal et al., 2006). Besides, earlier
studies on Hungarian surface water monitoring reports that 8 out of the examined 21
pesticide residues exceeded the 0,1 μg/l limit concentration required by EU directives
(Kárpáti et al., 1998).
From the aspect of food safety another important soil property is its element
content, primarily the heavy metal content. Excessive amounts of metals originated
from agricultural or industrial production often cause human actions (Biehl and Buck,
1987). It is important to monitor the cadmium, lead, mercury and arsenic contents in
both soils and the products from different locations, although the latter is not heavy
metal by original definition, but in several cases it poses similar risk. Besides limit
values in different national and international standards, the Ministry of Agriculture in
Netherlands published the so-called LAC values, which define the limit values that are
“the lowest level in soil that, when exceeded, may give rise to the emergence of
negative impacts on the yield and the quality of agricultural products, and with the
health of human beings and animals” (Brus et al., 2005). This recommendation gives
different limit values according to both soil type (as sand or clay) and grown plant (as
grassland, arable land with flowers or arable land with food for human consumption). In
Hungary the current limits are declared in 10/2000. (VI.2.) KöM-EüM-FVM-KHVM
ministry order in which the background (A) levels, contamination limit (B) levels and
environmental remediation contamination limit (D) levels are determined. Naturally, the
aim of production often raises a claim to decrease the maximum heavy metal level in
soils, e.g. in the case of producing drug or medicine raw materials.
In our case study we had an opportunity to evaluate and estimate three kinds of
cropping sites in the aspects of food safety; an intensively used (for wheat and maize
production), an extensively used (for meadow) and one used in bio production.
MATERIALS AND METHODS
Soil samples: Examined sites were the followings: two winter wheat production sites
near to Szentes and Sárrétudvari, two maize production sites by Nádudvar and
Hajdúböszörmény, a meadow near to Bakonszeg and a bio production area at the
Látókép Experimental Station of University of Debrecen, Centre of Agricultural
Sciences. Site samplings were made in the summer of 2005 from the upper 20
centimetres of soil and from the 100-130 cm depth and 200-235 depth as well. In 2006
370

we made a deeper vertical sampling to 300 centimetres depth of soil and we separated
10-12 layers in the profiles based on their physical properties.
Pesticide residue analysis: Pesticide residue analysis was made in the Soil and Plant
Conservation Service of Miskolc in 2005 and in the Institute of Food Science, Quality
Assurance and Microbiology in 2006. Sample preparation was carried out as follows:
after milling and drying, 50 g of the soil was accurately weighed and added 7 ml of 0.2
M ammonium-chloride (Sigma). After resting for approximately 15 minutes nhexane:acetone
(1:1) mixture was added, and the solution was shaked for 2 hours in
laboratory shaking machine. The mixture was filtered on Büchner funnel and the
acetone was separated with 700 ml of distilled water. The phase of n-hexane was dried
on sodium-sulphate and concentrated with rotation evaporating machine. The residue
was dissolved in 5 ml n-hexane. The extract was cleaned by column-chromatography,
the column filled with 8 g of Alumina (activity: V.). The evalution was carried out with
n-hexane, finally the solution was evaporated again and dissolved in 1 ml of n-hexane.
During the sample preparation we used organic solvents in the quality of GC-residue
analysis (Scharlau). The detection of the analytes was performed on a Varian CP-3800
gas chromatograph with ECD detector (Nederland). Operating conditions: CP 8907
Factor Four Capillary Column, 15 m * 0.25 mm, temperature program: 50 °C, 1 min;
50-180 °C, 25 °C min-1; 180-250 °C, 4°C min-1; 250°C, 1 min, helium carrier gas (1.0
ml min-1, 6.0 Linde), injected volume: 1 µl, temperature of the detector: 250 °C. The
standard materials and internal standards were prepared from MERCK Chlorinated
Pesticide Mix 40.
Heavy metal content analysis: Determination of heavy metal concentrations was made
in the Institute of Food Science, Quality Assurance and Microbiology. Samples were
dried at 60 o C, ground, sieved through a 2 mm mesh sieve and digested in HNO3-H2O2
according to Hungarian methodological standard procedure: MSZ 21470-50:1998
(Kovács et al., 2000). Merck and BDH standard solutions were used to prepare the
stock-solutions, and REANAL (Budapest, Hungary) solid chemicals were also used.
Ultrapure water was used to prepare the solutions (Millipore, Paris, France). The
determinations were made by a Perkin-Elmer Optima 3300 DV Inductively Coupled
Plasma - Optical Emission Spectrometry (ICP-OES) equipment.
RESULTS AND DISCUSSIONS
In 2005 by the analysis of element composition we found that the examined fields show
significant differences in macro nutrient concentrations (N, P, K, S), but show
surprisingly homogenous distribution in the case of several micro nutrients and toxic
elements (Table 1). We found no significant amount of pesticide residues in the
ploughed upper soil layer (0-20 cm layer), but in the case of maize and winter wheat
fields a small degree of accumulation was observed (Table 2). These results from the
pesticide residue analysis induced us to continue the sampling in wider soil profile.
We found as a result of analysis in 2006 that the bio production field at Látókép
did not contain detectable amounts of pesticide residues. In the case of other sites the
detected amount of pesticide residues was diverse in the case of different chemicals, but
in all case it was under the contamination limit value (A value) of 10/2000. (VI.2.)
KöM-EüM-FVM-KHVM ministry order in the whole soil profile (Figure 1). The peaks
371

visible at lower soil layers does not mean risks in production and in food safety in the
case of the examined cultivated plants, partly because of the relatively low
concentrations, partly because we made additional product (grain and tissues of animals
fed on these forages) analysis on residues and we have not found detectable amount of
pesticide residues in this products.
Table 1: Element content of upper ploughed soil layers of examined fields (mg/kg)
nationwide
element Nádudvar Hajdúböszörmény Bakonszeg Sárrétudvari Szentes average*
P 1032 887 881 1009 918 535
K 4443 2858 3265 3145 3658 2952
S 123 118 138 152 159 128
Zn 76,4 76,4 72 73 70 47
Cu 18 14 12 15 12 18
Ni 20 19 19 20 19 14
Mn 762 704 608 536 643 457
Cd 0,957 0,92 0,94 0,93 0,96 0,81
Pb 24 22 22 21 20 14
* average of TIM (Hungarian Soil Information and Monitoring System) samples
Table 2: Pesticide residues of different soil levels of examined fields, 2005 (μg/kg)
Depth Sum of HCH Sum of drins
Sum of DDT /
DDD / DDE
Hajdúböszörmény
(maize)
0-20
100-130
200-235
0, 024397
35, 75
24, 45
0, 002113
0, 72
0, 84
0, 071583
11, 77
8, 97
0-20 0, 025776 0, 00248 0, 009218
Nádudvar (maize) 100-130 32, 95 4, 94 2, 41
200-235 30, 69 3, 6 1, 92
0-20 0, 00764 0, 00017 0, 01774
Szentes (wheat) 100-130 10, 26 0, 24 0, 58
Sárrétudvari
(wheat)
Bakonszeg
(meadow)
Látókép (bio)
200-235 10, 12

Although the toxic element content of upper soil levels showed no high values (and
therefore risk in food safety) again we analysed the element composition of soil samples
besides the pesticide residues. In the case of the bio field of Látókép we measured lower
values than the limit value by Hungarian orders, in homogenous distribution in the more
or less deep soil layers. On the other hand, in the case of intensively cultivated fields the
cadmium concentration is near to the B value, suggesting contamination in certain
layers (Figure 2). Although the measured values are lower than the contamination level
(C value) currently not in arrangement and the environmental remediation
contamination limit (D value), it is recommended to analyse this parameter in the case
of those sensitive plants that are susceptible to accumulation of cadmium. The little
higher concentration (~10-15%) than the natural average value of copper in the case of
some samples is a consequence of intensive agronomy. From the results the near to limit
value of zinc concentration at Szentes in lower soil layers is also remarkable.
Nádudvar
(maize)
Szentes
(wheat)
Depth (cm)
Depth (cm)
0
50
100
150
200
250
300
0
50
100
150
200
250
300
0 5 10 μg/kg 15 20 25
sum of HCHs
sum of drins
sum of DDTs
0 5 10 μg/kg 15 20 25
sum of HCHa
sum of drins
sum od DDTs
Figure 1: Pesticide residues in the examined soil profiles, 2006 (μg/kg)
373

Látókép
Nádudvar
Szentes
Depth (cm)
Depth (cm)
Depth (cm)
0 20 40 60 80 100 120
0
50
100
150
200
250
300
(mg/kg)
0 20 40 60 80 100 120
0
50
100
150
200
250
300
(mg/kg)
0 20 40 60 80 100 120
0
50
100
150
200
250
300
(mg/kg)
Figure 2: Toxic element content in the examined soil profiles, 2006 (mg/kg)
374
Cd
Cu
Pb
Zn
Cd
Cu
Pb
Zn
Cd
Cu
Pb
Zn

ACKNOWLEDGEMENTS
This work was supported by ALAP1-00058/2004 GAK: “Farm to fork” for cereals for
animal food (2005-) and GVOP-3.1.1.-2004-05-0416/3.0: Application of modern bio
and food analytical methods in food raw material processing (2005-) research projects
REFERENCES
10/2000. (VI.2.) KöM-EüM-FVM-KHVM együttes rendelete. Magyar Közlöny, 53:
3156-3167
Biehl, M.L., Buck, W.J., (1987): Chemical contaminants: their metabolism and their
residues. J. Food Protect. 50. 12, 1058–1073.
Brus, D.J., de Gruijter, J. J., Römkens, P.F.A.M. (2005): Probabilistic quality
standards for heavy metals in soil derived from quality standards in crops.
Geoderma 128 (2005) 301– 311.
Kan, C.A., Meijer, G.A.L. (2007): The risk of contamination of food with toxic
substances present in animal feed. Animal Feed Science and Technology, 133, 84–
108.
Kannan, K., Tanabe, S., Giesy, J.P., Tatsukawa, R. (1997): Organochlorine
pesticides and polychlorinated biphenyls in foodstuffs from Asian and Oceanic
countries. Reviews in Environmental Contamination and Toxicology 152, 1-55.
Kárpáti, Z., Győrfi, L., Csanády, M., Károly, G., Krómer, I. (1998): Ivóvizek
növényvédő szer szennyezettsége. Egészségtudomány, 42, 143–152.
Kovács, B., Prokisch, J., Győri, Z., Balla Kovács, A., Palencsár, A.J. (2000): Studies
on Soil Sample Preparation for Inductively Coupled Plasma Atomic Emission
Spectrometry Analysis. Commun. Soil. Sci. Plant Anal., 31.11-14, 1949-1963.
Oldal, B., Maloschik, E., Uzinger, N., Anton, A., Székács, A. (2006): Pesticide
residues in Hungarian soils Geoderma 135. 163–178.
Shegunova, P., Klánová, J., Holoubek, I. (2007): Residues of organochlorinated
pesticides in soils from the Czech Republic. Environmental Pollution 146. 257-
261.
375

MICROCLIMATIC STUDY OF THE MAIN WINE-GROWING
REGIONS IN ROMANIA DURING 2006
DRD. Dorin POPA
* Universitatea Oradea, Facultatea de Protectia Mediului
ABSTRACT:
From a climatic point of view, Romania differs from other geographical regions, the result
being specific microclimates favourable for vine cultivation. Our microclimate influences
cultivated vine sorts and the wine quality.
Analyzing climatic particularities of five wine-growing regions during 2006, from a
point of view of thermic resources, pluviometric conditions and climatic events, we can
conclude that, from a climatic point of view, 2006 was a favourable year for vine culture,
securing premises for a good production, in quantity as well as in quality.
Key-words: climate, vine, wine-growing regions, quality.
1.1 CLIMATIC CHARACTERIZATION
1.1.1. Wine-growing region Moldavian Hills (Dealurile Moldovei)
Climate particularities during 2006 and their consequences over vine plantations will be
presented based on specific time periods, analyzing critical climate factors having impact
over vegetation status and production potential.
From a thermic point of view, 2006 was a year warmer than usual, the medium
temperature oscillating between 8,6 and 11,4 0 C. 2006 started with a cold winter, even frosty
at night and in the morning. In January, the snow was set on most of the land, except the
south of the region, where its absence resulted in damages. The lowest temperature in
January was between –19,9 0 C and –26,8 0 C. As for the highest temperature, this was
between 31,4 0 C and 36,1 0 C, during August.
Analysing the rainfall amount, there has been concluded that, overall, in Moldavia was
recorded in 2006 a surplus of 7,3 l/m 2 than the average multi-annual amount. The highest
values were recorded in the north of the region, where the rainfall amount had a surplus of
19,2 % than the average multi-annual amount. At the other side is the Central-South of the
region, where the deficit was of 10%.
On a monthly basis, the rainfall amount during 2006 was higher from January to May,
except for February and June. The highest amount was in August, the surplus being of 99 %
over the average amount. The daily water quantities were in most of the places over 30
l/m 2 . The deficit of buds due to frost was of 11 – 15 % in the South and 20 – 35 % in the
North-Central region.
Over January, February and the beginning of March, the soil stayed frozen, with a
depth of 1-10 cm on most of the agricultural region, even 20-35 cm on some parts.
Summer was cold at the beginning, with high thermic amplitudes and a high haziness,
delaying the growth and development processes at all the sorts. All along the summer there
were reported rainfalls, mainly sudden showers, but also torrential. All over the region, on
small areas, stormy winds and hail storms partially affected the vine plantations.
Autumn started with a nice and warm weather, with few rainfalls. The end of autumn
and the beginning of winter (December, 2006) was marked by a high thermic regime and
few rainfalls. It is important to point out that in December 2006 there was no snow layers in
the region.
376

l/mp
120
100
80
60
40
20
0
Fig. 3 Monthly evolution of rainfalls compared by average multi-annual amount
in Moldavia (situated East from Siret)
28,8 25,9
10
24,2
67,7
26,4
53,9
42,9
64,6
58,7
85,8
77,3
I II III IV V VI VII VIII IX X XI XII
58
66,6
105,4
52,8
19
43,4
2006 normal
1.1.2 Wine-growing region Podişul Transilvaniei
2006 was a year of quite reduced thermic resources, with an average annual temperature of
9.9°C and a pluviometric regime of 589.4 mm close to normal (644.6 mm) (Tabel 1).
During winter there were low temperature values, the lowest being -20.3°C (in January),
partially affecting buds viability of the sorts sensitive to frosty weather, requiring cutting
compensations in spring.
Vine started vegetation later, but the richer thermic resources during summer led to
recovery of this delay, the vegetation development being normal. During August the
haziness was high and there were massive rainfalls (169,4 mm), leading to a delay of grape
maturation and creating favourable conditions of Botrytis cinerea attack. The first frost was
recorded on 17.10 (-2.8°C).
Tabel 1: Main climate data for podgoria Tarnave Vineyard – Viticultural Center Blaj
Climatic Average monthly Extreme Extreme Total rainfalls ( mm)
indicator temperature temp. temp.
Month ( o C) lowest
( o highest
C) ( o realized normal diff.
C)
+/ -
January -5,6 -20,3 7,1 9,2 26,3 -17,1
February -0,4 -18,0 16,0 11,1 21,2 -10,1
March 4,4 -12,5 21,5 63,9 28,9 +35,0
April 12,1 -0,5 23,5 80,1 68,3 +11,8
May 15,5 4,4 31,5 54,3 80,2 -25,9
June 18,6 5,5 32,5 88,4 93,6 -5,2
July 21,5 9,5 33,2 17,0 99,0 -82,0
August 19,5 9,6 33,2 169,4 64,0 +105,4
September 16,6 4,5 27,1 27,8 56,7 -28,9
October 10,9 -3,5 28,0 31,7 36,6 -4,9
November 5,2 -5,5 17,1 12,8 36,5 -23,7
December 0,5 -11,5
377
9,1 23,7 33,3 -9,6
14,8
34
7,3
32,6
7
29

1.1.3 Viticultural region Banat Crisana
Annual thermic regime was slowly in excess (an annual average temperature of 13,0 o C) and
slowly reduced during vine vegetation period (April – September average values of
19,1 o C). Haighest thermic value was recorded in July ( 39,1 o C), from June till September
the temperature values being over 30 o C, slowly affecting the photosynthetical process at a
foliar level. The lowest thermic value was recorded in January ( –15,1 o C), negative
temperature being recorded from October till March, but the fruit buds were not affected.
Buds viability was slowly affected in depressionary areas, without significant losses.
Thermic accumulation from the standard vegetation period (April – September) favourised
in a certain amount sugars accumulation, stopped by the highest temperature values from
the vegetation period.
Insolation value was close to normal, the photosynthetic process and the crop quality
not being disturbed. From a hydrological point of view, there was a surplus of moisture
content, favourising cryptogamic diseases. Normal humidity values were recorded in July-
August, favourising grape maturation.
As for the rainfall regime, according to the real evaporation, 2006 had higher amounts,
a hydrological deficit being recorded in July, September and October.
The dangerous winter conditions recorded were hoar-frost, block frost and white frost.
Hoar-frosts were recorded from October (4 days) to March (1 day), block and white frost
being recorded in December (1 day) and February (3 days). Among the worm season
conditions there were recorded 2 cases of hail-storm in June and strong wind weather, even
stormy weather in April and June.
1.1.4 Viticultural region Colinele Dobrogei
Examining the thermic regime of 2005-2006 viticultural year, we can say that its values
were higher than the average multi-annual values. Thus, during the vegetative rest period,
November, December and January had an average monthly temperature higher than
normal, and during the vegetation period, from April to January, all months had higher
temperature values than normal, the difference ranging from 0,7 0 C in April (lowest) to
3,7 0 C in August (highest).
During summer the highest temperature recorded was between 33,0 0 C and 36,0 0 C,
stimulating the vegetative raise of the vines, on the background of the massive rainfalls
from the second part of summer.
We can conclude that the 2005-2006 viticultural year, vine took advantage from a
more effective thermic regime than the average, leading to a superior viticultural
production, mostly in quantity and less in quality, due to the reduced number of days with
sunshine. Regarding the pluviometric regime, we can mention the lower rainfall amounts in
June, a situation that created a strong hydrological pressure in the plantation, irrigations
being mandatory. But the amount of rainfalls in July, August and September solved the
problem. The relative air humidity was of 85%, higher than the average 73%, influencing
and favourising cryptogamic disease attacks on vine, especially mildew on sensitive sorts,
but also blast and botrytis on the other sorts.
1.1.5 Viticultural region of Muntenian and Oltenian Hills
Analysing viticultural year 2005-2006 (October 2005- September 2006) we find average
temperature values higher than normal and less rainfalls, leading to an extension of vine
shoot maturation and a delay of the relative rest of the vine (fig. 1).
The reduced amount of rainfalls in May (16,4 mm, compared to the average of 60,1 mm)
and the lower thermic regime continued to have a negative influence on plant development.
Blooming started late (16 th - 19 th of June) and lasted till the end of the month. The rains
378

during this period, though not so significant in quantity (43,6 mm), created most favourable
conditions for the development of pathogenic fungus.
Thermic balance during July (Σ°738,0) and August (Σ°710) and the normal level of
rainfalls led to a recovery of growing differences and a normal development of the
vegetative system and of the grapes.
Climatic conditions in September, total hours of insolation, rainfalls and the active
thermic balance favoured the accumulation of sugars, of coloured substances and of
specific fragrance of every sort.
ToC/ mm
60.0
50.0
40.0
30.0
20.0
10.0
0.0
Fig. 1 Evolution of the average daily temperature ( o C) and of rainfalls (mm),
during 01.04 -30.09.06
April
ToC Σmm
May
June
1 11 21 1 11 21 31 10 20 30 10 20 30 9 19 29 8 18 28
If we analyse the climatic profile (for the vegetation period) for 2006 (tabel 4) and we
compare it with the normal values of the viticultural center Valea Calugareasca and with the
optimal values for obtaining quality crops, we can see that all the indicators had optimal or
almost optimal values in order to obtain quality white and red wines, having controlled origin
denomination (DCO).
379
July
August
September

Tabel 4 Climatic profile during vine vegetation period in 2006 Valea Calugareasca Viticultural
Center
Indicator name
Normal
values
Values
recorded
in 2006
Optimal
period
Observations
Global thermic balance ( o C) 3635 3562
2700-
3500
Useful thermic balance ( o C) 1801 1716
1000-
1700
> 17 o 23,8
C table wines
> 19 o Average temperature in July (
C high quality wines
o C) 25,2
18.5-23.2 o
> 21 C quality wines having
controled
denomination
origin
Highest average temperature in
August ( o C)
29,0
26,8
24.5-30.0
Values higher than 34 o C are
restrictive, beacuse they lead
to physiological process
blocking.
Highest temperature in August
( o 34,4
C)
34,2 2300 favourability for
aromatic sorts.
Indicator of coolness at night IF 19,8 13,6
>18
< 12
Worm nights, favourable to
maturation
Cold nights, unfavourable to
maturation.
1.2. CONCLUSIONS
Each wine-growing region has distinctive features from a microclimatic point of view, but
we can make a general analysis, taking into account the common climatic elements during
the analysed period, as it follows:
• thermical resources were high, a little over the multinational average values;
380

• the pluviometric regime was also higher than the average multinational values, being
quite uniformly distributed throughout the whole year;
• climatic accidents as the hail-storms in Moldavia and Dobrogea did not severely affect
vine plantations.
Losses of buds due to low temperatures during winter (-18 0 C) were of 0 and 35%, a
normal situation, solved by compensation cuttings.
Autumn was long and warm, favourising sugars accumulation and colour for the red
grape sorts.
An exception from a microclimatic point of view for 2006 was Transylvannia Plateau,
where thermic resources were quite low, the annual average temperature being 9,9 0 C, the
pluviometric regime 589,4 mm, the average multiannual value being of 644,6 mm. Also,
the lowest temperature (-20,3 0 C) affected frost-sensitive buds. Haziness and massive
rainfalls during the ripening period favourised mouldiness on grapes.
On the whole, we can say that 2006 was a microcimatic faourable year for most of the
wine-growing regions, this leading to good productions, in quantity as well as in quality.
BIBLIOGRAPHY
* Ministerul Mediului şi Gospodăririi apelor
* Administraţia Naţională de Meteorologie www.inmk.ro
* Raport 2006 ICDVV Valea Călugărească
* Raport 2006 SCDVV Odobeşti
* Raport 2006 SCDVV Bujoru
* Raport 2006 SCDVV Murfatlar
* Raport 2006 SCDVV Blaj
* Raport 2006 SCDVV Iaşi
* Raport 2006 SCDVV Miniş Măderat
381

WHEAT GRAINS PROCESSING INFLUENCE AT QUALLITY PARAMETERS
Timar Adrian
University of Oradea, faculty for Environmental Protection
ABSTRACT
This study try to find if processing of wheat grains is important. Althrough we try to
find how parameters of grains is changed during proccesing and if this parameters are
significant improuved. Paper is a part of my Phd. Thesis and will be coroborate with
parameters evolutions during storage time in thre diferent systems.
Key words : wheat grains, gluten, starch, rheologichall parameters, wheat grains purity.
1. INTRODUCTION
For evaluation the importance of processing we use data imputs for following
parameters: Organolepticall analysis, Physical analysis (Purity, Hectolitric weight, 1000
grains weight, Absolut weight, Specific weight, Grains humidity), Chemical analysis
(Mineral contents, Ph, Proteins content, Gluten content, Starch content), Rheological
analysis (Elasticity of gluten, Extensibillity of gluten, Degradation of gluten). Methods
used for analysis are according with romanian standards and are quottation in latest
studys.
2. MATERIALS AND METHODS
Taking samples: We use to take samples cilindrical and electromechanical probes. From
serface and upper layers samples was taken with cilindrical probes. From deep layers
samples was taken with electromechanical probes. Procedure was according to Thierer
L.V. 1976 and Duda M. 2003.
Obteining working samples :We form successively elementar, brutto, homogenized,
laboratory and work samples according with Mureşan et al. 1986.
2.1.Organoleptical analysis
Was study colour, aspect, smell and tste of grains according Thierer L.V. 1966. If this
parameters was out of normal range grains was considered out of standards and study of
those samples was ended.
2.2.Physical analysis
We study follow parameters : Purity, Hectolitric weight, 1000 grains weight, Absolut
weight, Specific weight, Grains humidity.
1./ Purity was study according romanian method, Muresan T. 1986 and Duda M. 2003.
P% = mx100/M %
m – weight of elements in g;
M – initial weight of sample in g.
2./ Hectolitric weight was study with hectolitrical 20 l ballance according Muresan T.
1986 and Duda M. 2003.
3./ 1000 grains weight was study with analitical ballance 0,01 g precision range
accooording Muresan T. 1986 and Duda M. 2003.
4./ Absolut weight was study according Muresan T. 1986 and Duda M. 2003, using
follow formula :G = (100 – U)G/100
U - humidity of grains, %; G – 1000 grains weight, g.
382

1. Specific weight was study according Muresan T. 1986 and Duda M. 2003,
using follow formula: m = G/V
G – 1000 grains weight, g; V - volumme of 1 000 grains, cm 3 .
2.2.Chemical analysis
We study following parameters : Mineral contents, Ph, Proteins content, Gluten content,
Starch content.
1. Mineral contents was made according Leonte M. 2006 by calcination at
600 °C.
2. Ph was measured by titrating with NaOH according Thierer L.V. 1966.
3. Proteins content was study by mineralyzade and titrating this extract
according Leonte M. 2006. Results was using in following foemula :
P%=[(V1 – V)x0,007x5,70]x100/m
V1 - H2SO4 0,5 n volume in ml;
V - NaOH 0,5 n volume in ml;
m – samples weight, în g;
0,007 – N, g equivalent in 1 ml H2SO4 0,5 n;
5,70 - proteic coeficient of wheat, g for 1 g N.
4. Gluten content was study according Thiere L.V. 1966 and Leonte M.
2003.
5. Starch content was study by Ewers method according Thiere L.V. 1966
and Leonte M. 2003.
2.3. Rheological analysis
We study following parameters : Elasticity of gluten, Extensibillity of gluten,
Degradation of gluten.
1. Elasticity of gluten measured lenght until epruvets was tear according Thiere
L.V. 1966 and Leonte M. 2003.
2. Extensibillity of gluten was study according Thiere L.V. 1966 and Leonte M.
2003, and measured capacity of gluten sphere to regain initial shape after
ending deformation. Results was expressed in conventional value from 1 for
very elastic and 5 for unelastic sample.
3. Degradation of gluten was study according Auerman method quotattion by
Thiere L.V. 1966 and Leonte M. 2003.
2.4. Experimental Methodic
Samples was study according following schema :
V1 – raw materials;
V2 – processing with fanning maschine type TA 1215 according Costin I. 1983 and
Bucurescu N. 1992;
V3 – processing with fanning maschine type TA 1215 and selector type MCS 5
according Costin I. 1983 and Bucurescu N. 1992;
V4 – processing with fanning maschine type TA 1215, selector type MCS 5 and
gravitational typ G 3,3 equipment according Costin I. 1983 and Bucurescu N.
1992
2.5. Biological material
We study wheat, Triticum aestivum L. ssp. vulgare Host McKey, kind named „Dropia”.
Dropia, was created by I.C.C.P.T. Fundulea, in 1992 and registered in 1993.
383

Grain is big, ovoid, and colour is red.. 1000 grains weight is around 44 şi 48 g. and
hectolitrical weight is around 73 şi 74,5 g.
Dropia is in B1-A2 quallity groupe, protein content is 13 – 14,8 %, wet gluten 23,3
– 38,8 % and dry gluten 9,2 – 15,4 %.
This study has been done in 15 july 2005 at harvest moment.
2.6. Statistics methodic
We use Polifaact statistic processing software.
3. RESULTS AND DISCUSSION
3.1. Physical analysis
Table 1. Purity, P % results
Nr. crt. Variant P, (%) % Diference Semnification
1 V1 81,00 100,00 - mt.
2 V2 81,80 101,00 0,80 ***
3 V3 82,60 102,00 1,60 ***
4 V4 84,30 104,10 3,30 ***
DL (p 5%)= 0,01; DL (p 1%)=0,02; DL (p 0,1%)=0,02
After every kind of processing of grains purity grows very significant so in V4 we have
the best results.
Table2. Hectolitric weight MH % results
Nr. crt. Variant MH, (kg) % Diference Semnification
1 V1 77,47 100,00 - mt.
2 V2 77,87 100,50 0,40 ***
3 V3 78,17 100,90 0,70 ***
4 V4 78,47 101,30 1,00 ***
DL (p 5%)=0,10; DL (p 1%)=0,15 ; DL (p 0,1%)=0,24
After every kind of processing of grains Hectolitric weight grows very significant so in
V4 we have the best results.
Table.3. 1000 grains weight, MMB g results
Nr. crt. Variant MMB, (g) % Diference Semnification
1 V1 47,00 100,00 - mt.
2 V2 47,80 101,70 0,80 ***
3 V3 47,27 100,60 0,27 ***
4 V4 47,60 101,30 0,60 ***
DL (p 5%)=0,06; DL (p 1%)=0,09; DL (p 0,1%)=0,14
After every kind of processing of grains 1000 grains weight grows very significant so in
V4 we have the best results.
384

Table 4. Absolut weight MA g results
Nr. crt. Variant MA, (g) % Diference Semnification
1 V1 33,12 100,00 - mt.
2 V2 33,96 102,50 0,84 ***
3 V3 34,22 103,30 1,10 ***
4 V4 35,35 106,70 2,23 ***
DL (p 5%)=0,01; DL (p 1%)=0,02; DL (p 0,1%)=0,02
After every kind of processing of grains Absolute weight grows very significant so in
V4 we have the best results.
Table 5. Specific weight MS g results
Nr. crt. Variant MS, (g) % Diference Semnification
1 V1 1,40 100,00 - mt.
2 V2 1,44 103,10 0,04 ***
3 V3 1,49 106,20 0,09 ***
4 V4 1,50 107,10 0,10 ***
DL (p 5%)=0,01; DL (p 1%)=0,01, DL (p 0,1%)=0,02
After every kind of processing of grains Specific weight grows very significant so in V4
we have the best results.
3.2. Chemical analysis
Table 6. Mineral contents % results
Nr. crt. Varianta Mineral contents, (%) % Diference Semnification
1 V1 1,82 100,00 - mt.
2 V2 1,82 100,00 - -
3 V3 1,82 100,00 - -
4 V4 1,81 99,40 - 0,01 ooo
DL (p 5%)= - ; DL (p 1%)= - , DL (p 0,1%)= -
After every kind of processing of mineral content decrease very significant in V4
especialy but because DL is nule this is a insignificant variance.
Table7. Ph results
Nr. crt. Variant Ph
1 V1 2,5
2 V2 2,5
3 V3 2,5
4 V4 2,5
Is no variation in course of processing.
385

Table 8. Proteins content % results
Nr. crt. Variant Proteins content, (%) % Diference Semnification
1 V1 13,80 100,00 - mt.
2 V2 13,50 97,80 - 0,30 ooo
3 V3 13,45 97,50 - 0,35 ooo
4 V4 13,21 95,70 - 0,59 ooo
DL (p 5%)=0,01; DL (p 1%)=0,02, DL (p 0,1%)=0,02
Because external components are eliminated during processing after every stage protein
level is decreasing. Rate of decreasing is very significant.
Table 9. Gluten content % results
Nr. crt. Variant Gluten content, (%) % Diference Semnification
1 V1 28,80 100,00 - mt.
2 V2 28,87 100,20 0,07 mt.
3 V3 28,90 100,30 0,10 *
4 V4 29,27 101,60 0,47 ***
DL (p 5%)=0,09, DL (p 1%)=0,13; DL (p 0,1%)=0,21
Variant V4 is the first what improuve very significant gluten content. V2 is almoust the
same lice witness sample and V3 is a little improuvment just significant.
Table 10. Starch content % results
Nr. crt. Variant Starch content, (%) % Diference Semnification
1 V1 59,37 100,00 - mt.
2 V2 59,83 100,80 0,46 ***
3 V3 59,94 101,00 0,57 ***
4 V4 60,70 102,20 1,33 ***
DL (p 5%)=0,06, DL (p 1%)=0,09, DL (p 0,1%)=0,14
After every kind of processing of grains Specific weight grows very significant so in V4
we have the best results.
3.3.Rheological analysis
Table11. Elasticity of gluten % results
Nr. crt. Variant Elasticity of gluten % % Diference Semnification
1 V1 2,00 100,00 - mt.
2 V2 2,00 100,00 - mt.
3 V3 2,00 100,00 - mt.
4 V4 1,00 50,00 - 1,00 ooo
DL (p 5%)=0,10; DL (p 1%)=0,15; DL (p 0,1%)=0,24
386

Very significant decrease are in V4 because this stage reduce also proteins content. The
rest of variants are like witness.
Table 12. Extensibillity of gluten mm results
Nr. crt. Variant Extensibillity of gluten, (mm) % Diference Semnification
1 V1 15,30 100,00 - mt.
2 V2 15,30 100,00 - mt.
3 V3 15,32 100,00 0,02 -
4 V4 15,80 50,00 0,50 ***
DL (p 5%)=0,03; DL(p 1%)=0,04; DL(p 0,1%)=0,07
Just in V4 we find very significant value, but because DL are very small this is
especialy in V3 just theoreticaly.
Table 13. Degradation of gluten mm results
Nr. crt. Variant Degradation of gluten, (mm) % Diference Semnification
1 V1 42,03 100,00 - mt.
2 V2 41,43 98,60 - 0,60 ooo
3 V3 40,73 96,90 - 1,30 ooo
4 V4 40,50 96,40 - 1,53 ooo
DL (p 5%)=0,15, DL(p 1%)=0,23; DL (p 0,1%)=0,37
Decreasing very significant of gluten degradation in every variants is obvious and show
in V4 variant the best result.
4.CONCLUSIONS
After this study we find very significant improuvments for almoust all parameters and
is obvious why is necessary to process the wheat with complete technological line.
The improuvments are so important to justify the best and complex production line, full
scale plant.
REFERENCES
1. Ardelean M., Sestraş R., Cordea Mirela, Tehnică experimentală horticolă, Edit.
Academicpres, Cluj – Napoca, 2005;
2. Bucurescu N. ş.a. : Sămânţa şi pregătirea acesteia pentru însămânţări, Edit. Ceres,
Bucureşti, 1992;
3. Costin, I., Tehnologii de prelucrare a cerealelor în industria morăritului, Ed.
Tehnica, Bucureşti, 1983;
4. Duda M. M., Vârban D., Muntenu S., Fitotehnie, Îndrumător de lucrări practice,
partea I, Edit. AcademicPress, Cluj – Napoca, 2003;
5. Leonte M., Tehnologii şi utilaje în industria morăritului, Edit. Millenium, Piatra
Neamţ, 2003;
6. Mureşan T., Pană N.P., Cseresnyes Z, Producerea şi controlul calităţii seminţelor
agrocole, Edit. Ceres, Bucureşti, 1986;
387

7. Thierer L V. Tehnologia recepţionării, depozitării, condiţionării şi conservării
produselor agricole, Edit. Ceres, 1976;
8. Thierer L.V., Dumitrescu M., Huştiu I., Oprescu I., Tehnologia recepţionării,
depozitării, condiţonării şi conservării produselor agricole, Ed. Ceres, Bucureşti,
1971;
9. Thierer, L.V., Determinarea calităţii produselor agricole vegetale, Bucureşti, Edit.
Agro- Silvică, 1966;
388

OCEAN DYNAMICS: AIR-SEA INTERFACE GASES EXCHANGE 1
Speranta Coldea,
Faculty of Environment Protection, University of Oradea,
Oradea, Str. Gen. Magheru Nr.26, Romania
ABSTRACT
Having known the tracer conservation equation and discussed ocean transport, we will
consider now the processes at the air-sea interface that need to be specified in order to
solve the tracer continuity equation. The most important process that we must consider
here there is the gas exchange. This process has a major impact on the distribution of
gases within both the ocean as well as within the atmosphere. We will introduce an
overview of the major gases in the atmosphere and ocean and some important processes
in which they are involved. Further we will discuss the solubility of gases in seawater
and the processes at the air-sea interface that control the exchange between the
atmosphere and ocean.
Keywords: atmosphere – ocean interaction, gases exchange.
1. INTRODUCTION
The terrestrial atmosphere composition is a unique one in our solar system due to the
influence on long terms of biological processes upon al the basic components, with the
exception of noble gases (see Table.nr.1). The noble gases are near the equilibrium state
between ocean and atmosphere (with the exception of a supersaturation due to air
bubbles solving, bubbles that are injected in ocean by the braking tides, and by the
fluctuations resulted from mixing, heating and from the water-air fluxes). The
distribution of ocean noble gases is determined especially by their solubility. The
equilibrium distribution between the atmosphere and ocean of the basic gases is
determined by the chemical and biological processes that produce the gas consumption
in the ocean. From the eight reactive gases given in the Table nr.1, three of them, e.g.
O2, CO2 and N2O, are produced and consumed in great quantities by the oceanic
processes. The Ocean doesn’t produce methane, but the flux of this gas to the
atmosphere is negligible compared to that from terrestrial sources (∼ 2%). The ocean
produces and consumes also N2, but we don’t know if the ocean is a source or a sink, or
it is neuter for this gas. Some gases have an impact on the thermal state of our planet by
caption of long wave radiation emitted by the Earth, from so called greenhouse effect.
We must observe that CO2 and N2O are in bigger quantities compared with the
beginning of the industrial era and contribute semnificatively to the greenhouse effect.
We will explore partially the vertical and horizontal distribution of the gases, and we
will give a special attention to the concentrations at the ocean surface in contact with the
atmosphere. This distribution has some unusual particularities for O2, CO2, and N2O
that we will introduce in our analysis. The nonquilibrium state between ocean and
atmosphere of CO2 is larger than that of O2. For CO2 the curve seems to begin to be
larger and it is opposite to that of O2. The same biological processes which decay O2 in
the deep waters will enlarge the concentration of the total carbon. This fact explains
why the curve for CO2 at the equator is opposite to that of O2, but doesn’t explain why
389

the maximum of CO2 have the tendency to be larger than that for O2. We introduce the
following question at which we must give a response: which are the relative
contributions of the biological and physical processes to this distribution of gases.
From the measurements it can be seen that the N2O distribution is near to the
atmospheric equilibrium on the most latitudes, with the exception of those closed to the
equator, where N2O is in a big quantity that would imply the existence of an oceanic
source. We need to see which this source is. To answer such questions we need to know
the gases solubilities, that tell us at which concentrations are the gases. We must also
discuss about the gases exchange rates that determines how rapidly a corps immersed in
water will be in equilibrium with the atmosphere at the ocean surface. In a future paper
we will discuss also about the biological processes that take place inside the ocean,
processes that together with the physical ones force the gases to go out from the
equilibrium state with atmospheric air.
2. THE GASES SOLUBILITIES
We need to estimate the gas concentration in mmol ⁄ m 3 in sea water, which is not in
equilibrium with the gas from wet air above of ocean. The gases atmospheric
concentrations are given as partial pressures in physical atmosphere (atm) defined by
the relation:
pA = χA ⋅P (1)
where the molar fraction χA is the moles number of the component A in the unity of air,
P being the total atmospheric pressure in atm. The most suitable definition for the gases
solubility is given as:
CA = SA ⋅pA = SA⋅χA⋅P (2)
where CA is the solution concentration of the component A in mmol⋅m -3 , equal with 1
μmol⋅l -1 ; SA is the solubility in mmol⋅m -3 ⋅atm -1 . This parameter is related to Bunsen
coefficient β or to the parameter that is function of solubility, F. These parameters could
be related. In the Table nr.2 the equation and the coefficient for calculation of Bunsen
coefficients and of solubility fractions F for some gases are given. The solubility decays
with the increasing of the temperature for all gases.
3. THE GAS EXCHANGE
The vertical motions in the atmosphere and in the ocean are intensively suppressed near
the air-ocean water interface. There were been proposed a variety of models to explain
the effect of this phenomenon on the gas transfer transverse to the air-water interrface.
We will take into consideration the stagnant film model. This model supposes that the
turbulence suppression at the sea-air interface could be represented by molecular
diffusion inside a stable film of finite depth, both in ocean and in the atmosphere. This
model could be easier visualized and gives us the possibility of a direct discussion for
the relative importance of the atmospheric and ocean boundary layers. The model of the
stagnant film represents the transition from the turbulence inside the free surface fluid to
the molecular diffusion from inside the stable film by a simple structure made from two
layers, illustrated in the Fig.1. If we take into account the fact that doesn’t exist nor
sources nor sinks inside the film, it may be affirmed that fluxes in these films will be
given by the first Fick law:
390

∂ C
Φ = - ε
(3)
∂z
where Φ is in mmol⋅m -2 ⋅ s - 1 . Gas flux inside the atmospheric film is represented by the
expression:
)
0
0 k
Φ = − k ( p − p ) = − a ( C − C
(4)
a a a a S a a
and inside the ocean film we have the relation:
Φ = − k ( C
0
− C )
(5)
o o o o
Table 1: The atmospheric composition influenced by chemical and biological
processes
________________________________________________________________
(a)The atmospheric composition (b) Influenced by chemical
(molar fraction in dry air) and biological processes
___________________________ppm___________________Yes___________ No___
N2 0,78084 ± 0,00004 CO2 358 Ar N2
O2 0, 20946 ± 0,00002 Ne 18,18 ± 0,04 Ne O2
Ar 0,00934 ± 0,00001 He 5,24 ± 0,004 He CO2
CH4 1,72 Kr CH4
Kr 1,14 ± 0,01 Xe H2
H2 0,55 N2O
N2O 0,312 CO
Xe 0,087 ± 0,001 O3
CO 0,05 - 0,20
O3 0,01 - 0,50
See West and East, 1982, for all gases, with exception of CO2, CH4 and N2O, that are
given after Houghton, 1996.
The index 0 indicates the concentration and the partial pressure at the air-water
interface, where z = 0.
This partial atmospheric pressure of a gas was converted in the concentration
equivalent in water, C, by using the solubility S. In these equations ka and ko are the gas
exchange coefficients, having the units of velocity and respectively of solubility. ko is
also named transfer velocity. ka ⁄ S and ko have values of ε ⁄ Δz in the model of stagnant
film, where Δz is the depth of this film.
The measurements of pressures and of partial concentrations are made above the
air-water interface, never on this ocean surface. We will consider from the continuity at
this interface that Φa = Φo = Φ and Ca 0 = Co 0 , with the aim to eliminate the surface
concentrations from eq.(4) and eq.(5) . It will be obtained that:
Φ = − K( C − C )
(6)
a o
391

with
= +
K k
o
1 1
ka ⁄ S is much larger than 1000 cm ⁄ hour. As compared with the values of order of 10
cm⁄hour for ko, that permits us to ignore the stable film from the atmosphere, this will
give K = kO, and
Φ = − k ( C − C )
(8)
O a O
or, as a function of partial pressures, we have:
Φ = − k ( p − p )
(9)
s
with ks = kOS.
The measurements of the gas exchange constants were made in wind tunnels and
using in situ markers for a variety of gases, under various wind velocities and
temperature conditions. The measurements indicate a dependency of friction in water,
u*O, for kO of and also of Schmidt number, defined as
Sc = ν ⁄ ε (10)
where ν is the kinematic viscosity, ko of the form
kO = γ -1 Sc -n u*O (11)
and γ is a non-dimensional constant. The exponent n at the number Sc is included to
permit that the gas exchange do not comports as when it should suffer a transversal
transition to a stable film with kO = ε ⁄ Δz (e.g. with n = 1). n could be determined by
gases exchange measurements with more that a single gas and a heat exchange in the
same conditions:
log( k 1 / k 2)
n = − O O
(12)
log( Sc1
/ Sc2)
In Table nr.2 are given different models for the gases exchange coefficient. These have
a difference between them of more than a factor of 2.
4. CONCLUSIONS
There were made laboratory studies and also in situ experiments. The laboratory
studies were made in hydrodynamic tunnels, with which are analyzed the gases
exchanges and the exchange coefficients are measured by measurement of the
equilibrium for a marker mass in water. The exchange on a given period of time is the
flux. The exchange coefficient is obtained making the rapport between the flux and the
difference of concentration air-water, measured with eq.(5). Such experiments have
indicated that the gases exchange coefficients at the air-water interface varies as
functions of wind velocity, U, giving us the idea that of existence of three linear regimes
separated by the transition from a plate water surface to one with one with rugosities,
and from a rugous surface to one for which the tides begin to break up. The theory of
the micrometeorological boundary layer, that tell us that n = - 2 ⁄ 3 [1]-[4], seems to
be good when the water surface is plate. The Model of the substitution film, which
proposes the existence of a film that is periodicaly substituted by the fluid volume, gives
392
a
S
k
a
O
(7)

n = - 1 ⁄ 2 [1]-[4]. The later model is applied when the water surface is rugous (with
tides).
Table nr.2
___________________________________________________________Solubility
Symbol Conversion to SA
___________________________________________________________
For using with pA humid = pA dry (1 - pH2O/P)
−3
β A 6 mmol m
Bunsen coefficient βA SA = × 10 −1
VA mol l
Solubility function F
F
3
6 mmol m
−
SA =
× 10
( P − p ) −1
H2O
mol l
______________________________________________________________________
For using with pA dry
β
−3
6
Bunsen coefficient βA SA =
A mmol m
× 10 −1
V mol l
A
−3
F 6 mmol m
Solubility function F SA = × 10 −1
p mol l
It can be observed that a not so soluble gas as O2 has an exchange rate larger than a
more soluble gas as CO2. This fact is perfectly compatible with the important role of the
bubbles in the gas exchange process previously discussed. The not so soluble gases
exchange is increased by a bigger fractional quantity when the bubbles are developed,
due to the fact that the gases with a lower solubility have a lower transfer coefficient
transversal to the air-sea interface than the gases with bigger solubility. The bubbles
transfer could be separately discussed. A result of the laboratory studies is: it
demonstrates that the sensibility of gases exchange for a variety of conditions, that are
difficult to reproduce in a laboratory situation.
A series of techniques for studying the gases exchange in a more realistic mode
were developed, in the field conditions. There are three such techniques:
• The method of the radon deficiency
• The method of the oceanic radiocarbon inventory, and
• The dual method of a tracer.
We will refer to in situ (in field) studies, describing them and presenting here only the
method of the radon deficiency.
(a) The method of the radon deficiency is based on the Radon-222 distribution at
the ocean surface. This radioisotope, with the de half value life time of 3,825 days, is
formed by the decay of Radiu-226 at oceanic water surface. We know from
393

concentration measurements of the radium how much radon-222 is formed in a unit of
time. The measurements effectuated in the mixed layer present a deficiency relative
to the quantity at which we should expect. The flux to the atmosphere can be estimated
from this deficiency and the gas exchange coefficient is found from the rapport of the
flux to the air–water concentration gradient. We will present a simple estimation, but we
will firstly consider the process of a series of decays, which imply radium-226 and
radon-222. Radiu-226 has a half value life time much longer than radon-222. These two
isotopes are put in isolation until when they reaches equilibrium, where the exchange
rates for which the two isotopes are decayed are equal. It is better to give the
concentration of these radioisotopes by the activity, noted as A, which is equal with the
product between the decay constant λ and the atoms number that are present in a
volume unit:
A Rn-222 = λ⋅CRn-222 (13)
where CRn-222 represents the atom concentration per volume unit. In the case of our
considered isotopes the concentration is given in dpm ⁄100 Kg, where dpm is the
number of decays in a minute. The decay constant is given as a rapport between ln(2)
and the mean half value life time. The equilibrium activity of Rn-222 is of the form A Rn-
222 Ra-226
= A .
The radon deficiency due to the gas exchange is given by:
Φ = h(A Ra-226 - A Rn-222 ) (14)
where h is the Radon-222 layer depth and the activities are the values on this layer. The
flux will be of the form:
Φ = - kapă(CRn-222)aer - (CRn-222)water =
k
= −
water
[ A ) − ( A ) ]
(15)
λ Rn−222
air Rn−222
water
The concentration of the atmospheric radon Rn-222 is much lower than that from the
ocean. Then we can ignore the activity of Rn-222air and we make equal the last two
equations, obtaining the following result:
⎛ A ⎞
k = λ h⎜
Ra − 226
− 1⎟
(16)
water ⎜ A ⎟
⎝ Rn − 222 ⎠
From the made measurements it was obtained (in a mixed layer of 40 m in depth) that
the rapport of Ra-226 to Rn-222 is 1,6, giving a pushing velocity kappa of the form:
ln( 2)
−1
k
water
= 40 m(
1,
6 − 1)
= 4,
4 m d = 10 cm / hour (17)
3,
85d
(c) Applications
One of the advantages of in situ researches is that they have permitted a better
understanding of the role played by organic films for determination of the gases
exchanges. After obtaining such results more types of models for this phenomenon of
the gas exchange were been given. It is not the aim of this paper to develope models,
which were only used in the next discussion and could be given in the Table nr.3. We
wish that, on these models base to answer at some questions that were been not
394

elucidated yet. We must also imply the supersaturation of 3% oxygen observed any
place in the oceans., but also we must consider a lot of other processes: from the
sensibility to temperature of the oxygen solubility it can be observed that a water
heating, water that is in equilibrium with the atmosphere leads to a supersaturation in O;
an other mechanism which must be considered is the mixing.- a mixing of waters at two
different temperatures will also give a supersaturation. The third mechanism which
must be considered as contributing to the supersaturation is bules forming.
One of the known model for bule forming was done by Woolf and Thorpe (1991);
At a wind velocity of 7,5 m⁄s this model gives a supersaturation in oxygen of 0,1 %. The
theory of bule injection is probably the cause of a quart from the observed
supersaturation. The remaining suprasaturation in O may be given by some processes as
are the oxygen production by photosynthesis in the ocean surface layer. The
suprasaturation in O leads at a leak of oxygen to the atmosphere gas flux. Another
mechanism that must be considered is the oxygen leak to the ocean termocline by a
vertical exchange. The oxygen production by photosynthesis will equilibrate the leak
rate by these two mechanisms. We can now estimate the oxygen leak by gas exchange.
We will also include the bule injection for the temperature corrections and it is
obtained:
Φ = - kapă[(1 + ζ +Δe)⋅(CO2)aer – (CO2)apă] (18)
The oxygen concentration in water is suprasaturated by 3 % related to the oxygen
concentration in the air, e.g. (CO2)water = 1,03 (CO2)air , where (CO2)air = 0,241 molm -3 is
the saturation concentration, at the surface global mean temperature of 17,64 0 C and
with a water salinity of 24,78 %. The correction given by the process of bule injection is
of 0,007, at a wind velocity of 7,5 ms -1 .
From the oxygen solubility we can estimate a surface temperature lower with
0,3 0 C, that will give ζ = 0,004. For the gas exchange the coefficient obtained from the
model of Liss and Merlival is given in the table nr.3. By using the mean global
temperature it is obtained for the Schmidt number ScO = 663. This value gives a gas
exchange coefficient of 17,4 cm⁄hour from the model of Wanninkhof, and a value of
10,9 cm ⁄ hour from the model of Liss and Merlival. Converting these values in units of
m⁄hour and substituting all the obtained values the gas exchange flux will be
Φ = - 4,4 la - 7,0 m -2 yard -1 (19)
The oxygen production estimation is superimposed onto the typical estimations for the
oxygen biological production in the surface ocean layer that is of the order of 5 mol m-2
yard-1.
On the basis of the previously made analysis, which combines different
expressions from various model for the oxygen superasaturation with some in situ
obtained empirical data, we could give the conclusion that the superasaturation in O,
globally observed at the ocean surface could be explained by a combination of the
following processes: the bule injection, the effect of a lower temperature of the surface
water, and, the most important mechanism – the production of oxygen by
photosynthesis. The phenomenon of oxygen O2 suprasaturation conducted by the
biological processes and the net oxygen flux to the atmosphere which this implies must
be equilibrated in the subsaturation regions by a net flux of oxygen to the ocean. Other
way the level of O2 should decrease has not been observed yet. A zone where there is
395

oxygen subsaturation is that of equator, and we don’t discuss here this particular region.
As a conclusion the results obtained for the mean gases exchanges could be given in kg
and estimations for the increasing in radiocarbon obtained from different threedimensional
models are given in the Table nr.4. The last column in Table nr.4 gives the
radiocarbon increasing normalized to the global inventory estimated at 7,67 x 10 9 atoms
cm -2 .
Table 3: The expressions for the exchange coefficients kwater given as a convention in
cm⁄hour, the wind velocity U is in ms-1, obtained by the various models proposed for
the gas exchange
(1) Liss si Merlival (1986)
kwater = 0,17U(Sc ⁄ 600) -0,5 for U ≤ 3,6 ms -1
kwater = (U – 3,4)2,8(Sc ⁄ 600) -0,5 for 3,6 < U < 13 ms -1
kwater = (U – 8,4)5,9 (Sc ⁄ 600) -0,5 for U > 13 ms -1
(2) Wanninkhov (1992)
kwater = 0,39Umean 2 (Sc ⁄ 600) -0, 5 to be used with the mean wind velocity
kwater = 0,31U 2 (Sc ⁄ 600) -0,5 to be used with the mean wind velocity on short term
(3) Butin and Etcheto (1995)
kwater = 0,34(U 2 + σu 2 (Sc ⁄ 600) -0,5
(4) Broecker (1985)
kg = 0,0118 (U – 2)
(5) Maier-Reimer and
Hasselmann (1987)
kg = 0,050
Table 4:. The mean gases exchanges and the estimated in radiocarbon increasing
Formulation for kg global mean of radiocarbon increasing
the gas exchange (mol m -2 yard -1 ppm -1 )
The models results
(a) Brocker (1985) 0,061 1,000
(b) Liss and Merlival (1986) 0,029 0,607
( c) Maier-Reimer and Hasselmann 0,050 -
(1987)
(d) Broecker (1985 and 1995) 1,070 and 1,074
REFERENCES
396

RESEARCHES ON THE QUICK METHOD OF SUGAR PRODUCTIVITY
EVALUATION
Gheorghe Sarca
University of Oradea, Faculty for Environmental Protection
ABSTRACT
In a sugar factory it is necessary to be able to calculate very quickly the sugar
productivity possible to obtain by processing the beet having a certain technological
quality. That is the reason why, in the following lines, it is presented a very quick
method used by the Institute of Alimentary Chemistry, the laboratory for sugar research
and engineering, for such kind of quick calculations, but with a high theoretical and
practical precision.
INTRODUCTION
Table 1 is completed after the calculations.
Table1. 1 The quick calculation of sugar productivity
No Name of product
Quantity of: Purity Content of:
Product Dry Sugar % Dry Sugar
substance
substance
Kg/100 kg Kg/100 kg Kg/100 kg % ˚Brix %
beet beet beet
C SU Z O B P
0 1 2 3 4 5 6 7
1 Beet noodles 100,00 - 16,00 - - 16,00
2 Undeterminable
sugar losses, at
diffusion
- - 0,20 - - -
3 Moist beet pulp 80,00 - - - - -
4 Pressed beet pulp 31,10 - 0,28 - 18,00 0,9
5 Water from the
beet pulp’s
pressing
48,90 - - - - -
6 Total water for
diffusion
103,70 - - - - -
7 Fresh water for
diffusion
54,80 - - - - -
8 Diffusion juice 123,70 18,19 15,52 85,30 14,70 12,50
9 Mud from
carbonation
10,00 - 0,07 - - 0,7
10 Undeterminable
sugar losses, at
purification
- - 0,05 - - -
397

11 Thin juice 124,70 17,11 15,40 90,00 13,70 12,35
12 Thick juice 26,30 17,11 15,40 90,00 65,00 58,50
13 Undeterminable
sugar losses, at
steaming, boiling
and
crystallization in
three products
- - 0,30 - - -
14 Total products - 17,02 15,10 - - -
15 Molasses 5,66 4,80 2,88 60,00 82,00 50,85
16 Molasses type
50%
5,76 4,80 2,88 60,00 83,33 50,00
17 Moist crystal
sugar
12,300 12,24 12,22 99,85 99,5 99,35
18 Crystal sugar 12,246 12,24 12,22 99,85 99,95 99,80
19 Water
evaporated at the
sugar drying
0,054 - - - - -
For calculating this total the following method is recommended:
C1 has the value of 100.00 because the total is calculated for 100 kg beet noodles;
P1 is determined with the laboratory analysis. In the given example it was admitted the
value of 16.00%;
Z1=P1;
Z2 – the value recognized by the extractor’s supplier is accepted. In the given example
the value of 0.20% was admitted, specific to the extractor type DDS;
SU2 is considered 30% from Z2, because, by fermentation or enzymatic processes, the
sugar is not wholly changed into volatile substances and water, but it is particularly
changed in lactic acid. The recommendation is given by The Laboratory for sugar
research and engineering at the Institute of Alimentary Chemistry, Bucharest;
C3 is considered 80% of C1;
P4 is determined by laboratory analyses. In the given example the value of 0.9% is
accepted;
B4 is determined by laboratory analyses. In the given example the value of 18% is
accepted;
C4 is calculated from Muck’s diagram, according to the known methodology;
Z4 = (C4+P4)/100;
C5 = C3 – C4;
Z8 = Z1 – Z2 – Z3
P8 is determined by laboratory analyses. In the given example the value of 12.50% is
accepted;
B8 is determined by laboratory analyses. In the given example the value of 14.70˚Brix
is accepted;
Q8 = (P8/B8) – 100;
Z8 = ((Z1 – Z2 – Z4)/P8) – 100;
398

SU8 = (Za/QB) – 100;
C8 = (Z8/P8) – 100;
C6 = C8 + C3 – C1;
C7 = C6 – C5;
P9 is determined by laboratory analyses. In the given example the value of 0.70% is
accepted;
C9 is accepted as being equal with 10.00 kg / 100 kg beet;
Z9 = (C9 x P9)/100;
Z10 = 0.05;
B11 is determined by laboratory analyses. In the given example the value of 13.70˚Brix
is accepted;
P11 is determined by laboratory analyses. In the given example the value of 12.35% is
accepted;
Z11 = Z8 – Z9 – Z10;
SU11 = SU8 – SU12;
Q11 = (Z11/SU11) x 100;
C11 = (Z11/P11) x 100;
B12 is determined by laboratory analyses. In the given example the value of 65.00˚Brix
is accepted;
P12 is determined by laboratory analyses. In the given example the value of 58.50% is
accepted;
Q12 = (P12/B12) – 100;
SU12 = SU11;
Z3 is accepted with the value of 0.3 kg/100 kg beet;
Z14 = Z12 – Z13;
B15 is determined by laboratory analyses. In the given example the value of 82.00˚Brix
is accepted;
P15 is determined by laboratory analyses. In the given example the value of 50.85% is
accepted;
Q15 = (P15/B15) – 100;
Z15 = (SU11-Z11) x (Q15/(100-Q15));
C15 = (Z15/P15) x 100;
SU15 = (Z15/P15) - 100;
Q16 = Q15
P16 = 50;
B16 = (P16/Q16) x 100;
SU16 = SU15;
C16 = (Z16/P16) – 100;
Z17 = Z14 – Z15;
B17 is determined by laboratory analyses. In the given example the value of 99.50% is
accepted;
P17 is determined by laboratory analyses. In the given example the value of 99.35% is
accepted;
Q17 = (P17/B17) x 100;
SU17 = (Z17-Q17) x 100;
C17 = (Z17/p17) x 100;
399

SU18 = SU17;
Q18 = (Z18/SU18) – 100;
B18 is determined by laboratory analyses. In the given example the value of 99.95% is
accepted;
P18 is determined by laboratory analyses. In the given example the value of 99.80% is
accepted;
C18 = (Z18/P18) – 100;
C19 = C17 – C18.
CALCULATION FORMULAS OF SOME TECHNOLOGICAL PARAMETERS
Calculation of the water quantity for diluting the thick mass final product
The following relation is used in order to calculate the water quantity necessary for the
diluting of the thick mass final product before the additional crystallization through
cooling:
[1 l water/100 kg thick mass final product], in which:
B is the content of dry substance in the thick mass final product while discharging it
from the vacuum apparatus, in ˚Brix:
NZ1 – content of non-sugar in the thick mass final product while discharging it from the
vacuum apparatus, in kg/100 kg thick mass;
A1 – content of water in the thick mass final product while discharging it from the
vacuum apparatus, in kg/100 kg thick mass;
NZ2 – content of non-sugar expected to be produced by diluting the thick mass final
product, in kg/100 kg thick mass;
A2 – content of water in the thick mass final product after dilution, in kg/100 kg thick
mass
Relations between the values of different indicators determined in laboratory analyses
The following indicators are:
O – purity;
Z – content of saccharose polar-metrical determined from a product, expressed in %;
S – content of apparent dry substance of a product refractory-metrical determined,
expressed in ˚Brix;
A – content of water of a product, expressed in kg/100 kg product;
NZ – content of non-sugar of a product, representing the sum of the content of organic
substances and mineral substances in a product, expressed in kg/100 kg product;
Z/A – ratio non-sugar / water or solubility of sugar in water;
C – content of conductor-metric ash of a product, expressed in %;
SO – content of organic substances in a product, expressed in %;
RO – organic ratio that represents the ratio between the content of organic substances
and conductor-metric ash in a product;
CS – saline coefficient, expressed in the ratio between the content of sugar and
conductor-metric ash in a product
400

There are the following calculation relations between these indicators:
Z
Q = * 100 =
B
A = 100 – B [kg/100 kg product];
S / A*
( 100 − B)
B
Q*
B
Z =
100
Z
B = * 100 =
Q
=
= Z/A * (100 - B ) [%]
100*
S / A
S / A + NZ / A
Z / A*
( 100 − B)
* 100 [%]
Q
Z Q*
B
NZ = B – Z = * 100 - [%]
Q 100
Z
Z/A =
100 − B
=
Q*
B
[%]
100*
( 100 − B)
B − Z Z / Q − Z Z ⎛ 1 ⎞
RO = - 1 =
- 1 = * ⎜
C C
⎜
−1⎟
[%]
C ⎝ Q ⎠
But Z/C is the saline coefficient, noted CS. It results:
⎛1
− Q ⎞
RO = CS * ⎜
⎟ - 1,
⎝ Q ⎠
From where results:
Q
CS = (1 – RO) * .
1−
Q
REFERENCES
1. Banu C., 1992 – „Progrese tehnice, tehnologice şi ştiinţifice în industria
alimentară”, Ed. Tehnică, Bucureşti.
2. Beceanu Dumitru, Chira Adrian, 2003 – „Tehnologia produselor agricole”, Ed.
Economică, Bucureşti.
3. Sarca Gheorghe, 2004 – „Materii prime vegetale”, Ed. Universităţii din Oradea.
4. <strong>International</strong> Sugar Journal, 2006 december.
5. Popescu V., Stroia A., 1998 – „Tehnologii moderne de industrializarea sfeclei de
zahăr”, Ed. Ceres, Bucureşti.
401
[%]

RESEARCH REGARDING THE CONCENTRATION OF THIN JUICE AND
THICK JUICE
Gheorghe Sarca
University of Oradea, Faculty for Environmental Protection
ABSTRACT
The thin juice boils off in an installation working on the multiple effects principle. The
process the concentration is based on is the vaporization through boiling the thin juice at
boiling temperature.
INTRODUCTION
The highest temperature sugar is submitted to during the technological process of cane
processing is in the first body of the installation and it has values of 126-129 o C. Due to
the high temperature and the time these temperatures are maintained, losses of sugar
occur and they are presented in table 1.
Table 1. Quantity of decomposed sugar in the vaporization station
with multiple effect in mg/hour
Temperature Apparent content of dry substance of juices that boil off in the
vaporization station, in °BRIX
o C 15 25 35 45 55 65
80 0,0443 0,0300 0,0156 0,0178 0,0199 0,0232
85 0,0614 0,0420 0,0222 0,0261 0,0295 0,0330
90 0,0789 0,0540 0,0289 0,0343 0,0391 0,0450
95 0,0964 0,0660 0,0356 0,0426 0,0487 0,0555
100 0,1139 0,0780 0,0422 0,0507 0,0583 0,0675
105 0,1384 0,0936 0,0489 0,0587 0,0679 0,0792
110 0,1629 0,1092 0,0556 0,0666 0,0775 0,0891
115 0,1748 0,1186 0,0622 0,0747 0,0861 0,0991
120 0,2822 0,2340 0,1856 0,2268 0,2677 -
125 0,5329 0,5081 0,4838 0,5938 0,7043 -
130 2,0552 1,4609 0,8666 1,0234 1,1799 -
135 3,5775 - - - - -
140 5,0999 - - - - -
In figure 1 is presented the chart of a modern installation of thin juice concentration in a
sugar plant. This installation reduces to minimum the time for heating the juice,
ensuring the decrease of thick juice dyeing. The capacity of such an installation,
manufactured by the French company MAGUIN, is of 14000-20000 t cane/24 hours.
- entry of thin juice
- high pressure steam
- steams 124 o C
402

- preheaters
- towards condensation
- thick juice
- heating
- general services
Fig. 1 - Thin juice concentration installation
MATERIALS AND METHODS
Another concentration installation used in sugar plants is that of four effects and “0
body” whose thermal regime is presented in table 2. This installation is profitable
especially in the conditions of juice processing with no thermal stability, as well as in
the case of plants that don’t have electric vortex wheels.
Table 2. Research methods regarding the temperature regime at the station of
vaporization concentration with four effects and “0 body”
Basic indicators
Number of vaporization effect
0 1 2 3 4
Heating steam temperature 141 114 106 97 84
Heating steam pressure, ATA 3,89 1,71 1,91 0,95 0,56
Useful temperature difference, °C 25,5 6,2 6,4 10,4 22,4
Boiling temperature for juice, °C 116,3 108,6 100,4 87,4 61,4
Temperature loss due to
hydrostatic pressure °C
0,3 0,6 1,4 2,4 3,4
Secondary steam temperature, °C 115 107 98 84 57
Secondary steam pressure, ATA 1,77 1,35 0,99 0,58 0,184
Temperature losses in the steam
pipes °C
1 1 1 1 1
Condensate temperature, o C 139 112 104 95 81
403

The main technological and energetical tasks of a thin juice concentration installation
are the following:
• concentration of thin juice up to the limit of avoiding the danger of accidental or
uncontrolled crystallization of saccharose before vacuum apparatus. Generally, in
the conditions of Romania, the thick juice can concentrate up to 69,5-70,00 o Brix
• recovering the evaporated water from the thin juice and ensuring the requierement
of secondary steam necessary for technological purposes specific to the plant
• condensation of the steam received from the vortex wheel under the shape of
condensate, that returns to the Thermoelectric Power Plant of the plant.
The correct operation of the thin juice concentration station is ensured by the operating
way of the main users of secondary steam produced as an effect of juice concentration.
Among the main users, the preheaters of juices and syrups have an important place.
In tables 3, 4 and 5 are present the total coefficients of thermal transfer, in
W/(m 2 .grade), of diffusion juice, clear juice I, thin juice and thick juice or syrups.
Table 3.Total coefficients of thermal transfer, in W/(m 2 .grade) at diffusion juice
preheaters
Temperature of heating Total coefficients of thermal transfer at circulation
agent, speed of juice, in m/s of:
o C
0,1 0,2 0,3 0,4 0,5
Under 100 375 655 836 1017 1220
Over 100 466 815 1045 1275 1513
Table 4. Total coefficients of thermal transfer, in W/(m 2 .grade) at diffusion juice,
clear juice I and thin juice preheaters
Circulation speed
of juice through the
preheaters pipes
Temperature difference, Δt, o C
m/s 10 15 20 40 50
I. For diffusion juice (raw juice)
1,0 710 676 648 627 606 578
1,5 941 901 857 815 780 731
2,0 1163 1164 1101 1045 976 920
12,3 1303 1234 1164 1101 1045 976
2. For clear juice I and thin juice
1,0 2071 1931 1833 1743 1659 1589
1,5 2231 2126 2022 1917 1645 1722
2,0 2440 2280 2182 2043 1952 1645
2,5 2580 2931 2301 2182 2022 1882
404

Circulation speed of
the concentrated
syrup
Table 5. Total coefficients of thermal transfer, in W/(m 2 .grade)
at thick juice preheaters (concentrated syrup)
Thermal transfer
coefficients for content of
dry substance in °BRIX,
DE
m/s 60 65
0,3 404 299
0,5 676 466
0,7 752 543
0,9 927 676
Observations
In the case of calculation of
newly assembled preheaters we
will use values of juice or syrup
speed:for juices 1,5 m/s
- for syrups 0,7 m/s
RESULTS AND DISCUSSIONS
Results obtained in the principles of using the thin juice concentration installation
In the thin juice tank, placed in front of the concentration installation, there must be a
permanent stock of juice that ensures the feeding of the concentration installation for
10-15 min . If this stock of thin juice is lacking we have to introduce in the tank
condensate from the third level of the concentration station with multiple effect.
The functioning of the concentration station is based on the circulation of the juice
and of the forward flow steam, with the gradual reduction of pressure and temperature.
From every body of the concentration station, secondary steams are taken over and
directed for heating in various technological points of the plant, at various equipments
and installations. Generally, the last concentration effect of a concentration installation
works under vacuum, the equipment that represent this effect being connected to its
own barometric condenser.
The quantity of vaporized and removed water from the juice in the concentration station
depends on the following factors:
• content of sugar of the processed cane and the sutirage made at the sugar extraction
installation
• dilution grade of juice during the calco-carbonic purification process
• final concentration of thick juice that is evacuated from the last level of the
concentration installation.
The physical and chemical phenomena that occur during the thin juice concentration are
the following:
• saccharose decomposition under the influence of high temperature
• formation of coloring substances that determine the increase of the dyeing with 5
units ICUMSA/min – 100 g saccharose. Generally, during the concentration of the
juice, the dyeing increases with 50-100%
• decomposition of groups of non-saccharates, for example, amides and inverted
sugar existing in the juice, firstly of those quantities that have not been destroyed
in the specific phases of calco-carbonic purification. As an effect of amides
decomposition ammonia is given off;
• precipitation of insoluble salts, firstly carbonate, sulphite, oxalate and aluminiumcalcium
silicates. The phenomenon is due to the increase of the juice concentration,
405

ecoming an oversaturated solution for many calcium salts. Also, by the
decomposition of groups of non-saccharates, oxalic acid is formed and it produces,
through the reaction with calcium ions, insoluble salts or hardly soluble calcium
ones. The sediments produced in the concentration station, partially deposit
themselves on the thermal exchange surfaces and partially leave the installation,
being carried along by the thick juice flow;
• decrease in the alkalinity of the juice as compared to the dry substance content, as a
result of the carbonate neutralization by the formed acids, the juice giving off
carbon dioxide;
• in the case of juice overcarbonization at the second carbonization, in the steamers
of the concentration station, the decomposition of the calcium hydrocarbonate in
calcium carbonate hardly soluble and in CO2 occurs.
The thick juice resulted from the concentration of the thick juice in the concentration
station must have the following technical features:
1. the content of dry substance: 62-70 o Brix, according to its purity and the fulfilment
of the thermostability criteria
2. dyeing: 1500-2500 units ICUMSA/100 o Brix
3. content of calcium salts in the case of technological mature and healthy cane: 50-60
mg CaO/100 o Brix
4. alkalinity to phenolphthalein: 0,005-0,06 g CaO/100 ml.
5. In these conditions, the quantity of thick juice that results from 100 kg cane is 23-
28 kg.
REFERENCES
1. Banu C., 1992 – „Progrese tehnice, tehnologice şi ştiinţifice în industria
alimentară”, Ed. Tehnică, Bucureşti.
2. Beceanu Dumitru, Chira Adrian, 2003 – „Tehnologia produselor agricole”, Ed.
Economică, Bucureşti.
3. Sarca Gheorghe, 2004 – „Materii prime vegetale”, Ed. Universităţii din Oradea.
4. <strong>International</strong> Sugar Journal, 2006 december.
5. Popescu V., Stroia A., 1998 – „Tehnologii moderne de industrializarea sfeclei de
zahăr”, Ed. Ceres, Bucureşti.
406

RESEARCH ON SEEDING AGENTS TO EFFECT THE SUGAR
CRYSTALLIZATION
Gheorghe Sarca
Oradea University, Faculty of Environmental Protection
ABSTRACT
The quality of sugar is the decisive factor in ensuring the sugar commercializing
through the conservation of old markets and expansion towards new markets. The sugar
quality is also the key element which determines the price maintenance in according
with the expenditures performed or even the diminution of the price as a consequence of
the quality lessening.
INTRODUCTION
Securing the technological process efficiency is mainly done through the following:
• Decreasing to the minimum level the sugar amount re-circulated from the 1 st step
of crystallization to the 2 nd one;
• Decreasing to the minimum level the sugar amount re-circulated from the 2 nd step
of crystallization to the 3 rd one;
• Permanent preoccupation to avoid the drawing in the molasses of those small
crystals that can pas through the loops of the centrifugal sieves;
• Obtaining the top quality sugar mainly defined by:
- solution colouring, sugar-in-water solutions transparency
- polarisation, which shows the amount of saccharose content out of the crystal
sugar;
- humidity and the content of conductor-metrical ashes;
- the size the shape and the crystal homogeneity;
all these are being worldwide achieved by the way in which the crystallizing and
shaping technology regarding the emerged crystals is conducted.
MATERIALS AND METHODS
Methods for starting the saccharose crystallization
There are well-know 3 methods for saccharose crystallization starting:
• seeding in the field of supersaturating metastable by inserting desired quantity of
crystallizing germs in the concentrated syrup;
• crystallization centres formation by a thermal shock effect in the super-saturation
intermediate level;
• spontaneous or natural crystallization in the unstable field.
The main method for starting the saccharose crystallization out of its super-saturated
solutions takes place through the seeding method. The seeding is the preliminary
condition for the transition to the boiling and to the controlled and automatic
crystallization of the sugar as only in this way one can reach to the process
reproducibility to the
407

Seeding with powder sugar
The quality of the obtain sugar relies, and especially the homogeneity of the crystals
obtained, the shape of the crystals, the occurrence and the conglomerates content, all
depend on the way the seeding is done. In Romania, powder sugar has been used for a
very long time and it is still being used as seeding agent. The use of powder sugar in
alcoholic suspension presents certain advantages as against the use of only powder
sugar as such. The advantages of the seeding with powder sugar are the following:
• one can accurately establish the number of germs contained in the seeding solution
which leads us to the awareness of the total number of germs introduced in the
vacuum apparatus;
• one can ensure the uniformity of the germs from their size point of view;
• one can achieve the advanced dispersal of the sugar particles out of the alcoholic
suspension. This dispersal attracts itself an improvement of the sugar’ quality both
from its crystal aspect point of view and from chemical point of view. Due to the
advanced disposal degree, the conglomerate formation is avoided. The crystal
sugar conglomerates are the formations resulted from the sugar crystals bonding.
During their shaping these conglomerates include “mother” syrup. This fact leads
to the augmentation of the conductor-metrical and carbonic ashes in the crystal
sugar;
• achievement of a better homogenising of the crystallization germs within the
supersaturated syrup mass;
• the opportunity for preparing a higher amount of powder sugar alcoholic
suspension due to its long-lasting stability;
• replacing the sugar powder, consider “classical agent for seeding” which is due
also to the humidity contained in the sugar crystals which is freed through the
crystals’ milling which enhance the crystal sugar tendency to form in the lob run
of crystal lumps.
A method for sugar powder suspension preparation in isopropyl alcohol
In the specialised literature several method for the preparing the alcoholic suspension
for sugar crystallisation seeding are mentioned. These methods have been studied
within the Sugar Research and Engineering Laboratory from the Food Chemistry
Institute Bucharest, but one can adapt them to our working methods. The method
adopted consists in sugar milling directly in isopropyl alcohol.
The method is two-phased one, namely:
• obtaining the base suspension;
• the base suspension dilution and obtaining the seeding suspension.
On the basis of the experimental results obtained in the laboratory, which consist in
sugar milling in isopropyl alcohol in a 5.0 l milling machine on balls working on rolling
and having the a 46 rotations/min spin several working conditions have been established
which are presented bellow.
Obtaining the base suspension
• for a charge on employ:
• 100 grams of crystal sugar having a granulation of 0.2-0.8 mm;
• 200 ml. of isopropyl alcohol;
408

• about 160 porcelain balls having a diameter of 30 mm with a total mass of 1.0 kg;
• about 600 porcelain balls having the diameter of 12 mm with a total mass of 1.o
kg;
• the milling period is 1 hour.
Obtaining the seeding suspension Out of the base suspension thus obtained, the seeding
suspension is prepared through its dilution in isopropyl alcohol in a 1:200 ratio.
Maturing the seeding suspension The thus prepared powder sugar suspension in
isopropyl alcohol, called seeding suspension, is let to repose for a maturing period of 4
weeks. In the meanwhile, the germs smaller than 1 mm are being dissolved while the
sugar dissolved crystallizes on larger particles. Subsequent to this maturing period
equilibrium is established while the number of sugar particles from the seeding
suspension actually remains constant.
Criteria for ensuring the reproducibility of the seeding suspension quality In order to
always obtain a suspension with appreciatively the same number of particles on the
mass unit, one recommends the observance of the following measures:
• using the sugar with the same granulation for milling namely the sugar of 0.2-0.8
mm, deprived of powder or conglomerates;
• maintaining the ratio “sugar parts” against the “isopropanol parts”. As smaller
quantity of sugar is used on a isopropanol unit as brutish the sugar from the
obtained suspension will be. The best results are obtained using for a part of
granulation sugar of 0.2 mm-0.8 mm or a ratio of 2.0-2.2 parts of isopanorol;
• observing the milling period for the same mill machine functioning conditions
(criteria). The milling period prolonging does not imply a better milling.
A method for powder sugar preparation in ethyl alcohol
The permanent tendency to improve the preparation process of powder sugar suspension
preparation during the saccharose crystallization striking, in the sense to improve the
dispersal technique in solvent, led to a new preparation method of the seeding
suspension which is based on saccharose reduced solubility in ethyl alcohol. This
method consists in sugar precipitation from a saturated solution in ethyl alcohol. The
specialised literature recommends a similar method which consists in precipitating of a
21 grams of sugar which is found in a saturated solution at 55ºC, a solution of 28ºBrix
concentration or 28% in ethyl alcohol.
Preparing the base suspension The laboratory experiments accomplished on the milling
machine based on balls allowed the elaborating of the following working method:
• one introduce 21 grams ethyl alcohol in the milling machine on which one pour, in
a thin stripe, a 28% sugar solution which contains 420 grams of powder sugar;
• one activate the milling machine in rotating (spin) motion. The milling functioning
period is of 30 minutes during which the sugar crystallizing under the suspension
form;
• after the sugar crystallized under suspension, the suspension is pulled out of the
milling machines on balls and it is let for reposing for 10 up to15 minutes;
• one replace the supernatant liquid with a new fresh film (layer) of ethyl alcohol;
• the mixture of ethyl alcohol and sugar is again introduced in the milling machine
on balls which is activated for another 10 minutes.
409

• The sugar suspension thus prepared can be used after a minimum period of two
days, as n the meanwhile it becomes stable. In order to be preserved for a long
period of time, the film of supernatant alcohol is replaced one more time with fresh
ethylic alcohol.
Form the point of view of shape and size, the sugar crystals of the suspension thus
prepared are very homogeneous.
Preparing the seeding substance Out of the base suspension one prepare the seeding
suspension through the dilution of 1,200 in isopropyl alcohol. In comparison with the
method described, the current method which employs ethyl alcohol for base suspension
preparation is easier to be executed.
Establishing the number of germs introduced within a “decoction” The crystal sugar
obtaining of a certain granulation depends on the number of germs introduced at a
single seeding in the vacuum apparatus. Before reaching to this, one establish the
number of germs from the seeding suspension and then one calculate the required
quantity (amount) function of the size of the sugar crystals which is sought to be
obtained. For establishing the number of germs in the sugar suspension there are used
the following methods:
- the microscopically method;
- the sedimentation method;
- the sieving (sifting) method
In this purpose one can employ; the sedimentation balance, the Andreasen dropper or
the Coulter counter. Bearing in mind the number of germs which are contained in the
seeding suspension, we calculate the necessary amount to effect crystallization. In
practice, a surplus of germens is employed in experiment due to the different supersaturation
in the component parts of the vacuum apparatus, especially in the case in
which this is not endowed with a mechanical system for stirring up. The exceeding
quantity of crystallizing germs depends on other factors such as, foe example:
- the crystallizing step at which the striking (effecting) is been done;
- the constructive type (model) and the capacity of the vacuum apparatus;
- the pressure of the heating steam.
In the case of the A product the specialised literature recommends the use of a
crystallization germs surplus of 30-50% in comparison with the theoretical necessary.
The crystallizing efficaciousness (rated capacity) from a gross mass is theoretically
know as it can be calculated using the calculation formula based on gross mass purity
and of the inter-crystalline syrup and on the dry-mass content of the gross mass. The
calculation formula for the crystals extracted of gross mass’ efficaciousness (rated
capacity) is the following:
where:
SUmg is the content of dry-mass of the gross-mass în ºBrix:
Qmgg – purity of the gross mass in %
Qsi – purity of the inter-crystalline syrup, meaning the purity of the green syrup l in %
410

For the crystal mass the following formula is used:
p = d 3 X 1.1 [mg],
where:
p=is the desired individual mass of a single sugar crystal, in mg:
d – is the web dimension of the whole corresponding to the desired size of the sugar
crystal, in mm;
The specific mass of the crystal sugar in non-settled way
For example if we want to obtain sugar crystals with the web dimension of the whole
sized of 0,7mm, the calculation is done as following:
- the desired individual mass of a single sugar crystal which will be obtained
after crystallization is calculated using the following formula:
0.7 3 X 1.1 = 0.3773 mg;
a single tone of crystal sugar obtained through crystallization, formed through
crystallization and constituted from crystals with an individual mass of 0.3773 mg, will
contain:
1 000 000 000 : 0.3773 = 28 X 10 8 crystals
This number represents also the number of crystallization germs of which must be,
theoretically, introduced in the vacuum apparatus for crystallization striking (initiating)
and to obtaining of the tone of crystal sugar.
If we take into consideration:
- the vacuum apparatus capacity 50l;
- the crystal efficaciousness (output rate) 45% we can calculate:
- the amount of crystal sugar which must be obtained at a single decoction
namely: 50 X 0.45 = 22.5 tones
- the number of crystals contained by the decoction thus obtained is:
22.5 X (28 – 10 8 ) = 630 X 10 8 crystals
RESULTS AND DISCUSSIONS
This number, namely 630 X 10 8 represents also the number of germs which must be
introduced in the vacuum apparatus at crystal initiation (striking).
If we consider the fact that through the previous measurements is has been established
that in 100 ml of seeding suspension there are 325 X 10 9 crystallization germs, an
amount of 20 ml seeding suspension calculated according to the following formula
(630-10 8 ) : (325-10 9 ) = 20 ml will be required.
Taking into account the exceeding amount of crystallizing germs, 30 – 50% sugar the
real amount of seeding suspension, introduced in the vacuum apparatus at a single
decoction with 0.7 mm crystals, is calculated using the following formula:
20 X (1.30 – 1.50) – 20 ml
This is the way of seeding suspension preparation and also the way to calculate the
seeding suspension volume required to initiate (strike) the crystal sugar in the above
mentioned conditions and data.
411

REFERENCES
1. Banu C., 1992 – „Progrese tehnice, tehnologice şi ştiinţifice în industria
alimentară”, Ed. Tehnică, Bucureşti.
2. Beceanu Dumitru, Chira Adrian, 2003 – „Tehnologia produselor agricole”, Ed.
Economică, Bucureşti.
3. Sarca Gheorghe, 2004 – „Materii prime vegetale”, Ed. Universităţii din Oradea.
4. <strong>International</strong> Sugar Journal, 2006 december.
412

WATER DIFFUSION IN BREAD DURING BAKING
Ruska L.*, Chereji Rodica**, Purcărea Cornelia**, Timar A.**
* Bakery Bicaciu
** University of Oradea, Faculty of Environmental Protection
ABSTRACT
A study of heat and water transport in bread during baking was performed. Loaves of bread
were fermented twice and baked in a conventional ven at 225 0 C, with no forced convection,
for 35 min. The local water content and two or three different temperatures were measured
inside the bread during the baking process. Bread baked from wheat flour was used to
investigate the mechanisms of water transport inside a loaf during baking. The water
content was measured in the centre, 1 cm from the base, in the bottom crust, 1 cm under the
top surface and in the top crust. The temperature was measured on the bottom surface, in
the centre, and 1 mm from the optic fibre. The results indicate that up to 70±5 0 C, where a
structural change is taking place, the water content in the centre changes little. However,
after reaching this temperature the water content in the centre of the loaf rises due to
vapour transport from the warmer regions. The water appears to be moving towards the
coldest region, and not towards the geometrical centre.
Keywords: water; diffusion; transport; baking; bread
INTRODUCTION
During baking, starch is gelatinised and proteins are denaturated. The denaturation of
proteins releases water while the gelatinisation of starch absorbs water. These two
phenomena occur during the same temperature interval of 60-85 0 C, and contribute to the
change from dough to crumb (Marston, P. E. and Wannan, 1976). The temperature-rise at
the surface and heat transport towards the centre also involves water diffusion
(Thorvaldsson, K. and Skjoldebrand, 1996) and therefore a change in the water content
distribution.
The water content affects the gelatinisation of the starch Kokini, J. L., Lai, L. S. and
Chedid, L., 1992) and the final quality of the bread. During baking, the water content on the
surface of the loaf becomes lower than in the centre. This, in combination with the high
temperature, is one of the factors that makes the crust different from the crumb. An example
of a reaction that is affected by the water content and contributes to the crust formation is
the Maillard reaction, which results in the browning of the crust and the development of
bread aromas (Skjoldebrand C., 1986). Conversely, the starch gelatinisation and the protein
denaturation affect the diffusion of water by absorbing and releasing water. This makes the
baking process very complex. In 1972 Auerman suspected that the water content rose in the
centre of a loaf during baking, due to evaporation and condensation. Both Sluimer and
Krist-Spit, 1978 and de Vries et al. 1998 found an increase in the water content, with 3.5 g
water 100 g bread in the centre of the loaf immediately after baking.
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The objective of this study was to investigate whether the transport of water towards the
centre actually occurs during the baking process and to establish when the water transport
starts, ends, and what direction it takes. A second objective was to investigate whether the
rise in water content inside the loaf is centred at the coldest region or whether it is
geometrically centred.
MATERIALS AND METHODS
Bread recipe: A standard recipe for white French bread was used in this study with the
following quantities and ingredients for one loaf: wheat flour 370 g, water 200 g, salt 6 g,
sugar 6 g, oil 6 g, dry yeast 4.5 g.
Water content and temperature measurements: During the baking processes the optic fibre
was placed at five different sites in the loaf, in the centre, 1 cm from the base, in the bottom
crust, 1 cm under the top surface and in the top crust. The temperatures in the oven and in
the loaf were measured with copper-constantan thermocouples type T. The thermocouples in
the loaf were placed on the bottom surface, in the centre or 1 mm from the optic fibre. In
two of the loaves the temperature was measured at seven different places during the baking
to locate the coldest region and study the temperature gradient. The thermocouples were
placed, (a) on the top surface, (b) on the bottom surface, (c) on the side surface, (d) a third
of the way to the centre from the bottom surface, (e) half way to the centre from the top
surface, (f) in the centre, (g) two thirds of the way to the centre from the bottom surface.
Oven: The study used a conventional oven with a good temperature stability ±3 0 C and no
forced convection.
Baking procedure: The ingredients were mixed in a dough processor at 80 rpm for 2 min
and at 185 rpm for 3 min. The dough was left to rise for 35 min and mixed again at 185 rpm
for 2 min. It was then divided into three pieces which were rolled out into rectangular slabs.
The rectangular slabs were placed on top of each other in the baking tin, with the
thermocouples and the optic fibre between. The rectangular, 1.5 L baking tin had
dimensions of 9x25x6 cm. At one end of the tin there was an open slot, from the base to the
top, where the optic fibre and the thermocouples could protrude. The dough was allowed to
rise for 45 min in the tin before being placed into the oven, where it was baked for 35 min at
225 0 C.
Separation of crust and crumb: In one loaf the crust was cut off immediately after baking.
The crust pieces were separated into crust in direct contact with the baking tin, and crust in
contact with the air. The crust in contact with the baking tin was separated into side and
bottom crust. The crust was dried and weighed to see how much of the dough had become
crust and how much water had been present as dough. The water content of the crust in
contact with the air was compared with the total water loss of the loaf directly after baking.
The water content of the crust in contact with the baking tin was compared with the rise in
measured water content in the centre of the loaf.
414

RESULTS
Temperature distribution: When the loaf is placed in the oven the temperature rises on the
surfaces Figure 1. The temperature is approximately the same on all of the surfaces which
are in contact with the baking tin (i.e. on the base and sides of the loaf). The temperature of
the base rises slightly faster than the side temperatures due to heat conduction through the
base from the ceramic plate in the oven. At the top surface the temperature is slightly lower,
0-10 0 C, than the base temperature, up to approximately 80 0 C. The top surface temperature is
higher than the bottom temperature from 0 0 C to approximately 30 0 C, when removed from
the oven. Throughout the loaf there is a temperature gradient, with the coldest region a little
below the centre Figure 1. When the temperature reaches 100 0 C there is a plateau in the
temperature rise, until all of the water has evaporated. On the surface there is no plateau
whereas behind the crust, in the crumb of the loaf, the temperature remains at the plateau.
Water content distribution: The water content measured in the centre decreases when the
loaf is placed in the oven, until the temperature reaches approximately 70"5 8C. There is
then a change in the measured water content, which begins to increase. Figure 2 shows a
typical graph of the changes in temperature and water content in the centre during the
baking. At 1 cm underneath the top surface the changes in the measured water content are
identical, although the initial rise is approximately 3 g water 100 g bread and the decrease is
approximately 13 g water 100 g bread Figure 3. At the top and absolute bottom the decrease
in water content commences immediately and a crust is formed.
Figure 1: Changes in temperature inside the loaf during baking (a) top surface; (b)
bottom surface; (c) side surface; (d) 1 cm from the bottom surface; (e) 6.8 cm from the
bottom surface; (f) centre, 4.6 cm from the bottom surface; (g) 3.5 cm from the bottom
surface. Total heights 9.0 cm
DISCUSSION
Temperature distribution: During the fermentation of the dough the temperature distribution
in the loaf is fairly constant. The surface temperature of the loaf rises quickly during baking.
415

The high temperature on the surface leads to heat transfer into the loaf. The water in the
pores inside the loaf contributes to the transfer of heat through the material. This occurs due
to the evaporation of the water at the warm end of a pore and by condensation of the water
at the cold end, known as the heat pipe effect or the Watt principle. This causes a
temperature gradient inside the loaf. The partial water vapour pressure is directly related to
the temperature, and a vapour pressure gradient is built up from the surface to the centre of
the loaf. The results show that the temperature of the surfaces _ in direct contact with the
baking tin Figure 1, curves b. and c is generally lower than the temperature of the top
surface Figure 1, curve a . This is probably why the coldest region is centred slightly below
the geometrical centre of the loaf Figure 1, curves e, f and g.
Water content distribution: From the results it can be seen that the measured water content
in the centre of the loaf decreased up to a centre temperature of approximately 70±5 0 C. This
is probably due to the quantity of dough in the measuring volume in front of the optic fibre
decreasing as the total volume of the loaf increases. Earlier measurements have shown that
there is a good correlation between the rise of the loaf and the decrease in the measured
water content. The true water content is probably constant during this period as the pore
system is not continuous in the dough, which prevents the water from moving. When the
temperature reaches approximately 70 0 C the chemical changes commence, which make the
dough change into bread. The pore structure previously consists of discrete pores in which
the water vapour is captured.
Figure 2: Typical diagram of the changes in water content in the centre of the loaf. The
decrease in water content during the first period in the oven is a measured decrease only
and not an actual decrease. The water content starts to rise in the centre, when the
temperature has reached 70±5 0 C
416

During the chemical changes the pore structure is opened up and becomes continuous. In
this new pore system the water is more free to move. As can be seen from the results, the
measured water content in the centre starts to increase at approximately 70±5 0 C, which
confirms the theory that the water is not free to move until the structural change occurred.
The partial water vapour pressure gradient caused by the temperature gradient attempts to
reduce itself.
Figure 3: Typical diagram of the changes in water content under the top crust of the loaf.
The decrease in water content during the first period in the oven is a measured decrease
only, not an actual decrease. When the temperature has reached 70±5 0 C the water
content increases with approximately 3 g water/100 g bread and then slowly decreases
with approximately 13 g water/100 g bread
The water vapour therefore starts to move towards the centre and towards the surface. As
can be seen in Figure 2 the water content appears to increase in the centre when the
temperature has reached 70±5 0 C, and appears to decrease 1 cm from the bottom surface. At
1 cm under the top crust, Figure 3, the results show an increase in the measured water
content immediately after the temperature has reached 75 0 C. This is probably due to the
transport of water vapour towards the colder parts being prevented by the dough. Only when
the interior has changed to bread can the water move freely. Part of the rise could also be
due to compression of the crumb near the surface, as the crust has started to form and the
interior is still rising. Part of the rise could therefore be a measured rise only and not an
actual rise. Figure 4 shows the cross section of a loaf with a rough division into regions of
increasing and decreasing water content, at the time point when the loaf is removed from the
oven. The water contents of the regions have been estimated as a mean value from the
measured water contents. The water content of the dough was 41.8 g waterr100 g dough.
The crust is set at 0 g water/100 g bread, the region below the top crust, DV10 decreasing
417

volume 10 g water/100 g bread is set to a decrease of 10 g water/100 g bread, the region
around the centre, DV2 decreasing volume 2 g water/100 g bread is set to a decrease of 2 g
water /100 g bread and the centre volume, IV increasing volume, is increasing. In the crust
separation investigation, the crust weighed 80.9 g which corresponds to 139 g of the dough.
The remainder of the dough, 441 g, would then have become crumb. From Figure 4 the
volume of the crumb regions has been estimated as a percentage of the total crumb volume;
DV10=18.5 cm 3 /100 cm 3 , DV2=30.6 cm 3 /100 cm 3 and IVs50.9 cm 3 /100 cm 3 .
Figure 4: Cross section of a loaf with a rough division into regions of increasing and
decreasing water content, immediately after the loaf is removed from the oven. All figures
are in mm. The water content of the regions has been estimated to a mean value out of the
measured water contents. The crust is set to 0 g water/100 g, the region just below the top
crust, DV10, is set to a decrease of 10 g water/100 g bread, the region around the centre,
DV2, is set to a decrease of 2 g water/100 g bread and the centre, IV, is increasing.
On the surface in direct contact with the baking tin the water cannot diffuse into the air, so
all the water that evaporates in the warm parts must move towards the colder parts in the
centre of the loaf. If the water from DV2, the bottom crust and the side crust, except for the
12 g of water, moved towards the centre, this would lead to a mean value of the water
content in IV of 51.4 g water/100 g bread.
418

CONCLUSIONS
The water content in the centre of the loaf rises during the baking process. The water moves
towards the centre by the evaporation of water vapour near the surface, where the
temperature is high, and condensation closer to the centre, where the temperature is lower.
There is a change in the measured water content when the temperature reaches 70±5 0 C. This
implies that the chemical changes that cause the bread to change from dough to crumb and
open up the pore structure occur just prior to this temperature interval. The transport
towards the centre starts when the temperature has reached 70±5 0 C and stops when the
temperature reaches 100 0 C and there is no temperature gradient left. The coldest region
occurs just below the geometrical centre and the water vapour moves towards that region.
As can be seen in the objectives, the conclusions fulfil the aims of this research work.
REFERENCES
MARSTON, P. E. AND WANNAN, T. L. Bread baking the transformation from dough to
bread. The Bakers Digest, 50( 4 ), 24-49 ,1976.
THORVALDSSON, K. AND SKJOLDEBRAND, C. Water Transport in Meat during
Reheating. Journal of Food Engineering, 29, 13-21, 1996.
KOKINI, J. L., LAI, L. S. AND CHEDID, L. L. Effect of Starch Structure on Starch
Rheological Properties. Food. Technology, 46 (6) , 124-139, 1992.
SKJOLDEBRAND, C. Heat and Mass Transfer in Baking of Bread. SIK Service Series No.
752, 1986.
AUERMAN, L. J. Backprozess. In: Technologie der Brother stellung. Basel: VEB
Fachbuchverlag, pp. 237-278, 1977.
SLUIMER, P. AND KRIST-SPIT, C. E. Heat transport in dough during the baking of bread.
In: MORTON, I. D. Ed. Cereals in a European Context. Basel: Ellis Horwood Ltd., pp.
355-363, 1987.
DE VRIES, U., SLUIMER, P. AND BLOCKSMA, A. H. A quantitative model for heat
transport in dough and crumb during baking. In: ASP N. G. Ed. Cereal Science and
Technology in Sweden. Basel: Lund University Chemical Centre, pp. 174-188, 1988.
419

BREAD PROPERTIES AND CRUMB STRUCTURE
Ruska L.*, Chereji Rodica**, Purcărea Cornelia**, Timar A.**
* Bakery Bicaciu
** University of Oradea, Faculty of Environmental Protection
ABSTRACT
The relationship between bread-crumb cellular structure and many aspects of quality in a
loaf of white bread justifies investigations of how the structure arises during processing of
the dough. Following a brief overview of the development of bread cellular structure in the
dough, three parts of the literature pertaining to crumb appearance (visual texture) and
bread quality are reviewed, with emphasis on the mechanical properties (physical texture)
of the crumb. The importance of an objective segmentation of the two macroscopic phases
(crumb cells and cell walls solids) is emphasised in digital image analysis studies of breadcrumb
structure. A review of studies where mechanical properties have been measured in
fundamental units has sections on the mechanical properties of the composite structure and
on recent analyses of the mechanical properties of the solid phase. Finally, models which
have been used to relate structure to mechanical properties. Compared with the rule of
mixtures, these bounds represent a good (52%) improvement in the ability to predict values
for bread crumb moduli (crumb firmness).
Keywords: Bread;Crumb; Mechanical properties; Structure; Processing. 1
INTRODUCTION
The relationship between crumb structure and crumb appearance may be self-evident, but
this same crumb structure is also a determinant of loaf volume (Zghal, Scanlon, &
Sapirstein, 1999), the resilience of the loaf [the typical consumer purchasing decision
assessment test (Ponte & Ovadia, 1996)], the texture during eating and even the taste
(Baker, 1939). Therefore, some knowledge of the structure that defines crumb appearance
will permit us to predict many of the quality attributes of bread from knowledge of the
ingredients and how they are processed into the cellular structure on which bread quality is
established. The review addresses three parts of the literature pertaining to crumb
appearance, bread crumb physical texture and bread quality.
MATERIALS AND METHODS
1. Formation of the bread crumb
Bread: Bread as a solid is ‘‘soft’’ (Scanlon, Sapirstein, & Fahloul, 2000), and, like many
other foodstuffs (Campbell & Mougeot, 1999), is comprised, at a macroscopic level, of two
phases- a fluid (air) and a solid (cell wall material). When viewing a cross-section of bread
crumb (Fig.1), it is apparent that the solid phase is entirely connected (Torquato, 2000);
420

those portions that are not, are not bread crumb, but merely bread crumbs. In Fig. 1 it can be
seen that the air cells are isolated. However, in three dimensions, at least a portion (and
perhaps most) of the cells are connected. The volume fraction of the phases and the nature
of their connectivity (Warren & Kraynik, 1997) determines the structure, and consequently
the mechanical properties of the bread.
Fig. 1. Digital image of bread crumb.
Breadmaking: The mix ingredients that are used to create a basic formula dough are flour,
water, leavening agent (yeast or chemicals) and sodium chloride. In order to convert these
mix ingredients into the structure shown in Fig. 1 a number of processing operations are
performed. The operations are carried out in such a way that the dough can possess the
appropriate mechanical properties which will permit it to retain gas and thus produce a wellexpanded
loaf of bread with an even crumb structure. Three objectives are sought in the
processing operations :
1. mixing and development of the dough (mixing and fermenting);
2. formation of a foam structure in the dough (moulding, proofing and baking);
3. stabilization of a porous structure by altering the molecular configuration of the
polymeric components in the cell walls through the application of heat (baking).
2. Evaluating bread crumb cellular structure
Crumb cellular structure (or its grain) is an important quality criterion used in commercial
baking and research laboratories to judge bread quality alongside taste, crumb colour and
crumb physical texture. Bread crumb visual texture accounts for approximately 20% of the
weighting used in judging bread quality. Regardless of the weight assigned to it, crumb
grain is believed to have considerable importance in defining bread quality since the
accuracy in scoring other quality attributes in bread (e.g. loaf volume, loaf symmetry)
depends on the underlying crumb grain characteristics. In bread crumb scoring, the
examined parameters are crumb fineness (open versus closed cells), uniformity, cell shape,
and cell wall thickness (Pyler, 1988).
Image acquisition: In examining the visual texture of bread, particularly if one is trying to
obtain quantitative information on its structural organisation to determine the influence of
formula or processing conditions, the goal is to enhance the contrast between the two phases
421

in the image. In examining the structure of other cellular products, a number of image
acquisition methods have been used including:
1. transmission of light through thin sections, where the solid phase appears at the low end
of the grey level scale;
2. reflection of light from the surface of the specimen , where the shadows cast by cut cell
walls attenuate the light intensity that is reflected from the recessed cell walls, so that
the solid phase appears at the high end of the grey level scale;
3. x-ray tomography, where differences in absorption between the two phases permits
nondestructive evaluation of the internal structure.
3. Mechanical properties of bread crumb
3.1. Measuring crumb mechanicalp roperties
Compression: The most commonly used method to measure crumb physical texture is
indentation (AACC, 1983), a compressive loading test from which ‘‘modulus’’ values can
be obtained. Deformation of a crumb specimen between parallel plates in a uniaxial
compression test can also be used to measure the mechanical properties of bread crumb.
Fig. 2 shows a typical stress–strain curve for bread crumb under compression, to
indicate two important parameters used for characterizing the properties of the bread crumb:
Young’s modulus and critical stress. Bread crumb stressed parallel to the loaf’s longest axis
had values for Young’s modulus 2–3 times those of specimens compressed in the other two
directions, and values for critical stress 2–2.5 times those in the other directions. Uniaxial
compression of wheat starch bread (also tested fresh) gave a value of 18 kN m_2 for the
compressive modulus of crumb specimens compressed parallel to the long axis. Fresh
specimens of a soft wheat flour bread, compressed parallel to the long axis of the loaf,
yielded rather low values for the compressive modulus at 300 N m_2 (Piazza & Masi,
1995).
Fig. 2. Typical stress strain curves for bread crumb under tensile and
compressive loading. Young’s modulus (E), critical stress (_c), and failure stress (_f)
denoted.
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Tension: For bread prepared under laboratory baking conditions from the same wheats, but
loaded in tension parallel to the vertical axis of the loaf, values for Young’s modulus were
8.0 and 7.0 kN m_2 Considerably larger values have been obtained when very small strain
(0.02%) dynamic compressive/ tensile tests have been used to determine the mechanical
properties of bread crumb: a value of approximately 50 kN m_2 having been obtained for
the elastic part of Young’s modulus. The ability to propagate a crack in bread crumb
specimens with a long notch means that it is possible to derive a fracture toughness value
for bread crumb.
3.2. Measuring mechanical properties of cell walls
Although the mechanical properties of the bread are a function of the crumb cell wall
materials and the structure created by processing, changes in bread crumb physical texture
with shelf life (staling) are ascribed solely to cell wall property changes. A number of
factors account for this. Of the two phases in bread crumb, only the physical properties of
the solid phase will change. Most studies indicate that changes in the starch polymers in the
crumb walls are responsible for the firming of bread crumb over time (Zobel & Kulp, 1996),
although loss of moisture will evidently contribute to a change in the properties of the cell
walls which would be manifest as a firming of the crumb (Piazza & Masi, 1995). Changes
in bread crumb structure are not usually discussed in relation to staling (Zobel & Kulp,
1996), even though image analysis studies (Zghal et al., 1999) indicated substantial
structural changes arising from crumb moisture losses, with the magnitude of these changes
dependent on flour type used to make the crumb. Therefore, to eliminate structural
rearrangements within the crumb from affecting studies of firming mechanisms within
crumb wall components, direct measurement of the properties of the solid phase is desirable.
The most common method of measuring the mechanical properties of the solid phase is to
perform mechanical measurements on the compressed crumb (Scanlon et al., 2000).
RESULTS AND DISCUSSION
4. Relationship between structure and physical texture of cellular solids
In order to predict mechanical parameters that can be used to characterize bread physical
texture, physical models must be employed to relate the mechanical properties to the
structure quantified by imaging techniques. The degree of sophistication of the model is to a
certain extent determined by the amount and type of information provided by the imaging
system. Therefore, a number of factors that influence the mechanical properties of cellular
solids will be described along with descriptions of specific models and/or experiments that
have linked these structural properties to mechanical properties.
4.1. Measurements of structure related to volume fraction
Bulk density: Bulk density, like density, is defined as mass per unit volume (kg m_3). Bulk
density has been used to describe the density of the cellular solid. However, bulk density is
usually used to describe the packing density of commodities such as grains, where the space
423

etween discrete particles contributes to a reduction in density of the bulk collection
compared to the density of the individual grains. Given the discussion in Section 2.1 on the
nature of connectivity of bread crumb, bulk density will not be used to describe the density
of bread and other cellular solids. Where the term appears to have been used to describe the
density of the cellular solid, it will be discussed simply as density (_), as distinct from solid
or cell wall density (_s) which is used to define the density of the materials comprising the
solid phase.
Density: As for other foamed polymeric materials, density (or its inverse, specific volume,
m3 kg_1) has an enormous effect on the mechanical behaviour of bread crumb (Fig. 3).
Because of the large contrast in densities between the two macroscopic phases of bread
crumb, a simple measurement of bread density permits the volume fractions to be calculated
without recourse to image analysis for determining the distribution of the two phases in the
bread. Strong relationships between density and various mechanical properties have been
reported for bread and other cellular food products. For bread crumb differing in
composition (flours of 100% rye, 50% blend of wheat and rye, and
Fig. 3. Typical compressive stress strain curves for bread crumb created by proofing for
various times to generate different densities
100% wheat), crumb firmness linearly increased while crumb elasticity linearly decreased
with increasing density.
The rate of change in mechanical properties of bread crumb as a function of density was
dependent on bread composition, with rye bread having the highest slope.
Relative density: Because there is variation in cell wall density with changes in formulation
or processing conditions, then the density of the cellular solid would be expected to change,
and therefore the effect of the structure of bread on its physical texture would be partially
424

obscured. It has been stated that relative density is the dominant physical character
representing the three-dimensional structure of cellular solids, and it has been used in many
studies, including those of biological materials(Warbuton et al., 1990), to quantify the
dependence of mechanical properties on structure.
4.2. Predicting crumb properties with little or no imaging information
Information on the volume fractions of the individual phases can be obtained from imaging
data, but it is more readily gained from density measurements. The distribution of phases
within the crumb may be irregular (Fig. 1), but statistical homogeneity
Fig. 4. Illustration of statistical homogeneity in bread crumb. Thresholded
images are one-eighth sections taken from different regions of Fig. 1. Exact
structures are different, but statistical homogeneity is
apparent.
within the bread crumb is assumed. This refers to instances where a subregion of crumb can
be selected whose volume fractions and properties are essentially the same as those of the
bread crumb specimen as a whole, even though the actual connectivities may be different.
An illustration of statistical homogeneity in bread crumb is given in Fig. 4.
CONCLUSIONS
As for other composite cellular solids, density exerts a strong influence on the mechanical
properties of bread crumb. With information on the physical properties of the solid phase of
bread crumb, it is possible from a knowledge of bread crumb density to define a range of
modulus values which the bread crumb will definitely possess. Quantification of additional
structural information by image analysis will permit improvements in predictive capacity.
To take better advantage of models used for predicting the physical properties of industrial
and biomedical cellular solids from their structure, precise and systematic measurements of
the mechanical properties of breads created from various raw materials and processing
conditions is advocated.
425

ACKNOWLEDGEMENTS
We are grateful for research funding from The Backery Bicaciu.
REFERENCES
AACC. (1983). AACC method 74-09: Bread firmness by universal testing machine. In
Approved methods of the American Association of CerealChemists Vol II (8th ed.). St.
Paul, MN: American Association of Cereal Chemists, Inc. BAKER, J. C. (1939). The
permeability of bread by air. CerealChemistry, 16, 730–733.
CAMPBELL, G. M., & MOUGEOT, E. (1999). Creation and characterization of aerated
food products. Trends in Food Science & Technology, 10, 283–296.
PIAZZA, L., & MASI, P. (1995). Moisture redistribution throughout the bread loaf during
staling and its effect on mechanical properties. CerealChemistry , 72, 320–325.
PONTE, J. G. Jr., & OVADIA, D. Z. (1996). Instrumental methods. In R. E. Hebeda, & H.
F. Zobel (Eds.), Baked goods freshness (pp. 151– 170). New York: Marcel Dekker, Inc.
PYLER, E. J. (1988). Baking science and technology, volume II (pp. 850– 910). Merriam,
KS: Sosland.
SCANLON, M. G., SAPIRSTEIN, H. D., & FAHLOUL, D. (2000). Mechanical properties
of bread crumb prepared from flours of different dough strength. Journalof
CerealScience , 32, 235–243.
TORQUATO, S. (2000). Modeling of physical properties of composite materials.
<strong>International</strong>Journalof Solids and Structures, 37, 411–422.
WARBURTON, S. C., Donald, A. M., & Smith, A. C. (1990). The deformation of brittle
starch foams. Journalof Materials Science, 25, 4001–4007.
WARREN, W. E., & KRAYNIK, A. M. (1997). Linear elastic behavior of a low-density
Kelvin foam with open cells. Journalof Applied Mechanics, 64, 787–794.
ZGHAL, M. C., SCANLON, M. G., & SAPIRSTEIN, H. D. (1999). Prediction of bread
crumb density by digital image analysis. Cereal Chemistry, 76, 734–742.
ZOBEL, H. F., & KULP, K. (1996). The staling mechanism. In R. E. Hebeda, & H. F.
Zobel (Eds.), Baked goods freshness (pp. 1–64). New York: Marcel Dekker, Inc.
426

THE MATHEMATICAL MODELLING OF THE PRESSURE REGULATOR
AND OF THE ELECTROMAGNETIC INJECTION AT ENGINES WITH THE
REVOLUTION AND THE TEMPERATURE OF THE ENVIRONMENT
SURROUNDINGS
Vasile Blaga 1 , Nicolae Chioreanu 2 , Adriana Cătaş 3
1 University of Oradea, Faculty of Environment Protection,
2 University of Oradea, Faculty of Environment Protection
3 University of Oradea, Faculty of Science, Department of Mathematics,
acatas@uoradea.ro
ABSTRACT
The author proposes a personal model for the calculation of pressure regulator and
electromagnetic injection, the volume of fuel injected in cycle and the duration of the
injection with the number of rotation at the total full charge load. This kind of mode can
be used in the modeling system of electronic gasoline injection moonlit or multiunit. To
carry out this model it is necessary modeling engines with spark lighting cycle with
gasoline injection with a model helping cycle proposed by the author.
Keywords: pressure regulator, valve, pulverization, duration of injection.
THE OPTIMUM CALCULATION OF THE PRESSURE REGULATOR
Pressure regulator maintains constant pressure injection in supplying installation. [3; 5].
Calculation scheme of the pressure regulator is presented in figure 1.
Figure 1: Scheme of pressure regulator: 1-membrane; 2- springs; 3- valve; 4-
connection to the tank; 5- entrance connection; 6- exit connection; 7- housing.
427

Static balance equation of regulator membrane is given by the following relation:
F = F + F ,
(1)
a
ga
b
where:
2
2
π ⋅ Dr
π ⋅ Dr
Fa = Ka ⋅ f; Fga = ⋅ pga
; Fb
= ⋅ pb
;
4
4
and Ka is elastic constant springs; f- spring arrow; D-diameter of the regulator
membrane; Fa-power pressure of spring; Fga –pressure power from manifold; pga –
pressure from the manifold; pb –gasoline pressure in masterly of the injection; Fb –the
gasoline injection power.
π ⋅ D
4
π ⋅ D
4
2
2
r ⋅ pb
= Ka⋅
f +
r ⋅ pga
After the changing, relation (1) become :
4
p b = ⋅ Ka
f pga
;
2 ⋅ +
π ⋅ Dr
or:
p b = K r + pga
;
(2)
where:
Kr – is the constant regulator.
4 ⋅ Ka
⋅ f
= 2
π ⋅ D
;
K
Kr r
r
=
1..
4
THE OPTIMUM CALCULATION OF THE ELECTROMAGNETIC INJECTION
The injector proposed by the author are with electronic command Renix type, with
conic top needle with four holes of pulverization or Mono-Motronic with conic top of
needle with three holes of pulverization.
The volume of gasoline injected in cycle is proportional with the injection pressure
and the duration of injection. [3;6].
The section of the passing pulverization hole is determined by the following relation :
[6]
__
__
d d '
d 1
Aa
Ac
Ac
sa
sin
;
2 2
2 sa
sin ⎟ ⎡ ⎛ ⎞⎤
⎛ β ⎞⎛
− ⎞
= π ⋅ ⎢ + ⎜ − ⎟
⎟⎥
= π ⋅ ⎜ ⎟ ⎜
(3)
⎣ ⎝ ⎠⎦
⎝ ⎠⎝
⋅ β ⎠
where: AI-passing section offered by conic top needle; sa-raising up needle; dv-diameter
of the needle in top zone; β-cone angle tight; dp-sack diameter.
Raising up needle sa is considered to be constant.
428
,

It was marked with AI the area of the leaking section near the needle top of injector with
conic top.
Ai = f(sa, β, d) = ct;
Ai = π(dp - 0,5 sa sinβ)sa sin β/2;
sa = 0,15 mm; dp = 1…1,2 mm; β = 60 0 .
Figure 2: The calculation scheme of electromagnetic injector
Discharge of gasoline which passes the injector leaking section is calculated by the
relation:
2(
pb
− pga
)
Q = μ ⋅ A
; (4)
b i i
ρb
where μI coefficient of discharge in the section offered by the needle; μi = 0,8 - 0,93 ;
Taking in consideration relation (2):
Q
2K
= μ ⋅ A
(5)
r
b i i
ρb
where: Ai is the leaking section at the injector; pb – gasoline pressure at the entrance of
injector; pga – air pressure from the manifold; ρb – gasoline density; Kr - constant
regulator pressure; Qb- discharge of the gasoline.
In the other hands it is lanown from the relation of the discharge definition, that is
the leaking fuel volume during a time unit.
We obtain
Q
b
V
=
t
b
i
mcb
=
ρ ⋅t
b
i
From the equality of relation (5) and (6) results un duration of the injection, ti:
mcb
1
ti
= ⋅
ρb
μ ⋅ A ⋅
2K
r
mcb
=
;
μi
⋅ Ai
⋅ 2K
r ⋅ ρb
1
d =
λ ⋅ Lo
m
=
m
i
i
ρ
b
429
(6)
cb
aer
;

m
cb
maer
maer
mad
m
= = ⋅ = ξ ⋅d
⋅mad
; ξ =
λ ⋅ L m λ ⋅ L
m
o
ad
o
where ξ is the coefficient which represents the ratio between necessary air quantity for
burning moor and the mixed quantity fuel accepted mad. Results:
t
ξ ⋅ d ⋅m
ad
i = [s], [7]
μi
⋅ Ai
⋅ 2K
r ⋅ ρb
where: mcb is the volume of the fuel; maer – the volume of the air aspirated by the
engine; mad – quantity of mixed fuel admitted; d – proportioning.
In figure 3 is represented the variation of the volume of the fuel mcb with the revolution
and the temperature of the environment surroundings. In figure 4 is represented the
variation of the duration of injection ti with the revolution and the temperature of the
environment surroundings.
The working of the curled engine with lambda transmitter and catalyst, makes
coefficient λ to be mentioned as close as possible to λ=1 (stoechiometric dosage).
On the base of a personal model the author realized analytic calculation of pressure
in the manifold pga and the admission pressure pa, the calculation of engine pressure
regulator and the duration ti of the electromagnetic injector.
The equation of working engine by spark equipped with electronic engine injection
were determined and introduced in the calculator.
For the modeling cycle engines with spark lighting with engine injection are noticed
the initial dates, pressure expression from un manifold pga and the pressure of air at the
end of the admission pa, the thermal volume charging qcb, the filling efficiency ηv, the
raising ratio of the pressure in α, at volum constant burning, raising ratio of the after
burning volume δ, the temperature of the evacuated gasoline Te, the coefficient of the
residual burning gasoline γr, the temperature at the end of the admission Ta.
All these expression were analytic calculated and correlated among them in order
to be introduced in the calculator. The calculator gets the command to repeat this
operation till the getting of the imposed error of the engines with spark lighting
parameters.
It is calculated the mechanical theoretic work proposed Ltp, the average pressure
proposed ptp, theoretical proposed efficiency and the mechanical loosing of the cycle.
430
aer
ad
;

mcb
[g]
mcb
[g]
mcb
[g]
0,040
0,030
0,020
0,010
0,000
0,70
0,800,90
1,00
lambda 1,10
1,20
500
1625
2750
3875
5000
Speed [1/min]
0
0
0
0
0
0
0
0
0
0
500 1000 1500 2000 2500 3000 3500 4000 4500 5000 5500
0
0
0
0
0
0
0
0
0
Speed [1/min]
0,70 0,80 0,90 1,00 1,10 1,20 1,30
lambda
mcb
0,030-0,040
0,020-0,030
0,010-0,020
0,000-0,010
mcb,
lamda = 1
Series1
mcb
[g],
n = 5250
Series1
Figure 3: The variation of the volume of the fuel mcb with the revolution and the
temperature of the environment surroundings
431

ti
[ms]
ti
[ms]
ti
[ms]
6
5
4
3
2
1
8,00
6,00
4,00
2,00
0,00
0,70
0,800,90
1,00
lambda 1,10
1,20
5000
500
1625
2750
3875
Speed
[1/min]
0
500 1000 1500 2000 2500 3000 3500 4000 4500 5000 5500
7
6
5
4
3
2
1
0
Speed [1/min]
0,70 0,80 0,90 1,00 1,10 1,20 1,30
lambda
ti
6,00-8,00
4,00-6,00
2,00-4,00
0,00-2,00
ti,
lamda = 1
Series1
ti,
n = 5250
Series1
Figure 4: The variation of the duration of injection ti with the revolution and the
temperature of the environment surroundings
432

The calculation of engines with spark lighting parameter, was sectioned in 10
proceedings and functions. Near the declared constant values at the beginning of the
program, it is considered as initial dates, presumed to be lanown choosing arbitrary
from the statistical dates of engines with spark lighting cycle: Tao=322 K; Tzo=2530 K;
Tuo =2660 K and kco =1,3; kvo =1,3; kuo =1,2; kdo=1,3; keo=1,3; without them are not
possible to calculate in general way all others coefficients and the other temperature and
the physical seize which characterize the cycle. In this way the seize above will play the
parameter role, variable will the temperature in which thermal process evaluate. These
temperatures depend on the adiabatic coefficients which in fact are stability by going
though many times of the cycle until these coefficients become constant with 0,000009
error.
This exit decision from the cycle for a certain revolution is given by the diminish of the
constant error of temperature Ta under 1,5 K.
The calculation of algorithm is made up of the following steps:
1) The initial parameter relation Tao, Tzo, Tuo şi kco, kvo, kuo, kdo, keo for one revolution.
2) The cycle is going through, obtaining new values for k coefficients and Ta, Tz, Tu
temperatures.
3) It is made differences between old and new calculated values (∆k, ∆T).
4) If the differences between (∆k, ∆T) are lower than the imposed error, then the new
calculated values are valid and they are going to the next revolution value and 1-4
points are repeated.
5) If the differences (∆k, ∆T) are above the imposed errors then the cycle is going
through by the recalculation of the adiabatic coefficient values and temperature,
considered new value as initial parameter. This re-going through is realized until
the descend of the differences (∆k, ∆T) under the imposed errors.
This program calculate the characteristic seize of one thermodynamic cycle in general
way only with the restriction of the energetic efficiency of burning ηar and the using
coefficient of warmth during the burning stage at a constant volume ξo, which define as
a performant middle of study.
CONCLUSIONS
The engines with spark lighting modeling cycle with the gasoline injection proposed by
the author consist of the presentation of the initial dates of the calculation program,
calculation and the correlation between the engines with spark lighting parameter
expression with the gasoline injection for the achievement of the program calculation.
The engines with spark lighting cycle calculation proposed, with gasoline injection is an
helping cycle for the simulation on the calculator of the gasoline injection. The
simulation on the calculator permit the determination of the theoretical economicaltehnical
parameters proposed; mechanical work theoretical proposed corresponding to
the diagram, theoretical pressure proposed, theoretical efficiency, theoretical specific
consumption proposed. The loosing mechanical pressure and the pumping are
calculated, the theoretical economical-tehnical parameters of the engine.
Using the proposed model by the author the volume of the fuel is calculated which
reset of the engine cycle in function with the revolution of the total load, the debit of the
433

gasoline which goes through the electromagnetic section of the injection, reaching the
aim in view, the determination of the injection duration with the revolution in total load.
It is conceived the calculation of the pressure regulator and of the electromagnetic
injector. It was made the calculation of the injection duration ti with the revolution and
load which represents the model proposed by the author.
It was effectuated a engines with spark lighting program for the calculation of
parameters with the gasoline injection with the under- program:
• the calculation program of engines with spark lighting parameters (depending on n
and λ at to=-35...+45ºC and po=1·10 2 kPa) [3].
• the calculation program of engines with spark lighting parameters (depending on n
and to at λ=1 and po=1·10 2 kPa) presented in annex B;[3].
REFERENCES
1) Apostolescu, N. and Chiriac, R. , Process of burning in driving with inward
alight.The Technical Edition, Bucharest 1998.
2) Băţagă, N. , Driving with inward alight.The Didactic and Pedagogical Edition,
Bucharest 1996.
3) Blaga, V. , The contribution at modeling of gasoline injection at M.A.S. Work of
doctor`s. The Technical University of Cluj-Napoca 2000.
4) Blaga, V., The dynamic of car, The University Edition Oradea,2005.
5) Blaga, V.,Engine with gasoline injection, The University Edition Oradea,2005.
6) Delanette, M., Les Motors a injection. Edition Tehniques pour L’automobil et
L’industrie, iunie, 1989.
7) Negrea, V. D., Venetia Sandu The combating of medium poluttion in motor vehicle.
The Technical Edition, Bucharest, 2000
8) Turcoiu, T., Boncoi, J. şi Time, Al, Equipment’s from injection for engine with
internal burning. The Technical Edition, Bucharest 1987.
434

THE VARIATION OF EFFECTIVE POWER AND SPECIFIC EFFECTIVE
FUEL CONSUME OF 106-20 ENGINE WITH THE REVOLUTION AND THE
TEMPERATURE OF THE ENVIRONMENT SURROUNDINGS
Vasile Blaga 1 , Nicolae Chioreanu 2 , Adriana Cătaş 3
1 University of Oradea, Faculty of Environment Protection,
2 University of Oradea, Faculty of Environment Protection
3 University of Oradea, Faculty of Science, Department of Mathematics,
acatas@uoradea.ro
ABSTRACT
Working system of engine is definite by rotation and load, but besides those, it requires
knowledge about thermal system, that is machine part temperature situation, the
temperature of cooling fluid, admitted air temperature, evacuation gases temperature
height corrections.
Engine load suits effective power Pe delivered by engine to a certain rotation. The
load is regulated through speed pedal, controlling Mono-Motronic mixture quality
system and air quality of other systems. Outside engine action applied of consumer
(motor vehicle or brake trying on trial engine stand) represent engine attempt when
thermal system of the engine doesn’t vary in time, working system is stabilized, contrary
the engine is working in provisional system.
Keywords: gasoline, injection, lambda, effective power, revolution.
THE GENERAL MODEL FUEL INJECTION MODEL
Load remark may be realized through comparison between effective power Pe and
continued effective power Pec developed on the some rotation, named load coefficient
χ= Pe/Pec [2];[3].
Through continued effective power it can be understand maximum value of power,
which can be developed by an engine permanently, at a named rotation, without
modification of engine indexes and without appearance of some wear defections. For
short lapse of time (for example 15 min. for one hour interval) the engine can develop a
superior power, named intermittent superior power Pei. Maximum continuously power
settled by industrial unit engineering is the nominal power Pen, and adequate/suitable
rotation, nominal rotation.
The load according power Pec (χ=1) is named complete load the one according Pei
power absolute load and according idly running Pe=0 (theoretical χ=0) null load.
Between null and complete load (0

Mixture quality estimate is done frequently by air excess coefficient x, definite by report
χ=L/Lo, where L means air quantity which is available for burning one kilo fuel, and Lo
is minimum quantity of air available to burn one kilo fuel.
For theoretic dosage χ=1, for poorest one χ>1 and for the richest χ=1.
Block scheme of general model used modulate gasoline injection system of M.A.S. is
presented by figure 1 [3].
Figure 1: Block scheme of general model used modelling gasoline injection
system of engines spark ignition
It includes correlation system of aspirated air by engine with a quantity of gasoline
injected on cycle.
Entering magnitudes into the system are x load, n rotation, angular speed, position
of obdurate and the emergence one are effective moment, effective efficiency, effective
consummation, polluting emission. Engine process is influenced by thermal system of T
engine.
Following the study made for many different fuel injection systems it can be
noticed that every system no matter that it is a mechanic or electric one, they have basic
scheme, based on internal burning engine working. Mechanic systems have on their
principle scheme base two circuits: fuel circuit, air circuit and: suitable correction.
Electronic systems includes, beside that, the electric circuit and firing circuit for
Motronic system [4]. The main injection equipment load is correlating air quantity
aspirated by the engine with quantity of injected fuel on the cycle, such that may result
the best dosage every engine working system. That can be possible with correlation
between air and fuel quantities and a series of engine working parameters (air, pressure,
into the collecting input obturater position), engine rotation [1]; [5].
Figure 2 represents three dimensional variations effective power of 106-20 engine
with Bosh Mono-Motronic MA 1.7 gasoline injection and, figure 3 represents three
dimensional variation of specific effective fuel consume of the same typo of engine, fit
with the same injection system.
436

437

Pe
[kW]
60
50
40
30
20
10
0
n = 5250[rot/min]
lambda = 1.0
-40 -30 -20 -10 0 10 20 30 40 50
to
Figure 2: Variations effective power of 106-20 engine with the revolution and the
temperature of the environment surroundings
438

ce
[g/kW h]
400.019
350.019
300.019
250.019
200.019
150.019
100.019
50.019
0.019
319.83
n = 5250[rot/min]
lambda = 1.0
-40 -30 -20 -10 0 10 20 30 40 50
to
Figure 3: Variation of specific effective fuel consume of 106-20 engine with the
revolution and the temperature of the environment surroundings
Quantity of air aspirated by engine can be expressed after enumerated functional
parameters must to count on injection pressure, flowing section through injector and
injection length.
Often it is prefer that opening length of electromagnetic injector to depend on
depression from the input collector, because quantity of injection is correlating with
quantity of air aspirated on the cycle with, as it is known from injection mechanic.
Equipments are after depression from admitted collector dependence of rotation is
smaller in this case following to be done rotation corrections for working systems witch
demand such corrections. Flowing corrections of injected gasoline into cylinder are
439

demanded for a succession of transitory system of enfin working, cooling start, cooling
liquid temperature, lubrication oil temperature atmosphere pressure.
Accounting all those, to realize injection equipment’s it must be applied the
fundamental principle, graphically the gasoline from the tank and represses it to
electromagnet injectors. gasoline pressure upstream injectors are constantly mentioned
with a pressure regulator, which allows the return to reservoir of gasoline excess
suppressed by supply pump.
Electromagnetic injectors associated to each engine cylinder, are opened once a
cycle (of one cameo shaft rotation) by stream impulses proceeded from a computer.
Control impulse length depends on depression from impute collector, engine rotation
and a series of other correction dimensions [5].
CONCLUSIONS
Those dimensions are perceived with electro-mechanical translators’ help, being
transmitted to the computer like electrical dimensions.
On this basically principle can be modelled an important variety of respond to
demanding of performance increase of engines, dominating affective consumption of
fuel and polluting emission factors from evacuation gases as much as necessity
encumbering and adaptability.
REFERENCES
1. Apostolescu, N. and Chiriac, R. , Process of burning in driving with inward
alight.The Technical Edition, Bucharest 1998.
2. Băţagă, N. , Driving with inward alight. The Didactic and Pedagogical Edition,
Bucharest 1996.
3. Blaga, V. , The contribution at modeling of gasoline injection at M.A.S. Work of
doctor`s. The Technical University of Cluj-Napoca 2000.
4. Blaga, V.,Engine with gasoline injection, The University Edition of Oradea,2005.
5. Delanette, M., Les Motors a injection. Edition Tehniques pour L’automobil et
L’industrie, iunie, 1989.
6. Negrea, V. D., Venetia Sandu, The combating of medium poluttion in motor
vehicle. The Technical Edition, Bucharest, 2000.
440

THE EFFECT OF THE GENOTYPE AND EXPLANT UPON THE SOMATIC
EMBRYO INDUCTION IN THREE QUERCUS SPECIES
AI. Timofte*, M. Palada-Nicolau**, M. Ardelean***, D. Pamfil***
*University of Oradea, Romania
**Forest Research and Management Institute, Research Center Simeria, Romania
***University of Agriculture and Veterinary Medicine Cluj-Napoca, Romania
ABSTRACT
The somatic embryogenesis is an advanced method for clonal propagation and a useful
tool for ex situ conservation of genetic resources.
Three oak species were tested for the somatic embryogenesis ability: Quercus robur
(three provenances), Q.petraea (one provenance), Q.frainetto (two provenances).
The most important factor affecting the efficiency of somatic embryogenesis was
found to be the developmental stage of the zygotic embryos used as juvenile explants.
The high embryogenic ability of young zygotic embryos and low embryogenic ability of
maturing zygotic ones showed the strong positive relationship between somatic embryo
induction and accumulation of reserve nutrients in cotyledons.
Two nutritive culture media and two growth hormone combinations were tested.
The effect nutritive media was insignificant, but the effect of hormone combination was
either week, or masked by the effect of explant type or that of the genotype. As for
genotype, some differences were found among species, as well as among the
provenances of the same species, regardless the explants type. These differences were
analysed statistically and some week correlations were found.
The subsequent phases of the somatic embryogenesis process (multiplication,
germination and plant regeneration) were not related to the initial embryo efficiency,
but only to the quality of stabilized embryogenic lines.
Key words: oak species, somatic embryogenesis, explants, genotype, statistics.
INTRODUCTION
The in vitro clonal propagation in oaks is an important step in the breeding activity
using clonal strategies, and in the same time a tool for the application of somaclonal
selection procedures in the improvement of adaptability traits.
The use of somatic embryogenesis in oak clonal micropropagation is preferable in
the applications concerning somaclonal variation and selection, because the plants
regenerated from somatic embryos usually have monocellular origin and consist in
clones of long-term culture, submitted of in vitro selection procedures.
Generally, the somatic embryogenesis can be considered an efficient mean of
clonal propagation, when the following conditions are accomplished:
• the stable embryogenic cultures constantly produce a big amount of somatic
embryos
• the embryogenic ability of such cultures can be maintained for long time by serial
adventitious embryogenesis
• the somatic embryos can be efficiently converted into acclimatable plants.
• Starting with the first successful embryogenic inductions (Chalupa, 1987, 1990,
441

Gingas and Lineberger, 1989; Jorgensen, 1988, 1993), the method was
continuously improved. As a result of many experiments, nowadays is rather easy
to obtain a big amount of somatic embryos out of juvenile explants, and also to
conserve the embryogenic ability of the selected somaclones for long periods
(Wilhelm et al., 1996). The embryogenesis occurs mostly direct on the explant,
without a callus phase, and the stabilisation of embryogenic cultures is based on
the serial adventitious embryogenesis.
This advanced method of plant propagation was also applied in the ex situ conservation
of genetic resources, using the cryopreservation techniques (Jorgensen, 1991; Tutkova
and Wilhelm, 1999).
The somatic embryogenesis in oak, as an alternative to propagation by cuttings,
provides the possibility of mass production of cotyledonary embryos, that can be either
cryopreserved, or maintained by long-term culture as stable embryogenic somaclones
(Wilhelm et. al., 1996, 2000).
The aim of this paper is the improvement of somatic embryogenesis technique in
different oak genotypes, analysing the relationship among diverse factors affecting the
embryo induction efficiency
MATERIAL AND METHODS
Plant material: As a genetical background, the plant material was represented by three
oak species, in each of them one or more provenances being investigated: Quercus
robur (three provenances), Q. petraea (one provenance) and Q. frainetto (two
provenances).
The immature acorns harvested at 4 different dates were dissected and used as
sources of explants. The explants were represented by immature zygotic embryos in
different developmental stages or fragments of more advanced embryos. The
developmental stages of zygotic embryos have been defined and correlated with the
morphological characteristics of acorns. (Palada-Nicolau, Hausman, 2001)
Culture medium: Four media were tested for the induction of embryogenesis,
representing combinations between two nutrient recipes (1/2 MS – Murashige, Skoog,
1962) and DCR (Gupta, Durzan, 1985) and two combinations of growth regulators:
QE1: 1/2MS with 10μM/l 2,4-D and 2μM/l BAP
QE2: 1/2MS with 5μM/l NAA - 5μM/l BAP
QE3: DCR with 10μM/l 2,4-D and 2μM/l BAP
QE4: DCR with 5μM/l NAA - 5μM/l BAP
After two subcultures on this medium, a hormone-free ½ MS medium was used for the
proliferation of embryogenic structures
The protocol: Sterilisation of immature acorns by immersion for 10 min. in solution of
HgCl2 0,2 %, followed by abundant rinsing in sterile distilled water;
a) Dissection of acorns in aseptic conditions, isolation of embryos and inoculation on
agar solidified culture medium. Petri dishes (∅ = 10 cm) containing cca. 25 ml
medium were used. Five explants were deposed in each Petri dish. In the case of
the more advanced embryos (stages 3b and 4) around ¾ of the cotyledons were
removed, and the embryo with the basal part of cotyledons was planted on the
culture medium.
442

) The cultures were incubated at cca. 22 o C in darkness. The duration of subcultures
ranged between 21 and 30 days.
c) The observations were made at the end of first two subcultures. All reactions of the
explant to the culture conditions (embryogenesis, callogenesis, rhyzogenesis and
germination) were recorded. If necessary, a binocular stereomicroscope was used.
Structure of the experiment
3 oak species: Quercus robur, Quercus petraea, Quercus frainetto
6 provenances: 3 of Quercus robur, one of Q. petraea and two of Q. frainetto.
4 developmental stages of explants (4 explant types): stages 1, 2, 3 and 4
4 culture media, represented by:
2 variants of nutritive medium : ½ MS (Murashige-Skoog, 1962) and DCR (Gupta,
Durzan, 1985)
2 variants of growth regulator combinations, for each nutritive medium
In order to establish the effect of culture medium upon the efficiency of somatic embryo
induction, comparisons were made between nutritive media (QE1 + QE2, versus QE3 +
QE4), on one hand, and between the two combinations of growth regulators (QE1 +
QE3, versus QE2 + QE4), on the other hand. Three replications consisting of five to ten
explants have been counted for each parameter.
Statistical analyses
In order to validate the results of the oak somatic embryogenesis experiments, in which
the main parameters were genotypes, explant stages and culture media, statistical
analyses were made, using three ways ANOVA, lay/out with incomplete blocks, for the
character „embryogenic induction” to the species Q. robur and Q. frainetto. The results
have been presented as bilateral analysis tables, exclusively for the experimental factors
and interactions found in ANOVA table having the test F significant.
For the species Q. petraea, only two ways ANOVA in randomised blocks design was
used, for experiments in three replications.
'
Because some experimental data had values “0”, the transformation x = x + 1 was
used, all calculations being performed with transformed values (Ardelean et al., 2005).
All the results were reported to the number of viable explants.
The statistical validation of the differences among the experimental variants was made
by the multiple range test (Duncan or Turkey). These tests allowed the objective
comparison of all differences between every two variants.
RESULTS AND DISCUSSION
1. Somatic embryogenesis experiments
The somatic embryo induction was mainly direct. The embryos and embryo clusters
were visible on the explant surface after 30 to 45 days (Fig. 1).
They were detached from the explant tissue and cultivated on media with the same
composition to stabilisation.
The stable embryogenic cultures were cultivated on media with small
concentration of cytokinines or on hormone-free medium, in order to obtain
proliferation by serial adventitious embryogenesis (Fig. 2).
443

Fig. 1 – Formation of somatic embryos and
embryo clusters on the cotyledonnary explant
Fig. 2 – Stable oak embryogenic
culture
Besides embryogenesis, other reactions of the explants to the culture conditions were
observed: callogenesis, rhyzogenesis and germination.
The viability of explants after two subcultures ranged between 55 to 85% at the young
explants (stage and 2), and 71 to 100% in the case of more developed explants (stages 3
and 4). The differences between species and provenances, concerning the viability were
due mostly to technical reasons (distance to the field, number of days in conservation
before dissection, etc.).
All records concerning the efficiency of somatic embryo induction and the
frequency of other reaction of the explants were reported only to the viable explants.
It was found that the most important factor affecting the somatic embryo induction
efficiency was the developmental stage of the explant. In all oak species, the
embryogenic ability decreased constantly from the youngest to the more advanced
zygotic embryos used as explants (Fig. 3). The same trend was recognised in the case of
the provenances inside the species, regardless the culture medium used for the induction
(Fig. 4 and 5).
% embryogenic induction
50
45
40
35
30
25
20
15
10
5
0
Stage 1 Stage 2 Stage 3 Stage 4
explant type (developmental stage of zygotic embryo)
Q. robur
Q. petraea
Q. frainetto
Fig. 3 - The effect of explant type (developmental stage of zygotic embryo)
upon the embryogenic ability, in three Quercus species
444

% embryogenic inductio
60
50
40
30
20
10
0
Stage 1 Stage 2 Stage 3 Stage 4
explant type (developmental stage of zygotic embryo)
Târgu-Mureş
Dumbrava
Stefanesti
mean
Fig. 4 - The effect of explant type (developmental stage of zygotic embryo)
upon the embryogenic ability, in three provenances of Quercus robur
As for the genotype, there were differences among both species (Fig. 3) and
provenances inside species (Fig. 4, 5, 6).
% embryogenic induction
40
35
30
25
20
15
10
5
0
Stage 1 Stage 2 Stage 3 Stage 4
explant type (developmental stage of zygotic embryo)
Griva
Pad.Sarului
mean
Fig. 5 - The effect of explant type (developmental stage of zygotic embryo)
upon the embryogenic ability, in two provenances of Quercus frainetto
445

Regarding the species, the best embryogenic ability was found in Q. petraea and the
worst, in Q. frainetto. The embryogenic ability of Q. robur (average, 17,8%) and Q.
petraea (av. 18,5%) was similar, but that of Q. frainetto was significantly inferior
(average 10,4%).
Concerning provenances, in Q. robur, the best results were obtained in the
provenance Tg. Mures (av. 22,9%) and the worst, in Dumbrava Sibiului (av. 12,9).
Between the Q. frainetto provenances, Griva was superior (av. 12,5%), and Padurea
sarului was the worst (average 8,3%).
Statistical analyses in two ways Anova system were made in order to validate the
correlations between the effect of genotype and developmental stage.
% embryogenic induction
25
20
15
10
5
0
genotypes
average
Tg.Mureş Dumbrava Sib. Stefanesti Mihaesti Griva Pad.Sarului
Q. robur Q. petraea
Q. frainetto
genotypes
Fig. 6 - The effect of the genotype (species, provenances) upon the
embryogenic ability in oak
The effect of culture medium proved to be totally insignificant, besides the differences
observed among the variants cultivated on different media. The situation was the same
as well for the nutrients as for growth regulators
In order to discriminate the role of each factor upon the efficiency of embryogenic
induction, three-ways Anova statistical analyses were performed.
2. STATISTICS
2.1. Trifactorial experiments
2.1.a. The effect of provenance, stage of explant and culture medium upon the embryo
induction efficiency in Quercus robur
The three way Anova for the effect of provenance, stage of explant and culture medium
upon the embryo induction efficiency in Quercus robur is presented in the Table 1.
446

Table 1 The three way Anova type 3x4x4 for the effect of (provenance x stage of
explant x culture medium) upon the embryo induction efficiency in Quercus robur
Source of variation MSD DF MS F-test
TOTAL 4.569 143
Big plots 0.198 8
Replications 0.003 2
Factor A (PROVENANCE) 0.164 2 0.082 10.52* > 6.94; 18.00
Error (a) 0.031 4 0.008
Mean plots 2.895 27
Factor B (STAGE) 2.453 3 0.818 44.75** > 3.16; 5.09
Interaction A x B 0.112 6 0.019 1.02 < 2.66; 4.01
Error (b) 0.329 18 0.018
Small plots 1.476 108
Factor C (MEDIUM) 0.037 3 0.012 0.97 < 2.74; 4.07
Interaction A x C 0.045 6 0.008 0.59 < 2.23; 3.06
Interaction B x C 0.058 9 0.006 0.50 < 2.01; 2.66
Interaction A x B x C 0.419 18 0.023 1.83* > 1.74; 2.21
Error (c) 0.917 72 0.013
Looking in the table, one can notice that the calculated F is bigger than the theoretical F
for the transgression probability of 5% and 1% only in the cases of provenance, explant
stage and the threefold interaction provenance x stage x culture medium, that means the
variants variability is due to real causes, true differences existing among the
experimental variants.
The results showed a significant action of the provenance and a highly significant
action of the stage of explant upon the embryogenesis. The threefold interaction of
above-mentioned factors was also significant.
The existence of the significant triple interaction shoved the utility to study the averages
of all combination of parameters (Săulescu et al.., 1967), presented in the following
three tables of test Duncan.
The data from the table 2 present the effects of provenance and stage of explant
upon the embryogenic ability in Q. robur.
Looking in the table 2, one can notice that the provenance Dumbrava Sibiului
presented a rate of embryo induction significantly inferior to the other two provenances
that are similar and that the provenance Tg. Mures was the best. As for the explant
types, the early developmental stages of the zygotic embryos 1 and 2 had a significant
positive effect upon the embryo induction efficiency, but the more advanced stages (3
and 4) had a negative effect, comparing to the average.
447

Table 2 The effect of provenance and developmental stage of explant upon the rate of
embryogenic induction in Q. robur (test Duncan)
Nr.
crt.
Stage of explants
(zygotic
embryos)
Provenance
Embryogenic induction and its significance*
S1 S2 S3 S4
Average on
provenances
1 Tg. Mureş 0.88a 0.66abc 0.58bc 0.47c 0.65 A
2 Ştefăneşti 0.78ab 0.66abc 0.61bc 0.45c 0.63 A
3 Dumbrava Sibiului 0.73abc 0.64abc 0.48c 0.42c 0.57 B
Average on stages 0.80M 0.65N 0.56NP 0.44P
*Differences among any variants followed by one common letter are not significant.
DS 5% for the interaction between provenance and stage: 0,225-0,261.
DS 5% for provenance: 0,050-0,051.
DS 5% for the stage of explants: 0,134-0,145.
We can conclude that it is very important to harvest the acorns for somatic
embryogenesis experiments in early stages, and when it is not possible, to plant on
culture medium a big number of explants (5 fold more than usually), in order to ensure
the somatic embryo induction, despite of the very low efficiency.
It is also useful to preview a big number of explants when new provenances are tested,
because of their effect upon the embryogenesis.
Table 3 The effect of provenance and culture medium upon the rate of embryogenic
induction in Q. robur (test Duncan)
Nr.
crt.
Provenance
Culture
medium
Rate of embryo induction(%) and significance*
QE1 QE2 QE3 QE4
Average on
provenances
1 Tg. Mureş 0.64ab 0.65ab 0.65ab 0.65ab 0.65 A
2 Ştefăneşti 0.60ab 0.59ab 0.62ab 0.69a 0.63 A
3 Dumbrava Sibiului 0.57b 0.57b 0.55b 0.58b 0.57 B
Average on culture media 0,61M 0,60M 0,61M 0,64M
*Differences among any variants followed by one common letter are not significant.
DS 5% for the interaction between provenance and medium: 0,092-0,110.
DS 5% for provenance: 0,050-0,051.
DS 5% for culture medium: 0,184-0,200.
As it can be seen in the table 3, the effect of culture medium upon the rate of embryo
induction was insignificant, despite of some differences among provenances.
We can conclude that the composition of culture medium was not a limitative
factor, media with different nutritive compositions and auxin/cytokinin ratios of 5/1
(QE1, QE3) and 1/1 (QE2, QE4) being equally suitable for the induction of somatic
embryogenesis.
448

Table 4 The effect of developmental stage of explant and culture medium upon the
rate of embryogenic induction in Q. robur (test Duncan)
Nr.
crt.
Stage of explants
(zygotic
embryos)
Culture medium
Rate of embryo induction(%) and significance*
Average
S1 S2 S3 S4
on
culture
media
1 QE1 0.76abc 0.65cde 0.57defg 0.45ghi 0.61 A
2 QE2 0.79ab 0.67cde 0.51fghi 0.43i 0.60 A
3 QE3 0.82a 0.61def 0.56efghi 0.44hi 0.61 A
4 QE4 0.83a 0.69bcd 0.59def 0.46ghi 0,64 A
Average on stages 0.80M 0.65N 0.56NP 0.44P
*Differences among any variants followed by one common letter are not significant.
DS 5% for the interaction between culture medium and stage: 0,106-0,129.
DS 5% for culture medium: 0,184-0,200.
DS 5% for the stage of explants: 0,134-0,145.
In the table 4 it is shown that the best results were obtained in the stage 1, on the media
QE3 and QE4, and the worst, in the stage 4 on the medium QE 2.
2.1.b. The effect of provenance, stage of explant and culture medium upon the embryo
induction efficiency in Quercus frainetto
The three way Anova for the effect of provenance, stage of explant and culture medium
upon the embryo induction efficiency in Q.s frainetto is presented in the Table 5.
Table 5 The three way Anova type 3x4x4 for the effect of (provenance x stage of
explant x culture medium) upon the embryo induction efficiency in Quercus frainetto
Source of variation MSD DF MS
F-test
TOTAL 2.524 95
Big plots 0.081 5
Replications 0.010 2
Factor A (PROVENANCE) 0.002 1 0.002 0,053.49; 5.95
Interaction 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
actions are insignificant (table 6).
Table 6 The effect of developmental stage of explant upon the rate of embryogenic
induction in Q. frainetto (test Duncan)
Stage of explants (zygotic Average on stages Significance of the
embryos)
differences*
S1 0.746 A
S2 0.548 B
S3 0.488 BC
S4 0.424 C
*Differences among any variants followed by one common letter are not significant.
DS 5% for stage: 0,110-0,118.
The high embryo induction rate in the stage 1 proved to be highly significant, as well as
the very low embryo induction efficiency obtained in the stage 4.
2.2 Bifactorial experiments in Quercus petraea: the effect of stage of explant and
culture medium upon the embryo induction efficiency
(4 stages x 4 culture media)
All differences between the results obtained on different culture media and also the
interactions stage x medium were insignificant, but the differences among stages were
found to be highly significant (Table 7)
Table 7 The effect of developmental stage of explant upon the rate of embryogenic
induction in Q. petraea (test Duncan)
Stage of explants (zygotic Average on stages Significance of the
embryos)
differences*
S1 0.814 M
S2 0.590 N
S3 0.422 P
S4 0.311 Q
*Differences among any variants followed by one common letter are not significant.
DS 5% for stage: 0,086-0,093.
CONCLUSION
1. All oak species tested Quercus robur (three provenances), Q. petraea (one
provenance) and Q. frainetto (two provenances) and all provenances tested inside
each species showed embryogenic ability: the possibility of somatic embryo
induction out of very juvenile explants (cotyledons of immature zygotic embryos).
2. The efficiency of somatic embryo induction varied between the variants of the
experiment from 50 % (Q. robur, provenance Tg. Mures, stage 1) to 1,7 % (Q.
frainetto, stage 4), excluding the variants giving negative results, especially those
with explants in the stage 4.
3. The most important factor affecting the efficiency of somatic embryo induction
450

was the explant type (the developmental stage of the zygotic embryo used as
explant); the early developmental stages of the zygotic embryos 1 and 2 had a
significant positive effect upon the embryo induction efficiency, but the more
advanced stages (3 and 4) had a negative effect, comparing to the average.
4. We can conclude that it is very important to harvest the acorns for somatic
embryogenesis experiments in early stages, and when it is not possible, to plant on
culture medium a big number of explants (5 fold more than usually), in order to
ensure the somatic embryo induction, despite of the very low efficiency.
5. The second factor affecting the embryogenic ability, as importance, was the
genotype. The best embryogenic ability was found in Q. petraea and the worst, in
Q. frainetto. The embryogenic ability of Q. robur (average, 17,8%) and Q. petraea
(av. 18,5%) was similar, but that of Q. frainetto was significantly inferior (average
10,4%).
6. Concerning provenances, in Q. robur, the best results were obtained in the
provenance Tg. Mures (av. 22,9%) and the worst, in Dumbrava Sibiului (av. 12,9).
Between the Q. frainetto provenances, Griva was superior (av. 12,5%), and
Padurea sarului was the worst (average 8,3%).
7. The effect of culture medium proved to be totally insignificant, besides the
differences observed among the variants cultivated on different media. The
situation was the same as well for the nutrients as for growth regulators
8. The composition of culture medium was not a limitative factor, media with
different nutritive compositions and auxin/cytokinin ratios of 5/1 (QE1, QE3) and
1/1 (QE2, QE4) being equally suitable for the induction of somatic embryogenesis.
BIBLIOGRAPHY
1. Ardelean, M., Sestraş, R., Cordea, M., 2005 - “Tehnică experimentală horticolă”,
Ediţie revizuită şi adăugită - Editura Academicpres, Cluj-Napoca, pg.18
2. Chalupa, A.V., 1987 - "Somatic embryogenesis and plant regeneration in
Picea,Quercus, Betula, Tilia, Robinia, Fagus and Aesculus" - Comm. Inst. Forest.
Cehosloveniae, Vol. 15: 133-148.
3. Chalupa, A.V., 1990 - "Plant regeneration by somatic embryogenesis from cultured
immature embryos of oak (Quercus robur L.) and linden (Tilia cordata Mill.) -
Plant cell. Rep., 9: 398-401
4. Gupta, P.K., Durzan, D.J., 1985 - “Shoot multiplication from mature trees of
Duglas fir (Pseudotsuga menziesii) and Sugar pine (Pinus lambertiana)”- Plant Cell
Rep., 4: 177-179.
5. Gingas,V.M, Lineberger, R.D., 1989 - "Asexual embryogenesis and plant
regeneration in Quercus" - Plant Cell, Tissue and Organ Culture, 17: 191-203.
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Plant physiol. 132: 638-640.
7. Jorgensen, J., 1991 - "Cryopreservation of haploid embryos of Quercus petraea,
Fagus sylavtica and somatic embryos of Aesculus hippocastanum" - In: Ahuja
(Ed.), Woody Plant Biotechnology, Plenum press: 355-356.
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Suppl.1: 344s-350s.
451

9. Murashige, T., Skoog, F., 1962 - "A revised medium for rapid growth and
bioassays with tobacco tissue cultures" - Physiol. Plant. 15: 473-497.
10. Palada-Nicolau, M., Hausman, J-F, 2001 -Comparison between somatic and
zygotic embryo development in Quercus robur L. - Plant Biosystems, 135 (1) x-xx,
2001, pg. 1-9.
11. Săulescu, N. P., Săulescu, N.N., 1967 – „Câmpul de experienţă”, ed. a II-a- -
Editura Agro-Silvică, Bucureşti
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embryos" In vitro Cell. Dev. Biol. Plant, 35: 180
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regeneration via somatic embryogenesis and investigations on Agrobacterium
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(Eds.), Somatic Cell Genetics and Molecular Genetics of Trees, Kluwer press: 125-
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& Developmental Biology – Plant series, Volume 36: Issue 5
452

ABSTRACT
COMPARATIVE TESTS FOR SEPARATION METHODS OF
ORGANOCHLORINE PESTICIDES IN MILK
Adriana Chiş, Vasile Bara
University from Oradea, Faculty for Environmental Protection
Oradea, Romania
In the case of food that contains more than 2%, OCP, is determined through separation,
together with fat. This is why the comparison of various methods presented by the
specialised literature through which it can be separated from milk fat, is very useful.
The validity of the methods has been tested through the comparison of the determined
values with the ones determined through the reference method indicated for the
determination of milky fat. The application of various methods shows that there are not
major quantitative differences; however, some of them are closet to the reference
values. This thing is good to know in order to be able to apply in safety conditions, the
fastest method, the less expensive or, simply, the most accessible in the laboratory that
performs the determination.
Keywords: organochlorine pesticides (OCP), fat, milk
1. INTRODUCTION
Fat separation from milk is the first phase in OCP determination, from milk. Despite the
fact that OCP usage has been limited due to the toxic effects and bioaccumulation,
research on this kind of food contaminant is necessary because they occur under
sanitary - veterinary surveillance (Savu C-tin and Georgescu Narcisa, 2004). The
monitoring of food chemical contamination, is an essential compound of food safety
and, according to Codex Alimentarius, it supplies valuable information on the identity
of chemical contaminants for the selection of priorities in what regards their follow-up
and limits. The program GEMS/FOOD (Global Environment Monitoring System/Food
Contamination Monitoring and Assessment Program) of evaluation of the chemical
contaminants level in foods, is essential to reach this purpose. According to this
program we can observe that the determinations do not refer only to the primary foods
such as milk and diary but also to the ones along the food chain, that can be responsible
of contamination. In this regard we can state exemplify the cereals. The organochlorine
compounds are followed to the total diets, as shown in Table 1
Table 1: Chemical contaminants under sanitary – veterinary surveillance –
extract (after Hura Carmen, 2005 )
Contaminants Food
Chlorine compounds: DDT (metabolites),
HCH (isomers); polychlorine bi-phenyls,
dioxins, etc.
453
Milk, powder milk, butter, eggs, animal
and vegetal fat, fish, cereals, oil, mother
milk, total diets (menu)

2. MATERIALS AND METHODS
OCP are liposoluble substances, which leads to their accumulation in fats. For this
reason, in the qualitative and quantitative determination of this kind of toxic the first
phase is the separation of fat from the matrix elements, in this case, cow milk. (Hura
Carmen, 2006) However, for this kind of food product the OCP-kind residues content,
is expresses in mg/kg fat. So in order to be sure of an accurate extraction of the toxic for
the purpose of its quantitative determination is useful to know the efficiency of fat
separation methods. This is the preliminary phase of quantitative determination of
pesticides. (Guidelines on Good Laboratory Practice in Pesticide residue Analysis,
1993)
2.1 Determination of fat content
2.2.1– Reference method
Fat content of tested probes has been determined using as reference method, the etherchlorhydric
method (Weinbull method), applied to 100g of test, working with two types
of whole milk. We have worked with a Soxhlet extractor with four posts. After the
mineralization, the fat was extracted in p.a. oil ether, which has been divided in sand
bath, under the niche. For each test there were three determinations plus the witness test.
2.2.2 Application of alternative methods of milk fat separation
For the extraction of fat from milk, we used the following methods:
- Extraction with AOAC method
- Column extraction
- Partition extraction
Extraction through the AOAC method modified:
(Association of Analytical Communities)
The method has been modified (Sprecht, W. in 1987 and Cuniff, P. in 1997) in the
proportional diminution (1:10) of all reactive quantities used, due to the volume of the
test tubes for centrifugation at our disposal. So, in a 50 ml centrifugation test tube, put
10 lm ethanol, 0.1 g potassium oxalate and 10 ml milk. Mix it very well. Add 5 ml
ethylic ether and agitate it thoroughly for 1 minute, then we add 5 ml oil ether and
agitate it thoroughly for 1 minute. Afterwards, centrifuge it for 5 minutes at 1500
rot/min; afterwards transfer the solvent layer in a 100 ml separation funnel with 50 ml
water and 3 ml solution of saturated sodium chlorine. Extract the residue twice agitating
thoroughly with 5 ml ethylic ether: oil ether 1 : 1 (v/v) Centrifuge it and transfer the
solvent layer in the separation funnel after each extraction. Carefully mix combined
extracts with water. Drain and remove water layer. Wash the solvent layer, twice, with
two portions of water, of 100 ml each, remove water each time. For dehydration, pass
the solvent solution through a column of anhydrous sodium sulphate of 50 mm and
exterior diameter of 25 mm; collect the eluate in a Berzelius glass. Wash the column
with small portions of ether (three times, 5 ml each time). Gather all etheric extracts in a
weighted vase and evaporate the solvent from the combined extracts at steam bath
temperature under a current of air, to obtain the fat.
454

Column extraction (Beck, H. and Mathar W. 1985)
Mix 10 ml of liquid milk sample in a mortar with see sand and sodium sulphate (1 + 1
mix) to obtain a dry friable product. Transfer the mix in the extraction glass-tube
(interior diameter 12 mm and 300 mm length). Before this, introduce in the tube a tap of
“glass cotton” and a layer of 2 cm of sodium sulphate, for dehydration. Eluate the
column with a mix 2:1 (v/v) n-hexane and acetone. Collect the eluate and evaporate it in
a spinning evaporator at approximately 50 ºC under reduced pressure, using balloons of
constant weight to determine fat.
Partition extraction (SR EN 1528-2, 2004)
In a 1000 ml glass, add 100 g milk, 500 ml 2:1 (V/V) n-hexane and acetone mix and
homogenize for 4 minutes. Allow phases to separate. Decant the superior organic layer
in a separation funnel which contains 500 ml sodium sulphate solution. Add in the glass
50 ml 2:1 n-hexane-acetone mix and decant in the separation funnel to ensure the
qualitative transfer of the organic phase. Agitate the separation funnel for 30 seconds.
Allow phases to separate and remove inferior, watery phase.
Agitate the organic layer with another 500 ml sodium sulphate solution. Remove
inferior layer as before but keep about 2 ml in the funnel. Spin the separation funnel
around its axis to remove all the water from the vase’s walls. When all water settled,
drain the remaining watery phase and remove it. Put approximately 20 g sodium
sulphate with frit glass and pass the organic phase through the sodium sulphate in a
round bottom balloon which has been weighted first. Evaporate the solution in a
spinning rotator at approximately 50 ºC under reduced pressure.
RESULTS AND DISCUSSIONS
For each separation method, the final phase is the one when the solvent is removed
through evaporation and the fat is determined gravimetrically.
MG = (M2 –M1) / M1
% GR = MG /M X 100
MG - FAT MASS
% GR – FAT PERCENTAGE
M – MASS OF THE WORKING TEST
M1 – mass of the recipient where extracts are collected, empty
M2 – mass of the recipient where the extracts are collected, after removing the solvent
Note : for the column extraction method we determined the density of the test because
the methods implies the introduction of a certain volume, while the calculus refer to the
masses.
The results obtained are presented in the tables 2 and 3 for the two types of test in work.
455

Table 2: Determination results for test L1
Fat content %
Methods
Reference
method
AOAC
Column
extraction
Partition
extraction
Determination 1 4.02 3.87 3.99 4.04
Determination 2 4.22 4.08 4.15 4.28
Determination 3 4.15 3.93 4.12 4.33
Nr 3 3 3 3
Average 4.130 3.960 4.087 4.217
Standard deviation 0.101 0.108 0.085 0.155
t 1.9913 0.5642 -0.8155
Table 3: Determination results for test L2
Fat content %
Methods
Reference
method
AOAC
Column
extraction
Partition
extraction
Determination 1 3.9 3.73 3.81 4.02
Determination 2 3.85 3.62 3.98 4.09
Determination 3 4.07 3.89 4.12 4.15
Nr 3 3 3 3
Average 3.940 3.747 3.970 4.087
Standard deviation 0.115 0.136 0.155 0.065
t 1.8414 -0.2692 -1.9274
The statistic analysis was performed through the t – student test (Ardelean M. 2005) and
it shows the fact that the differences are not significant between the reference method
and the three applied methods, which are used for pesticide separation with fat in milk
tests that are supposed to be contaminated.
However the AOAC method values are the most far away from the reference one
and the “t” value for 4 degrees of freedom are very close to the value that make the
distinction from un-significant values to significant values. But, since we worked at this
case on a reduced scale, as shown from the description of the work methods, it is
possible that the deviation is influenced by this aspect.
However, the extraction method and the partition method have results very close to
one another in respect to the reference method and can be used for the fat separation in
the case of pesticides determination from milk tests, which can be seen in the graphics
from figure 1 and 2. Later, the separated fraction is submitted to qualitative and
quantitative purification and determination through instrumental methods.
456

Fat Percentage
Fat percentage
4.4
4.3
4.2
4.1
4
3.9
3.8
3.7
3.6
3.5
4.2
4.1
4
3.9
3.8
3.7
3.6
3.5
4.22
4.15
4.02
4.08
3.93
3.87
4.15
4.12
3.99
1 2 3 4
Ref AOAC Extr Part
Figure 1: Variation %Gr for L1
4.07
3.9
3.85
3.89
3.73
3.62
4.12
3.98
3.81
1 2 3 4
Ref AOAC Extr Part
Figure 2: Variation %Gr for L2
457
4.33
4.28
4.04
4.15
4.09
4.02
Series1
Series2
Series3
Series1
Series2
Series3

6. REFERENCES
1. Beck H, Mathar W, 1985, Bundesgessundheitsdl. 28, pag 1 -12
2. Cunniff, P. (Ed): Official Methods of Analysis of the AOAC INTERNATIONAL
[Metode Oficiale de Analiză AOAC Internaţional], 16 !h edition, Arlington VA USA
1995, Voi 1, Chapter 10 pp 1-10, Method No 970.52
3. Guidelines on Good Laboratory Practice in Pesticide residue Analysis [Ghid de
Bună Practică de Laborator la analiza reziduurilor de pesticide], Codex alimentarius
commission. In Codex Alimentarius Volume Two, Pesticide residues in food -
Rome; Food and Agriculture Organization of the United Nations (FAO); World
Healh Organization (WHO) 1993 Part 4.3, pp 417-455
4. Hura Carmen, 2005, Chemical contamination of food in Romania (in orig.
Contaminarea chimică a alimentelor în România) 2005, Editura Cermi, Iaşi
5. Hura Carmen, 2006, Lab guide – Analysis methods for food products (in orig. Ghid
de laborator – Metode de analiză pentru produse alimentare), Editura Cermi, Iaşi
6. Savu C-tin, Georgescu Narcisa, 2004, Food safety (in orig. Siguranţa alimentelor),
Editura Semne, Bucureşti
7. Specht, W.1987,:Organochlorine and organophosphorus pesticides [Pesticide
organoclorurate şi organofosforice]. In: Deutsche Forschungsgemeinschaft, Manual
of Pesticide Residue Analysis, VCH Verlagsgesellschaft Weinheim, Voi. 1, pp 309-
319, Method S 10.
8. *** SR EN 1528-2, octomber 2004, Fat food products. Determination of pestidices
and polychlorbiphenyls (in orig. Produse alimentare grase. Determinarea
pesticidelor şi policlorbifenililor (PCB) ), Part 2 : Extraction of fat and PCB and
determination of fat content (in orig. Extracţia grăsimii, pesticidelor şi PCB-urilor
şi determinarea conţinutului în grăsime)
458

HIGHLIGHTING THE STATE OF FRESHNESS OF THE FRESH RAW
MATERIAL FISH ON THE BASIS OF THE PHYSICAL-CHEMICAL
PROPERTIES
Purgea Ramona
University of Oradea,Faculty for Environmental Protection
ABSTRACT
The assessment of the state of freshness of the raw material fish may be achieved beside
the sensorial analyses (smell, colour, consistency, etc.) also based on the physicalchemical
ones: the biological particularities of the fish, the structure and chemical
composition of the tissues, the technology of capturing and life suppression impart
certain characteristics to the biochemical processes in the fish flesh (Eftimie V.M.,
2001).
Keywords: fish, easily hydrolysable nitrogen, ammoniac, H2S and pH.
1./ INTRODUCTION
After the death of the fish, the biochemical processes from the fish flesh are carried out
with an increased dynamics that leads to the faster installation of the specific
modifications. The fish flesh is rich in protein and non-protein nitrogen and poor in
carbon hydrates (Ward, D.R. et al., 1991, Constantiniu S., 1999).
2./ MATERIALS AND METHODS
178 samples of whole fresh fish were drawn, from the two units, A and B, seeking the
verification of 16 batches (8 batches from unit A and 8 batches from unit B). The draw
of samples was performed after classifying the fish raw material in quality classes
(based on species, size, corporal weight, state of freshness, etc.) from the batches of
indigenous fish, which were verified from sanitary-veterinary point of view at the
reception in the two fish processing units.
91 samples from 8 batches from unit A and 87 samples from 8 batches of unit B
were drawn. After collection, the fresh fish samples were introduced in sterile packages
(polyethylene one time use bags), in order to prevent the subsequent contamination and
were sent to the laboratory with a view to performing the physical-chemical
examination for determining: the easily hydrolysable nitrogen, the ammoniac in freestate,
the sulphurated hydrogen and the pH. The whole fresh raw material fish must
correspond from physical-biochemical point of view to the conditions of quality
foreseen in STAS 5386-86 (Zaika,L.L. et al., 2000).
According to STAS 5386-86 (Romanian standardisation) , the conditions of
admissibility for the fresh fish are:
• easily hydrolysable nitrogen, mg NH3/100 g = 30
• ammoniac in free-state (Nessler reaction) = absent
• H2S = absent
• pH = maximum 6,2.
459

3./ RESULTS AND DISCUSSIONS
From the 16 analyzed batches (8 in unit A and 8 in unit B), in 8 of them (3 batches from
unit A and 5 batches from unit B) one highlighted exceedings of the maximum admitted
limits of the physical-chemical parameters, according to the stipulations of STAS
5386/86.
From the 178 draws performed in the 2 units, a number of 37 of them responded
positively as a result of the physical-chemical examination to the following parameters:
the easily hydrolysable nitrogen, the ammoniac in free-state, the sulphurated hydrogen
and the pH.
In table 1 the results obtained as a result of performing the physical-chemical
examination of the samples of whole fresh raw material fish.
As a result of the performed physical-chemical examinations one obtained 37
positive samples from the 178 analyzed (20.78% in the two units with a contamination
rate of 17.58% in 16/91 samples) in unit A and 24, 13% (21/87 samples in unit B (Table
1).
Predominant were exceedings of the maximum limits for the easily hydrolysable
nitrogen, which were recorded in 12 of the 178 analyzed samples (6, 74%) in the two
units, followed by values above the allowed limits of the pH in 9 of the analyzed
samples (5, 05%) and positive results of the Nessler reaction in 10 samples (5, 61%) and
of the reaction for highlighting H2S in 5 samples (2,8%).
In 6 of the 16 batches one did not record exceedings of the maximum allowed
limits for the physical-chemical parameters, according to the stipulations of STAS
5386/86 (batches 1A, 4A, 6A, 7A, 4B and 6 B).
The inadequate results of the physical-chemical parameters of freshness obtained
as a result of performing the laboratory examinations corroborate with the results of the
organoleptic examination of the batches of fresh raw material fish.
Table 1: The results of the physical-chemical examination of the batches of fresh raw
material fish
Crt. Batch 2A Batch 3A Batch 5A Batch 1B Batch 2B Batch 5B
No. NH3 pH NH3 pH NH3 pH NH3 pH NH3 pH NH3 pH
1. 39 7,1 41,9 7,3 51,9 7,6 52,3 7,7 52,0 7,6 51,4 7,5
(the values above the maximum allowed limits of the NH3 and pH parameters)
4./ CONCLUSIONS
As a result of the physical-chemical determinations performed for the samples of fresh
raw material fish one recorded exceedings of the maximum allowed limits for the easily
hydrolysable nitrogen, pH, positive results of the Nessler reaction and of the reaction for
highlighting H2S (sulphurated hydrogen).
460

REFERENCES
1. Eftimie V.M. Particularitatile morfo-structurale, consideratii biochimice,
microbiologice si parazitologice ale pestelui materie prima, destinat procesarii.
Referat 2 in cadrul tezei de doctorat, Facultatea de Medicina Veterinara Bucuresti,
iunie 2001.
2. Zaika,L.L., J.G.Phillips,J.S.Fanellli and O.J.Scullen. 2000. Revised model for
aerobic growth of Shigella flexneri to extend the validity of prediction at
temperatures between 10 and 19ºC.Intl. J. Food Microbiol. 41; 9-19.
3. Ward, D.R. and C.R. Hackney (eds). 1991. Microbiology of Marine Food
Products. Van. Nostrand Reinhold, New York.
4. Constantiniu S.-Yersinii. Biologie si diagnostic de laborator, Ed. Corson, Iasi, 1999
461

ASSESSMENT OF THE HYGIENIC QUALITY OF THE FISH FRESH RAW
MATERIAL BASED ON THE BACTERIOLOGICAL EXAMINATIONS
Purgea Ramona
University of Oradea, Faculty for Environmental Protection
ABSTRACT
Immediately after catching it, the flesh and some internal organs of fish are normally
sterile. There are however, numerous microorganisms, especially bacteria, on the skin
(103-109/g), on the gills(103-107/g) and in the digestive tube(103-108/g). The number
of bacteria from different parts of the organism vary very much and reflect, mostly, the
degree of contamination (Barzoi D.,Apostu S., 2002, Yasumoto, T. et al., 1993).
Keywords: fish, microorganisms: sporulated gram-positive bacteria and gram-negative
bacteria
1./ INTRODUCTION:
The pathogenic germs present in fish may multiply, reaching contamination levels
capable of determining the occurrence of illnesses in man.
2./ MATERIALS AND METHODS
With a view to highlighting the microbial contamination, 178 samples of whole fresh
fish were drawn, from 2 units, A and B, one monitored the verification of 16 batches (8
batches from unit A and 8 batches from unit B). The draw of samples was performed
after classifying the fish raw material in quality classes (based on species, size, corporal
weight, state of freshness, etc.) from the batches of indigenous fish, which were verified
from sanitary-veterinary point of view at the reception in the two fish processing units.
91 samples from 8 batches from unit A and 87 samples from 8 batches of unit B were
drawn. After collection, the fresh fish samples were introduced in sterile packages
(polyethylene one time use bags), in order to prevent the subsequent contamination and
were sent to the laboratory with a view to performing the microbiological examination.
The whole fresh raw material fish must correspond from bacteriological point of view to
the conditions of quality foreseen in STAS 5386-86 (Barzoi D. et al., 1999, Huss H.H.,
2004).
According to STAS 5386-86 (Popa G. et al., 1991) the whole fresh fish must not
contain bacteria in the depths of the muscular mass; gram positive bacteria are allowed
(cocci, bacilli), which do not produce in cultures gases, indol, sulphurated hydrogen
under the condition that the fish does not present organoleptic and physical-chemical
modifications.
3./ RESULTS
From the 16 analyzed batches (8 in unit A and 8 in unit B), in 15 of them one
highlighted the presence of bacteria in the depths of the muscular masses.
From the 178 draws performed in the 2 units, a number of 124 of them responded
positively as a result of the bacteriologic examination for the presence of the bacterian
flora in the depths of the muscular masses.
462

In table 1 the results obtained as a result of the direct isolation on the 3 culture
environments used in parallel for the highlighting of the mesophile aerobe bacteria and
respectively of the anaerobe bacteria or optionally anaerobe from the samples of whole
fresh raw material fish are shown.
Table 1: Evaluation of the bacterian contamination of the fresh raw material fish at
reception in the two processing units
Crt. Unit/batch
Isolation environments Total positive
No.
M1 M2 M3 samples/ batch
1. 1A 10/10 10/10 9/10 10/10(100%)
2. 2A 10/12 10/12 9/12 10/12(83,33%)
3. 1B 8/10 8/10 7/10 8/10(80%)
4. 2B 12/13 12/13 11/13 12/13(92,3%)
5. 3B 7/10 7/10 7/10 7/10(70%)
6. 4B 10/11 9/11 9/11 10/11(90,9%)
7. 5B 4/10 2/10 2/10 4/10(40%)
8. 6B 10/12 10/12 9/10 10/12(83,33%)
9. 7B 11/11 9/11 8/11 11/11(100%)
10. 8B 2/10 0/10 0/10 2/10(20%)
11. 3A 0/11 0/11 0/11 0/11(0%)
12. 4A 9/12 9/12 9/12 9/12(75%)
13. 5A 4/13 4/13 3/13 4/13(30,76%)
14. 6A 9/10 9/10 9/10 9/10(90%)
15. 7A 8/12 8/12 8/12 8/12(66,66%)
16. 8A 10/11 9/11 9/11 10/11(90,9%)
T1
15 contaminated 124/134 115/134 109/134 124/134
T2
batches
16 Analyzed
batches
(92,53%)
124/178
(69,66%)
(85,82%)
115/178
(64,60%)
(81,34%)
109/178
(61,23%)
(92,53%)
124/178
(69,66%)
M1=Nutritive glucosed bullion. M2=Nutritive glucosed agar, M3=Nutritive glucosed
bullion with liver, T1= Total samples + from the contaminated batches,
T2=Total samples + from the draws of the two units
As a result of the performed bacteriologic examinations one obtained 124 positive
samples from the 134 analyzed (92.53% in the two units with a contamination rate of
65.93% in 60/91 samples) in unit A and 73,56% (64/87 samples) in unit B
The anaerobe bacteria of type Bacillus were predominant, these being isolated in
85 samples from the 178 samples analyzed in the two units(47.75%), followed by the
type Clostridium, found in 25 samples of the 178 samples analyzed(14,04%); the aerobe
bacteria(cocci an bacilli) were isolated in proportion of 7,86% of the total of
contaminated samples.
In 1 of the 16 batches the presence of the bacterian flora was not confirmed in the
depths of the muscular masses(batch 3A).
463

For the culture environments that allow the growth and development of the aerobe
mesophile bacteria(glucosed bullion respectively glucosed nutritive agar) one obtained
rates of positivity of 92,53% in the contaminated batches and respectively 85,82% and
on the environment of culture used for cultivating anaerobe bacteria(the glucosed
nutritive bullion with liver), the positivity rate was of 81,34%.
One of the analyzed batches proved to be contaminated in proportion of 100% with
aerobe mesophile bacteria: batch 1A; other batches presented a contamination of over
90%; the batch 2B(92,3%); the batches 8A and 7B(90.9%); the batch 6A (90%),
according to the table 1. In the other batches, the contamination rate observed was more
reduced: 66,66% for batch 7A, 70% for batch 3B, 75% for batch 4A, 81,8% for batch
7B etc.
Table 2: Distribution of the bacterian stems isolated as a result of the analysis of the
batches of whole fresh fish in the units A and B
Crt. No. Unit Sporulated gram-positive bacteria Gram-negative
Bacillus Type Clostridium Type bacteria
1. A 44/91 (48,35%) 10/91 (10,98%) 6/91 (6,59%)
2. B 41/87 (47,12%) 15/87 (17,24%) 8/87 (9,19%)
3. A+B 85/178 (47,75%) 25/178 (14,04%) 14/178 (7,86%)
4./ CONCLUSIONS
1. The fresh raw material fish presented as a result of the evaluation of the bacterian
contamination at the reception in the two units A and B a contamination of 65,93%
in unit A and of 73,65% in unit B.
2. The results obtained from the samples of fresh raw material fish drawn from the
two units A and B reveal a level of contamination with sporulated gram positive
bacteria of type Bacillus of 47, 75% followed by the type Clostridium 14,04%; the
mixed gram negative microbial flora (cocci and bacilli) represented a level of
contamination of 7,86% from the total of analyzed samples.
REFERENCES
1. Barzoi D.,Negut M., Meica S.Toxiinfectiile alimentare. Ed.Diacon
Coresi,Bucuresti,1999.
2. Barzoi D.,Apostu S. Microbiologia produselor alimentare. Ed. Risoprint, Cluj-
Napoca, 2002
3. Huss H.H. Ababouch L., Gram L. 2004. Assesment and management of seafood
safety and quality. FAO Fischeries Technical Paper, Rome,444.
4. Popa G.,Stanescu V. Controlul sanitar veterinar al produselor de origine animala.
Ed. Didactica si Pedagogica, Bucuresti, 1991
5. Yasumoto, T.. and M.Murata. 1993. Marine toxins. Chem. Rev. 93;1897-1909
464

FORENSIC ASPECTS REGARDING DISASTER MANAGEMENT
Carmen Radu*, Camelia Buhaş*,Nicolae Csep**, Radu Gabriela* Andrei Csep*
* University of Oradea – Facultyof Medicine and Pharmacy
** University of Oradea – Faculty for Environmental Protection
ABSTRACT
Recent events, especially international, but also some that happened in our country,
such as the Mihăileşti ammonium nitrate truck explosion, led us to believe that we have
to be prepared for a rapid, correct and professional reaction. Therefore, establishing the
correct identity of each body represents an extremely important stage within the disaster
approach. Key words: disaster, identification center, identifying the bodies, morgue,
autopsy.
INTRODUCTION
The term disaster (catastrophe) can be approached according to the initial event:
• the accident is a sudden and unpredictable event, causing in the organism a injury
or a functional disorder, permanent or temporary (Robert)
• the catastrophe is a phenomenon of a greater extension producing a complete
disorder or even a violent end in the existence of an individual. It is synonymous to
the calamity, cataclysm, disaster or drama.
• the calamity is a public misfortune, including starvation, war, and epidemic.
• the word cataclysm comes from the Greek „kataklusmos” which means deluge and
can disturb the Earth’s crust through earthquakes, flooding, etc.
• the disaster comes from „disastro” – astrological term meaning a fatal event, a
serious misfortune and therefore destruction and ruins. It signifies the influence of a
star ceasing to be favorable, on the other side it is a misfortune produced by fate.
• the scourge represents all the big calamities to fall upon the population: avalanches,
flooding, huge tides.
• the sinister constitutes a natural catastrophical event, caused by bad weather and
which produces destructions and losses (fires, flooding, shipwrecks, earthquakes,
etc.)
As a conclusion, a catastrophe may be defined as a sudden and unusual, of great
ampleness, hitting a community and causing important damages both as concerns the
humans (great number of victims), and as concerns the materials (deteriorations and
destructions of the environment).
According to art. 2 of the Ordinance 47/1994 (approved by Law 124/1995) in Romania
disaster means:
• natural destructive phenomenon of geological or meteorological origin or the
sickening a great number of persons or animals suddenly occurred as a mass
phenomenon.
• event with extremely serious effects on the environment, caused by an accident
The mass disaster represents an event which is not in a direct relationship with an act
of war and which produces the loss of at least 10 human lives or when the number of
bodies is bigger they could investigate by means of local resources.
465

From their origin’s point of view, the disasters are divided in:
• natural, being represented by massive ecological destructions, which can
overwhelm a community needing help from outside. E.g. earthquakes, flooding,
hurricanes (typhoons) and tornadoes, land sliding, volcanic eruptions.
• artificial, which are also divided in: technological (events caused by human errors
or omissions such as: inadequate transport of dangerous substances, toxic gas
leaks), social (represented by deliberate human acts, such as wars, revolutions,
terrorists’ acts, events followed by huge life and material losses constituting the
social disasters) and ecological (caused especially by people and which affects the
Earth, the flora and fauna on multiple levels. E.g. destroying the planetary forest
and some animal species.
The disasters can have the following consequences:
• injured people needing treatment and urgent transportation to the hospital;
• deceased who need an identification as accurate as possible;
• survivors who had not been injured and do not need medical care;
• victims of disasters who lost their houses and urgently need shelter, hence the
importance of creating a center for receiving such victims,
• relatives or friends who ask for information about the persons involved in the
accident;
The sanitary infrastructure is composed of: local medical dispensaries, regional
hospitals, reference hospitals, specialized clinics. All public resources must be
integrated in this system so that they could meet the national plan’s objectives.
MATERIALS AND METHODS
The forensic’s attributions in case of disasters:
• to search the place, to record the deaths, to assist the body packing and labeling
together with the Police and the Prosecutor (a map of he place will be drawn on
which each body or segments of the body will be marked)
• when in situations involving contamination risks when researching the place
(chemical, biological accidents) the forensic will instruct the team involved in
recuperating the bodies and they will wear protection suits.
• he has a very important role in supervising the operations at the identification
center and especially in the actual identification of the bodies (a mixed team
composed of prosecutors, police officers, emergency ambulance representatives,
firemen, forensics, dentists, anthropologists, technical experts, etc.)
• will supervise the persons involved in the incident, who have no injuries, but who
can develop the post-traumatic stress disorder.
The operations at the Identification Center (the center will be organized at the nearest
morgue, but a temporary morgue can be also arranged in a warehouse, hangar, military
base, etc.)
I will now mention the necessary steps for a good functioning of the center:
• the reception of the bodies at the morgue (by a team formed of forensics, police
officers, medical students);
466

• depositing the bodies and the evidences (which will be packed, labeled and sealed);
• the identification center’s structure (the center’s leader, usually a prosecutor, will
collaborate with other institutions in order to obtain the necessary equipment,
scientists);
• identifying the bodies;
• the autopsy;
• the relatives’ reception;
• the cooperation with other agencies (essential for a correct investigation).
Methods of investigation:
• the visual investigation is relative method due to the circumstances of the
identification done by a family member of a close friend of the victim. The
forensic takes part at the accomplishment of the spoken portrait based on the
Bertillon method; a modern way of identifying is represented by the robot portrait.
• the palmar or plantar prints: are based on the existence of the ante mortem pints
for comparison or they can be collected on the personal items.
• the odontological identification – the dental structures are very resistant, even in
case of severe mutilations, advanced decompositions or even burns, but this
method also is based on the existence of a ante mortem material for comparison
(declarations, dental files, dental radiographies)
• the genetic identification – a sample of spleen or muscle is collected and
represents the source for the DNA procedure. In this case too one needs ante
mortem data or the genetic profile can be determined using biological samples
from the close relatives.
• For identification the classical methods can be used, represented by the serological
methods (blood categories, Rh) or modern techniques of genotypicity: the DNA
analysis by the methods RFLP and STR. (the most common one)
• the anthropological identification by comparing the anthropological characters of
the group or individuals (note the importance of comparing the ante mortem
radiographies with the post mortem ones.)
• another way of identification is the facial reconstruction by special techniques.
RESULTS AND DISCUSSION
Identification conclusions, may be placed within one of the following categories:
• positive identification;
• probable identification;
• unidentified ( situation when the gathered information regarding the body will be
kept in case of a future identificaton).
The forensic has to make sure of the sustained efforts of the team in order to establish
the correct identification of each body. The difficulties in identifying the bodies are
according to the nature of the disaster, with special difficulties in case of body
carbonization, decomposition, and disintegration (plane accidents, explosions).
Unfortunately in some cases it is impossible to do the identification, in which case the
body will remain unidentified. In an investigation, the decision to abandon the
467

identification does not have to be taken quickly, but when taken the information about
the body will be kept for a possible future identification.
ACKNOWLEDGEMENTS
In order for the identification team’s activity to develop in time and systematically, both
on site and at the Identification Center I propose the introduction of some forms to be
filled in by the coroner. Each coroner will follow 10 cases at most, thus receiving 10
forms numbered from 1 to 10; the second coroner gets the forms from 11 to 20, etc.
According to the number of available coroners and according to the collective
accident’s complexity (number of victims) the repartition of the cases / coroner is
variable, but not more than 10 cases / coroner.
THE PROCESSING FORM: attached to each case, filled in and signed by the chief of
each compartment.
FORM NO. 1 IDENTIFICATION PROTOCOL
Reception
-body
-body fragment
Pictures
Personal items
Fingerprints
Radiological
investigations
Dental
investigations
Laboratory
Mortuary
services
Identification
number
Place
on the map
DATE SIGNATURE
FORM NO. 2 BODY FRAGMENTS IDENTIFICATION
Body fragment
(identification number)
1.
2.
Place
on the
map
Type of
tissue or
organ
Type of
lesion
DATE SIGNATURE
468
Signature Date
Biological traces and
biological condition

FORM NO. 3 ITEM IDENTIFICATION
Personal objects
(identification number)
Place
on the map
DATE SIGNATURE
Type Content Traces
REFERENCES
1. Matei V., Actualităţi în medicina legală- Note de curs , Timişoara ,2001;
2. Beliş V., Tratat de medicină legală, vol. I şi II, Ed. Medicală, Bucureşti, 1995;
3. Proceedings of the 1st Asian-Pacific Congress on Legal Medicine and Forensic
Sciences, Singapore, September, 1983;
4. Dogaroiu C., Drăghici V., Investigaţia dezastrelor cu multiple victime, Revista
română de medicină legală, vol.X, nr.1, martie 2002;
5. Şteiner N., Mănăstireanu D., Curs practic de urgenţe medico chirurgicale –
Introducere în medicina de dezastre, Ed. Didactică şi Pedagogică, Bucureşti 1996;
6. Radu C. C., Rolul medicului legist în accidente colective, Editura Universităţii
Oradea,2006.
469

ENERGY RECOVERY FROM ANIMAL PRODUCTS
László Nagypál
Tessedik Sámuel College Faculty of Agricultural Water and Environmental
Management Institute of Environmental Sciences
5540 Szarvas, P.O. Box 3, Hungary
ABSTRACT
In the 21 st century Hungary there will not be sufficient traditional sources of energy so
that new kinds of energy will be required. The main aim of the research is to identify
renewable sources of energy which cannot be utilized in the places where they become
are available. Possible sources are the waste products of some industries which could
be recycled in the economy. For example, waste products which cannot be processed or
sold since are unwanted by-products which typically the owners seek to dispose of. The
utilization of renewing energy sources as an alternative to the fossil fuels is one of the
main efforts. (Barna 2002.)
In agriculture various types of product and waste are produced which could be a
source of energy. For example, plant residues, (energy) grass, animal manure and the
carcasses of dead animals. One possibility for cheap energy is the burning of the meat
meal and fat produced in the animal protein processing plants. The results of this
research suggest that these materials could be used as a supplementary energy source in
a cement works or coal-fed power-station. (Boros, Csokonay 2002.)
INTRODUCTION
The animal carcasses from farms are transported in a special lorry to the Animal Protein
Processing Plants, where they are treated. The process involves grinding the carcasses
into granular form which are then sterilised at 133 °C.
The 3 categories of animal wastes
EU regulations identify three categories for animal carcasses.
Category 1: carcasses with BSE prion or other diseases which can be transferred by
cross contamination;
Category 2: carcasses without risk of BSE or other diseases - manure, blood, bowels,
mud of sewage of slaughter-house;
Category 3: carcasses without risk of BSE or other diseases skin, blood (but not bovine
blood) bones, fur, hooves.
Possibility of treatment or utilization of animal carcasses
Category 1:
– burning in an incineration plant,
– burning with another material (e.g. with coal-dust in a coal-fed power-station, or
cement in the cement works) (These materials can not be used in animal feed.)
Category 2:
– used for manure or a soil enriching material,
470

– utilised in a biogas processing plant,
– utilised in a composting prism,
– burned in an incineration plant,
Category 3:
– utilised in the animal feed industry,
– utilised in biogas processing plants,
– utilised in a composting prism,
– burned in an incineration plants,
In Hungary there are six agro-processing plants which produce bone and meat meal
from different types of carcass (e.g. cat, dog, cow, pig, chicken), and there are a further
six plants which produce meat meal from the wastes of nearby slaughter-house. The
first group of plants incinerates the carcasses of cows infected by BSE. The pathogenic
prion is harmful in the meat meal, so must be burned. EU regulations don’t allow the
use of meat meal from cattle. (Connett 2005.)
The twelve plants manufacture 300.000 tons of products and burn about 60.000 tons
meat meal and fat. The European Waste Catalogue numbers are (EWC) 020102 or
020202 or 020203.
MATERIAL AND METHODS
Samples were taken form the meat meal and the components important from point of
view of burning were analyzed. The chemical composition of flue gas produced in the
course of burning was calculated by computer modelling and using the method of
approximation. Any parameter of the flue gas has to be lower than the legally approved
values. Based on the identified composition of the flue gas the optimum technology of
burning as well as the optimum technical parameters could be determined.
Measurements
Samples weighing a total of four thousand grams were collected from industrial fat and
meat meal. The chemical composition of the samples was analysed in two independent
laboratories (a/1-a/3, b/1-b/3) and the parameters are presented in the tables below.
Table1: Analyses of a wet meat meal sample
Samples Carbon (%) Hydrogen (%)
a/1 42,0 4,5
a/2 41,8 2,7
a/3 40,6 2,3
Table 2: Analyses of a dry meat meal sample
Samples Carbon (%) Hydrogen (%)
b/1 42,2 1,6
b/2 38,8 1,9
b/3 41,3 3,3
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After the laboratory analyses the heat energy produced during the burning of a 100 g
sample was calculated in the meat meal using the two main components (neglecting
heat loss and the creation heat of pollutants).
The calculation
The enthalpy of reaction was calculated from the measured parameters.
ΔH – enthalpy from reaction (thermal energy will rise from burning of meat meal)
ΣΔHkproduct-ΣΔHkstart= ΔH
I. C + O2 = CO2
II. 2H2 + O2 = 2 H2O
For 1 mol:
I. ΔH = ΔHCO2 – (ΔHC +ΔHO2) = -394 kJ
II. ΔH = 2ΔHH2Ogaz– (ΔHO2 +2ΔHH2) = -484 kJ
For 100 g dry sample:
The arithmetical middle of b/1-b/3 samples:
Content of carbon: 41,3 %, the content of hydrogen 2,27 %.
The heat from the reaction: ΔH = 1335,66 + 274,42 =1610,08 kJ.
For 1000 g sample:
Later the quantity of heat for 1000 g of material was calculated at 16,1 MJ/kg.
The viscosity of fat and water was analysed and the results at different temperatures are
presented in the next table.
Viscosity of
fat (°E)
Viscosity of
water (°E)
Table 3: Viscosity and temperature of fat and water
23,4 40
Temperature (°C)
60 80 98,3
28,15 3,89 2,35 1,78 0,51
1,00 0,65 0,61 0,35 0,21
Diagram 1 shows the rising viscosity of fat and water at different temperature.
472

Viscosity [◦E]
14
12
10
8
6
4
2
0
0 50 100 150 200 250
Temperature [◦C]
Diagram 1. The viscosity and temperature of fat ♦ viscosity of fat ■ water of fat
RESULTS AND DISCUSSIONS
• It is possible to store the fat before incineration only above the solidification point.
To ensure its proper state of aggregation at the time of spraying into the burner, the
temperature must be maintained at 90 ºC.
• Before beginning the industrial use of meat meal, the composition ash content of
the meat meal needs to be analysed to prevent corrosion. According to the
laboratory analyses, the ash content, should be 3% (m/m) residual, or less than that
after the burning, is 2.02 % (m/m).
• The meat meal should be burned together with another material, or by pelleting.
• In the 2 nd laboratory the meat meal after burning at 500 ºC was light grey which
indicates that the carbon content was destroyed at this temperature. In Hungary the
permissible burning temperature is 850ºC for wastes free of halogens.
• Based on the calculations a good fuel material is gained if the meat meal is burned
as released energy during the reaction is ~16 MJ/kg.
REFERENCES
Barna, Gy. (2002): Energy recovery from animals carcass in the rendering plants,
Recycling of wastes and second raw materials, 2: 53-61 p.
Boros, T. (2002): The BSE and energy recovery of animal waste, Recycling of wastes
and second raw materials, 4: 61-64 p.
Csokonay, J. (2002): The German industry of meat meals, Recycling of wastes and
second raw materials, 7: 49-59 p.
Connett, P. (2005): Zero Waste 2020 versus incineration, St. Lawrence University, 1-22
p.
473

THE IMPLICATIONS OF MATERNAL TOXOPLASMOSIS INFECTION IN
REPRODUCTION
Sonia-Maria Drăghici, A. Csep, Ildiko Lenard, Adriana Jarca, Nicoleta Negruţ
University of Oradea,Faculty of Medicine and Pharmacy
ABSTRACT
Toxoplasmosis is a zoonosis of parasitic etiology produced by the protozoan
Toxoplasma gondii that accidentally affects the human, most often it is latent in the
immunocompetent adult and child, but turns into a severe and redoubtable disease, with
grave complications in fetus, newborn, and immunodepressed persons.Owing to these
complications, sustained and consistent prophylactic measures are necessary.
Key words: toxoplasmosis, biological cycle, immunodepression, chronic endometritis,
chorioretinitis, congenital, prophylaxis.
1./ Epidemiology
The biological cycle of the parasite comprises the passage from the definitive host (in
most of the cases the cat, but also other mammals, especially felines) -where the
intestinal cycle takes place with sexed and gametogonic multiplication- to the
intermediary host (human, mouse, pig, sheep, etc.) -where the extraintestinal, asexual
cycle takes place.
The intestinal cycle in the definitive host results in the production of oocysts,
which are eliminated with the feces in the external environment, where, through
maturation, sporocysts and sporozoites result, capable of infesting a new host.
The extraintestinal cycle in the intermediary host consists of the transformation of
the sporozoites in tachyzoites, of which some can persist in the organs insufficient in
antibodies (brain, marrow, skeletal muscles), others are spread in lymph and blood,
being destroyed in immunocompetent organisms. The critical state of balance between
the host and the parasite can modify in the immunosupressed. Immunosupression can
occur during lifetime through the appearance of either external causal factors (physical,
chemical, medicinal, cortisonic or cancer therapy), or internal (chronic
immunodepressive diseases, immunologic, neoplasic, dysmetabolic, endocrinologic,
etc.)
The ways of transmission of toxoplasmosis from animals to human are multiple,
sometimes complex, human intertransmission is possible parenterally (transplant of
infected organs or transfusion of infected blood), or vertically from mother to fetus (by
the passage of sporozoites through the placenta directly into the fetal blood). Enteral
transmission by the consumption of aliments contaminated with oocysts and
insufficiently or unprepared thermally, or by sporozoites present in the environment
surrounding the source (sick animal) is the most frequent means of transmission of
disease from animals to human.
2./ Clinical features of acquired toxoplasmosis in immunocompetent pateints
Immunocompetent persons can be infested as early as childhood and adolescence.
474

As in any other cyclic, infectious, self-limiting disease, there can be observed a period
of incubation of 10-21 days, after consumption of infested meat, or even shorter, 5-18
days, after ingestion of oocysts.
The state period comprises the acute infection, which in 80-90% of cases in
asymptomatic. Thus can be explained the determination in populational studies from
many European countries (e.g. France) of an 80% of IgG antitoxoplasma antibodies
possessors, indicators of them having previously acquired the infection.
In 10-20% of cases, even in immunocompetents, the acute infection can manifest
symptomatically, with lymphoganglionic, febrile or mononucleosic syndrome
(adenopathy, fever, asthenia, myalgia, fugitive exanthema, hepatosplenomegaly without
angina, lymphocytosis persistent for days or weeks). During this stage of acute
symptomatic disease, clinical confusions can be made with other diseases that develop
the so-called „mononucleosis-like syndrome” (rubella, HIV infection, Epstein-Barr
infection). Extremely rarely, immunocompetents develop myocarditis, pericarditis,
hepatitis, encephalitis or meningoencephalitis. The presence of acute infection, either
asymptomatic or symptomatic is proven by the IgM antitoxoplasma antibodies.
Chronic (or latent) infection exists in all infested persons after the resolution of
acute phase, for a shorter or longer period of time (weeks-years), sometimes generating
ocular manifestations of recidivating chorioretinitis, at other times chronic toxoplasmic
endometritis. The appearance probability of clinical symptomatology of chronic
infection in immunocompetents is also owing to external or internal factors which
generate a degree of immunodepression. Acutization of chronic infection is marked by
the presence of IgM antitoxoplasma, together with elevated titers of IgG
antitoxoplasma.
The production mechanism of ocular toxoplasmosis in immunocompetents is the
reactivation of congenital infection or acquired acute infection which eventually
cronicizes. The forms of manifestation can be: chorioretinitis or focal necrotic retinitis
(vision disorders, scotomas, local pains, photophobia, epiphora), glaucoma with vision
loss, posterior uveitis or panuveitis, papilitis with optic atrophy.
Endometrial toxoplasmosis resulted after chronicized acute infection, can
generate sterility, abortion, premature birth and infection transmission to the conception
product, resulting in congenital malformations compatible or incompatible with life.
Fig. No.1 – Toxoplasmic Encephalitis Fig. No.2 – Toxoplasmic Chorioretinitis
475

Fig. No.3 – Acute
Toxoplasmic Endometritis
Fig. No.4 − Chronic
Toxoplasmic Endometritis
3./ Clinical manifestations of toxoplasmosis in the immuno-depressed patients
The production mechanisms are the same as in immunocompetents: acute infection with
all its consequences and the reactivation of chronic infection, in the latter the
appearance rate of chronic forms of disease being much higher than in
immunocompetents.
The categories of patients exposed to the infection due to a higher or lower degree
of immunodepression are: homosexuals, users of intravenous drugs, patients with
chronic diseases, diabetics, alcoholics, and patients treated with immunosuppressing
agents (neoplasm, lymphoma, leukemia, collagenosis, organ transplant).
HIV infected patients are the category with maximum risk of toxoplasmosis with
fulminant and severe evolution, neurological, pulmonary, ocular affection, sometimes
with infectious shock.
Cerebral toxoplasmosis may evolve with diffuse encephalopathy,
meningoencephalitis with disorders of the conscience state, headache, convulsions,
focal neurological lesions or myelopathy with motor and/or sensorial disorders of limbs,
urinary bladder and intestine. Chorioretinitis is usually concomitant with CNS
infection.
Pulmonary affection comprises severe interstitial pneumonia with acute
respiratory insufficiency and decease in case of unrecognized etiology or absence of
specific treatment.
Toxoplasmic myocarditis is manifested with cardiac rhythm disorders, cardiac
failure, at times in association with hemorrhagic pericarditis which poses great problems
of differential diagnosis.
Immunodepressed persons may develop less common clinical manifestations:
panpituitarism, insipid diabetes, inadequate secretion of antidiuretic hormone, orchitis,
gastrointestinal, musculoskeletal and hepatic affection, chronic endometritis and
secondary sterility.
4./ Clinical aspects of congenital toxoplasmosis
Vertical transmission of toxoplasmic infection from the mother with acute infection or
chronic acutized toxoplasmosis, generate a severe disease of the embryo or fetus,
476

usually incompatible with life or conducive to grave congenital malformations and
ulterior infantile mortality.
Transplacental transmission of mother’s acute infection has a high incidence up to
50% of cases. Chronic maternal infection may generate a lower rate of transplacental
transmission (up to 25% of cases).
In the transplacental transmission during acute maternal infection, it is very
important to determine the moment of infection related to the stage of pregnancy:
• In the first trimester the disease seldom occurs (10-25% of cases) but is severe,
leading to spontaneous abortion, stillbirth or newborn with severe forms of disease.
• In the second trimester of pregnancy, toxoplasmosis occurs in 30-45% of cases,
with intermediary graveness
• In the third trimester, the disease is more frequent (60-65% of cases), but often
benign, with functional manifestations, without macroscopic organic
malformations. During the life of the newborn, he can manifest disorders of
somatic, psychological, intellectual development or functional-enzymatic defects
There are no certain data referring to the toxoplasmosis manifestation of the conception
product according to the stage of pregnancy, in the case of mothers with chronic
reactivated toxoplasmosis by the aforementioned immunodeppressive factors.
The risk of fetal affection is conditioned by the immune response of the mother at
the moment of infection, the number of parasites transmitted to the fetus, and the age of
the fetus at the moment of mother’s sickening. Hence, the lower the gestational age, the
graver the fetal affection, but the risk of transmission is higher.
In the untreated mother the transplacental transmission rate is 50%, and in the
treated mother 25%. The period with a high risk of developing congenital toxoplasmosis
is considered to be between the 10 th and 24 th weeks of pregnancy, and low risk between
the 26 th and 40 th weeks of pregnancy.
If toxoplasmosis occurs in mother in the first trimester of pregnancy, it mostly
results in the death of the fetus and spontaneous abortion, and if it occurs later, it will
manifest in the infant as symptomatic congenital toxoplasmosis either since birth (1 in
10 cases), or only after months/years (9 in 10 cases).
Fig. No.5 – Chronic Toxoplasmic Encephalopathy
Clinical forms of disease are:
• Severe forms manifested as grave neonatal infections with multivisceral affection
(fever or hypothermia, icterus and hepatosplenomegaly, hydro- and/or
microcephaly, intracerebral calcifications, micro-ophtalmy, nystagmus, strabismus,
477

glaucoma, cataract, chorioretinitis, optic atrophy with cecity, deafness, encephalitis,
pneumonia, myocarditis, hemorrhagic syndrome). These forms are incompatible
with life; newborns survive, but exhibit severe sequelae: blindness, epilepsy,
psycho-motor retardation (they may appear within weeks, months or years since
birth)
• Moderate forms manifested as hepatosplenomegaly and lymphoadenopathy, in the
first two months of life, then with CNS and ocular affection (after months or years
since birth)
• Light forms manifested as ocular affection (isolated micro-ophtalmy, strabismus,
chorioretinitis), neurological affection (hypotonia, transient somnolence), or hepatic
affection (persistent icterus for several months afterbirth)
Children with congenital toxoplasmosis may exhibit at birth: prematurity, low weight,
icterus, petechiae, ecchymoses by thrombocytopenia, anemia, hepatomegaly,
splenomegaly, pneumonia, different degrees of ocular affection, mental retardation,
micro- or macrocephaly, hydrocephaly, cerebral calcifications, epilepsy, encephalitis.
Newborns without clinical signs of toxoplasmic infections may ulteriorly exhibit
ocular affection (chorioretinitis, strabismus, blindness), CNS affection (neurological and
psychological deficiencies, mental retardation, convulsions, learning disability),
ponderal deficit. The risk of ocular affection in these children is 1 in 3 cases.
5./ General prophylactic measures of toxoplasmosis
The acute toxoplasmic infection is difficultly prevented in the circumstances of the
existence of extrahuman natural focality (sick felines). A proof of this fact is the high
number of persons found positive at the IgG antitoxoplasma test. The importance of
acute infection prophylaxis is not evident at first glance, since it only leads to
symptomatic sickness or chronic infection in a small percent of cases. However, the
epidemiologic research demonstrates the severity of pathological consequences of
infection in the circumstances of immune response modification of the intermediary
host, due to the numerous immunodeppressive factors existing in the environment
which intervene at unexpected times.
Therefore, the following general measures are necessary:
• Neutralization of infective sources (sanitary-veterinary measures, with periodic
parasitological surveillance of domestic cats, hygienization of animals’ food, oocyst
pollution prevention)
• Measures of alimentary hygiene: consumption of aliments (meat, milk, etc.) only
after an adequate thermal processing, correct washing of fruit and vegetables
• Measures of individual, environmental hygiene (active epidemiologic surveillance
of risky professional domains)
• Prevention of infection transmission by blood transfusion or organ transplant
(exclusion of seropositive donors)
• Detection of acute infection and prompt treatment
• Sanitary education of pregnant women
• If the infection has occurred in the first trimester of pregnancy, therapeutic abortion
is highly recommended, if the infection has occurred after the third month of
pregnancy, the newborn is to be supervised and treated.
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The family doctors must be informed about the families where premature births,
repeated abortions, malformed children or cases of sterility occurred, in order to
perform an epidemiologic investigation regarding Toxoplasma gondii infection.
Family planning offices as well as gynecologists must evaluate the possible
toxoplasmic infection in future mothers.
In Romania it is imperative to make the etiologic diagnosis by evidencing not only
the specific IgM and IgG antibodies but also the antigenic structures which indicate the
presence of living parasite inside the organism.
REFERENCES
1. Jones JL et all – Congenital toxoplasmosis a review, Obstet Gynecol Surv., 2001
mai, 56(5): 296-305.
2. Johnson LL. SCID mouse models on acute and relapsing chronic Toxoplasma
gondii infections. Infect Immun. 1992;60:3719-3724.
3. Kmiec K et all – Congenital toxoplasmosis in own studies, Wiad Parazytol,
2001,47 Suppl 1:51-7
4. Leport C, Remington JS. Toxoplasmose au cours du SIDA. Presse Med.
1992;21:1165-1171.
5. Mandell et all – Principles & Practice of Infectious Dieseases, 5 th edition 2004,
Toxoplasma Gondii, 2859-2882.
6. Sinibaldi J, De Ramirez I. Incidence of congenital toxoplasmosis in live
Guatemalan newborns. Eur J Epidemiol. 1992;8:516-520.
7. Subauste CS, de Waal Malefyt R, Fuh F. Role of CD80 (B7.1) and CD86 (B7.2) in
the immune response to an intracellular pathogen. J Immunol. 1998;160:1831-
1840.
8. Swartzberg JE, Remington JS. Transmission of Toxoplasma. Am J Dis Child.
1995;129:777-779.
9. Tirard V, Niel G, Rosenheim M, et al. Diagnosis of toxoplasmosis in patients with
AIDS by isolation of the parasite from the blood. N Engl J Med. 1991;324:632.
479

BOTULISM: CLINICAL – EPIDEMIOLOGICAL STUDY
Ildiko Lenard*, Sonia Draghici*, Viorica Coldea**, Mirela Indries*,
Nicoleta Negrut*, Andrei Csep*, Timea Lenard***
* University of Oradea, ** County Clinical Hospital of Oradea,
*** County Clinical Hospital of Cluj-Napoca
ABSTRACT
Botulism is an alimentary toxiinfection caused by exoneurotoxina, clinically
characterized through bilateral and symmetric motional paralysis, secretor troubles,
asthenia. OBJECTIVE: clinical – epidemiological assessment of patients with botulism
hospitalized in the Clinical Hospital of Infectious Diseases, Oradea. PATIENTS AND
METHODS: retrospective study on 29 patients with botulism hospitalized between
January 1997 – December 2006, in whom there were analyzed epidemiological and
clinical parameters. RESULTS: There was observed: an increased incidence of botulism
in Bihor county, because of culinary customs, being predominate the cases in the
months of February and May (both at the end of winter and Easter period); there
weren’t evidenced significant differences as concerns the sex and area of provenience
distribution of cases, with a morbidity level on the group of age of 15-24 years; most of
the clinical forms were moderate , without being noticed particular forms of botulism;
68,96% of cases were formally hospitalized in other medical services, because of the
non – recognition of the clinical symptomatology; the favorable evolution of the disease
under treatment with antibotulinic serum.
Key words: botulism, symmetrical paralysis, antibotulinic serum.
1./ Introduction
Botulism is a systemical disease, making the clinical table of alimentary toxiinfection,
through the action of an exoneurotoxina of protean nature, delivered by Clostridium
botulinum (positive, anaerobe, sporuled and telluric gram bacillus which persists in the
soil, marine bank, ponds or rivers banks ). Clinically there is observed a bilateral and
symmetrical motional paralysis of the ocular muscles, pharynx +/- respiratory muscles
(by blocking the parasympathic colinergical synapses and the neuromuscular junctions
by the botulinic toxin ) (Rummel et al, 2007), associated with secretor troubles and
marked asthenia (Mandell et al, 2005; CDC 2005).
Botulinic toxin is the most popular toxic biological substance, supposing that 200g
of botulinic toxin could be enough to destroy the globe population (Villar et al, 2007).
At present there are known 7 types of botulinic toxin (A, B, C, D, E, F, G), the
frequency of human decays being caused by A, B and E types (Garcia-Rodriguez et al,
2007). Botulism is a rare disease, with different spread around the world, its seriousness
being in accordance with the type of botulinic toxin (A type from the vegetals and E
type from the fish being the most virulent ) and the infecting doze. The transmission is
made through contaminated aliments: vegetables, fruits ( under juice form or tinned
food ), meat and prepared meat (mostly pork), fish (Kirk, 2007). Depending on the
transmission way there are described three particular forms of botulism, but with similar
clinical tables Mandell et al, 2005; Mayo Clinic, 2006; Rebedea et al, 2000):
1. botulism of ingestion-the most frequent way, being resulted the toxiinfection;
480

2. botulism of inoculation-through the toxin’s penetration straight in the general flow
from the level of a wound contaminated with clostridies;
3. infantile botulism-the botulinic toxin being produced in vivo, by colonizing the
intestinal tract of the suckling, with Clostridium botulinum ( Arnon et al, 2006;
Brett et al, 2005).
2./ Objective:
Assessment of clinical-epidemiological parameters on patients with botulism,
hospitalized in the Clinical Hospital of infectious Diseases, Oradea between January
1997 – December 2006.
3./ Patients and methods
The retrospective study in 29 patients with botulism hospitalized in the Clinical
Hospital of Infectious Diseases, Oradea between January 1997 – December 2006, was
made on the base of registered data in the clinical observation papers of the patients.
There were analyzed the following parameters:
• demographic: age, sex, provenience areas;
• epidemiological: way of transmission, source of contamination;
• clinical: incubation, clinical forms of the disease, evolution under etiological
treatment.
The paraclinical confirmation of botulism was made through the test of specific
seroneutralization in the mouse.
3./ Results
In the period taken to be studied there were hospitalized in the Clinical Hospital of
Infectious Diseases Oradea, a number of 912 patients with alimentary toxiinfection, of
whom 29 patients were diagnosed with botulism (3.17% of the total cases of alimentary
toxiinfection). The incidence of the cases of botulism was as follows:
6
11
nr. Cases
0 5
Fig. nr.1. Incidence of cases
481
2
3
1
1
0
1
2
3
4
5
6
7
8
9
10

At the diagnosis moment, the patients had the average age of 32.1 years (limits 3-69
years), with the median of 33 years. From this group, there were 15 men (52%), with
the average age of 33.66 years and 14 women (48%) with the average age of 30.42
years.
48% male
52%
Fig. nr. 2. Repartition of the sex cases
female
The highest level of morbidity was described for the group of age of 15-24 years with a
total of 9 cases (32%).
10%
14%
14%
3%
0%3% 0%
7%
0-1year
1-4 years
17%
32%
Fig. nr. 3. Repartition on the age group
482
5-14years
15-24year
25-34year
35-44year
45-54year
55-64year
65-74year
75-84year

The repartition of the provenience areas cases was dominated by the rural area with 16
cases (55% ), versus the urban area with 13 cases ( 45% ), without being noticed any
significant difference.
45%
55%
Fig. nr. 4. Repartition of the provenience areas cases
rural
urban
The months with the most cases of botulism were February with 12 cases (41.4%) and
may with 7 cases (24.1% ).
The way of the transmission was digestive: in 22 cases (75.86% ) there were
incriminated prepared meals from pork meat (smoked bacon, smoked sausages home
prepared ), 4 cases (13.79 % ) were correlated with the consume of the vegetal tinned
food and only in 3 cases (10.34% ) the epidemiological survey was negative. There
were described 5 familial hotbeds (each of them with 2-5 persons ), the rest of cases
evolving individually.
The average period of time from the possible infection till the appearance of the
first signs was of two days.
All of the cases evolved with ocular paralyses (diplopia, troubles of adaptation,
palpebral ptosis, midriasis ), paralyses of the pharynx muscles (disphagia, disartry,
disphony, queasiness reflex diminution and the abolition of the velopalatin reflex) with
secretor troubles (mucosa dryness) and marked asthenia. In 6 (26.8%) cases there were
described paroxistical abdominal pains with meteorism, 7 (24.13%)cases developed
urine acute retention, in one case (3.44%) it was associated the paralysis of the
respiratory muscles and in one case (3.44%) it evolved with coma. 27 (93.10%) patients
developed average forms of disease, in 2 (6.89%) cases the evolution being severe with
respiratory paralysis, respectively coma.
The evolution was favorable under the etiological treatment with polyvalent
antibotulinic serum (in doze of 20-50ml/day on adult, and 10-25ml/day on child),
antibiotherapy (in the 2 severe cases) symptomatic alimentation through the gastric
probe.
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4./ Discussions
Botulism is a serious disease which not being treated leads to demise (Arnon et al, 2006,
Mandell et al, 2005, Mayo Clinic, 2006). The historical study of the decays hotbeds
prove the decrease in time of the number of the collective hotbeds, with the increase of
the individual cases. In the present study 14 (48,27%) persons belong to the familial
hotbeds, the rest of 15 (51,72%) ill people being individual cases of decay (Mandell et
al, 2005; McLauchlin et al, 2006; WHO, 2006). It is also observed an increased
incidence of botulism in Bihor county in the last 10 years, the maximum incidence
being registered in 2004 with a total of 11 (37,93%) cases, fact that reveals the customs
of preparing meat, fish and many other tinned food, frequently, incorrectly in precarious
conditions of hygiene.
A feature of the present lot consists of the non-recognition of the initial clinical
table, 20 (68,96%) patients being hospitalized with different diagnoses in other medical
services, being transferred in our service after a rigorous neurological exam and the
exclusion of the other neurological diseases; thus: 7 (24,13%) patients were formally
hospitalized at Neurology and 1 (3,44%) case at Pediatry, digestive signs, marked
asthenia and specific neurological signs, as well as the existence of one or more
members of the same family with characteristic symptomatology missing the initial
examiners.
The data from the literature point out a bigger frequency of botulism on men
(60%) with the highest level of morbidity at 30-39 and 50-59 years (Mandell et a, 2005;
Rebedea et al, 2000), unlike our lot where the report men/women tries to become equal
with the highest level of morbidity at the group of age 15-24 years (from the category of
pupils/students, with the frequency of the alimentary packs incorrectly preserved).There
weren’t observed peculiar differences as concerns the provenience area of the patients
or their level of education.
Most of cases evolved as average clinical forms, the evolution being severe only in
two cases, with the appearance of respiratory insufficiency because of respiratory
muscular paralysis on a young woman patient of 20 years and of coma on a 3 year old
girl, the clinical evolution being further favorable in both situations. The predominant of
the average forms of the disease, as well as of the eradicators of pork meat involved in
most of diseases, lead us to the association with the B type of exotoxin, frequent in
Europe (Mandell et al, 2005; Rebedea et al, 2000), although the toxinopatia of the
botulism cases couldn’t be made.
A bigger frequency of the diseases in the months of February and May is also
correlated with the local customs ( the pig butchery in December, meat preservation and
that of other pork products being smoked during months) and with the culinary customs
for Easter.
5./ Conclusions
1. Botulism is a severe alimentary toxiinfection which, not being treated leads to
demise; the prevention of the decay consists of a harsh alimentary hygiene.
2. The increased incidence of the decay (both under familial hotbeds and individual
cases) in Bihor county because of the culinary customs specific to this geographic
area.
484

3. The patients hospitalizations most frequently in other medical services; through the
non-recognition of the clinical table.
4. Almost equal distribution on sexes and area of provenience of cases with the
predominant affection of the group of age 15-24 years.
5. Favorable evolution of cases under treatment with antibotulinic serum promptly
introduced.
REFERENCES
1. .Arnon SS, Schechteck R, Maslanka SE, et al. Human botulism immune globulin
for the treatment of infant botulism. N Engl J Med 2006 feb 2; 354(5): 462-71;
2. Brett MM et al. 2005: A case of infant botulism with a possible link tom infeant
formula milk powder: evidence for the presence of more than one strain of
Clostridium botulinum in clinical specimens and food.
www.pathema.tigr.org/pathema/botulism.shtlm;
3. Coordinating Center for Infectious Diseases/ Division of Bacterial and Mycotic
Diseases. 2005 oct 6. www.cdc.gov/ncdidod/dbmd/diseasesinfo/botulism
4. Garcia-Rodriguez C, Leoy R, Arnat JW, et al. Molecular evolution of antibody
croos-reactivity for two subtypes of type A botulinum neurotoxin. Nat Biotechnol
2007 jan; 25(1): 107-16;
5. Kirk M, Chan Tack MM. Botulism. 2007 mar 30. www.emedicine.com/med/topic
238.htm;
6. Mandell GL, Bennett JE, Dolin R: Clostridium botulinum. Principles and Practice
of Infectious Diseases 5 th ed, 2005(2): 2822-26;
7. Mayo Clinic Staff. Botulism – Comprehensive overview of causes, symptoms,
treatments in adults and infants. 2006 mar 28.
www.mayoclinic.com/botulism/article.htm;
8. McLauchlin J, Grant KA. Food-borne botulism in the United Kingdom. J Public
Health (Oxf). 2006 dec; 28(4):337-42;
9. Peter PT, Kim J. Botulism. 2006 jun 6. www.emdicine.com/med/topic.htm;
10. Rebedea I et al. Botulism. Boli Infectioase, 2000; 219-23;
11. Rummel A, Eichner T, Weil T, et al. Identification of the protein receptor binding
site of botulism neurotoxins B and G proves the double receptor concept. Proc Natl
Acad Sci 2007 jan 2; 104(1):359-64;
12. Villar RG, Elliot SP, Davenport KM. Botulism: the many faces botulinum toxin
and its potential for bioterrorism. Infect Dis Clin N Am 2006; 20(1):313-27;
13. World Health Organization (WHO): Outbreak news. Botulism, Thailand. Whly
Epidemiol Rec 2006 mar 31; 81(13):118.
485

STUDIES REGARDING THE INFLUENCE OF SALICYLIC ACID ON THE
GERMINATION OF MAIZE CARYOPSIS (Zea Mays) AND ON THE
PEROXIDASE ACTIVITY IN MAIZE PLANTLETS
Cornelia Purcarea*, Dorina Cachita-Cosma **
University of Oradea, Faculty for Environmental Sciences*
Uiversity of Oradea, Faculty of Sciences**
ABSTRACT
In this paper we studied the influence of exogenous salicylic acid administration in
different concentrations (0.1mg/l, 1.0mg/l, 5.0mg/l), on maize caryopsis germination in
comparison with the germination of control lots, wich was not treated with salicylic
acid., and on the peroxidase activity in the maize plantlets resulted after 72, 96 and 120
hours of germination. The results showed that the exogenous salicylic acid reduced the
germination capacity of the maize caryopsis germination as compared to that of the
reference lots, and intesified with 6.8 and 9.06% the peroxidase activity in the roots and
with 5% in coleoptiles at 1.0 mg/l concentration of the maize plantlets. The exogenous
salicylic acid influences was dependent on the concentration wich was used.
Keywords: salicylic acid, maize, germination, peroxidase activity.
1./ INTRODUCTION
Salicylic acid (SA) that was named after Salix plant (willow), was first discovered as a
major component in the extracts from willow tree bark that had been used as a natural
anti-inflammatory drug from the ancient time to the 18th century. The name SA, from
the Latin word Salix for willow tree, was given to this active ingredient of willow bark
by Raffaele Piria in 1838 (Raskin, 1992)
The salicylic acid or o-hydroxibenzoic acid, nowadays considered to be an
important signal molecule, involved in the development process of plants (Ryan and
Farmer, 1992). Actually, SA is considered a plant growth regulator, which influences
many of the vegetal processes, such as: germination (Cutt and Klessig, 1992), stomatal
behaviour (Larque-Saaveda, 1979), membrane permeability (Barkosky and Einhellig,
1993), rate of photosynthesis (Khan and Prithiviraj, 2003), etc. In 1974, Cleland and
Ajami, proved that the SA has an endogeneous regulator of flowering.
In the opinion of Handro et al., (1997), the SA supports flowering and tuberization
of plants. They studied the in vitro effect of SA of flowering of Streptocarpus nobilis,
but also of in vitro tuberization of Ullucus tuberosus. In the presence of 0,5-2,0 μM SA
the number of Streptocarpus nobilis floral buds significantly increased and floral
scapes were significantly longer as compared with the controls cultured without SA..
Picomolar levels of SA, enhance cell growth and the embryogenesis in the cell
suspension cultures of Coffea arabica,as compared with entreated control cultures
(Quiroz-Figueroa et al. 2001). They reached the conclusion that SA can activate as a
morphoregulator signal, wich supports the hypothesis that SA is functioning as a growth
regulator. The authors also suggested that it is possible that this phenolic compounds act
as a signal to induce the differentiation processes. An alternative explanation is the
486

possibility that, due to the chelating properties of these compounds, some inhibitors
present in the embryogenic cultures are inactivated.
The process of generating free radicals can be minimized by adding SA, which can
chelating transitional metals, thus reducing the production of hydroxyl radicals (HO - ) or
it can act directly as an antioxidant against these types of free radicals of oxygen
(Antofie et al., 2003).
Prachi et al. (2002) have found that addition of 104 M SA, to callus cultures of
Zingiber officinale produced the increased of peroxidase and β-1,3-glucanase activity,
as well as the appearence of two new proteins in the calli.
Some enzymes, such as guaiacol-peroxidase and glutation-reductase, are activated
by the treatments with SA solutions and others, such as catalase are inhibited as a result
of SA treatments and changes in the patterns of its isoensimes can be noticed (Horvath
et al., 2002).
The aim of this work was to study the influence of the exogenous SA on maize
caryopsiss germination (Zea Mays) and on the peroxidase activity in maize plantlets
(roots and coleoptiles) derivates from its..
2./ MATHERIALS AND METHODS
Sample preparation: SA treatments were applied to lots of 150 maize caryopsis
sample, germinated into plastic recipients. The maize caryopsis used in this study have
90% germination faculty.
The germination was maded in plastic recipients, on a filter paper, moistened with
20 ml:
- tap water , for the control sample,
- salicylic acid solution 0.1mg/l
- salicylic acid solution 1mg/l,
- salicylic acid solution 5mg/l.
The germination was maded at room temperature. Every day, the quantity of
solution from the recipients was brought to the level of 20 ml. The germination
temperature was around 22 ± 2˚C.
Germination energy, represents the speed with which the germination process begins.
It is estimated by the number of germinated grains in 72 hours. The number of normally
germinated grains over the three repetitions is divided by 100 for each plastic recipient
to determine the germination energy (Andrei et al. 1981).
Preparation of enzyme extract: 1g for each samples separatelly (roots and coleoptiles)
were collected from each variant, at 72, 96 and 120 hours of caryopsis put on
germination, and were blended with 4ml phosphate buffer solution, pH 7.0, diluted 1:9
with distilled water, cooled at 4˚C.
The samples were centrifuged at 10000 rpm, for 15 minutes at 4˚C, and supernatant
were separated. The extract is kept in the refrigerator, for 2 hours for stabilizing and
expressing enzyme activity.
Enzyme assay: peroxidase activity (PA) was determinated at 483nm wavelenght using
phosphate buffer of 7.0 pH, hydrogen peroxide as substrate and p-phenilendiamine as
chromogen (Hans-Luck, 1970 modified by Sipoş et al. 2003). The extension for each
sample was noted after 30 seconds of reaction period. For each extract were made four
determinations.
487

The data obtained at the influence of the SA on PA were. The data were statistically
evaluated using the standard deviation (SD) and t-test methods.
The values of the probabilities were determined from tables using the values of the
“t” distribution and the freedom degrees based on which the variance of the empiric
series was calculated.
3./ RESULTS AND DISCUSION
Where germination energy is concerned, the data obtained after counting the germinated
of 300 maize caryopsis, for each concentration of SA, were mathematically processed
obtaining the arithmetical mean of the three values. The obtained results were written
in the diagram in figure. 1.
Germination energy %
100
80
60
40
20
0
90
80.5
Control SA 0.1mg/l SA 1.0mg/l SA 5.0mg/l
SA solution
Figure.1– Percentage differences in the germinative energies of maize caryopsiss
(Zea Mays) after 72 hours of germination
The germination energy (figure.1) of the maize caryopsis was lower when were used
SA solutions of different concentrations, as compared with the control sample, a fact
also emphasized in the reference literature by Cutt and Klessig (1992). When using
1.0mg/l concentration SA solution, the germination energy was 55% from germination
energy of control lot. 5mg/l SA solution inhibit 100% the germination of maize
caryopsis.
Figure 2 shows an overview of the percentage differences in the extinction which
reflect the intensity of the peroxidase activity in roots and in coleoptiles of the maize
plantlets originated from the maize caryopsis germination on SA solutions of 0.1mg/l,
1.0mg/l and 5.0mg/l concentrations, for 72, 96 and 120 hours in comparison with the
same parameter determined in the roots and coleoptiles of the control sample, for which
the extinction was considered to be 100%.
488
55
0

difference of extinction %
120
100
80
60
40
20
0
72 hour
root
96 hour
root
120
hour
root
72 hour
Cp
time of germination
96 hour
Cp
120
hour Cp
control
SA 0.1mg/l
SA 1.0mg/l
SA 5.0mg/l
Figure.2 – Percentage differences in extinction which reflect the intensity of the
peroxidase activity in the roots and coleoptiles of maize plantlets.
Table 1. Results synthesis of peroxidase activity measurements in the vegetal extract
obtained from the roots and coleoptiles of the plantlets after 72, 96 and 120 hours of
germination on the substrated moisted with SA solutions of different concentrations
Parameter Hour of
germination
Peroxidase
extinction in
roots
Peroxidase
extinction in
coleoptiles
72
Salicylic acid mg/l concentration
Average± SD
Control SA 0.1mg/l SA 1.0mg/l
0.844±0.029 0.889±0.007 0.924±0.011
*
***
0.917±0.057
n.s.
n.s.
0.741±0.014 0.758±0.006 0.791±0.005
n.s.
***
0.931±0.039 0.992±0.0515 1.006±0.0105
n.s.
*
0.908±0.044 0.929±0.034 0.977±0.015
96 0.864±0.028 0.894±0.04
120
72
96
120
n.s.
0.761±0.021 0.788±0.0059
n.s.
*
0.799±0.0139
*
Studying the peroxidase activity intensity separately in the roots and coleoptiles of the
plantlets obtained from the germination of the maize caryopsis in SA solutions, after 72
489

hours of germination, we observed an increase of the PA in the extract obtained from
the roots as compared with the same parameter determined in the control sample, the
increase being very significant in the case of the germination in SA solution of 1.0mg/l,
and significant when using a SA solution of 0.1mg/l concentration (table 1).
Studying the variation of the same parameter after 96 hours, an insignificant
increase of the peroxidase activity was observed as compared with the control sample at
concentrations of 0.1 and 1.0 mg/l SA solution (table 1).
After 120 hour of germination, we observed a very signifiant increase of the PA in
the extract of the roots in the case of germination in 1.0mg/l SA solution, and an
insignifiant increase for 0.1mg/l SA solution. (table1.)
For the coleoptiles extracts, the intensity of the peroxidase activity after 72, 96 and
120 hours of germination has insignificantly modified as compared with the same
parameter of control sample when we used 0.1mg/l SA solution, and significantly
increase for 1mg/l SA solution for 72 and 120 hour of germination (table 1).
4./ CONCLUSIONS
It can be observed that the exogenous SA inhibits the germination energy of the maize
caryopsis with 10 and 45% from control lot, using SA solution with 0.1 and 1.0 mg/l
concentration and 100% in case of 5.0mg/l SA solution.
The peroxidase activity of the vegetal extracts obtained from the roots of the
plantlets obtained after germination on filter paper moistened with SA solution was
very significantly increase after 72 and 120 hours from germination for 1.0 mg/l
concentration SA solution and significantly increase for 0.1mg/l SA solution, as
opposed to the peroxidase activity of the extracts obtained from coleoptiles, where its
activity changed insignificantly after 72 and 120 hours of germination for 0.1mg/l SA
solution, and significantly for 1.0mg/l SA solution.
In conclusion exogenous salicylic acid are influenced different physiological
plant process.
REFERENCES
Andrei M., Anghel I., Popescu I., Stoica E., (1981): Lucrări practice de biologie
vegetală, Editura didactică şi pedagogică Bucureşti.
Antofie M., Carasan ME., Brezeanu A., (2003): Salicylic acid effects on two
Gardenia Jasminoides cell lines. Acta Horti Bot. Bucureşti, 30:135-145.
Barkosky RR., Einhellig FA., (1993): Effects of salicylic acid on plant water
relationship. Journal of Chemical Ecolology, 19:237-247.
Cleland FC., Ajami A., (1974) – Identification of the flower-inducing factor isolated
from aphid honey dew as being salicylic acid. Plant Physiol. 54:904-906.
Cutt JR. Klessig DF., (1992): Salicylic acid in plants. A changing perspective.
Pharmacetical Technology, 16:25-34.
Handro W., Mello CM., Manzano MA., Floh ES.,(1997): Promotive effects of
Salicylic acid. R. Bras. Fisiol.Veg. 9(2): 139-142.
Hans-Luck, (1970): Methdos of Enzymatic Analysis Verlag Chemie Gmb H.
(Ed).Weinheim Bergstr. Academic Press, New york and London.p.885-888.
490

Horváth E., Szalai G., Pál m., Páldi E, Janda T., (2002): Differences between the
catalase isozymes of maize (Zea mays L.) in respect of inhibition by various
phenolic compounds. Acta Biol Szeged 46 (3-4): 33-34.
Khan W., Prithiviraj B., (2003): Photosynthetic responses of corn and soybean to
foliar application of salicylates. Journal of Plant Physiology, p. 18
Larque-Saaveda A., (1979): Stomatal closure in response to salicylic acid treatment.
Pflanzenphysiology, 93: 371-375.
Prachi-Sharma TR., Singh BM., (2002): Salicylic acid induces Resistance to
pathogen. European Journal of Plant Pathology 108 (1): 31-39.
Quiroz-Figueroa F., Mendez-Zeel M., Larque-Saavedra., Loyola-Vargas VM.,
(2001): Picomolar salicylate levels enhance cell growth and embryogenesis. Plant
cell reports 20:679-684.
Raskin I., (1992) – Salicylate a new plant hormone. Plant Physiol. 99: 799-803.
Ryan CA., Farmer EE, (1992): Oligosaccharide signals in plants: a current
assessment. Ann Rev.Palnt Physiol. Molec.Biol., 42:651-674.
Şipoş M.,Floriş C.,Chiriac C.,(2003): Activitatea peroxidazică în embrionii şi
plantulele de grâu (Triticum aestivum L.soiul Turda) rezultate prin germinaţia
cariopselor submersate, în prealabil, în azot lichid -196˚C. Analele Univ. Oradea,
Fasc.Biologie, Tom X., 315-320; 333-342.
491

THE INFLUENCE OF SALICYLIC ACID ON CONTENT OF ASSIMILATORY
PIGMENTS IN THE PRIMARY LEAVES OF WHEAT (Triticum aestivum)
PLANTLETS
Cornelia Purcarea * , Dorina Cachiţă-Cosma **
*University of Oradea, Faculty for Environmental protevtion
**University of Oradea, Faculty of Sciences
ABSTRACT
Salicilyc acid (SA) is a phenolic compound recently recognized as plant growth
regulator which is involved in many physiological processes including photosynthesis.
In the present investigation we were studied the influence of exogenous SA solution on
the content of assimilatory pigments in the primary leaves of the wheat plantlets. The
wheat grains were presoaked 6h with 0.01 mM, 0.1 mM, or 1.0 mM SA solutions and
in water for the control lots. After that, we planted the plantlets in sand and sprayed
their coleoptiles and primary leaves, each day for an additional 7 days, with the same
solutions. In the 14 th days of germination we determined the content of assimilatory
pigments extracted with N,N-dimethylformamide (DMF). The results showed that
exogenous 0.01 mM, 0.1mM, or 1.0 mM SA solutions, enhanced clorophyll a,
clorophyll b, and carotenoid pigments in the wheat plantlets primary leaves.
Keywords: wheat, salicylic acid, primary leaves, clorophyll a, clorophyll b, carotenoid
pigments.
INTRODUCTION
In adition to the classical plant hormones, new natural growth substances with
regulatory roles in tissue culture have been discovered in the last few years (Gross and
Parthier, 1994). One of these substance is salicylic acid and its derivatives. Salicylic
acid could be raised to the status of the above phytohormones because it has significant
impact on the various aspects of the plant life (Hayat and Ahmad, 2007).
Salicylic acid or ortho-hidroxibenzoic acid belongs to a diverse group of plant
phenolics. These are compounds with an aromatic ring bearing a hydroxyl group or its
functional derivative (Raskin, 1992). Salicylic acid is a natural signaling molecule
involved in the regulation of different physiological processes including photosynthesis.
The metabolic aspect of plants, supplied with SA solution or its derivatives shifted
to a varied degree depending on the plant type and the mode of application of SA
solution. The application of SA solution (20 mg/ml) to the foliage of the plants of
Brassica napus, improved the chlorophyll contents (Ghai et al., 2002). Similarly,
soakink the grains of wheat in 10 -5 M of SA solution determinated in the plants a higher
pigment contents wich declived as the concentration of SA was increased above that
concentration (Hayat et al., 2005). Moreover, 30 days old plants of Brassica juncea
sprayed with 10 -5 M of SA solution possessed chlorophyll 20% higher than those
sprayed by water only, however the maximum concentration (10 -3 ) decreased the
492

chlorophyll contents and the values were below that the water sprayed control at 60
days stage (Fariduddin et al., 2003).
Soaking the seeds of Vigna mungo in aqueous solutions of SA (10-150μm) lead a
decrease in the content of chlorophyll and carotenoid in the leaves of subsequent plants,
but supplementing SA throught irrigant did not prove as severe as seed treatment
(Anandhi and Ramanujam, 1997).
Salicylic acid activated the synthesis of carotenoids, xanthophylls and the rate of
de-epoxidation but decreased the level of clorophyll pigments, both in wheat and moong
plants also the ratio of clorophyll a/b, in wheat plantlets (Moharekar et al., 2003).
Jianping Xue et al., 2006, studied the effects of different concentration of SA
solution on the growth of Pinellia ternate. When the height of plant was about 10cm,
sprayed them with different concentration of SA solution and measured height, total
chlorophyll content, activity of SOD, MAD content, photosynthesis speed, intercellular
CO2 concentration, the transpiration speed and the leaf temperature. The results
indicated that intercellular CO2 concentration was increased, leaf temperature were
decreased and photosynthesis speed was well in 0.5 mM SA solution. In conclusion, the
concentration of 0.5 mM SA solution was suitable for the growth of P. ternate.
MATERIALS AND METHODS
For study the action of SA treatments under laboratory conditions, the wheat caryopsis
were soaked for 6 hours in 0.01 mM, 0.1 mM, or 1.0 mM SA solutions, and in water
for the control lot. Then the grains were germinated for 7 days in plastic boxes. The
germination was made on moistened filter paper with distilled water, at 22±2 °C.
After 7 days of germination we planted the plantletss in sand, leaving them there
for an additional 7 days, and sprayed theire coleoptiles and primary leaveas each day
with 0.01 mM, 0.1 mM, or 1.0 mM SA solutions or with water for the control lot.
In the 14 th days we determined the content of chlorophyllian and carotenoid
pigments of the wheat plantlets primary leaves, using N,N-dimethylformamide, 99.9%,
(Moran and Porath, 1980) for the extraction. The extraction of chlorophylls in higher
plant tissue using N,N-dimethylformamide (DMF), expedits the process and enables the
determination of small samples with low pigment level (Moran, 1982). For extraction,
50 mg fresh weight of primary leaves, were collected separately from each samples, and
were blended with 5ml DMF and then cooled at 4˚C for 72 hours. The supernatant was
separated and the content the pigment was determined using a Spekol 11 (Carl Zeiss
Jena) spectrophotometer, at 664nm wave length for chlorophyll a, 647 nm for
chlorophyll b, and 480 nm for carotenoids. For each sample we made 3 determinations.
The data obtained after the spectrophotometrycal determination, were
mathematically processed using formulae proposed by Moran and Porath (1982). For
the determination of the specific extinction coefficients (SEC), they made pure
chlorophyll a, chlorophyll b and carotenoid pigments solution similarly prepared from
DMF extracts. The SEC were determined by the equation:
Aλ =ε cl
Where Aλ is the absorbance at a given wavelength, ε is the SEC of the solution at
wavwlength λ, c is the concentration g/l, and l is the beam-path (1cm) in the measuring
493

cuvette. Solving equation for A664, A647 and A480 wavelength Moran obtained the
formulae for determination of chlorophyll a , chlorophyll b and carotenoids contents.
Clorophyll a (mg/g SP) = (11.65 A664 – 2.69 A647) • v/SP
Clorophyll b (mg/g SP) = (20.81 A647 – 4.53 A664) • v/SP
Carotenoids (mg/g SP) = (1000 A480 – 0.89 clorophyll.a – 52.02 clorophyll.b)/ 245 •
v/SP
Where: A480 – the value read with a 480 nm filter
A647 – the value read with a 647 nm filter
A664 – the value read with a 664 nm filter
v – ml of solvent used
SP – mg of material used for one extraction/sample
Chlorophyll a and b – quantity in mg calculated in the first two formulas
The results obtained after the content of assimilatory pigments determination are
averages of 3 determination and were statistically processed using the “t- test”. The
values of the probabilities were determined from tables using the values of the “t”
distribution and the freedom degrees based on which the variance of the empiric series
was calculated.
RESULTS AND DISCUSSION
Studying the content of chlorophyllian pigment (chlorophyll a and b) and carotenoids
on the primary leaves of the wheat plantlets obtained from each experimental variants,
we observed that the influence of the exogenous SA solutions treatment was dependent
on the concentration which was used and the type of pigment wich was analised. The
results obtained were presented in table 1 and graphically represented in figure 1.
The content of chlorophyll a (table 1, figure 1.) increased significantly (with 17%
from control lot considered 100%) after treatment with 0.01 mM SA solution. A very
significant increased of chlorophyll a contents was observed, with 40% from the control
lot in the case of treatment with 0.1 mM SA solution, and with 23% from the control lot
in the case of treatment with 1.0 mM SA solution.
In the case of the chlorophyll b contents (table 1., figure 1.) it could be observed a
significant increased, with 20.9% from control lot when using a 0.01 mM SA solution, a
very significant increased, with 36% from the control lot, in the case of treatment with
0.1 mM SA solution and with 24.3% from the control lot in the case of treatment with
1.0 mM SA solution.
Studying the carotenoids pigments content (table 1., figure 1.), the results show
that the accumulation of these pigments in the leaves of wheat plantlets on the 14 th days
of germination, increased significantly in comparison with the same parameter
determined from the control lot, in the case of treatment with 0.01 mM SA solution
(with 38.8% from the control lot), with 50% from the control lot when the treatment
was made with 0.1 mM SA solution and with 38.3% from the control lot for treatment
with 1.0 mM SA solution.
494

Table 1.Estimative mean values for the content of assimilatory pigments of the wheat
plantlets (Triticum aestivum) leaves after treatment made with SA solutions of
different concentrations.
Parameters
Chlorophyll
a mg/g
Chlorophyll
b mg/g
Carotenoid
pigm. mg/g
Control
lot
0.65±0.03
0.44±0.02
0.18±0.01
Salicylic acid solution
(mM concentration)
0.01mM 0.1mM 1.0 mM
Average ± standard deviation
0.76±0.01 0.91±0.22 0.801±0.012
** *** ***
0.532±0.02
**
0.25±0.02
**
0.60±0.014
***
0.27±0.021
**
0.547±0.016
**
0.249±0.01
***
p

and their coleoptiles and primary leaves were sprayed with the same concentration SA
solution or with water in the case of control lot. The value for the control lot was
considered 100%.
CONCLUSION
The exogenous 0.01 mM, 0.1 mM and 1.0 mM SA solutions treatments significantly
increased the assimilatory pigments contents in leaves of wheat plantlets (Triticum
aestivum) when the leaves and the coleoptiles was sprayed with this solution, in
comparison with the same parameters measured in leaves of wheat plantlets of the
control lot treated with water. The 0.1mM SA solution produced the highest increased
(with 40% from the control lot for chlorophyll a, with 36% from the control lot for
chlorophyll b and with 50% from the control lot for carotenoid pigments).
Salicylic acid, in the used concentration, activated the synthesis of carotenoids,
xanthophylls and the level of clorophyll pigments, in primary leaves of wheat plantlets.
REFERENCES
Anandhi, S., Ramanujam, M. P., 1997, Effect of salicylic acid on black gram (Vigna
mungo) cultivars, Ind. J. Plant Physiol., 2, p138-141..
Fariduddin, Q., Hayat , S., and Ahmad, A., 2003, Salicylic acid influences net
photosynthetic rate, carboxylation efficiency, nitrate reductase activity and seed
yield in brasssica juncea, Photosynthetica, 41, p 281-284.
Ghai, N.,Setia, R.C., 2002, Effect of paclobutrazol and salicylic acid on chlorophyll
content, hill activity and yield components in Brassica Napus L (cv. GSL-1),
Phytomorphol., 52, p 83-87.
Gross,D.,Parthier, B., 1994, Novell natural substances acting in plant growth
regulation, J.of Plant growth regulation, 13, p 93-114.
Hayat, S., Fariduddin, Q., Ali, B., and Ahmad, A., 2005, Effects of salicylic acid on
growth and enzymes activities of wheat caryopsislings, Acta.Agron.Hung., 53, p
433-437.
Hayat S., Ahmad A., 2007, Salicylic Acid: A Plant Hormone, SPRINGER (Ed)
Dortrecht, The Netherlands.
Khan, W., Prithiviraj, B., Smith, D.L, 2003, Photosynthetic responses of corn and
soybean to foliar application of salicylates, Journal of Plant Physiology, 160, p
485-492.
Moharekar, S.T., Lokhande, S.D., Hara, T., Tanaka, R., Tanaka, A., and Chavan,
P.D., 2003, Effect of salicylic acid on clorophyll and carotenoid contents of wheat
and moong caryopsislings, Photosynthetica, 41, p 315-317.
Moran, R., Porath, D., 1980, Clorophyll determination in intact tissue using N,N-
dimethylformamide, Plant Physiol, 65, p 478-479.
Moran, R., 1982, Formulae for determiantion of chlorophyllous pigments extracted
with N,N- dimethylformamide, Plant Physiol. 69 (6), p 1376-1381.
Raskin I., 1992, Salicylate, a new plant hormone, Plant Physiol., 99, p.799-803.
Xue Jianping, Zhang Aimin, Fang Zhongming, Sheng Wei, 2006, The effect of
Salicylic acid on the growth of Pinellia Ternate, The 11 th <strong>International</strong> Congress of
Plant Tissue culture & Biotechnologie, China, p 96.
496

TREE GROWTH EQUATIONS IN DIFFERENT SITES FOR MIXED STANDS
OF SESSILE OAK WITH BEECH FROM THE MIDDLE BASIN OF THE
CRISUL REPEDE RIVER
Sorin Dorog
Faculty of Environmental Protection, University of Oradea
ABSTRACT
The determining of some growth equations, which comprise simultaneously increase
factors and diameters in relative values, is of interest because these can be used
afterwards for stands which are similar in structure and vegetate in the same
environmental conditions.
The paper presents three equations for three types of sites mostly encountered in
mixed stands of sessile oak with beech from the middle basin of the Crisul Repede
river. These equations are representative for the types of sites presented, their applying
in other areas of the country and eventually, in other types of sites, presents verrifyings
and ulterior adjustments of the mathematical pattern. The paper also presents the
intervals of the diameter categories for which these equations are viable..
Key words: equations, tree growth, mixed stands, beech, sessile oak.
1. INTRODUCTION
The dendrometrical characteristics of the even-aged stands present a larger or smaller
variability in relation with the species and the environmental conditions. The diameters
have according to the experts in this field variotion factors comprised between 20 – 30
%, this variability being caused by radial increases (Giurgiu V., 1967).
The stands formed by light demanding species are characterised by high
correlation coefficients compared to shade and half – shade demanding species (Leahu
I., 1994). Relatively even – aged stands have a decrease of the radial growth rythm for
the superior diametre categories (Giurgiu V., 1979). In such circumstances there is a
difference between radial growth average on each stand and the radial growth of the
medium diameter category.
The liniarity after which the variation of the radial growth can be ajusted in relation
with the diameters at even – aged stands, was demonstrated an emphasized by the F test.
The determining of some growth equations, which can comprise at the same time
growth factors and diameters in relative values, shows interest because of the easy way
with which these can be used afterwards at stands, which are not only structurally
similar, but also having similar environmental conditions (Cook E., 1985).
Once these equations have been established, they provide the users accurate results
concerning the growth and samples are taken with the boring bit from the medium
category of diameters (Cook E., 1990).
This paper was based upon the analysis of mixed even – aged and relatively even –
aged stands of sessile oak and beech, in three types of sites, representative with regards
to the offered conditions and percentage surface. In the studied stands trial squares of
2500 m 2 were placed and in these places all the trees were inventoried, starting with the
8 cm diameter. For the medium diameter tree category, samples were taken with the
Pressler’ s borer (Dorog S., 2006).
497

2. WORKING METHODS
Radial increases have different characteristics, which have had an impact upon what
working method was used. Thus, in order to establish the diameter categories from
which samples were taken with the help of the Pressler’s borer, the following issues
were taken into account:
• For the same category of diameters, the variation coefficients of growth have
reduced values, comprised between 20-30%;
• In the same diameter category, the species has little influence upon the variation
factors;
• The variation coefficients of the radial growths decrease from the inferior diameter
categories to the superior ones, as a size order, these are situated between 20-25%;
• Within the same category of diameters, the variation amplitude of the growths is
comprised somewhere in between 0,1 and 1,9;
• The variation coefficients of growth for the whole stand are larger, with regards to
values, than the variation factors established for the diameter categories, superior
to the medium diameter of the stand (Giurgiu V., 1972).
The selection of the trees, for the taking of the growth sample, was made as
follows(Dorog S., 2006):
• all the trees from the trial square were inventoried, established from the 8 cm
diameter upwards;
- the medium diameter of the trees from the test area was established;
- the trees, from which samples were to be taken, were selected; these trees
were chosen so that they would be a part of diameter categories, with values around the
arithmetic average.
The samples obtained with the Pressler’s borer have necessitated preliminary
procedures, before their actual measurement. The preliminary procedures were the
following:
• the conversion through grinding with abrasive paper (granulation 180) of the
samples;
• establishing by visual analysis which parts from the sample come from the inside
and which from the outside of the radial section.
The conversion and measurement of the samples included the following steps (Dorog
S., 2006):
• the scaning of the samples with a specialised program, secondary to the scaner and
saving he pictures in a *bmp format.
• the annual rings were established by means of a specialised programme of picture
analysis;
498

Image no.1 ImageTool programme was used to get the width of the annual rings
• the width of the annual rings was obtained with the Distance option from the
ImageTool programme;
• saving the results and transforming the size of the annual rings from dpi into
milimeters.
For this process of transformation of the pixels into milimeters, the following pattern
was used: Ring width (mm) = 0,042333*Ring width (dpi)
The equation coefficients (Leahu I., 1990) were established by the following steps:
• The obtaining of the medium growth for each particular tree;
• Establishing the growth average for all the samples taken from a type of
environment;
• The establishing of the medium diameter for the diameter categories and within
these, there were trees from which samples were taken with Pressler’s borer;
• The calculating of the relative values for the entire string of diameter values and of
the relative values of the medium growths, respectively.
3. RESULTS AND CONCLUSIONS
On the basis of the algorythm presented previously, after the calculations, the following
values of the regression equation coefficients were obtained. These coefficients are
presented in the table below.
Table nr. 1Regression equation coefficients and the string of values which they are
suitable for
The linear regression equation coefficients
(y = ax+b)
Type of site
Coefficients Limit diameters
a b Minimum maximum
5153 0,0021 0,8486 16 40
5242 0,0462 0,7431 34 66
5243 0,0059 0,8274 24 64
499

In the linear equation y represents the radial relative increases and x represents the
relative values of the diameters. The relative values for both characteristics were
established in relation with their medium values.
The images corresponding to each type of environment were made simultaneously with
the values of the a and b coefficients.
Picture nr.2. The variation of the relative increases in relation with the relative
values of the diameters
relative increases /
relative diameters
Valorile cresterilor
relative
The variation of the relative increases and of
the relative diameter patterns
3
2
1
0
1 8 15 22 29 36 43 50 57 64
number of observations
Variatia cresterilor relative radiale in raport cu
Valorile diametrelor relative
2,5
2
1,5
1
0,5
0
1
12
23
34
45
56
67
78
89
100
111
Valorile diametrelor relative
Adatsor1
Adatsor2
Picture nr. 3. The variation of the relative increases and of the relative diameter
patterns at stands from the 5243 type of site
500
122
133
144

Following the results obtained, these conclusions occur:
• The present equations go with the pattern of variation of the previously presented
diameters
• The most significant variations of the radial growths occur in mixed stands
• The equations can be used in the previously established conditions to establish
relative radial growths at other stands, which vegetate in similar environmental
conditions in the middle basin of the Crisul Repede river
• The equations are viable only locally, in other areas, they need to be verified by
further research
• These equations were elaborated for even-aged stands, their applying to relatively
even-aged stands must be verified.
REFERENCES
1. Cook E., 1990, A conceptual linear aggregate model for tree rings, In Cook, E.R.
Kairiukstis L.A. (eds). Methods of dendrochronology. Applications in the
environmental sciences, Kluwer Academic Publishers, Dordrecht 98-104
2. Cook E., 1985, A time series analysis aproach to tree ring standardization, School
of renewable natural resources, The university of Arizona
3. Dorog S., 2006, Consideration upon the increment of sessile oak and commun
beech forests in the middle watershed of Crişul Repede river, Natural resources and
sustainable development pag. 857-864 ISBN (10) 973-759-158-5, ISBN (13) 978-
973-759-158-6, HU ISBN-10: 963-9274-99-2, HU ISBN-13: 978-963-9274-99-0
4. Dorog S., 2006, Determinarea unor ecuaţii pentru creşterea arboretelor amestecate
de gorun cu fag din bazinul mijlociu al Crişului Repede, Analele Universităţii din
Oradea, Fascicula Silvicultură vol XI, pag. 129-134 ISSN. 1453-9489
5. Dorog S., 2006, Compararea creşterii goruneto-făgetelor din bazinul mijlociu al
Crişului Repede cu tabelele de producţie româneşti, Analele Universităţii din
Oradea, Fascicula Silvicultură vol XI, pag. 135-144 ISSN. 1453-9489
6. Giurgiu V., 1979, Dendrometrie şi auxologie forestieră, Editura Ceres, Bucureşti
7. Giurgiu V., 1972, Metode ale statisticii matematice aplicate în silvicultură, Editura
Ceres, Bucureşti
8. Giurgiu V., 1967, Studiul creşterilor la arborete, Editura Agro-silvică, Bucureşti
9. Leahu I.,1990, O modalitate de exprimare a relaţiilor interspecifice în raport cu
particularităţile auxologice ale arboretelor, în: Metode şi tehnologii moderne în
cultura şi exploatarea pădurilor, Universitatea din Braşov
10. Leahu I., 1994, Dendrometrie, Editura Didactică şi Pedagogică, Bucureşti
11. Leahu, I.,1994, Metode şi modele biometrice aplicate în dendrometrie,
Universitatea “Transilvania” din Braşov
501

THE SIMULATION AND EVALUATION OF A BREAK IN MIXED
STANDS OF BEECH WITH SESSILE OAK
Sorin Dorog
Faculty of Environmental Protection, University of Oradea
ABSTRACT
The work discusses the simulation and evaluation of a break for mixed stands of sessile
oak(Quercus petraea) with beech(Fagus sylvatica). Through this break, the structure of
the stand is modified and, to this extent, the work wants to establish the optimal
structure through successive simulations.
The structure of the stands offers complex information with regards to the
programme and the future development of the stand, thus, knowing the structure and the
way the break influences it, becomes very important. This paper analysed the
experimental distributions, obtained on the basis of the field data , in comparison with
those recommended in other works of speciality (normal distribution, Charlie’s
distribution and Beta distribution).
Key words: structure, distribution, mixed stands, beech, sessile oak
1. INTRODUCTION
The structure of the stands is the one that gives the most complex information about the
programme of the stand and about its future development, if essential
changes(unexpected) do not occur, changes that can modify entirely the initial
programme. In this way, knowing the structure of the stands becomes extremely
important, also from the perspective of the future breaks. In order to have a point of
refference, this paper analysed experimental distributions, obtained from the field data,
in relation to the distributions recommended in other works.
Applying different silvotechnical measurement systems influences not only the
size of the growth, but also the quality of the wood production, that is the relationship
between the volume of thick or thin trees, belonging to different species – which are
harvested and of various quality and also the efficiency of the social and ecological
functions, that the stands have to fulfill (Leahu I.,2001). Considering as representative
for this area the even-aged mixed stands or relatively even-aged stands of beech with
sessile oak, we will try to define the stands in connection with the proportion of the
species and also with the structure of the stands on diameter categories.
The simulation of the break offers information about the direction upon which the
stand has to be taken in order to improve its composition, structure, the optimal
functioning of its given attributes and its guidance towards the domestic aims,
established through a management plan.
2. WORKING METHOD
To evaluate the structure of mixed stands of sessile oak with beech, some stands were
selected, stands which vegetate in proper conditions with regards to environmental
conditions for the two species mentioned above.
The stands were chosen so that situations, in which they were partially or totally
derrived, would be avoided. The field was devised in rectangular test areas of 2.500 sq.
502

ms. The placement of the areas was selected in such a way so as to render the structure
of the stand as accurately as possible.
Within each area the diameters of all the trees from different species were
measured and also the heights of the trees at the medium central diameter, for each
particular species. After collecting the data from the field, the structure of the stand, for
every species and diameter categories were discussed. With the help of the data, the
structure of the stands was evaluated according to the diameter categories, using
distributions recommended by specialists in this field: Gauss distribution, A type
Charlier distribution and Beta distribution (Giurgiu V., 1979, Giurgiu V., 1972).
Hi-square criterium was applied to assess the quality of the adjustments on the
experimental distribution, after the theoretical ones. After the structure of the stands was
revealed, there was a simulation of some breaks(thinnings), through which it was
intended to promote the most valuable pieces of stand, in what species and quality were
concerned, to improve the quantitative production and especially the qualitative
one(Florescu I., Nicolescu N., 1998).
The simulation of a break has been accomplished relying on the data from the
field and also by using data, obtained on the basis of the adjustment of experimental
distributions with those recommended by the specialists. The quality of the simulated
break was verified by calculating the dispersion indicators, the indicators of the form of
the distribution, the intensity of the breaks connected to the number of trees, the basic
area and the volume, Hart-Becking index (spacing factor) before and after the break.
The volume was calculated with the help of the relative volume method, a method
which is recommended by the specialised literature.
The structural differentiation in a stand is a consequence of the integrality state and
leads, eventually, to the self-thinning of the trees, following the self-improvement trait
of the biological systems.
3. RESULTS AND DISCUSSIONS
To describe the simulation and the evaluation of a break in a stand, some synthetical
data is presented in the following lines, data which has been calculated before and after
the simulated break in a.u. 78C U.P.III, F.D. Dobreşti.
Table no. 1 The calculating of the medium diameter, dispersion indicators and of the
distribution form case 1
Calculated characteristics Before the break After the break
Medium diameter(cm) 22,04188482 26
Variance 67,44612783 65,92
Standard – Deviation 8,212559152 8,1191133
Variation factor 0,372588788 0,3122736
Asimetry -0,142394868 -0,9295828
Excess 1,89798843 2,5502924
From the above data one can notice that the simulated break consisted of the extraction
of the trees from the inferior diameter categories, mostly white beech trees, but also
503

sessile oak and dying oak trees, which have remained in the inferior content. The tree
number intensity is close to the value of 32%, fact which indicates a moderate to strong
thinning , while the volume intensity is somewhere around 11% , which points to a low
intensity thinning. Thus it can be concluded that a low thinning was simulated in the
stand. The spacing factor modifies itself also, meaning that the experimental value
increases after the break by 3%.
Table no. 2 Species proportion case 1
Basic area propotion Volume proportion Tree number
Species
Before After Before After
proportion
Before After
break break break break break break
FA 33,60 34,98 35,32 39,91 27,96 41,27
GO 42,29 43,69 43,74 49,06 26,88 38,89
CA 20,51 7,23 17,37 7,01 43,01 12,70
ST 3,60 3,75 3,56 4,02 2,15 3,17
CE 0,00 10,36 0,00 0,00 0,00 3,97
CI 9,95 0,00 8,30 0,00 2,69 0,00
Table no. 3 The calculation of the break intensity and of Hart-Becking index case 1
Calculated characteristics Values
Tree number intensity 32,25806452
Basic area intensity 3,924714353
Volume intensity 11,48726754
Spacing factor ( Hart-Becking index)
Before break
14,273994
After break
17,3427
Numar de arbori (buc)
Distributia experimentala, distributia dupa
simularea interventiei si ajustarea acestora dupa
distributia normala, Charlier tip A,Beta
20
18
16
14
12
10
8
6
4
2
0
d8
d12
d16
d20
d24
d28
d32
d36
d40
d44
Categorii de diametre (cm)
Distrib exp. Distrib. Sim interv..(ni) ni distrib. Norm.
ni Charlier tip A ni Beta
Image no 1 – Experimental distribution, the distribution after the simulation of the
break and adjusting after the normal law, Charlier type A and Beta, case 1
504
d48
d52
d56

Another possible simulated break in the stand from the a.u. 78C is the one in which
action is taken upon the trees from the inferior diameter category, as well as upon the
trees from the medium and superior diameter categories. Thus, the obtained data is
shown as follows:
Table no. 4 The calculation of the medium diameter, dispersion and distribution
form indicators, case 2
Calculated characteristics Before break After break
Medium diameter(cm) 22,04188482 21,338583
Variance 67,44612783 50,105542
Standard – Deviation 8,212559152 7,0785268
Variation Factor 0,372588788 0,3317243
Asimetry -0,142394868 0,1765171
Excess 1,89798843 2,2001468
Table no. 5 Species proportion case 2
Basic area proportion Volume proportion Tree number
Species
Before After Before After
proportion
Before After
break break break break break break
FA 33,60 33,26 35,88 35,15 27,96 35,43
GO 42,29 43,40 44,43 45,70 26,88 29,92
CA 20,51 20,46 16,08 16,23 43,01 33,07
ST 3,60 2,88 3,62 2,92 2,15 1,57
CE 0,00 0,00 0,00 0,00 0,00 0,00
CI 9,95 0,00 8,43 0,00 2,69 0,00
Table no. 6 The calculation of the break intensity and of Hart-Becking index case 2
Calculated characteristics Values
Tree number intensity 31,72043011
Basic area intensity 36,4963218
Volume intensity 36,99021667
Spacing factor ( Hart-Becking index)
Before break
14,273994
After break
17,274286
In this simulated break it can be noticed that the medium diameter does not grow
artificially, as seen in the first case, which emphasizes that by breaking in all diameter
categories, the medium diameter of the stand remains aproximately consistent. Further
more, in this case, the diffferences related to tree number, basic area and volume
intensities are not too big.
505

Table no. 7 The calculation of the medium diameter, dispersion and distribution form
indicators , case 3
Calculated characteristics Before break After break
Medium Diameter(cm) 22,04188482 23,173913
Variance 67,44612783 65,020477
Standard deviation 8,212559152 8,0635276
Variation factor 0,372588788 0,3479571
Asimetry -0,142394868 -0,2028422
Excess 1,89798843 2,0031626
Table no. 8 Species proportion case 3
Basic area proportion Volume proportion Tree number
Species
Before After Before After
proportion
Before After
break break break break break break
FA 33,60 34,62 35,32 35,73 27,96 36,23
GO 42,29 47,54 43,74 48,55 26,88 34,78
CA 20,51 13,51 17,37 11,49 43,01 26,09
ST 3,60 4,33 3,56 4,23 2,15 2,90
CE 0,00 0,00 0,00 0,00 0,00 0,00
CI 9,95 0,00 8,30 0,00 2,69 0,00
Table no. 9 The calculation of the break intensity and of Hart-Becking index case 3
Calculated characteristics Values
Tree number intensity 25,80645161
Basic area intensity 16,79057756
Volume intensity 15,80365993
Spacing factor ( Hart-Becking index)
Before break
14,273994
After break
16,571523
The last case describes a strong thinning(5), which is recommendable in the actual
situation, because of the fact that an intensity, that is too big, could destablize the
internal structure of the stand.
This pattern for the simulation and evaluation of the break puts at the user’s disposal
also the comparison of experimental distributions with those recommended as normal
by the speciality works(1,2). In continuation, the distribution formulas of the tree
number with regards to quality grades, for the above examples, are presented:
- Experimental distribution after Beta
, 3
0446 , 1 2
0, 01592 × 10 x − 7 57 − x
y = ( ) ( ) 7517
- distribution after break case 1
0153 , 2 2
436, 064 × 10 x − 7 57 − x
y = ( ) ( ) 9197 , 3
506

- distribution after break case 2
y = ( ) ( ) 1281 , 2
0706 , 1 2
10, 3337 × 10 x − 7 43 − x
- distribution after break case 3
y = ( ) ( ) 3354 , 2
0882 , 1 2
2, 9207 × 10 x − 7 49 − x
in which x represents the diameter categories and y the number of trees corresponding
to each diameter category.
Numar de arbori (buc)
Distributia experimentala, distributia dupa
simularea interventiei si ajustarea acestora dupa
distributia normala, Charlier tip A,Beta
20
18
16
14
12
10
8
6
4
2
0
d8
d12
d16
d20
d24
d28
d32
Distrib exp.
Categorii de diametre (cm)
Distrib. Sim interv..(ni) ni distrib. Norm.
ni Charlier tip A ni Beta
Image 2 – Experimental distribution, the distribution after the simulation of the
break and the adjusting after the normal law, Charlier type A and Beta, case 2
4. CONCLUSIONS
From the converted data in the mixed stands of beech with sessile oak there are
interesting aspects in relation to both the interspecific competition between the two
species and between these and the common hornbeam, the lime-tree and other species
of mixed hardwood. In order to develop the stand according to the management, aims
established through a working-plan, the silvotechnical breaks will follow the change in
the composition and structure of the stands in the direction given by the established
social-economical and ecological objectives.
The pattern for the evaluation and the simulation of the break in a stand has the
advantages that, before the break is performed, information related to the tree number
structure, the basic area, volume, species proportion, Hart-Becking coefficient values is
available, which allows for the possibility of analysing and improving of the break
according to the objectives established by the management plan.
507
d36
d40
d44
d48
d52
d56

The value of the spacing factor increases with each thinning, its value can modify with
up to 5% at each break (Florescu I., 1998). The data presented as examples in the
previous pages support our point.
The simulation of a low thinning is reckognised by the fact that tree number
intensities of over 25-30% occur, while in what the basic area and the volume are
concerned, they are situated between 5-10%, which is normal, if we take into account
that trees from the inferior categories are removed, trees left behind with regards to their
growth and placed in the lower content. The remaining sessile oak or Turkey oak trees
in the lower content are declined at the tree-foot and they are the first to be
recommended for extraction through tending processes. The combined thinning implies
the removal of the trees from all diameter categories, this can be noticed by the fact that
number of trees, basic area and volume intensities, respectively, are aproximately equal,
or differing by no more than 4 or 5%.
REFERENCES
1. Chiţea Gh., 1997, Biostatistică, Editura Universităţii Transilvania, Braşov
2. Chiţea Gh., 2001, Biostatistică, Editura Universităţii Transilvania, Braşov
3. Cristea Maria, 2004, Riscurile climatice din Bazinul hidrografic al Crişurilor,
Editura Abaddaba Oradea
4. Florescu I., Nicolescu N., 1998, Silvicultura, vol. I – Silvobiologia, Editura
Universităţii Transilvania, Braşov
5. Florescu I., Nicolescu N., 1998, Silvicultura, vol. II – Silvotehnica, Editura
Universităţii Transilvania, Braşov
6. Giurgiu V., 1979, Dendrometrie şi auxologie forestieră, Editura Ceres, Bucureşti
7. Giurgiu V., 1972, Metode ale statisticii matematice aplicate în silvicultură, Editura
Ceres, Bucureşti
8. Leahu I., 1994, Dendrometrie, Editura Didactică şi Pedagogică, Bucureşti
9. Leahu I., 2001, Amenajarea pădurilor, Editura Didactică şi Pedagogică, Bucureşti
508

THE FREQUENCY OF DAYS WITH DIFFERENT TEMPERATURES IN THE
CRISUL REPEDE HYDROGRAPHICAL BASIN
Ana Cornelia Moza
University of Oradea, Faculty for Environmental Protection
ABSTRACT
The particularities of the air temperature’s regime are indicated by a series of elements,
calculated or extracted graphically from the four stations histograms. Some of these
elements are: frequency of frosty nights, frequency of frosty days, winter days, summer
days and tropical days. These elements were analyzed during 1970-2005 at all 4
stations.
INTRODUCTION
The air temperature presents a series of particularities: depending on the altitude,
described by the decrease of the air temperature with the rise of altitude, and depending
on the orientation and the gradient of the slopes, described by their different warming.
While describing the climate of a region, the focus is on monthly and annual
frequency of days and nights over or under certain characteristic limits, represented in
number of days.
Over the year, the air temperature drops below or rises above certain limits,
according to which there were assigned frosty nights, frosty days, winter days, summer
days and tropical days.
MATERIAL AND METHOD
In order to point out the characteristics of the temperature, the data during 1970-2005
were used, obtained from the observations made at the four meteorological stations
(Oradea, Săcueni, Borod, Stâna de Vale), situated both in plain areas and mountain
areas, so that they cover the entire researched area.
The number of days with certain temperatures is extracted from the meteorological
tables for every month. Adding the respective values and dividing the result to the
number of months in the investigated years, we obtain the average number of days in a
month with characteristic temperatures, which is graphically represented by bars.
RESULTS AND DISCUSSIONS
1. The frequency of frosty nights
In the cold season, the nocturnal cooling reaches the highest values, so that the
minimum air temperatures may drop below -10ºC, situation in which the nights are
considered frosty.
In the Crisul Repede hydrographical basin, the frequency of frosty nights rises
directly proportional to the altitude from 11-12 cases in Săcueni, Oradea, up to 52 cases
in Stâna de Vale ( because of the flow of cold air from surrounding peaks and its
stagnation on the bottom of the depression for several days in a row) (fig.1).
Through out a year, frosty nights are recorded at Stâna de Vale meteorological
station from October until April, while at the other stations (Borod, Oradea, Săcueni)
509

the frosty nights begin in November and last until March. The maximum number of
frosty nights is recorded in January: 4.7 and 4.9 days in Săcueni and Oradea, 6.4 days in
Borod and 14.9 days in Stâna de Vale (fig.1).
55
50
45
40
35
30
25
20
15
10
5
0
Figure no.1: Variation of the number of days with
minimum temperature ≤10ºC (frosty nights)
I II III IV V VI VII VIII IX X XI XII An
Borod
Oradea
Sacueni
Stâna de Vale
2. The average frequency of freezing days (minimum temperature≤0ºC)
The freezing days have a higher share from October up to April, but low values (0.1
days) are recorded too from September to May in plain areas and depression, with
freezing days all year long in mountain areas.
The frequency of freezing days increases with the rise of altitude, reaching the
maximum value at Stâna de Vale (1108m) with 182 freezing days per year, while
lower, at Borod (333m) the value is 107.8 days per year, dropping to 91.4 days of
freezing days per year at Săcueni (fig.2).
In the studied area, over a 36 years period, under the influence of the
atmosfere’s general circulation and the local conditions, during the year the highest
number of freezing days appear in January 23-24 days in the plain areas, 25.4 days at
Borod, while at Stâna de Vale the maximum value of 29.6 days of freezing are
recorded. At Stâna de Vale station, freezing days are found all year long, but with a
lower frequency of 0.3 days in August and only 0.1 days in July.
510

200
190
180
170
160
150
140
130
120
110
100
90
80
70
60
50
40
30
20
10
0
Figure no.2: Variation of the number of the frosty days
(minimum temperature ≤ 0ºC) in the Crisul Repede
hydrographical basin during the period 1970-2005
I II III IV V VI VII VIII IX X XI XII An
Borod
Oradea
Sacueni
Stâna de Vale
3. Average frequency of winter days (maximum temperature≤0ºC)
Winter days are recorded from November to March, in the cold season, their frequency
rises along with the increasing activities of the Scandinavian, Greenland and especially
the Siberian anticyclone, which conditions the arctic air masses movement to the South.
511

50
45
40
35
30
25
20
15
10
5
0
Figure no.3: Variation of the medium number of the
winter days (maximum temperature ≤0ºC) in the
Crisul Repede hydrographical basin during the
period 1970-2005
I II III IV V VI VII VIII IX X XI XII An
Borod
Oradea
Sacueni
Stâna de Vale
Throughout the year, the winter days can be produced from November until March at
low altitudes and from October to April at high altitudes. The number of winter days
from the Crisul Repede hydrographical basin reaches the highest value of 48.8 days at
Stâna de Vale meteorological station and the lowest value of 22.9 days in Oradea.
Throughout the year, the highest frequency of these days is recorded in January 13.9
days at Stâna de Vale, when the entire year’s highest value is reached, while the lowest
number of winter days is recorded in March at Oradea 0.8 days. Winter days also take
place in large numbers in February and December, while starting with March, along
with the general warming of the atmosphere, to drop significantly at plain stations and
starting with April to decrease almost totally at Stâna de Vale (1.1 days) (fig.3).
4. Average frequency of summer days (maximum temperature.≥25ºC)
High temperatures are usually recorded in case of anticyclone regime, described by the
invasion of dry continental air from medium latitudes or tropical air from Central Asia
or Northern Africa. In these conditions a very hot weather sets in, accompanied by dry
winds. Usually the summer days settle in during the April-October period.
An average number of 80-90 days per year are recorded in plain meteorological
stations, 61.1 days in depressions, while as the altitude increases, the average number of
days drops, reaching at 9.3 days at Stâna de Vale (fig.4).
Over the year, the maximum number of summer days is reached in July (17.8 days
at Borod, 22.8 days at Oradea, 21.7 days at Săcueni) at low altitudes and in August, at
high altitudes, (4 days at Stâna de Vale) (fig. 4). The favourable period for summer days
decreases with the rise of altitude, so that at low altitudes it lasts from March until
512

October, and at higher altitudes (0.3 days at Stana de Vale) until September (0.4 days at
Stâna de Vale).
90
85
80
75
70
65
60
55
50
45
40
35
30
25
20
15
10
5
0
Figure no. 4: Variation of the medium number of
summer days (maximum temperature ≥25ºC) in the
Crişul Repede hydrographical basin during the period
1970-2005
I II III IV V VI VII VIII IX X XI XII An
Borod
Oradea
Sacueni
Stâna de Vale
5. Average frequency of tropical days (maximum temperature.≥30ºC)
High temperatures (≥30ºC) are conditioned by the movement of dry tropical air masses
within the country’s territory. The tropical days are days in which the maximum air
temperature is greater or equal with 30ºC.
The maximum number of tropical days in the Crisul Repede hydrographical basin,
as an average, is recorded at Oradea, 24.3 days, and at Săcueni, 20.3 days, then they
begin to drop as the altitude rises, 10.3 days at Borod, while at Stâna de Vale they
disappear (fig.5).
The favourable annual interval for producing tropical days takes place between
May and September, with a maximum number of days in August at Borod 4.2 days, in
July at Oradea 9.1 days and 7.6 days at Săcueni (fig.5).
513

25
24
23
22
21
20
19
18
17
16
15
14
13
12
11
10
9
8
7
6
5
4
3
2
1
0
Figure no.5: Variation of the medium number of tropical
days (maximum temperature ≥30ºC in Crişul Repede
hydrogaphical basin during the period 1970-2005
I II III IV V VI VII VIII IX X XI XII An
Borod
Oradea
Sacueni
Stâna de Vale
CONCLUSIONS:
1. The frequency of frosty nights rises directly proportional with the height in 11-12
cases/year in the plains and up to 52 cases/year in the mountain area. Frosty nights
appear from November until March in the plains and from October to April in the
mountains, with a maximum value in January, when the highest value of 14.9 days
is recorded at Stâna de Vale, and the lowest value is 4.7 days at Săcueni.
2. The frequency of freezing days raises proportional with the height in 91-96 cases in
the plains, in about 108 cases in the depression area of Borod and up to 182 cases in
the mountains. Throughout the year the highest number of frosty days is produced
in January, 23-24 days in the plains, 25 days in the depression area of Borod and
almost 30 days at Stâna de Vale.
3. The frequency of winter days increases as the altitude rises, starting with 23
days/year at Oradea, up to approximately 50 days/year at Stâna de Vale. Over the
year, winter days are found from November to March at Borod, Oradea, Săcueni
and from October to April in the mountains. The highest frequency of winter days
is in January, when the records show about 14 days in the mountains, about 9 days
in the depression area of Borod and 10-12 days in the plains.
4. The frequency of summer days is between 80-86 days/year in the plains, about 61
days/year at Borod, while in the mountain area the frequency is only about 9 days/
year. Throughout the year the highest number of summer days is recorded in July
(between 22-23 days) in the plains, about 18 days in the depression area of Borod,
while in the mountains the maximum number of only 4 days is recorded in August.
514

5. The frequency of tropical days ranges between 20-24 days in the plains, is about 10
days in the depression area of Borod, while in the mountains they lack. Over the
year the favourable interval for tropical days is set between May and September.
The highest frequency is recorded in July in the plains and in August, about 4 days,
in the depression area of Borod.
REFERENCES
1 Berindei O., Pop Gr., Măhăra Gh., Posea Aurora, (1977), Câmpia Crişurilor, Crişul
Repede, Ţara Beiuşului, Cercetări în geografia României, Editura Ştiinţifică şi
Enciclopedică, Bucureşti.
2 Cristea Maria (2003), Temperatura aerului în bazinul hidrografic al Crişurilor,
Analele Universităţii din Oradea, Seria Geografie, Tom.XIII, pag.77-80.
3 Cristea Maria (2004), Riscurile climatice din bazinul hidrografic al Crişurilor, Edit.
Abaddaba, Oradea, ISBN 973-8102-19-7
4 Gaceu O.(2002), Elemete de climatologie practică, Editura Universităţii din
Oradea, ISBN 973-613-216-1.
5 Gaceu O.(2005), Clima şi riscurile climatice din Munţii Bihor şi Vlădeasa, Editura
Universităţii din Oradea, ISBN 973-759-008-2.
6 Măhăra Gh. (1974), Variaţiile neperiodice ale temperaturii aerului în spaţiul
Câmpiei Crişurilor, Lucr.Ştiinţifice Seria A, Matematică-Fizică-Geografie, Oradea,
pag.37-45.
515

THE RISK OF INUNDATIONS INDUCED BY THE FLOODS ON THE
RIVERS FROM ALMAS-AGRIJ DEPRESSION
AND CLUJ - DEJ HILLS
Alina-Daciana Dumitra
Faculty of Geography, Babes-Bolyai University – Cluj-Napoca
ABSTRACT
The aim of this study was to analyse several aspects of the floods on the rivers from
Almas-Agrij Depression and Cluj and Dej Hills, because they often generated
catastrophic inundations.
In this work it were used series of data for 28 years within the interval 1976-2004
of nine hydrometrical stations: Almaş, Hida, Românaşi, in the Almaş-Agrij Depression,
and: Căpuşu Mare, Mera ( Rădaia), Aghireş, Maia, Borşa and Luna de Jos, in the Cluj
and Dej Hills, on the: Almaş, Agrij, Căpuş, Nadăş, Olpret, Borşa and Lonea rivers. The
main purpose of the study was to distinguish some aspects of floods that would
constitute a very important help for the prediction, the prevention and the diminishment
of the hydrological risks. Thus, the work presents the floods formation, analyses their
frequency of occurrence and describes some characteristics of flash floods (debits,
duration) from the area. The study represents a preliminary stage for further analysis
regarding the hydrological risks in the Almaş-Agrij Depression and the Cluj and Dej
Hills.
Key words: floods, risk of inundations
INTRODUCTION
From the beginnings the human societies were attracted by the riversides and took the
risk of living there (Pandi, 2002).
The inundations represent the most often spread hasard on Earth, with numerous
losses of human lives and material damages of huge proportions. Inundation means the
temporary cover of an area with stagnating or moving waters, as a consequence of the
raise in levels of a river, lake or other mass of water (Sorocovschi, 2002).
Catastrophic inundations are often generated by river floods. Floods represent the
significant growth and decrease of water discharge in the river bed (Mustăţea, 2005),
which, along with large waters, are phases of the flood flow that could take place in any
period of the year. They represent natural phenomena that are part of the chain of
normal events concerning water discharge.
The spatial allocation and amplitude of the floods are determined by natural and
also by antropic factors, depending on which their effects are different.
The climate has the main role in the formation of floods. The general circulation
of the atmosphere is the genetic factor which determines the non-periodical variation of
hydrological regime. Rainfall play the decisive part in producing the highest discharges.
Heavy rains usually affect small areas and determine floods in small catchement areas.
Morphology exerts its influence indirectly upon the discharge by: determining the
vertical zonality, the configuration, fragmentation and exposition of slopes.
516

At the same time, the genesis of floods is indirectly linked to the geological factor: the
lythology and the structure both influence the movement of water and the development
of the type of drainage.
Because of its hydrical and physical properties, the soil is an important factor
regarding water flow on its surface. The vegetation acts directly upon discharge by
influencing the regime of soil humidity, and indirectly by protecting against erosion .
Antropic activity acts against the genetic factors of flow (by clearance, plantation,
pasture etc.) on the one hand, and against the flow from the riverbed on the other hand
(by the mitigation of debits, embankments, water consume usages).
MATERIALS AND METHODS
To emphasize the territorial and temporal differences between the floods of the Almaş-
Agrij Depression and the floods of the Cluj and Dej Hills, there were analysed and
processed data that originate from nine hydrometrical station: Almaş, Hida, Românaşi
(Almaş-Agrij Depression), Căpuşu Mare, Mera (Rădaia), Aghireş, Maia, Borşa and
Luna de Jos (Cluj and Dej Hills) on the: Almaş, Agrij, Căpuş, Nadăş, Olpret, Borşa and
Lonea rivers. The data are resulted from the observations and measurements performed
over a period of 28 years (between 1976 and 2004), and they are archived in the Annual
Hydrological Studies of the Somes-Tisa Water Department of Cluj-Napoca.
The time frequency of floods within the studied time interval on the rivers of
Almas- Agrij Depression and Cluj-Dej Hills has been analysed based on these data. The
seasonal frequency of floods has been graphically represented, calculating the total
number of floods in each season at each hydrological station. The monthly frequency of
floods has been represented the same way using the same method, the calculation being
made for each month.
Then, also based on these data, it were described two types of parametres of flood
(the maximal debit and the duration). The main characteristic parameters of a flood are:
debits (maximum debit, base debit), duration (total duration or total time, increasing
time, decreasing time), water volumes, the form coefficient of flood, the medium flowed
layer, etc. Maximum debit ( Qmax m 3 /s) represents the parameter of the maximum
interest, because this is the moment of the biggest danger, the maximum overflow. The
duration of floods is an important element of which depends the size of the effects
which could be generated (Sorocovschi, 2005). The whole duration or total time is
composed by increasing time and decreasing time
The maximal debits ( Qmax ) of floods on the rivers of Almas-Agrij Depression and
Cluj-Dej Hills have been graphically represented for the period between 1976 and
2004. The years with highest and lowest risk of inundations have been clearly
underlined.
The total duration (Tt) of the flood has been obtained by calculating the number of
hours, from the time when the debit overpassed the basic debit, until the time it came
back to the value of the basic debit (of course, after reaching the maximal debit - Qmax).
The increasing time of flood has been obtained by adding the number of hours
from the time the debits started to grow until the time the maximal debit (Qmax) has been
recorded.
517

RESULTS AND DISCUSSIONS
In Almaş-Agrij Depression and in Cluj and Dej Hills, on the majority of rivers, the
registered floods were originated in pluvial (50 – 55% of the analyzed floods), followed
by a much smaller number of floods originated in pluvial-nival.
floods
winter spring summer autum
seasons
Almasu
Hida
Romanasi
Figure 1. The seasonal flood frequency in Depresiunea Almaş-Agrij (1976-2004)
floods
35
30
25
20
15
10
5
0
winter spring summer autum
season
Capusu Mare
Aghires
Mera(Radaia)
Figure 2. The seasonal flood frequency in Cluj - Dej Hills (1976-2004)
Figure 1,2 and 3 shows that, regarding the seasons, the maximum frequency is observed
in spring (35-55% of the entire number of floods), followed by summer (17 – 33%). In
518

winter the floods frequency has smaller values (included within the 14 – 25% of total
floods). The most reduced frequency of the floods (with values between 14 and 25 % of
the total floods) is recorded in the Almaş-Agrij Depression but also in the Cluj - Dej
Hills in autumn.
floods
35
30
25
20
15
10
5
0
winter spring summer autum
season
Borsa
Luna de Jos
Maia
Figure 3. The seasonal flood frequency in Cluj and Dej Hills (1976-2004)
It is noticed, regarding the floods frequency during the seasons, a spatial differentiation
on the Cluj and Dej Hills, thus: on the rivers from the east of the unit (Olpret, Borşa,
Lonea) the number of floods during the summer has a very close value to that number
which represents the floods during the winter (34 floods during the summer, and 32
floods in winter); on the Nadăş and Căpuş Streams (Cluj Hills) the difference is
remarkable, the number of summer floods being much higher than the number of winter
floods (20 floods, 46 floods respectively). More over, this considerable difference is
found again in Almaş-Agrij Depression (29 floods, 45 floods respectively).
The monthly frequency of floods is maximum in March on rivers of the Cluj and
Dej Hills (Olpret, Lonea, Borşa, Nadăş, Căpuş), representing 17–23 % of all selected
floods. High values of the monthly frequency of floods formation have been recorded in
April and May. The lowest frequencies of floods took place on Olpret, Borşa and Lonea
rivers in October and November. There were no floods on Căpuş and Nadăş rivers in
November, within the studied interval, and in December there were recorded minimal
values of the floods frequency (1.9% of total) (Fig.4, Fig. 5)
519

%
floods
14
12
10
8
6
4
2
0
14
12
10
I II III IV V VI VII VIII IX X XI XII
months
Capusu Mare
Aghires
Mera(Radaia)
Figure 4. Monthly floods frequency in Cluj- Dej Hills (1976-2004)
8
6
4
2
0
I II III IV V VI VII VIII IX X XI XII
months
Borsa
Luna de Jos
Maia
Figure 5. Monthly floods frequency in Cluj- Dej Hills (1976-2004)
The monthly frequency of floods formation presents a maximum point in May, on the
Almaş and Agrij rivers, in the Almaş-Agrij Depression (15 – 23% of the entire number
of sorted floods), followed by March and June (with the same number of floods). The
minimal frequency of floods or even their absence was recorded in November (Fig. 6)
520

%
14
12
10
8
6
4
2
0
I II III IV V VI VII VIII IX X XI XII
months
Almasu
Hida
Romanasi
Figure 6. Monthly floods frequency in Almas- Agrij Depression (1976-2004)
Figure 7 shows the maximum debits on rivers in Almaş-Agrij Depression. Here the
maximum debits had values between 150m 3 /s on Almaş river, in Hida, and 220 m 3 /s, on
Agrij river, in Românaşi. The highest risk of inundations took place in the year 1989
and ten years after, in 1999, when the maximal debits of floods have been recorded. In
1982 and 2001 the risk was serious, too. However the maximal debits of floods had
minimal values in some years (such as in 1984).
250
200
150
100
50
0
1981
1982
1983
1984
1985
1986
1987
1988
1989
1990
1991
1992
1993
1994
1995
1996
1997
1998
1999
2000
2001
2002
2003
2004
Almas
Hida
Romanasi
Figure 7. Maximum debits of floods on rivers in Almas-Agrij Depression (1981-
2004)
The maximum debits determined in Cluj and Dej Hills within years 1976 and 2004 had
values between 44.1 m 3 /s on the Nadăş river, in Aghireş, and 146 m 3 /s on Lonea river,
in Luna de Jos.
In the Almaş-Agrij Depression and in Cluj and Dej Hills the multiannual maximum
debits have varied between very large limits, depending on numerous factors
(climacteric conditions, receiving basin surface etc.) The maximum debits were
521

ecorded with a high level frequency on the streams in the Cluj and Dej Hills and in the
Almaş-Agrij Depression in May and June.
On most rivers in Almaş-Agrij Depression and Cluj and Dej Hills the most
considerable frequency represented those floods that maintained their total duration
between 5 and 15 days (30 – 45 % of all cases). The streams that had more frequent
floods with duration over 20 days (Borşa river) were excepted. The floods with a less
duration than 5 days had the most considerable frequency in Agrij, Almaş and Nadăş
basins, being determined by the torrential nature of the rainfalls, the reduced surface of
the basin (Nadăş to Aghires, 39.1 km 2 ) and by the relative large slope.
The increasing time of the floods is different, depending on the nature of rainfalls,
varying, in the analysed rivers, between 4 and 130 hours.
The maximum debit, increasing time, total time and the volume of floods are the
main elements of the flood hydrograph. They constitute the bench mark for flood.
CONCLUSIONS
The results presented above let us come to the following conclusion: the knowledge
regarding floods would help solving several issues like: the understanding of the
nature, the development of some protection or adaptation programmes etc. Establishing
the frequency and the amplitude of floods constitutes very important aspects
concerning the assessment of risk of inundations.
After the floods, it is well known that the set back to normal conditions and the
rehabilitation of the damaged environmental factors involves material effort and
important collective actions, sometimes difficult to perform (Stanescu, Drobot, 2002).
REFERENCES
1. Mustăţea, A., (2005), Viituri Excepţionale pe teritoriul României, Bucureşti;
2. Pandi, G., (2002), Riscul în activitatea de apărare împotriva inundaţiilor, Riscuri şi
Catastrofe, vol. I, Editura Casa Cărţii de Ştiinţă, Cluj-Napoca, p. 131-142;
3. Sorocovschi, V., (2002), Hidrologia uscatului, vol II, Editura Casa Cărţii de Ştiinţă,
Cluj-Napoca;
4. Sorocovschi, V., (2005), Câmpia Transilvaniei – Studiu hidrogeografic, Editura Casa
Cărţii de Ştiinţă Cluj-Napoca;
5. Stănescu, V. Al., Drobot, R., (2002), Măsuri nestructurale de gestiune a inundaţiilor,
Editura HGA Bucureşti;
6. Studii Hidrologice Anuale, 1976-2004, Arhiva Direcţiei Apelor Someş-Tisa, Cluj-
Napoca.
522

RESEARCH REGARDING MAIN CHEMICAL REACTIONS IN
PREDEFECATION OF DIFFUSION JUICE
Gheorghe Sarca
University of Oradea, Faculty for Environmental Protection
ABSTRACT
The quality of sugar is the decisive factor in ensuring the sugar commercializing
through the conservation of old markets and expansion towards new markets. The sugar
quality is also the key element which determines the price maintenance in according
with the expenditures performed or even the diminution of the price as a consequence of
the quality lessening.
INTRODUCTION
The main chemical reactions occurred in the predefecation of diffusion juice are the
following:
• Neutralisation of free acids, especially the neutralisation of the organic acids of
sugar beet and of the mineral acids contained in the extraction water:
2 R COOH + Ca(OH)2→ (R COO) 2Ca + H2O
• Ions exchange between magnesium and calcium which can be mainly described by
the intermediate of a chemical reaction such as:
2 R COOMe + Ca(OH)2+ Ca(OH)2→ (R COO) 2Ca+2MeOH
At pre-defecated juice defecation the following chemical reactions take place:
R CONH2 + H2O → R COONH4
H2O + R CONH2 → R COOH + NH2
H2O + CO2 → H2CO3
H2O + CaO → Ca(OH)2
Reduction substances + Ca(OH)2 → Colouring Substances + Organic Acids
During defecated juice carbonizing the following reactions take place:
CO2 + H2O → H2CO3
CO2 + OH → HCO?
H2Co3 + Ca(OH)2 → CaCO3 + H2O
An important role in providing the adequate conditions for the development of these
reactions is taken by the calcium oxide solubility in water and in the sugar-in-water
impure solutions. As higher level of calcium oxide solubility is as better the purification
effect would be. In the chart 1 the solubility of the oxide calcium solved in water and in
sucrose solutions is presented.
523

Table 1.The solubility of the calcium oxide in water
and in the in sucrose solutions in water
Sucrose The solubility of the calcium oxide in the in sucrose solutions in
water, solution expressed in g CaO/100 mg, at temperature: [ºC]
g/100 ml
water
0 20 40 60 80
0,0 0,135 0,126 0,109 0,088 0,071
1,5 0,252 0,175 0,132 0,118 -
3,0 0,476 0,280 0,200 0,145 -
6,0 1,172 0,660 0,310 0,212 0,130
12,0 2,082 1,969 0,936 0,422 0,204
18,0 4,140 3,553 1,942 1,168 0,356
MATERIALS AND METHODS
In order to obtain the largest amount of high-quality crystal sugar from the diffusion
juice its purification and removing the non-sugar are required. Actually, the current
systems designed for diffusion juice purification ensure a non-sugar parts removal
reaching at max. 45% of the non sugar of diffusion juice.
With the purpose of diffusion juice purification one must make use of cheap
methods and of those substances which would not alter the sugar and are not to cause
sugar loses while non sugar precipitated with these substances are to be easily separated
out of the juice mass and subsequently eliminated. Nowadays, for diffusion juice
purification on utilise calcium oxide and carbon dioxide. That is the reason for which
the current technological process of purification is called “calco-carbonic purification”.
During the technological process of sugar beet processing, the aim of calco-carbonic
purification of the diffusion juice is well settled and established. The amount of non
sugar eliminated during the purification process favorably influences the subsequent
development of the technological process in particular those operations accomplished at
high-level temperatures e.g. the concentration through vaporization, boiling and
crystallization but also the outturn and the quality of the sugar obtained.
The diffusion juice purification may be accomplished through other methods e.g.
ion exchange, ultra-filtration, ion-exclusion, electro-dialysis and inverse-osmosis.
Nevertheless, due to some reasons of economic effectiveness, the diffusion juice
purification obtained by the help of these methods is not used at industrial scale yet.
By definition, diffusion juice purification is the operation which aims the optimal
development of the subsequent phases of the technological process. The phases and the
operations which constitute the calco-carbonic purification process and the
technological parameters at the level of which these operations are executed are all
established function of the technological quality of the sugar beet and of the diffusion
juice processed. The purification of the diffusion juice must always have a positive
influence upon the entire production process’ effectiveness, an influence materialized as
following:
524

• Ensuring the juice’ thermal stability at concentration through vaporization, a
stability materialized by the avoidance of Ph dropping, by preventing juice intense
coloring and by the sucrose hydro-dialysis avoidance;
• Avoiding the intense frothing which may lead to great losses of sucrose along with
the froth which is produced;
• Minimal losses of sucrose as a consequence of its instability and its thermal
decomposition during its concentration through vaporization;
• Minimal values for the sugar remained in molasses;
• Ensuring the quality sugar production in order to maintain its stability during
depositing;
• Lending itself for subsequent use in the industries employing crystal sugar.
In the chart 2 the influence of the calco-carbonic purification upon the diffusion juice
non sugar is sketchily presented.
Non sugar of the diffusion juice
Non sugar Non sugar Non sugar Non sugar
non influenced precipitated decomposed absorbed
by by by and adsorbed
Calco-carbonic calcium oxide Ca(OH)2 and by limestone
Purification temperature
Sugar
remained
Sugar non-absorbed and non-adsorbed by Sugar reabsorbed
in thin juice the calcium carbonate (limestone) out of the
limestone
Chart 2 The action of the calco-carbonic purification upon the diffusion juice’
non-sugar and the non-sugar circulating during the diffusion juice purification
The amount of sucrose remained in molasses depends upon the composition and
concentration of the thin juice’ non-sugar. That is why, the elimination of the non-sugar
from the diffusion juice is a condition sine qua non for obtaining some crystal sugar’
good outturns.
525

The classical calco-carbonic purification of the diffusion juice consists in the juice’
treatment with lime milk and the subsequent precipitation of the excess (surfeit) of
Ca(OH)2. The resulted mud’ separation raises some special problems. That is the reason
for which, in practice, there have been introduced some purification methods to which
some compromises are being made between the purification effect and the separation
capacity, through decantation or through mud filtering.
RESULTS AND DISCUSSIONS
The technological role of the diffusion juice’ calco-carbonic purification in the process
crystal sugar obtaining consists in the elimination of a part of the diffusion juice’ nonsugar
substance employing some operations specific to each non-sugar groups and
which would provide ,at the given momentum, some optimal conditions for the
destruction and elimination of a certain sugar type. Punctually, the role of the calcocarbonic
purification consists in:
• Neutralizing the acidity of the diffusion juice and precipitating the calcium salts;
• Precipitating and separating diffusion juice’ colloids;
• Decompounding the reducing substances (agents) and the amides and obtaining the
heat-proof (thermo stable) juices;
• Non-sugar absorbing and adsorbing on the calcium carbonate (limestone)
• Mud decanting (separating) out of the purified juice;
• Sweetening the mud;
• Thin juice decalcifying and preventing the concentrating station’ peeling through
vaporization.
In order to obtain the diffusion juice purification one employ calcium oxide under the
lime milk shape and carbonadoing gas containing carbon dioxide.
In the chart 3 there are represented the characteristics of the lime milk used for
purification in sugar factories.
Table 3. The characteristics of the lime milk used for
calco-carbonic purification in sugar factories
Concentration in dry Specific CaO content Ca(OH)2 content
substance mass
ºBe ºBx g/cm 3 g/l g/100g g/l g/100g
1,49 2,69 1,0084 9 0,98 13,1 1,30
4,69 8,39 1,0314 39 3,87 52,7 5,12
7,59 13,69 1,0534 69 6,64 92,4 8,78
10,34 18,69 1,0749 99 9,29 132,0 12,28
15,54 28,19 1,1184 159 14,29 211,3 18,89
20,34 37,09 1,1614 219 18,93 290,6 25,02
25,49 46,79 1,2124 289 23,91 383,1 31,60
For an accurate evaluation of the function of the diffusion juice purification’ station
from a sugar factory it is necessary to be aware of the exact amount of carbonadoing
526

mud which is evacuated out of the process. This quantity depends upon the calcium
dioxide dose actively employed for purification and it has a direct influence upon the
following several indicators:
1. The sugar losses in the mud;
2. The calco-carbonic purification effect of which the efficiency of the purification
depends;
3. Providing the adequate conditions for separation thorough decantation and mud
filtering out of the turbid juice from 1 st (I) saturation.
REFERENCES
1. Banu C., 1992 – „Progrese tehnice, tehnologice şi ştiinţifice în industria
alimentară”, Ed. Tehnică, Bucureşti.
2. Beceanu Dumitru, Chira Adrian, 2003 – „Tehnologia produselor agricole”, Ed.
Economică, Bucureşti.
3. Sarca Gheorghe, 2004 – „Materii prime vegetale”, Ed. Universităţii din Oradea.
4. <strong>International</strong> Sugar Journal, 2006 december.
5. Popescu V., Stroia A., 1998 – „Tehnologii moderne de industrializarea sfeclei de
zahăr”, Ed. Ceres, Bucureşti.
527

DETERMINATION OF NITROGEN-CONTAINING POLYNUCLEAR AROMATIC
HYDROCARBONS IN SMOKED FOODS BY LIQUID CHROMATOGRAPHY
Lucian Bara
University of Oradea, Faculty of Environmental Protection
26, General Magheru Street, Oradea
ABSTRACT
Several smoked foods were analyzed for basic nitrogen-containing polynuclear aromatic
hydrocarbon (NPAH) content by a relatively rapid liquid chromatographic (LC) technique.
The analyzed products included both domestic and imported market basket commodities.
Nanogram quantities of NPAH standards were detected by UV and fluorescence detectors
connected in series. The NPAHs were extracted from basic aqueous ethanolic solution into
cyclohexane, extracted from cyclohexane into 6N HCl, and extracted back into cyclohexane
after neutralization of the acid.
The NPAHs were then purified by filtering the extract through deactivated basic
alumina. The eluate from this step was concentrated to dryness, and the residue was
dissolved in 95% ethanol and analyzed by LC, using a Vydac C-18 column and acetonitrilewater
(9 + 1) as the mobile phase. Recoveries of 3 NPAHs, 5,7-dimethylbenz acridine,
dibenz acridine, and dibenz acridine, each added to salmon and sausage at the 5 ppb level,
ranged from 62 to 101% by fluorescence measurement and from 64 to 106% by UV
measurement. None of the NPAHs used, as standards were found by either fluorescence or
UV detection at levels greater than or equal to 5 ppb in any of the foods analyzed.
INTRODUCTION
Interest in the production of petroleum substitutes from shale oil and coal has increased. The
increasing use of coal and synthetic fuels may significantly increase the levels of
heterocyclic nitrogen compounds in the environment because of the charasterically higher
level of organic nitrogen in coal and its by-products, compared to other fossil fuels. Many
nitrogen bases, thought to be products of incomplete combustion of organic matter that
contains nitrogen or to be derived from hydrolytic processes similar to those producing
polynuclear aromatic hydrocarbons, have been shown by various types of skin tests to be
toxic, carcinogenic, and mutagenic to terrestrial and aquatic organisms.
Many azaarenes have been identified in sources such as tobacco, automobile exhaust,
high boiling petroleum distillates, crude oil, shale oil, and lake sediments.
The purpose of our investigation was to develop an analytical method for the
extraction, cleanup, and LC analysis of the basic NPAH fractions obtained from some
smoked foods. The foods analyzed included both domestic and imported products.
528

MATERIALS AND METHOD
Apparatus
Clean all glassware with chromatic acid/sulfuric acid solutions, rinse thoroughly with
distilled water, oven-dry at 120 o C, and solvent-rinse before use to minimize interferences.
a) Meat chopper – Launders, Frary and Clark Co. or equivalent.
b) Digestion apparatus – 1) Heating mantle – 500 mL; 2) Friedrich condenser; 3)
Boiling flask
c) Separatory funnels
d) Flasks
e) Flash evaporator
f) Filter funnel
g) Sample clarification kit
h) Liquid chromatograph
i) Recording data processors
Reagents
a) Solvents – Ethanol, 190 proofs, distill before use; cyclohexane, distilled-in-glass;
acetonitrile, LC grade; double-distilled water.
b) NPAH standards – 5,7-Dimethylbenze acridine, dibenz acridine. Each standard
was purified by reverse phase LC, and the identity of each was verified by GC/MS
before use in this study.
c) Standard solutions – 1) Stock solutions – 0.1 mg/mL. Dissolve 1 mg of each
NPAH standard in 10mL ethanol. 2) Working standard solution dilute to volume with
ethanol.
d) Alumina – aluminum oxide 90, active basic, activity stage 1
e) Deactivated alumina – Prepared from alumina above
f) Solutions – 6N HCI, 10N KOH, ethanol-methylene chloride.
Extraction
Place 25 g homogenized smoked food into 500 mL boiling flask. Add 100 mL ethanol, 4 g
KOH, and boiling chips. Insert Friedrich condenser and reflux at rapid rate for 2 h. let digest
cool for 20 min, remove condenser, and filter contents of flask through ethanol-rinsed glass
wool pad into 1 L separatory funnel. Shake funnel vigorously for 2 min. let layers separate;
then draw lower aqueous layer into second 1L separatory funnel. Repeat extraction with 50
mL cyclohexane.
Add 20 mL 6N HCI and shake funnel for 2 min. let layers separate and draw off
bottom acidic layer into second 1L funnel. Repeat extraction in first funnel with 20 mL
portions of 6N HCI, and combine each extract with first extract in second funnel. Discard
cyclohexane layer remaining in first funnel.
Carefully add 50 mL 10 N KOH to acidic extracts while cooling funnel under cold tap
water. Add 50 mL cyclohexane and shake funnel for 2 min. Draw off lower aqueous layer
into second funnel and repeat extraction with 50 mL cyclohexane.
529

Liquid Chromatography
Inject 20 μL concentrate or standard solution onto Vydac 201TP54 column and run mobile
phase consisting of acetonitrile-water at flow rate of 1 mL/min. Between injections, flush
injection loop and port passages with 3 mL acetonitrile-water to prevent crosscontaminaton.
For tentative identification, compare retention times of any peaks observed
with those of known NPAH standards chromatographed under same conditions.
RESULTS AND DISCUSSIONS
All of the domestic smoked foods analyzed in this study were previously analyzed for their
polynuclear aromatic hydrocarbon content; ham, bacon, sausage, whiting, salmon, and trout
represent this group. Some important smoked foods were also analyzed; oysters, clams,
sardines, red snapper, and brislings represent this group. The 3 NPAHs used as standards for
this study were chosen primarily because they were readily available and easily purified.
These compounds represent some of the carcinogenic basic NPAHs, which might be found
in contaminated smoked foods.
CONCLUSIONS
A relatively short, straightforward LC method has been developed for the analysis of several
smoked foods for NPAHs. Although no NPAHs were isolated from any of the smoked foods
analyzed, the recovery data indicate that the method is reliable. A study of the distribution
ratios of DMBaAc, DbajAc, and DbahAc between cyclohexane and the aqueous ethanolic
saponification digest indicates that the best recovery of the azaarenes occurred when the
water-ethanol ratio was 1:1. Future studies will include oil-contaminated foods.
REFERENCES
1. Buchanan, M. V., Ho, C., Guerin (1981) in Chemical Analysis and Biological Fate:
Polynuclear Aromatic Hydrocarbons, Battelle Press, Columbus, OH, p. 133-144.
2. Hertz, H. S., et al. (1980) Anal. Chem. 52, 1650-1657.
3. Colin, H., Schmitter, J., & Guiochon, G. (1981) Anal. Chem. 53, 625-631.
4. Joe, F. L., Jr., Salemme, J., & Fazio, T. (1984) J. Assoc. Off. Anal. Chem. 67, 1076-
1082.
530

THE IMPORTANCE OF VOLATILE N-NITROSAMINES IN FOOD-
PRODUCTS
Lucian Bara, Camelia Bara, Vasile Bara, Dana Marele
University of Oradea, Faculty of Environmental Protection
26, General Magheru Street, Oradea
ABSTRACT
A variety of food products containing nitrite were analyzed for 14 volatile N-nitrosamines
by using a method demonstrated to be sensitive to 10 ppb. A total of 121 food samples were
screened for volatile N-nitrosamine content. N-Nitrosopyrrolidine was confirmed in fried
bacon at levels up to 139 ppb. N-Dimethylnitrosamine, N-nitrosopyrrolidine, and Nnitrosopiperidine
were also confirmed in spice-cure mixtures at levels ranging from 50 to
2000 ppb.
INTRODUCTION
The many publications in the literature during the last years pertaining to the study of
volatile Af-nitrosamines in the environment and in food supplies attest to the concern of the
scientific community with these potent carcinogens. It has been shown that nitrosamines are
formed by the action of nitrite on secondary and tertiary amines. The widespread use of
nitrite as a preservative and color fixative, coupled with the natural occurrence of amines in
foods, prompted the surveillance of products so preserved to determine the extent of these
compounds in our food supply. The food products investigated to date include alcoholic
beverages, bacon, fish, frankfurters, ham, spinach, bologna, prepared meat products, and
meat spice-cure mixtures. This report continues the study of volatile N-nitrosamines in our
food supply. The results of the analysis of 121 samples are given.
MATERIALS AND METHOD
Apparatus and Reagents: The silica gel and the solvents, methylene chloride, pentane, and
ethyl ether, were purified and tested prior to use to ensure the absence of interfering peaks.
Procedure: Fourteen compounds can be determined: N-nitroso-dimethylamine, -
methylethyl-amine, -diethylamine, -methylpropylamine, -ethyl-propylamine, -
dipropylamine, -ethylbutylamine, -propylbutylamine, -methylamylamine, -dibutylamine, -
piperidine, -pyrrolidine, -morpholine, and -diamylamine.
Briefly, the comminuted food sample is digested in methanolic potassium hydroxide
and the nitrosamines are extracted with methylene chloride in a liquid-liquid extractor,
distilled from alkaline solution, further isolated by extraction and column chromatographic
techniques, and determined by gas-liquid chromatography (GLC), using a modified
thermionic detector.
531

When a peak was observed at the retention time of one of the 14 nitrosamines, the identity
of this compound was confirmed by mass spectrometry (MS). To ensure adequate cleanup
for this analysis, the concentrated methylene chloride eluate from the silica gel column was
passed through an acid-Celite column.
The identities of the nitrosamines isolated were confirmed with a combined GLC-MS
system. A Varian Model 5600X, 2-stage membrane separator was interfaced to an Atlas
CH-4 mass spectrometer. The separator is an integral part of the Varian Model 1700 gas
chromatograph. This unit has the facility of venting the solvent to the atmosphere and
maintaining the MS vacuum. Accordingly, large quantities of solvent can be injected onto
the GLC column, vented, and prevented from entering the mass spectrometer. In this study,
30 /A of the final methylene chloride extract (equivalent to 100 ng nitrosamine) was injected
onto the GLC column.
RESULTS AND DISCUSSION
Before the recovery studies, the individual food products were analyzed for the 14 volatile
N-nitrosamines. In instances where a nitrosamine was suspected, recovery values were ad
justed appropriately. As shown in Table 1, recoveries of the 14 volatile N-nitrosamines
added at 10 ppb (/µg/kg) to 25 g of the food products ranged from 67 to 100%, with an
overall average of 84%.
A total of 121 food samples were analyzed during this survey, including bacon, lard, baby
food, total diet samples, miscellaneous pork products, spice-cure mixtures, imported
cheeses, and Icelandic national foods.
Table 1. Recoveries of nitrosamines a added at levels of 10 ppb to 25 g food product
Rec., %
Sample Product Range Av.
1 Bacon, hams, other pork
products (liver) 77-95 82
2 Baby foods 76-95 83
3 Cheeses (imported) 71-94 85
4 Fats and oils 67-100 83
5 Total diet products 75-100 86
6 Spice-cure mixtures 74-87 81
7 Icelandic foods 84-100 91
a 14 volatile A/-nitrosamines as described in text.
All bacon samples were purchased at local retail markets. Bacon was analyzed raw and also
after frying at 340 °F in an electric frypan for 3 min on each side. Excess fat was removed
from the bacon by blotting with paper toweling. As shown in Table 2, N-nitrosopyrrolidine
was found in fried bacon at levels ranging from 7 to 139 ppb.
532

The fat cooked out of some of the bacon samples was analyzed and was found to contain
from 19 to 92 ppb N-nitrosopyrrolidine. All values in Table 2 were confirmed by combined
GLC-MS, except Sample 16, which had a low concentration.
Table 2. N-Nitrosopyrrolidine (ppb) found in commercial bacon
Sample Raw Fried Fat cooked out
1 0 67 92
2 0 91
3 0 74, a 64 a 111," 129*
4 0 30
5 0 119, 139
6 0 129, c 104 c
7 0 33, 33, 33 47
8 0 34 48
9 0 10
10 0 16, 13 24
11 0 10
12 0 28, 29
13 0 42, 51
14 0 28,30 55,44
15 0 22, 20 19
16 0 7, d 7 d
17 0 23, b 19 c
18 0 13, 12, 13
19 0 8
20 0 42,34 49,44
21 0 36, c 29 c
22 0 65, 59, 72, 64
a
Held at 72°F for 8 hr.
b
Held at 72°F for a total of 34 hr.
c
Held at 72°F for 36 consecutive hr.
d
Not confirmed.
No nitrosamines were found in the raw bacon. One sample of dry-cured bacon (not shown
in the table) was analyzed; no nitrosamines were found in either the fried or the raw product.
It is evident that pan-frying does induce the formation of N-nitrosopyrrolidine in bacon, but
the actual mode of formation during the cooking process has not been definitely established.
It may be through the formation of nitrosoproline from proline and nitrite and subsequent
decarboxylation to nitrosopyrrolidine, or by direct interaction of pyrrolidine, which could
arise from proline or putrescine and nitrite.
Four bacon samples were also analyzed after having been kept at room temperature
(72°F) for 34-36 hr, before frying. It has been suggested that bacterial action on meat tissue
could result in higher concentrations of polyamines. In the presence of nitrite these could, in
533

turn, react to produce larger concentrations of N-nitrosamines upon frying. Samples 3, 6, 17,
and 21 in Table 2 were held at 72°F 34 hr above
that of Samples 2 and 16, while Samples 6 and 21 showed little or no increase. A reason for
these apparent inconsistencies could be the amount of residual nitrite present in the bacon.
Producers use different concentrations of nitrite in their cures. Bacon with a low residual
nitrite does not show an increase in nitrosamine formation after storage at room temperature,
since there is an insufficient quantity of nitrite available to react with the additional amines
formed. On the other hand, bacon with a high residual nitrite, which can continue to react
with the amines as they form at room temperature, may show a greater formation of
nitrosamines. After frying, the bacon was found to contain increased levels of Nnitrosopyrrolidine.
Table 3 shows the results of N-nitrosamine analyses on a variety of food products. Two
types of baby foods that contained meat cured with nitrite were analyzed, but no
nitrosamines were found (Samples 27 and 28). Of the 5 variety meat samples analyzed
(Samples 29-34), only the fried canned bacon (Sample 32) showed N-nitrosopyrrolidine to
be present. A peak at the retention time of N-nitrosodimethylamine (DMNA) was also
observed but the concentration was too low to confirm its identity.
Six total diet samples were analyzed (Samples 35-40) comprising (1) grain and cereal
products; (2) meat, fish, and poultry; (3) leafy vegetables; (4) root vegetables; (5) oils, fats,
and shortenings; and (6) dairy products. (These composites represent the typical diet of an
American male teenager between 15 and 19 years of age.) Because of the physical nature of
the oils, fats, and shortenings sample (Sample 40) and the dairy products sample (Sample
35), they were analyzed by the procedure for fat cooked out of bacon. None of the 6 total
diet samples analyzed was found to contain N-nitrosamines.
Since lard is produced from rendered pork fat (in some instances a cured product) and
high levels of N-nitrosopyrrolidine have been found in fried bacon and fat cooked out of
bacon, it was decided to analyze a sample of lard (Samples 41 and 42). No N-nitrosamines
were found, even after heating at 340°F for 10 min.
Several workers have reported finding DMNA and N-nitrosodiethylamine in cheese,
which had been processed with nitrite. However, none of the data had been confirmed. In
order to verify these findings we surveyed 10 imported cheeses (Samples 43-52). None of
the 14 volatile N-nitrosamines was found in any of these products.
Lamb is prepared by soaking it in a similar salt solution and further treated by smoking
for 6-8 hr over sheep manure. The high concentration of amines normally found in fish
coupled with the added nitrite and unusual mode of food processing should have provided
an ideal environment for nitrosamine formation. As shown in Table 3, 18 different samples
were analyzed (Samples 53-70). Despite the apparently ideal environment, no nitrosamines
were found in any of the samples.
534

A problem concerning the formation of nitrosamines in meat spice-cure mixtures has
recently been investigated. Salts, including nitrite, are added to a variety of spices in the
formulation of meat spice-cure mixtures. Amines present in spices could react with the
added nitrite and form nitrosamines. It was confirmed DMNA, N-nitrosopyrrolidine, and Nnitrosopiperidine
in meat curing mixtures at levels up to 25 ppm.
Until recently spice-cure mixtures have been marketed in several forms and formulations,
such as single packaging of spices and curing salts, "piggy-back" (separate packaging of
spices and curing salts), with and without sodium carbonate buffers, and with and without
protective coating of the nitrite crystals (zein corn protein).
A survey of 32 spice-cure mixtures was undertaken to determine the effectiveness of
the 3 marketing processes and also to determine the effect of storage time and temperature
on nitros-amine formation. The 9 samples found to contain nitrosamines are shown in Table
4. All data in the table were confirmed by MS.
Three samples were part of a time-temperature stability study. Each was buffered with
sodium carbonate. The spice-cure mixtures were analyzed at room temperature and 100°F at
0, 30, and 60 days storage. Samples 1 and 2 in Table 4 were nitrosamine-free when first analyzed,
but after heat and storage, both contained N-nitrosopiperidine from 150 to 420 ppb
and N-nitrosopyrrolidine from 50 to 95 ppb. The third sample (a control not shown in Table
4) did not contain nitrite and, as expected, no nitrosamines were found.
Table 3. Food products analyzed: no nitrosamines detected
Sample No. of analyses Product
Pork Products
23
24
25
26
27
28
29
30
31
32a
33
34
2
1
1
1
5
6
2
2
1
1
1
1
pork liver (raw)
pork liver (fried)
Polish style sausage (raw)
Polish style sausage (boiled)
Baby Foods
beans, potatoes, and ham
casserole creamed potatoes and ham
Variety Meats
Crumbled bacon
crumbled ham
bacon (imported canned; raw)
bacon (imported canned; fried)
beer sticks
smoked pepperoni
Total Diet Samples
35 1 dairy products
535

36
37
38
39
40
41
42
1
1
1
1
1
1
1
meat, fish, and poultry
grain and cereal
leafy vegetables
root vegetables
fats, oils, and shortenings
Fats and Oils
lard (uncooked)
lard (heated) b
Cheese
43 2 Edam
44 2 Gouda
45 2 Samsoe
46 2 Danbo
47 2 Gouda (baby)
48 2 Blue
49 2 Tilsit Sq. Havarti
50 1 Port Salut
51 1 Tybo
52 1 Tilsit Havarti
53
54
55
56
57
58
59
60
61
62
63
64
65
66
69
70
1
1
2
2
1
1
1
3
1
2
2
1
1
1
1
1
Icelandic National Foods
smoked lamb's leg
salted lamb
smoked mutton sausage
singed lamb's head
smoked salmon
smoked lamb's leg
singed lamb's leg
smoked mutton sausage
smoked trout
putrescent skate
singed lamb's head (oil flame)
smoked lamb (birchwood)
smoked lamb (beechwood)
smoked salmon
salted lamb
smoked mutton sausage
a Contained 6 ppb A/-nitrosopyrrolidine, confirmed by GLC-MS. b Heated at 340°F 10 min.
536

Sample 3, found to contain 343 ppb DMNA, 730 ppb N-nitrosopyrrolidine, and 2000 ppb
N-nitrosopiperidine, was a spice-cure mix being used at the time in a cured meat product.
From the data shown in the table it is evident that buffering with sodium carbonate does not
block nitrosamine formation. Samples 4, 7, and 9 were buffered and still DMNA, Nnitrosopiperidine,
and N-nitrosopyrrolidine were found. Ascorbate was also found to be
ineffective in blocking nitrosamine formation as shown by Samples 7 and 8, which
contained this compound and yet had levels of N-nitrosopiperidine and N-nitrosopyrrolidine
up to 232 ppb. Similar results with ascorbate were obtained. Two samples of spice-cure
mixtures utilizing the piggyback method of packaging were analyzed. No nitrosamines were
found. A limited study of 4 samples of spice-cure mixtures containing nitrite crystals coated
with zein were analyzed and all 4 were found to be free of nitrosamines. Our studies have
indicated that piggy-back packaging is the most effective method of preventing nitrosamine
formation, since the react ants do not come in contact until they are mixed with the
comminuted meat. Federal laws in the United States and Canada now prohibit the marketing
of meat spice-cure mixtures in any way other than the piggy-back method.
Table 4. Analysis of spice-cure mixtures for nitrosamines (ppb)'
Buffer 0 days 30 days 60 days
Sample 72°F 72°F 100°F 72°F 100°F
1 Na2C03 0 0 0 0 50 NPYR
2 Na,C03 0 0 150 NPIP 0 420 NPIP 95
3 Na2C03 343
730
2000
DMNA
NPYR
NPIP
4 Na4P207 72 DMNA
5 none 106
314
28
6 none 768
144
DMNA
NPIP
NPYR
NPIP
NPYR
7 b Na2C03 232 NPIP
d b none 160 24 NPIP
9 Na2C03 29 91
49
NPYR
DMNA
NPIP
NPYR
a DMNA, N-nitrosodimethylamine; NPYR, N-nitrosopyrrolidine; NPIP, A/nitrosopiperidine.
b With ascorbate added.
537
NPYR

REFERENCES
1. Castegnaro, M., Pignatelli, B., & Walker, E. A. (1974) Analyst 99, 156-162
2. Collins-Thompson, D. L., Chang, P. C, Davidson, C. M., Larmond, E., & Pivnick, H.
(1974) /. Food Sci. 39, 607-609
3. Kushnir, I., Feinberg, J. I., Pensabene, J. W., Piotrowski, E. G., Fiddler, W., &
Wasserman,
4. Sen, N. P., Iyengar, J. R., Donaldson, B. A., & Panalaks, T. (1974) J. Agric. Food
Chem. 22, 540-541
5. Sen, N. P., Schwinghamer, L., Donaldson, B. A, & Miles, W. F. (1972) J. Agric.
Food Chem. 20, 1280-1281
6. Fiddler, W., Doerr, R. C, Ertel, J. R., & Wasserman, A. E. (1971) JAOAC 54, 1160-
1163
7. Heisler, E., Siciliano, J., Krulick, S., Fein-berg, J., & Schwartz, J. H. (1974) J. Agric.
Food Chem. 22, 1029-1032
8. Palumbo, S. A., Smith, J. L., Gentilcore, K. M., & Fiddler, W. (1974) /. Food Sci. 39,
1257-1258
9. Sen, N. P, Miles, W. F., Donaldson, B. A, Panalaks, T., & Iyengar, J. R. (1973) Nature
London 245, 104-105
538

LUNG CANCER CAUSED BY SMOKING AND CADMIUM AND CHROMIUM IN
HUMAN LUNG TISSUE
Camelia Bara
University of Oradea, Faculty of Environmental Protection
26, General Magheru Street, Oradea
ABSTRACT
The pulmonary Cd and Cr content were determined from 53 lung cancer patients operated
for cancer and from 39 patients who died of non-malignant diseases. The results were
correlated with smoking habits, pulmonary emphysema and occupational history. Both the
pulmonary Cd and Cr increased with the amount of smoking. In ex-smokers the Cr content
in lung tissue did not diminish with the time since stopping smoking, but the Cd did follow
the half-life of about 9 years. The pulmonary Cd, compared with smoking habits, behaved
similarly in both the lung cancer and control patients, whereas Cr in the lung cancer patients
could not be explained solely by smoking, but some of the cancer patients may have been
occupationally exposed to Cr. Speciation was not studied.
Keywords: Cadmium, chromium, smoking, occupational hygiene, lung cancer, pulmonary
emphysema, lobe samples, sample preparation, direct current plasma atomic emission
spectrometry (DCP-AES).
INTRODUCTION
Lung cancer patients have usually been exposed to tobacco smoke for several decades. In
addition to organic compounds, tobacco contains carcinogenic metals such as As, Cd, Cr
and Ni. These metals may function as an indication of tobacco smoking in lung tissue, and
in this respect especially Cd 2 and Cr 3 have proved promising. The main source of Cd in
tobacco is considered to be phosphate fertilizers. Cr is a common environmental pollutant
originating from the burning of coal and from metallurgical chromium and cement
producing plants, and concentrating in tobacco leaves from the contaminated soil. In this
work the pulmonary Cd and Cr content was compared with the smoking habits and
occupational history of patients with and without lung cancer.
Table 1. Age and smoking habits of the lung cancer and control patients
Cancer Control
Number 53 39
Age 61 ±10 67±9
Smoking time (years) 39+11 37± 12
Pack years 41 + 15 36+17
Time since stopping smoking (years) 4 + 7 8±11
539

MATERIALS AND METHODS
The material consisted of 53 male lung cancer patients operated for the cancer, and of 39
control patients who died of non-malignant diseases and were autopsied. Because
pulmonary emphysema is a good indicator for smoking, the control material was collected
to include cases with various degrees of emphysema. The smoking habits and occupational
history were required from the patients or next of kin and from the hospital records (Table
1). The severity of emphysema was graded in the radiographs of excised air-inflated lungs
or lobes of lungs.
Fresh lung tissue for analyses was taken from the (apico) posterior and anterior
segment of the upper lobe and from the superior (apical) segment and basal part of the lower
lobe. The samples, weighing 0.5-2 g, contained no pleural surface or cancer tissue. Ninety
percent of the Cd analyses were made from the anterior segment and 80% of the Cr analyses
from the (apico) posterior segment of the upper lobe. The samples were prepared under a
laminar flow hood avoiding contamination. The fresh, vacuum-dried samples were ashed in
glass cups with a blank and NBS Bovine Liver (SRM 1577) control for 48-72 h in a low
temperature oxygen plasma asher (BIO-RAD Plasma Asher E 2000, 100-120 °C). The
tissue residue was dissolved in a total of 3 ml of 50 % ethanol and transferred into a Teflon
cup and dried at 80 °C for 24 h. The residue was digested in a mixture of 3 ml of
concentrated nitric acid and 0.2 ml of percloric acid. The solution was allowed to stand
overnight and dry-heated at moderate temperature (100-150 °C) in a sand-bath. Finally the
ash was diluted in 3-6 ml of high quality water. The Cd and Cr contents were determined on
a direct current plasma atomic emission spectrometer (DCP-AES) (SpectraSpan IIIB
coupled with a Hewlett-Packard 85 data processor) not considering speciation. A standard
addition method was employed for Cr to eliminate matrix interference. The analytical
characteristics are shown in Table 2.
Table 2. Analytical characteristics
Cd Cr______
Accuracy
NBS 1577 Bovine Liver 0.36 0.25
(0.274 ±0.086) a (0.125±0.057) a
Lanonorm-Metalle 2 0.028
(0.021-0.030) b
RSD (0.02 ppm) % 3.9 2.4
Detection limit (ppm) 0.010 0.017
_________________________________________________________
RESULTS AND DISCUSSION
Both the total pulmonary Cd and Cr content were higher in the lung cancer (Cd 3.0 ± 2.4
and Cr 6.1 ± 4.2 µg/g dry weight) than in the control patients (Cd 2.1 ± 1.9 and Cr 4.1 ± 4.0
540

µg/g dry weight). The pulmonary Cd increased with the amount of smoking measured in
pack years (r = 0.32, p = 0.005) in both the lung cancer and control patients. Cr correlated
positively with the smoking time in the control (r = 0.41, p

FORMATION OF AFLATOXIN M- TRIFLUOROACETIC ACID DERIVATIVE IN
OPTIMUM CONDITIONS
Lucian Bara
University of Oradea, Faculty of Environmental Protection
26, General Magheru Street, Oradea
ABSTRACT
Because thin-layer chromatographic (TLC) confirmation of identity and reverse-phase
liquid chromatographic (LC) determination with fluorescence detection of aflatoxin M1,
both require the derivative formed in the reactions conditions were studied to obtain
complete derivative formation. Of the various organic solvents tested, the reaction between
M1 and TFA preceded best in the nonpolar solvents hexane and isooctane. Other parameters
investigated were reaction temperature and time, aflatoxin M1 concentration, and solvent
volume. The following procedure is considered optimum: 200 µL each of hexane and
trifluoroacetic acid are mixed with M1 standard in a silylated glass vial or with milk residue
in a regular glass vial with a Teflon-lined screw cap and heated 10 min at 40ºC. The mixture
is evaporated to dryness under N2, and the derivative is saved for TLC or LC. No unreacted
aflatoxin M1 was detected by reverse-phase LC after this procedure was incorporated for
analysis of milk samples.
INTRODUCTION
The hemiacetal derivatives of aflatoxins B1 and G1 (B2a and G2a) have been used for thinlayer
chromatographic (TLC) confirmation of identity since 1971. The derivatives are
highly fluorescent and used extensively for the quantitation of these aflatoxins by reversephase
liquid chromatography (LC), because B1 and G1 are only weakly fluorescent in
aqueous mobile phases. Derivatives B2a and G2a are readily prepared by the trifluoroacetic
acid (TFA)-catalyzed hydration of aflatoxins B1 and G1 are at room temperature. This is not
true with aflatoxin M1 because the reaction rarely goes to completion. The first reversephase
LC method for determining aflatoxin M1 in milk and milk products, which measured
the fluorescence of the M1-TFA derivative (designated M2a), was reported by Beebe and
Takahashi in 1980. No formal evidence has been published that definitely identifies this
derivative as the hemiacetal; however, for consistency with the authors, it is reffered to here
as M2a. Beebe and Takahashi were successful in the reproducible derivatization of aflatoxin
M1 to M2; however, other scientists have encountered incomplete derivatization and have
incorporated different reaction conditions. Consequently, some of these workers have
chosen to quantitate aflatoxin M1 directly, even though M1 is less fluorescent than M2a.
In this report, the reaction parameters for derivatization of aflatoxin M1 or M2a are
examined, and the procedure that produces complete conversion of M1 to the M2a derivative
is given.
542

MATERIALS AND METHOD
Apparatus and Reagents
(a) Solvents. – All are reagent grade.
(b) Trifluoroacetic acid (TFA). – Purity 99 + %.
(c) Dichlorodimethysilane (DDS). – Prepare 5% (v/v) solution in toluene.
(d) Aflatoxin standard solutions. – Aflatoxin M1 standards were 0.5, 7.04, and 8.37
μg/mL in acetonitrile-benzene (1 + 9). Aflatoxin B1 and G1 standard solution was 8.0
μg each/mL in acetonitrile-benzene (2+98).
(e) Mobile phase. – Mix water-isopropyl alcohol-acetonitrile (80+12+8). Degas with
ultrasonic probe.
(f) Heating block. – Reacti-therm heating module or equivalent.
(g) Silylated vials. – Add 5% DDS solution to 1-1.5 dram vials and heat ca 40 min at
45-55°C. Discard solution and rinse vials 3 times with toluene and then 3 times with
methanol. Heat vials at 75°C for 20-30 min to evaporate methanol. Cap vials (with
Teflon liners) and store.
(h) LC system. – Spectra-Physiscs Model 8700, equipped with injector (Rheodyne
Model 7125) with 2.0 mL loop, recorder/integrator (Spectra-Physics SP4270),
fluorescence detector (Kratos FS970), set at 365 nm for excitation and 418 nm for
emission, and 4.3 mm id X 25 cm Zorbax ODSLC column. Mobile phase, waterisopropyl
alcohol-acetonitrile (80 + 12 + 8), 1.0 mL/min.
Initial Derivative Formation
The following were mixed in a Teflon-lined screw-cap vial: 200 µL solvent, 50 µL TFA,
and 50 µL aflatoxin M1 standard. The mixture was allowed to react at room temperature for
15 min and then was evaporated under nitrogen. The residue was dissolved in 2000 µL
water-acetonitrile (75 + 25) for LC. The injection volume was 50 µL.
RESULTS AND DISCUSSION
This study was initiated by duplicating the reaction conditions of Beebe and Takahashi.
They used n-hexane as a base solvent to dissolve the dairy extract residue containing
aflatoxin M1. Although hexane is an excellent solvent for this purpose, it is a poor solvent
for the polar aflatoxin M1, when present in pure form (e.g., aflatoxin M1 standard).
Therefore, other solvents were substituted for hexane to determine if any improvement
colud be obtained. Solvents were chosen that scanned the elutropic series, with values of -
0.25 to 0.95. The data indicate that the solvents that produced n-hexane, isooctane, and
1,1,2-trichlorotrifluoroethane. Generally, as the solvent polarity increased, the M1,
conversion decreased. Unreacted aflatoxin M1 was detected with all solvents. When water
was added to the reaction mixture, poor yields of M2a were obtained. Even though aflatoxin
M1 is less soluble in nonpolar solvents, the reaction was more complete with them. In this
study, aflatoxin M1 standard was added to the solvent-TFA mixture in 50 µL aliquots. This
technique prevented adsorption of M1 to the glass vial – a problem encountered with
aflatoxin standard solutions and discussed in more detail later.
543

Next, the optimum reaction temperature was determined for hexane and isooctane as base
solvents. 1,1,2-Trichlorotrifluoroethane was not tested because it is not a common
laboratory chemical. The reaction conditions were identical to those given in the text and for
the data in Table 1, except the temperature was varied from 10 to 50°C. As the temperature
increased, aflatoxin M2a was produced in greater yield until a decrease was detected at 50°C.
The optimum reaction temperature was 40°C. No unreacted M1 could be detected at
temperatures above ambient; however, some must have been present at 30 and 50°C
because less M2a was present. Aflatoxin M1 is not as fluorescent as M2a in polar mobile
phases, so small, undected amounts of M1 could be present.
The reaction temperature was held at 40°C, and hexane was selected as the residue
solvent because most aflatoxin laboratories use it routinely in their assays. No unreacted M1
was found after 15 min, and maximum aflatoxin M2a peak areas occurred at 30-45 min. This
was a longer reaction time than desired, so the TFA volume was increased from 50 to 200
µL, and the experiment was repeated. With equal volumes (200 µL) of hexane and TFA, the
reaction was complete at 10 min, and only M2a, was detected. On the basis of these data, it
appears that the best conditions for forming M2a, from M1 are mixing equal 200 µL portions
of hexane and TFA with the dairy extract residue and letting the mixture react at 40ºC for 10
min.
These reaction parameters were tested with increasing quantities of aflatoxin M1 t
determine the maximum amount of toxin that would react. Only aflatoxin M2a was visible
when quantities of less than or equal to 150 ng or 300 200 µL M1 standard were treated with
TFA. The data suggest that this volume (300 µL) of standard solution diluted the reaction
mixture excessively. To determine if this was true, a concentrated M1 solution (1675 ng in
200 µL) was treated, and a very small unreacted M1 peak-63 area counts – was detected.
Therefore, the quantity of aflatoxin M1 is not a critical factor in the derivatization, but
dilution of the reactants is to be avoided. With most of the current methods, the dairy
product extract is evaporated before it is treated with hexane and TFA; therefore, the
proposed technique will successfully form the M2a derivative.
If aflatoxin M1 standards are evaporated in a glass vial, M1 is irreversibly adsorbed to
the glass. Glass vials should be silylated to avoid adsorption of M1 during evaporation.
Standard solutions (benzene-acetonitrile) of less than 200 µL can be added to the hexane-
TFA mixture directly and the reaction will go to completion.
The derivatization procedure was tried also with standard solutions containing
aflatoxins B1 and G1 and it converted them completely to B2a and G2a. In a recent
international collaborative study, four collaborators obtained incomplete reaction of
aflatoxin M1 with TFA, probably because the specified procedure did not use heat. Since
then, the 4 collaborators have successfully used the procedure given in this paper (private
communications).
In summary, the optimum conditions for converting aflatoxin M1 to aflatoxin M2a are to
add equal 200 µL volumes of hexane and TFA to dry dairy extract in a screw-cap vial, mix
well, heat the vial at 40ºC for 10 min, evaporate the mixture under N2, and save the residue
544

for either TLC or reverse-phase LC determinations. This procedure also will successfully
form aflatoxins solutions should be transferred to silylated vials, prior to forming the
derivatives, to prevent irreversible adsorption of the aflatoxins to the glass; however, sample
extracts do not need silylated vials.
REFERENCES
1. Przybylski, W. (1975) J. Assoc. Off. Anal. Chem. 58, 163-164
2. Beebe, R.M. &Takahashi, D.M. (1980) J.Agric, Food Chem.28, 481-482
3. Tuinstra, L.G.M., Th., &Haasnoot, W. (1982) Fresenius Z. Anal.Chem.312, 622-623.
545

DETERMINATION OF “CARCINOGENESIS” BY ALTERNATIVE BIOASSAYS
Camelia Bara, Lucian Bara
University of Oradea, Faculty of Environmental Protection
26, General Magheru Street, Oradea
ABSTRACT
Chhabra et al. (1988) describe a “carcinogenesis” bioassay that appeared to show the
inhibition of spontaneous tumors by 4-hexylresorcinol in rats and mice. This is not an
unusual finding. It is even possible to find consistent patterns among these
“antitumorigens”. All four of the antidiabetic drugs tested in the program and all but one of
the antioxidants belong to the group with statistically significant dose-related reductions in
spontaneous tumors only; however, the group also includes a dioxin.
Chhabra et al. suggest that their test compound might be investigated as an
antineoplastic agent. Since there are more than 40 other compounds with the same property,
should they also be investigated as antineoplastic agents? If we think of the
“carcinogenesis” bioassay in a symmetric fashion (as capable of showing both tumorigenic
and antitumorigenic activity), a finding that a compound is a potential “carcinogen” have the
same validity as the finding that a compound is a potential “antineoplastic” agent.
In the following sections, an argument is made that, with the current design (maximum
tolerated dose and 18-24 months exposure), it can be expected that every compound tested
will produce an effect on the incidence of tumors, regardless of whether the compound is a
potential “carcinogen”, “antineoplastic” agent, or neither. Next, a statistical procedure is
outlined that can be used to test this hypothesis, and the procedure is applied to six
bioassays that were run on similar compounds in the same laboratory during the same
period. Finally, the implications of this hypothesis for interpretation of these bioassays are
discussed.
INTRODUCTION
The philosophical foundation of toxicology as a science rests on a thought experiment
attributed to the Renaissance alchemist Paracelsus (1493-1541). In this thought experiment,
a potential victim is given ever smaller doses of a “poison” until a dose is found that has no
apparent effect. Then, the potential victim is given ever-larger doses of some “nonpoison”
until serious biological damage results. The conclusion is the aphorism, “Solely the dose
determines that a thing is not a poison”.
There is a modern version of this thought experiment. Let as consider giving a rat or a
mouse a dose of a biologically active drug, such as a B-adrenergic receptor-blocking agent.
Let the dose be large enough to produce almost 100 % blockade. The catecholamines that
the body produces in the normal course of life will fail to have the effects expected, and
homeostasis will be affected, perhaps calling forth greater output of catecholamines. These
agents will then bind to secondary receptor sites, producing responses, which, in turn, will
546

trigger still other responses. In this way, the animal may eventually reach some state of
equilibrium but in the presence of a changed biochemical milieu.
If, now, we continue treating this animal with the 100 % blocking dose for most of its
natural life span, we can expect this changed biochemical milieu to be reflected in the
patterns of lesions that are associated with the aging process. Thus, the treated animals will
tend to have a different syndrome of chronic lesions than untreated controls. If the strains of
rat or mouse chosen are prone to tumor, untreated, then we can expect that this shift in
senile lesions will involve a shift in neoplastic lesions.
Suppose we used some other drug, like e calcium channel blocker, or a monoamine
oxidase (MAO) inhibitor, or any other drug at a dose sufficiently high to produce a
sustained biological effect. The same argument holds, that the resulting change in
biochemical milieu will produce a shift in senile lesions (including tumor patterns).
Suppose, instead of a drug, we use an insecticide, or a dye precursor, or any other agent, but
at a dose sufficiently high ti produce a discernible biological effect (like a 10 % reduction in
weight). The same argument holds.
Thus, we can expect, just from the design of these studies, that the “carcinogenesis”
bioassay will show a treatment-related shift in neoplastic lesions for every compound tested.
MATERIALS AND METHOD
A. Using Data from the “Carcinogenesis”Bioassay
The standard statistical analyses of the “carcinogenesis” bioassay require that we compare
counts of animals with specific tumor types between treatment groups. As a result, the
unsual NTP study involves formal hypothesis tests comparing tumor incidences for 10-15
tumor types, each comparison involving a test of controls versus low dose; controls versus
high dose; two dose-response tests, adjusted for early deaths; and nonlinearity hypothesis
tests per sex per species.
The problem of multiple testing and inflation of the false-positive error rate has had
considerable discussion in the literature. However, there is another side of the problem, the
false-negative error rate. There are only 50 animals per treatment, sex, species, and a
statistical test that compares the number of animals with the a given lesion cannot detect a
slight shift in the probability of lesion. For instance, if the background incidence of a
particular tumor is 20 % and if the treatment reduces that to 7%, half of the studies would
show a “significant” difference and half would not. Thus, the failure to show “significant”
differences between treatments and controls for some of the compounds that have been
tested is no proof that there was no effect. In fact, a rather dramatic increase or reduction in
a particular tumor type could have occurred without producing a “significant” result.
There are ways of strengthening the power of statistical hypothesis tests. One way is to
allow each animal to provide more information. When we compare the counts of animals
with a specific tumor type, each animal provides only 0 or a 1, the least amount of
information available from a single experimental unit. The fact that we look at several
547

different tumor types does not help. In fact, because of multiple testing we have to adjust the
nominal p value and make the test even less powerful. Since the individual animal is the unit
of experimentation, the greatest power can be gained by using the entire pattern of lesions
seen in a given animal and by using only one or two hypothesis tests for the entire sex X
species experiment.
The appendix describes a method of multivariate analysis that does it. General
multivariate methods foe analyzing chronic toxicity studies can be found in Salsburg. The
specific method described in the Appendix was applied to six AZO dyes, whose bioassays
took place at the same site during the same period of time, and these analyses are discussed
in the next section.
If the Paracelsean thought experiment is correct, and with it the hypothesis that any
treatment has an effect, then a powerful statistical test should detect a significant treatment
effect in almost all studies – “almost all” because no statistical test has 100 % power.
B. Constructing Additional Studies to Test the Hypothesis
Statistical tests that show a shift in tumor patterns for all or almost all compounds, sexes,
and species in these studies provide strong evidence in favor of the hypothesis. However,
scientific hypotheses are not necessarily proven by the reanalysis of old data. We usually try
to construct new experiments that provide an opportunity to refute the hypothesis.
We can construct such studies by taking compounds that appear to show a complicated
shift in tumors and trying to find the biochemical or tissue changes that occur as a result of
treatment. We have to be careful to recognize that this is a very general hypothesis, and we
should not restrict the search for changes in milieu to specific hormones or to specific
histopathological events. We also have to be careful that such studies are used primarily to
examine the validity of the Paracelsean hypothesis. If, for instance, we find a set of changes
that seem to be associated with one class of chemical compounds, and if that class tends to
be mutagenic or ‘carcinogenic” in other models, we cannot assume that we have found
marker or proximate events unique to that class, and far more work will be needed to
establish such findings as indicative of that class. I propose a set of experiments of
following type: We pick 10-20 of the compounds that have shown paradoxical shifts in
tumors in the NCI/NTP program (increase in tumor incidence for one type of tissue
associated with decreases in tumor incidence of another type of tissue, where the
paradoxical events may be in different sexes or species). Using the same doses and the same
strains of animal, we redo the feeding study for a small number of animals over a relatively
short period (1-3 months). At the end of this time, we sacrifice the animals and examine the
target tissues (tissues in which paradoxical tumor increases or decreases have occurred).
The hypothesis predicts that we will find differences in “milieu”. There are rough tools
for comparing general patterns of proteins or fragments of active agents between two sets of
tissue, which have been used in the primary screening of undifferentiated biological
products where we are looking for something “new”. It would not be necessary to identify
these differences. It should be sufficient to show that there are discernible differences. If
548

these general patterns appear to be identical between treatment groups, then that would be
evidence against this hypothesis.
SIX AZO DYES
As part of its ongoing program applying a “carcinogenesis” bioassay to chemical
compounds now in use, the NTP has reported on studies involving six AZO dyes. These
assays were conducted at a single site. Each dye was subjected to the same protocol, in
which male and female rats and mice were divided into three groups each: controls, low
dose, and high dose. There was an exception to the standard protocol. For three of the dyes,
single control groups of 90 animals each were used in the male and female rat portions of
the assay. Otherwise, there were separate controls for each sex/species/compound of 50
animals each).
This provides us with a repeated experiment against which to test the hypothesis
proposed above, using the method described in subsection A in the preceding section.
Three of the six dyes were declared “carcinogenic” for at least one speciesor sex. The
marginal counts of animal with tumors in specific sites (the usual NTP analysis) suggested
dose-related changes for all but 3 of the 48 sex X species comparisons. As indicated above,
however, multiple testing for tumors in specific sites raises the question of how to adjust the
nominal p values to keep the false-positive rate from going too high. Other suggestions have
been made based on the use of Bonferonni bounds and the statistical characteristics of the
tests used. Most of this amount to requiring a lower level of significance (usually around
0.01) for frequently occurring tumors. When this is done, all six dyes have significant doserelated
shifts in tumors for at least one sex or species and 36 of the 48 sex X species
combinations still show significant dose-related changes.
Analyses like these have been criticized because they are based upon multiple testing
and because the p-value adjustments tend to be of an ad hoc nature. To get around such a
criticism, I ran overall tests of shifts in tumor patterns as described in the preceding section
for all six studies.
Using methods described in the Appendix, data for each animal were converted to a
vector of 25 numbers, representing the maximum degree of dysplasia discovered in specific
organ systems, along with the number of weeks the animal was on study. Organized this
way, the 150 animals in a given sex X species X compound study can be thought of as a
cloud of points in multidimensional space. Animals with similar patterns of neoplastic
lesions will tend to provide points close together.
If there were no effect of treatment on neoplastic lesions, then the cloud of points
would be such that animals in a given treatment group would be randomly scattered thought
the space. The points closest to a given animal’s point would belong to animals associated
with any of the three treatment groups with equal probability. However, if treatment had an
effect on neoplastic lesions, we would expect animals in a given treatment group to be
clustered together more so than might be expected at random.
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The appendix describes two tests for homogeneous scatter, one along the first principal
component of the data, the other along the second principal component. The principal
components are vectors that identify the directions of greatest spread among the points. The
linear sum of elements that defines the projection of a point along a principal component is
uncorrelated with the sum that defines the projection along another principal component.
Because of this, the nominal significance test associated with two such tests can be adjusted
to ensure a false-positive rate of 5%. In this situation, a nominal p value of 0,0253 or less for
either of the tests implies an overall significance level of 5 %.
For each of the six tested compounds, there are significant treatment-related clusters in
at least one sex or species.
The results of this analysis support the contention that all tested compounds will have
an effect, in the current design of these studies.
DISCUSSION
If the hypothesis that every compound tested will cause some sort of a shift in neoplastic
lesions is true, then it will be necessary to reconsider the interpretation of these studies. A
treatment-related increase in tumors of one type is no more indicative, by itself, of
“carcinogenesis” than a decrease in tumors of another type is indicative of an
“antineoplastic” effect. The cause of these shifts is a complex biological interaction of a
living animal seeking homeostasis in the face of a biochemical derangement of its milieu.
The several hundred chronic toxicity tests that have been run in the past 10-15 years
provide the science of toxicology with an unprecedented opportunity to investigate the
nature of biological homeostasis and the response of a living animal to toxic pressure.
Regardless, of the eventual use of these studies to identify “carcinogens”, there is a wealth
of useful information in them, waiting to be uncovered.
In the meantime, can these studies be used to identify “carcinogens” (or even
“antineoplastic” agents), if the hypothesis of universal effect is true?
The answer, clearly, is no. In fact, the hypothesis of universal effect brings the problem
of finding “carcinogens” back among the traditional problems of toxicology. In traditional
toxicology, we recognized that chemical compounds cannot be divided into two classes,
poisons, (or toxic chemicals) and nonpoisons (or nontoxic chemical). All chemical will
cause biological damage at sufficiently high doses, and the dual task of toxicology is to
identify the nature of that damage as a function of dose and determine dose levels that have
a good chance of being “safe”. So, if all chemicals will cause a derangement of senile
neoplastic lesions, then it mekes no sense to divide the universe of chemicals into
“carcinogens” and “noncarcinogens”. This derangement of neoplastic lesions is more
complicated than the usual type of biological damage seen in subchronic and acute studies.
There are problems due to latency of expression and irreversibility of some of the lesions.
But, the problems of “carcinogenic” effects still belong in the general class of toxicological
problems. We need to find parallels to the two tasks described in the previous paragraph.
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The question of determining a “safe” dose has been badly distorted in the statistical
literature. Traditionally, toxicologists did not determine a “safe” dose by considering doses
at which no animals had a particular lesion. It was not a case of naively assuming that
seeing nothing in a small number of animals meant that nothing would occur. Instead, the
toxicologist examined the high-dose animals to determine a pattern of lesions that could be
associated with the compound at test. Then, he or she projected a set of putative precursors
of this syndrome of lesions. The traditional “maximum safe dose” was the dose at which
none of the lesions nor any of their putative precursors were seen in any animal. The
statisticians assumed that each animal was given a 0 (no lesion) or 1 (lesion). In fact, each
animal is scored in a complicated but biologically meaningful way. A “safe” dose was one
in which all the animals had scores less than the least score that could be associated with a
treatment effect.
The first toxicological problem, describe the “toxic” effect as a function of dose, is a
much more difficult one. We have addressed this problem elsewhere but only by suggesting
multivariate statistical methods that might be useful. If we do not use maximum tolerated
doses and reduce the length of studies to 12 months, then we may be able to describe a dose
response for frank carcinogens like the nitrosamines. But, when we use very high doses and
follow the animals for most of their natural life span, so that the “effect” is to shift lesion
patterns, increasing the incidence of one and decreasing the incidence of another, there is no
clear “direction” to the graph of response versus dose. To “solve” this problem, we will
need to think of dose response in some different fashion or we will have to abandon the
current design of these studies because they produce uninterpretable results.
APPENDIX: STATISTICAL METHODOLOGY
The individual animal tabulations in many of the NTP studies define organ and organsystems
that were routinely examined. In addition, there is a category of any organ not
routinely examined but for which histopathology was run because of a grossly observable
lump or bump. In the tabulation, individual neoplastic lesions are described in terms of
tissues type, degree of malignancy, and (possible) source. Tissue type designations tend to
be pathologist specific, so these were ingnored in this analysis. Instead, for each organ or
organ system a given animal was scored as follows:
0= no neoplastic lesion
1= worst neoplastic lesion was benign
2= worst neoplastic lesion was malignant but in situ
3= worst neoplastic lesion was invasive or metastatic
This produced a vector of 20-25 numbers for each animal (depending on sex and
species). An additional component was added by including the number of weeks on trial for
that individual animal.
All the animals in a given sex X species X AZO dye group, including the controls, the lowdose
animals, and the high-dose animals, were combined, and the first and second principal
components were computed for the 150 points.
551

The vector for a given animal was converted to the linear combination that represented the
projection of that vector onto the first principal component. These projections were then
ordered; a Kruskal-Wallis test was run the ordered projections. The same thing was then
done for the second principal component.
REFERENCES
1. Chhabra, R.S., Haseman, J., Hall, A., and Baskin, C. (1988), Inhibition of some
spontaneous tumors by 4-hexylresorcinol in F344 rats and B6C3F mice, Fundam.
Appl.Toxicol.11, 685-690
2. Haseman, J.K. (1983), A Reexamination of false positive rates for carcinogenesis
studies, Fundam. Appl. Toxicol.3, 334-339.
3. Salsburg, D. S. (1983), The lifetime feeding study in mice and rats – An examination of
its validity as a bioassay for human carcinogens, Fundam. Appl. Toxicol. 3, 63-67.
552

CLINICAL AND EPIDEMIOLOGICAL ASPECTS OF
HEPATIC HYDATID CYST
Mirela Indrieş, Viorica Coldea, Cristian Brisc, Ildiko Lenard, Sonia Drăghici,
Andrei Csep, Daciana Sabău, Nicoleta Negruţ
University of Oradea, Faculty of Medicineand Pharmacy Oradea
ABSTRACT
Human echinococcosis is a zoonotic infection caused by the tapeworm of the genus
Echinococcus. Human disease is a „dead end” which interrupts the life cycle of the
parasite. Hepatic localization is the most frequent (63%), because the infestation occurs by
digestive way, and liver is the first filter on this way. Hydatid disease may have any
localization. Cystic echinococcosis has public health importance not only in areas of
endemicity but also in countries or regions without endemicity due to the migration of
infected people and livestock exchanges.
Keywords: Echinococcus granulosus, hepatic hydatic cyst (HHC).
INTRODUCTION
Human echinococcosis is a zoonotic infection caused by the tapeworm of the genus
Echinococcus. Of the 4 known species of Echinococcus, 3 are of medical importance in
humans. These are Echinococcus granulosus, causing cystic echinococcosis; Echinococcus
multilocularis, causing alveolar echinococcosis; and Echinococcus vogeli. Echinococcus
granulosus is the most common of the three. Echinococcus multilocularis is rare, but is the
most virulent and Echinococcus vogeli is the most rare (Table 1) (Brunetti E, 2006).
Table 1: Echinococcus Species
Echinococcus Species Echinococcosis Diseases
1. Echinococcus granulosus Cystic Echinococcosis (cystic hydatid disease)
2. Echinococcus multilocularis Alveolar Echinococcosis
3. Echinococcus vogeli Polycystic Echinococcosis
AS 4. Echinococcus oligarthus Polycystic Echinococcosis
AS=mal Species
Incidence
Echinococcosis is a very rare disease in northern Europe. The disease is endemic in the
Balkan Area and in Eastern Europe. The endemic areas are the Mediterranean countries and
the Middle East. The intensive endemic areas are the southern part of South America,
Iceland, Australia, New Zealand and southern parts of Africa. The incidence of Cystic
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Echinococcosis in endemic areas ranges from 1-220 cases per 100,000 inhabitants. In USA,
echinococcosis remains a very rare disease (

Figure 3.-Life Cycle of Echinococcus granulosus.
Structure
Adult Echinococcus granulosus tapeworms are relatively very small, consisting of 3 to 5
segments, and usually are less than 1 cm long. Dogs and wild canids are the only final hosts
in which the adults are found, often adhering in great numbers to the small intestinal
mucosa. The scolex has four suckers and is crowned with a circle of hooks as in Taenia
solium. It is followed by a germinative neck region, one developing segment, and usually
one gravid segment containing several hundred eggs (Figure 2 A, B).
Figure 2. A- Adult Echinococcus granulosus worm, B- scolex showing hooks, C-
hydatid brood capsule with protoscolices
The hydatid larva is found in sheep, in many other herbivores and in humans. In humans
the cyst is slow-growing, but in a period of years may reach a diameter of 30 cm with a 1
mm thick, laminated sheath surrounded by fibrous reactive host tissue. The cyst is usually
fluid-filled and, if viable, has a germinative inner lining from which many thousands of
555

scoleces are budded off into the lumen or remain attached to the germinative wall (Figure 2
C). The floating scoleces often enlarge, become vesicular, and develop into daughter
floating colonies within the parent cyst. These in turn may bud off a third generation of
cysts within themselves. The result is an enclosed sac containing myriads of future adult
worms ready to infect a dog or other susceptible carnivore that feeds on the hydatid-infected
animal and the scolex-filled cyst fluid (Adamson ML, 1994).
Clinical Manifestations
Echinococcosis (hydatid disease) results from the presence of one or more massive cysts, or
hydatids, which can develop in any tissue site, including the liver, lungs (25%), heart, brain
(1%), kidneys (2%), spleen (1%), tyroid and long bones (3%). Hepatic localization is the
most frequent (63%), because the infestation occurs by digestive way, and liver is the first
filter on this way. Hydatid disease may have any localization. The clinical manifestations of
this infection therefore vary greatly, depending on the site and size of the cyst, but resemble
those of a slow-growing tumor that causes gradually increasing pressure. Infections in the
liver, lungs, or subcutaneous tissue sites may be asymptomatic for many years, but pressure
effects eventually develop. In sensitive or vital areas, hydatids produce a panoply of
symptoms, chiefly owing to mechanical compression or blocking effects but also include
collapse of infected long bones, blindness, and epileptiform seizures. The rupture of a
hydatid cyst may induce sudden anaphylactic shock. Secondary complications may occur as
a result of infection of the cyst or leakage of the cyst. Unilocular cysts are frequently single
(75%). However, they can also have a multifocal or multiorgan localization. The frequency
of multiple localization ranges between 11% and 30% from 2 % to 14%. Lung involvement
is solitary in 70% of the cases, and multiple in 30% Unilocular cysts can remain
asymptomatic for 5-20 years. The diagnosis is established, with few exceptions, by
paraclinical investigations, especially by imaging techniques (ultrasonography, conventional
radiology, computed tomography) as well as by immunological studies (ELISA). Although
drug treatment has been followed by many failed attempts, today we can witness its revival
due to the new drugs (mebendazol, albendazole) which completely destroy the univesicular
hydatid cyst in 50-80% of the cases after 3 months of treatment. Some authors reported a
sterilization rate of 95%. Nevertheless, the treatment of choice remains surgical.
Patients and Methods: We studied a lot of 18 cases with age between 0-69 years,
hospitalized at the Clinical Hospital Infectious Diseases of Oradea, between years 1998-
2006, with a hepatic hydatid cyst, established by clinical and paraclinical data. The study
was performed on: the patients observation files; the biochemical methods (a complete
blood count), the serologic methods (detection of the anti-Echinococcus antibodies with
ELISA method) and imaging techniques (ultrasonography, conventional radiology).
The Epidemiological Particularities:
The Number of cases with Hepatic Hydatid Cyst
From analyzing the annual incidence of the Hepatic Hydatid Cyst (HHC) between years
1998-2006, hospitalized the Clinical Hospital Infectious Diseases of Oradea, we observed
556

that the values are 1 case at 600.000 inhabitans of Bihor county in 1998 and it grows at 4
cases at 600.000 inhabitans in even years 2002, 2004, 2006 (Figure 4).
Nr. de cazuri
25
20
15
10
5
Incidenţa MNI între 1987-2003
0
1986 1988 1990 1992 1994 1996 1998 2000 2002 2004
Ani
Incidenţa MNI între 1987-2003
Figure 4 -The annual distribution of HHC cases in Bihor county, Romania.
It is specified that the real annual incidence of HHC could not be proper for the following
reasons: the most part of infections evolves as subclinic or inaparent infections, in this way
remaining undiagnosed and unreported.
The Distribution of Cases on Age Groups.
The highest incidence of HHC infection is to be found the age group of 61-70 years old
patients (33,33%), followed by the age group of 51-60 years old patients (22,22%), 49-50
years old patients (22,22%) and 31-40 years old patients (16,67%).
33,33%
22,22%
0,00%
Age Groups
5,56%
16,67%
22,22%
Figure 5 -The distribution of cases on age groups.
0-10 years
11-20 years
21-30 years
31-40 years
41-50 years
51-60 years
61-70 years
The Distribution of the Cases According to Sex
From the total of 18 patients with HHC, only 22,22% are men patients and 77,78% are
women patients (that is 14 of them).
557

Men
22,22%
Women
77,78%
Rural
77,78
%
Urba
n
22,22
%
Figure 6 -The distribution of casesaAccording to sex and to geographic origin.
The Distribution of Cases According to Geographic Origin
The majority of HHC cases come from the rural area 77,78% (that is 14 patients), most
probably due to a precarious accessibility to diagnosis and treatment units as well as to the
education of the patients.
Clinical-biological particularities of the Hepatic Hydatid Cyst hospitalized at Clinical
Hospital Infectious Diseases of Oradea between 1998-2006
On the first places have situated asthenia, hephatalgia and unappetite and only in 2 cases
have presented vomiting and fever (Table 2).
94,44
77,77
66,66
Asthenia Hepathalgy Headache Unappetite Vomiting Fever
Figure 7 -Clinical symptoms of HHC
In the majority of the cases the patients presented hepathomegaly and in 3 cases appeared
jaundice and hyperchrome urinary emissions.
558
50
33,33
11,11
%

Paraclinical data
From the laboratory analyses we detected the followings: leucocitosis appears in 2 cases,
eosinophilia (≥ 4 %) in 13 cases, anemia (Hg

3. The HHC is more frequently at women (77,78 %) than men, is more frequently in rural
area than in urban area and is more frequently at domestic animal holder/farmer.
4. Hepatic localization is the most frequent (60%), because the infestation occurs by
digestive way, and liver is the first filter on this way.
5. In the majority of the cases the patients presented asthenia, hephatalgia, unappetite,
hepathomegaly and only in 3 cases appeared jaundice and hyperchrome urinary
emissions.
6. In the majority of the cases HHC were single and only in 2 cases HHC have evoluated
with multiple intrahepatic localisations.
7. The lethal risk results in case of HHC breake followed by anaphylactic shock.
8. The true prophilaxy of domestic animal illness should prevent the existence of the
Echinoccocosis disease at human and should decrease the costs of treatment .
9. With a universal outspread, the HHC remains permanently an open problem.
REFERENCES:
1. Eckert J., Deplazed P.: Biological, epidemiological and clinical aspects of echinooccosis,
a zoonosis of increasing concern, Clin Microbiol Rev. 2004;17: 107-125.
2. G. Mandell, J. Bennett, & R. Dolin (Eds.) Principles and practices of infectious diseases
(5th ed.); 3205-3207.
3. Brunetti E, Filice C, Meroni V: Comment on percutaneous treatment of liver hydatid
cysts. AJR Am J Roentgenol 2006 Apr; 186(4): 1198-9.
4. Rogan MT, Hai WY, Richardson R: Hydatid cysts: does every picture tell a story?
Trends Parasitol 2006 Sep; 22(9): 431-8.
5. T. Vaida: Parazitologie medicală, Universitatea Oradea, 1993: 76-80.
6. M J Kumar, K Toe and R D Banerjee: Hydatid cyst of liver, Postgraduate Medical
Journal 2003;79:113-114.
7. Sréter T, Széll Z, Egyed Z, Varga I. Echinococcus multilocularis: an emerging
pathogen in Hungary and central eastern Europe. Emerg Infect Dis [serial online] 2003
Mar . Available from: URL: http://www.cdc.gov/ncidod/EID/vol9no3/02-0320.htm
8. Ammann RW, Eckert J.: Cestodes. Echinococcus.Gastroenterol Clin North Am. 1996
Sep; 25(3):655-89.
9. Radulescu S. Parazitologie medicala, Ed. All Educational, Bucuresti, 2000: 282-95. 10.
Zanc V. Parazitologie clinica, Ed. Sincron, 2001, Cluj-Napoca: 108-155.
10. WHO - Grupul neoficial de lucru OMS in problema hidatidozei. Directive privind
tratamentul hidatidozei chistice si alveolar la om. Buletin OMS, 1996.
11. Codreanu R., Simona R.: Orientari actuale in tratamentul bolii hidatice, Revista romana
de parazitologie, 2002(XII): 30-38.
12. Ş. Georgescu1, L. Dubei and al: Minimally Invasive Treatment of Hepatic Hydatid
Cysts, Romanian Journal of Gastroenterology, September 2005 (14 ); 249-252.
13. Adamson ML, Caira JN. Evolutionary factors influencing the nature of parasite
specificity. (Review) Parasitology, 1994 (109);85-95,
560

THIN LAYER CHROMATOGRAPHY OF STERIGMATOCYSTIN IN CHEESE
Bara Lucian
University of Oradea, Faculty of Environmental Protection
ABSTRACT
A one-dimensional thin layer chromatographic method has been developed for determining
sterigmatocystin in cheese. Cheese is extracted with acetonitrile-4% KCl (85 + 15). A
simplified liquid-liquid partition cleanup is used, and the sample extract is passed through a
cupric carbonate column for final purification. Sterigmatocystin is visualized by spraying
the plate with aluminium chloride. The fluorescence of the spot is enhanced 10-fold by
additional plate spraying with silicone-either mixture, enabling sterigmatocystin detection
and quantitation at 2 and 5 μ/kg, respectively. Average recoveries were 88.3 and 86.4% at
the 10 and 25 μ/kg levels, respectively.
Keywords: cromatography, cheese
INTRODUCTION
Sterigmatocystin is a mycotoxin that has potent toxic and carcinogenic effects. This
metabolite is produced by several species of Aspergillus and at least one species of
Bipolaris. Sterigmatocystin occurs naturally in grains, green coffee, fruit juices and fruitbased
infant foods, and cheese. Since sterigmatocystin-producing fungi are so ubiquitously
distributed, this metabolite must be considered as an important contaminant in these
commodities as well as in other foods and feeds. Thus, interest was aroused in analyzing
cheese for sterigmatocystin to determine levels and incidences of contamination.
Methods have been published for the determination of sterigmatocystin in grains and cereals
and have been used for other commodities; however, these methods were found to be
unsatisfactory for this project. The method of van Egmond et al., which is essentially their
earlier method with modifications, has been developed for the determination of the
sterigmatocystin in cheese, reportedly with a 5 μ/kg limit of detection. I tested this method
and concluded that it was complex and time consuming, and had a relatively high limit of
detection along with poor spiked sample recoveries. Experimental data collected during the
method evaluation indicated the potential for the development of a more satisfactory
procedure. The thin layer chromatographic (TLC) method described here resulted.
MATERIALS AND METHODS
Apparatus
a) Chromatographic tube – Plain, 22 x 250 mm
b) TLC plates – precoated, Sil G-25 HR No. 66-14-600-6. Do not activate plates.
c) TLC apparatus – Developing tank with cover for use with 20 x 20 cm glass
plates;spotting template; 10 μL syringe; longwave 15 watt UV lamp (use with
absorbing eyeglasses) or Chromato-Vue cabinet equipped with one or two 15 watt
561

lamps; if available, a thin layer plate densitometer with fluorescent capability,
excitation wavelength 365 nm and emission wavelength > 400 nm.
d) TLC plate sprayer – Kontes, K-422540
e) Blender – waring, equipped with 1 L jar and lied
f) Filter paper – Whatman 2V folded circles, 24 cm
g) Vials – 5 mL, conical, tapered inside, with Teflon-lined screw cap
Reagents
a) Solvents – reagent grade acetonitrile, hexane, methylene chloride, benzene, methanol,
acetic acid, and anhydrous ethyl ether. (Caution: Benzene is a suspected carcinogen.
Use necessary safety measures when handling this chemical)
b) Silicone DC-200 – 12 500 centistokes
c) Silicone-ether spray – silicone-anhydrous ether (18 + 82). If weighing silicone is more
convenient, 1g = 1mL
d) Potassium chloride solution – dissolve 4g KCl in 100 mL water
e) Aluminium chloride solution – Dissolve 15 g AlCl, in ethanol and dilute to 100mL
f) Calcium chloride – anhydrous, granular (8mesh). Prepare in advance CaCl2 –water (1 +
2, w/v). Hasten solution by placing in ultrasonic bath. Remove slight cloudiness by
filtering through Whatman 2V paper
g) Curpic carbonate, Cu (II) – Reagent grade powder
h) Diatomaceous earth – Hyflo Super- Cel
i) Sterigmatocystin standards – Stock standard – Prepare according to sec. 26.133.
Working standard – Dilute stock standard to ca ng/ μL benzene
j) Boiling chips – Si C. Float off fines and extraneous particles with water, wash with
acetone, and dry.
Extraction
Weigh 36 G composited sample into 1L Waring blender jar. Add 170 mL acetonitrile and
30 mL KCl solution; secure lid and blend 3 min at high speed. Decant extract through
Whatman 2V paper. Transfer 50 mL filtrate to 250 mL separatory funnel, add 50 Ml CaCl2
solution, vigorously shake ca 30 s, and let stand until layers separate; separation usually
occurs within 5 min.
Discard bottom layer. Remove excess water from inside the stem of the separatory
funnel by drying with laboratory wiping tissue, such as Kimwipes. Place a small wad of
glass wool into the stem of the separatory funnel to serve as a filter and drain the remaining
phase into a second separatory funnel. Add 50 Ml water to remaining phase and mix. Add
50 mL hexane, vigourously shake about 1 min, and let layers separate. Transfer bottom
layer to another 250 mL separatory funnel. Extract, using 50 and 25 mL portions of
methylene chloride, by shaking first extraction 1 min and second extraction 10 s. Again,
complete separation usually occurs within 5 min. Collect methylene chloride extracts
separately and save for column chromatography.
562

Column Chromatography
Loosely place glass wool plug in bottom of chromatographic tube and add ca 15 g
anhydrous sodium sulfate. Add 4 g cupric carbonate-diatomaceous earth (1 + 2); lightly
pack column by holding in upright position and by allowing it to drop gently on its tip onto
book or writing pad. Add 20 g sodium sulfate to top of column.
Sequentially pass methylene chloride extracts through column, beginning with 50 mL
portion. Collect that eluate and each successive eluate in 250 mL Philips beaker. Add 25 mL
portion to column when first portion has drained to below top layer of sodium sulfate. Rinse
column with 10 mL methylene chloride.
Add 4-5 SiC boiling chips to Philips beaker and evaporate solvent nearly to dryness on
steam bath under gentle stream of nitrogen. Cool to room temperature and quantitatively
transfer with small portions of methylene chloride to 5 mL vial. Evaporate solvent to
dryness under gentle stream of nitrogen. Save sample residue for TLC.
This Layer Chromatography
Add 100 μL benzene (screen < 10 ppb) or 200 μL benzene (screen1
h before using.
Spot plate as previously described and develop using benzene-methanol (95 + 5).
Solvent front travels 13-15 cm in 45 min.
563

Table 1. Recovery of sterigmatocystin added to Gouda cheese
Det. No. Found, μg/kg
10μg/kg Level
Rec., %
1
0, control
2
10.0
100
3
9.9
99
4
8.4
84
5
8.6
86
6
7.9
79
7
8.2
82
Mean
8.8
88.3
Std. Dev.
0.89
Coeff. Of var., %
10.2
25 μg/kg Level
1
0, control
2
27.6
110
3
21.4
86
4
19.7
79
5
20.1
81
6
20.0
80
7
20.8
83
Mean
21.6
86.4
Std. Dev.
2.99
Coeff. Of var., %
13.9
Calculation
Calculate concentration of sterigmatocytin:
μg/kg = ng/g = (S x Y x V) / (X x W)
where S = μL standard equal to unknown, Y = concentration of standard (ng/ μL), V = final
volume of sample extract (μL), X = μL sample extract spotted giving fluorescent intensity
equal to S, and W = g sample in final extract (9g).
RESULTS AND DISCUSSIONS
Table 1 shows the percentage recovery of sterigmatocystin from spiked Gouda cheese after
extraction and cleanup, as measured by one-dimensional TLC. The mean recoveries of
sterigmatocystin added at levels of 10 and 25 μg/kg were 88.3 and 86.4%, respectively. The
coefficients of variation were 10.2 and 13.9%.
While trying to resolve the problem of sterigmatocystin spots fading on TLC, I noticed
that spots, particularly those of 30 ng and less, were very visible, especially when the plate
was still warm; however, the spots faded rapidly upon cooling of the plate. I also observed
564

that spot intensity could be regenerated by reheating the plate. This fading was unlike fading
shown by aflatoxins, in that aflatoxin fading is minimized when plates are protected from
light and, in some cases, from atmospheric contaminants such as ozone. With
sterigmatocystin, I surmised that the activity of the silica gel was causing the observed
behaviour. I concluded that the problem would be solved if the silica was held in an
activated state, which could be accomplished by sealing the plate with paraffin or silicone.
Various mixtures of silicone-anhydrous ether were applied by spraying before arriving at the
mixture described here.
In addition to sealing the plate, the silicone also enhanced the fluorescence of the
sterigmatocystin spots approximately 10-fold. This type of fluorescent spot enhancement
had been noted earlier by other researchers (17), who used viscous organic solvents such as
mixtures of liquid paraffin and n-hexane as a spray.
No significant loss in fluorescent intensity of sterigmatocystin was observed in plates
that were stored unprotected from light and moisture for 2 weeks.
A sterigmatocystin standard curve gave a linear response in the range from 3 to 106 ng, with
a correlation coefficient (r) of 0.9994. One ng sterigmatocyctin standard is easily detected,
while 3 ng represents the lowest amount quantitated at a signal-to-noise ratio of 3:1.
The proposed method is simple, practical, rapid, economical, and precise. Incorporation
of calcium chloride solution into the sample extract before hexane defatting and methylene
chloride partitioning solved emulsion problems previously encountered when using other
methods. The elimination of emulsion formation during separatory funnel cleaup and
partitioning has possibly provided the breakthrough necessary to improve analytical
methods used for the determination of other mycotoxins. The concentration of calcium
chloride used may vary among commodities. The minimum concentration required to be
effective in this method has not been thoroughly investigated. Remaining TLC interferences
are eliminated by passing the methylene chloride extract through the cupric carbonate
column. Further work is in progress on the use of the method to determine other
mycotoxins.
REFERENCES
1. 1.Purchase, I., F., H., & van der Watt, J. J. (1970), Food Cosmet, Toxicol. 8, 289-296.
2. 2.van der Watt, J.J. (1974) Mycotoxins, Elsevier Scientific Publishing Co., Amsterdam,
The Netherlands, pp. 369-382
3. 3.Bartos, J. & Matyas, Z. (1983) Vet. Med. (Prague) 28, 189-192
4. 4.Nowotny, P., Baltes, W., Kroenert, W., & Weber, R. (1983) Lebensmittelchem.
Gerichtl. Chem 37, 71-72
5. 5.Official Methods of Analysis (1984) 14 th Ed., AOAC, Arlington, VA
565

TISSUE LEVELS AND BIOLOGICAL EFFECTS OF N-
NITROSODIMETHYLAMINE IN MICE DURING CHRONIC LOW OR HIGH
DOSE EXPOSURE WITH OR WITHOUT ETHANOL
Bara Camelia
1 University of Oradea, Faculty of Environmental Protection
ABSTRACT
In a study of the metabolism, disposition, and hepatotoxicity of the environmental
carcinogen N-nitrosodimethylamine (NDMA), as a function of dose in the drinking water
and of concomitant administration of ethanol, outbred Swiss mice were given NDMA for 1-
4 weeks at levels of 50-0.5 ppm, with or without 10, 20, or 30% ethanol. NDMA, assayed in
blood, liver, kidney, lung, and brain by thermal energy analysis after methylene chloride
extraction, was detectable (.0.5 ppb) in tissue levels of NDMA (1-4 ppb) near the detection
limit of 0.5 ppb; this was also found to be the minimal concentration causing significant
numbers of lung tumors in strain A mice after treatment for 16-18 weeks. Co-administration
of ethanol caused an increase in blood and tissue levels of NDMA at all levels of both
chemicals, often by a factor of 10 or more. Ethanol also partially alleviated the
morphological hepatotoxic effects of NDMA at 50 ppm (centrilobular hemorrhage and
necrosis). These results are consistent with competitive inhibition of metabolic activation of
NDMA by ethanol. Ten per cent ethanol did not induce liver NDMA demethylase activity
significantly and did not prevent loss of this activity from the livers of mice receiving 5-50
ppm NDMA. Thus, inhibition, rather than induction, of NDMA metabolism was the
predominant effect of ethanol, with increased levels of NDMA in blood and other tissues as
a consequence.
Keywords: N-nitrosodimethylamine, dose exposure
INTRODUCTION
The potent carcinogen NDMA is present in a variety of human contact sources.
Furthermore, low levels of NDMA, usually in the 0.1-1-ppb range, have been reported in
human blood and urine. Since the meaning of these human blood NDMA levels for cancer
risk is of interest, we have measured amounts of NDMA in blood and other tissues of mice
exposed chronically to various concentrations of NDMA in drinking water or liquid diet,
and correlated these values with incindence of tumors in the lung.
A second objective of the study was examination of the effect of chronic coadministration
of ethanol on tissue levels and hepatotoxicity of chronic NDMA, of concern
because of the occurrence of NDMA in some alcoholic beverages, frequent concomitant
exposure of humans to nitrosamine-containing cigarette smoke along with ethanol and
NDMA, as related to carcinogenicity and toxicity, have revealed that this and other alcohols
have complex effects on the metabolism, pharmacokinetics, and biological effectiveness of
NDMA and other nitrosamines. When present simultaneously with NDMA, ethanol inhibits
566

its metabolism by oxidative pathways, in vivo, in perfused liver, in intact liver cells in vitro,
in liver slices, and in cell-free preparations of microsomes. The inhibition is competitive in
nature. Conversely, chronic treatment of animals with ethanol results in induction of the low
Km form of NDMA demethylase. With regard to the biological consequences of these
effects of ethanol, liver cells or microsomes from rats pretreated with ethanol were more
effective than controls in catalyzing mutagenesis or DNA damage by nitrosamines in vitro,
but similar effects were not seen in vivo. Pretreatment of rats with isopropanol 24 hours
before NDMA led to potentiation of hepatotoxicity, whereas rats given ethanol in a liquid
diet for 3 weeks before NDMA experienced a reduction in hepatotoxicity compared to
controls. Effects on neoplasia are similarly complex. Ethanol given simultaneously with Nnitrosodiethylamine
or N-nitrosomorpholine caused an increase in number and size of
preneoplastic liver foci, but ethanol given simultaneously with various nitrosamines reduced
the number of liver tumors, sometimes with a concomitant increase in tumors of distal
targets such as esophagus or nose. Ethanol administered with N-nitrosonornicotine to rats
resulted in fewer esophageal but more nasal cavity tumors than in rats given the carcinogen
alone.
No studies have been reported on the short-term toxic effects of ethanol given
continuously and simultaneously with NDMA, even though this is presumably an exposure
mode commonly encountered by the human. Under these conditions, both induction and
competitive inhibition of NDMA demethylase would be possible, and either action could
influence the activation and / or detoxification of the chemical. The nature of the biological
phenomena resoving from this complexity of possibilities required empirical determination.
In the experiments reported here, we inquired as to whether co-treatment with ethanol
during exposure of mice to a low but tumorigenic dose of NDMA in the drinking water, or
to a high, hepatotoxic dose of this agent, would influence expression of hepatotoxicity or
levels of NDMA demethylase in the liver and, concomitantly, circulating levels of NDMA
in the blood and in organs distal to the liver.
MATERIALS AND METHODS
Lung Tumorigenesis Assay. Male strain A/J mice were housed in plastic cages with
hardwood shavings as bedding at 24+2 C, 40-60% humidity, and with a 12-hour fluorescent
light / dark cycle. Solutions of 500 ppb NDMA were diluted daily in distilled water from a
stock solution of 10 ppm NDMA; the later was kept at 4C in the dark and
prepared monthly. The drinking water solutions of NDMA were contained in amber bottles.
All bottles and sipper tubes were rinsed with distilled water before use. Control animals
received distilled water.
567

Expt. Diet Treatm
. time
(weeks)
A/J
mice
A/JCr
mice
Purina
5002
Purina
5002
Purina
5015
Table 1. Lung tumorigenesis in strain A mice by 500 ppb NDMA
12
14
16
18
18
Treatmen
t
NDMA
Control
NDMA
Control
NDMA
Control
NDMA
Control
NDMA
Control
Tumor
Beares /Total
8/49
8/50
14/50
8/49
23/50 a
10/50 a
27/99 a
18/100 a
33/100 a
17/100 a
Average No. of
Tumors (+ SD)
0.16+0.37
0.16+0.37
0.28+0.45
0.18+0.44
0.48+0.54 b
0.22+0.46 b
0.33+0.61
0.20+0.45
0.43+0.70 b
0.18+0.41 b
a 2
Significant differences between NDMA-treated and control mice, p

Exposure
time
weeks
1
2
3
Table 2. Effects of 50 ppm NDMA in the drinking water: water consumption, body weights,
NDMA demethylase, and pathological change
Treat-
ment
None
10% EtOH
50ppmNDMA
NDMA+EtOH
None
10% EtOH
50ppmNDMA
NDMA+EtOH
None
10% EtOH
50ppmNDMA
NDMA+EtOH
Average
Water
Consumption
(ml)
3.6+0.6 (4) a
4.5+0.4 (4)
2.4+0.1 (4)
2.6+0.3 (4)
4.0+0.3 (4)
4.0+0.8 (4)
2.3+0.6 (4)
1.9+0.4 (4)
4.9+1.7 (4)
2.9+0.3 (4)
2.5+0.6 (4)
1.8+0.3 (4)
Average Body
Weight
(g)
26.2+1.7 (10) a
26.8+1.9 (10)
23.4+1.9 (20)
22.4+2.4 (20)
26.8+1.5 (10)
25.9+1.9 (10)
24.6+2.3 (20) d
20.1+2.3 (20) d
27.9+1.7 (10)
28.6+1.7 (10)
24.6+2.3 (10) d
22.4+2.0 (10) d
NDMA
Demethylase
(Nmol
CH2O/mg
Pathological Change
protein/20 min)
Gross Histological
ND b
ND
1.0+0.7 (6) c
2.0+1.0 (6)
ND
ND
1.8+0.7 (6)
1.6+0.8 (6)
ND
ND
0.9+0.6 (6) c
0.5+0.4 (6) c
a
All values are + standard deviation. Numbers of parantheses are numbers of determinations
b
ND, not done
c
significantly different from value in untreated mice, 2.4+1.0 (N=7), p

The concentration of the chemical was confirmed by chemical analysis (extraction with
methylene chloride followed by measurement by gas chromatography).
In experiment 1, the mice were fed Purina Certified Rodent Chow. Treatment was
started at 4 weeks of age and continued for 12 , 14 or 16 weeks. For experiment 2, two diets
were formulated to be of defined composition with regard to essential nutrients, so that these
could be reproducibly manufactured if desired. One of these was similar to Chow 5002, as
used in experiment 1. The other was similar to Purina Mouse Chow 5015, a high fat, low
fiber diet. Treatment was started at 4 weeks of age and continued for 18 weeks. Lungs and
stomachs were fixed in Bouin’s solution and were examined under a dissecting microscope
for tumors. Lungs were examined grossly and after hand-sectioning into 1-mm slices. Any
questionable lesions were subjected to histological verification (7- μm sections, hematoxylin
and eosin staining). Statistical analyses included the x 2 test for differences in percentages of
tumor-bearing mice and the two-tailed Student’s t test for differences in average number of
tumors per mouse.
Subchronic Exposure of Mice. The mice were housed in polycarbonate cages with wire
inserts and fed NIH autoclavable diet 31. Environmental conditions included a temperature
of 25+2 C, humidity of 50+10% and 12 changes of room air / hour. Fluorescent lights were
automatically timed at 12 hours on / 12 hours off. Treatment was started when the mice
were 5 weeks of age. Drinking water was shielded from room lighting with aluminium foil
and was changed and measured twice weekly. NDMA and ethanol were diluted with
deionized water.
At kill, blood was collected after decapitation into cryotubes containing a few crystals
of ascorbic acid and 50 units of ammonium heparin. The carcass was placed on ice and the
livers, kidneys, lungs, and brains were quickly removed to cryotubes. The tissues were
frozen in liquid nitrogen and stored at –20 C until analyzed. In some experiments sections of
liver were fixed in 10% formalin for histopathology. Portions of liver to be analyzed for
content of NDMA demethylase activity were stored at –80 C prior to analysis.
Gross pathology was estimated semiquantitatively at kill by scoring on the basis of 0-3
(0 = normal, 1 = focal change in liver appearance and / or slight peritoneal fluid; 2 = general
liver change (pits, yellowing, or darkening) and / or moderate peritoneal fluid; 3 =
pronounced change in liver morphology and extensive peritoneal fluid). Similarly,
histopathological change was estimated semiquantitatively on a basis of 0-4 (0 = normal or
minimal focal change; 1 = minimal centrilobular hemorrhage; 2 = mild centrilobular
hemorrhage; 3 = moderate centrilobular hemorrhage with focal necrosis; 4 = moderate
centrilobular hemorhhage with massive necrosis).
In order to assess the effects of ethanol on tissue levels of NDMA under conditions of
controlled nutrition, Lieber-DeCarli Liquidiets were employed. Weanling female mice were
allowed to become accustomed to the diet for 5 weeks, until their weights stabilized. The
mice were then divided into two groups and those of one group were offered diet with
gradually increasing amounts of ethanol, according to the instructions of the supplier. The
mice accepted the ethanol-containing diet only poorly. After an additional 6 weeks of
570

attempted accommodation to the diet, it was decided to carry out the experiment with twothirds
of the recommended ethanol level (4.3 % of the diet, 24% of total calories). NDMA
was included in both ethanol and control diets at 10 ppm, and the mice were pair-fed for 2
weeks so that mice were given NDMA-control diet in amounts equal to that consumed by
the mice given NDMA –ethanol diet.
Estimation of Tissue Content of NDMA. The amount of NDMA in samples of blood and
other organs (pooled from 5-10 mice) was estimated by methylene chloride extractionthermal
energy analysis as described previously. The limit of detection of NDMA by this
procedure was 0.5-1 ppb, depending on the amount of starting material. Water blanks and
tissues from control mice were repeatedly assayed and did not contain measurable levels of
NDMA (

RESULTS
Tumorigenesis by 500 ppb NDMA. These experiments were designed to test whether a
significant increase in lung tumors could be reproducibly obtained after exposure of male
strain. A mice to 500 ppb NDMA in the drinking water for 16-18 weeks as previously
reported. The results presented in table 1 answer this question affirmatively: in three
separate trials involving strain A mice from different sources and fed widely differing diets,
exposure to 500 ppb NDMA resulted in significant increases in incidences of lung tumors
compared with controls. In experiment 1 it was found that 16 weeks of exposure was the
minimum time required for a significant difference to be manifested. The strain A mice in
experiment 2 exhibited somewhat fewer tumors after NDMA treatment than did those in
experiment 1. In spite of the large differences in diet composition for the two groups in
experiment 2, resulting in those given the 5002 diet weighing on average 19% less and
drinking 34% more than those given the 5015 diet (both differences significant; data not
shown), the diet composition did not significantly affect the incidences of lung tumors in the
control or NDMA-treated mice.
Forestomach papillomas were countered in experiment 1 as a possibly useful indicator of
tumorigenesis. The incidences of these tumors were not treatment related (table 1)
Effects of Subchronic Administration of 50 ppm NDMA with and without Ethanol. In
order to test whether exposure to a high, hepatotoxic dose of NDMA would result in
measurable tissue levels of NDMA, and whether this level might be influenced by the
simultaneous presence of ehanol, mice were given 50 ppm NDMA with or without 10%
ethanol for 1, 2, or 4 weeks. The NDMA was clearly hepatotoxic, as expected, giving clear
gross and histopathological signs of centrilobular liver damage and leading to a decline in
liver NDMA N-demethylase activity between 1 and 4 weeks of exposure (table 2). NDMA
was measurable in blood and all organ samples (table 3). Inclusion of 10% ethanol alone in
the drinking water had no effect on water consumption, body weight, or liver histopathology
(table 2). However, the ethanol significantly affected the interactions of NDMA with liver.
The mice who received ethanol along with the NDMA experienced noticeably less liver
damage, as indicated by both gross appearance and histopathology, especially after 1 or 4
weeks. Furthermore, the amounts of NDMA recovered from blood and organs were
consistently greater, by factors of 2-40, in the mice given ethanol as well, at all three time
points (table 3).
Subchronic Administration of Low Doses of NDMA with and without Ethanol. In male
and female mice given 5 ppm NDMA in the drinking water for 2 weeks, low levels (1-4
ppb) of NDMA were detected in the blood samples and in one sample each of liver and
brain (table 4). Co-treatment with 30% ethanol resulted in a dramatic increase in amounts of
the NDMA in blood and organs, to about 20 ppb in most cases; livers, especially of the
treated males, were the only exception to this pattern. Similar results were obtained in
experiment 2 (table 4) when 10% ethanol was used. In this case the amounts of NDMA
appearing in the blood and tissues of the mice co-treated with ethanol were less than after
30% ethanol, indicating dose dependency of the effect of ethanol. In experiment 3 of this
572

series, a dose of 0.5 ppm NDMA was given with or without ethanol. In the absence of
ethanol, NDMA was detected at low levels in half of the samples analyzed. When ethanol
was present, all samples contained NDMA at a rather uniform concentration of about 2-4
ppb.
Effects of Ethanol on NDMA Tissue Levels in Mice Pair-Fed a Liquid Diet. In order to
ascertain whether the effects of ethanol in the drinking water might be due to nutritional
differences between these mice and those not receiving ethanol, two groups with or without
4 % ethanol. Inlcusion of ethanol resulted in the appearance of significant levels of NDMA
in all tissues sampled, whereas, in the absence of ethanol, NDMA was detected at a low
level only in kidney.
NDMA Demethylase Activity in Livers of Mice Given Various Doses of NDMA with or
without Ethanol. The results presented in table 2 suggested that 10% ethanol in the
drinking water had little inductive effect on NDMA demethylase activity in the presence of
50 ppm NDMA, and that chronic treatment with this high dose of NDMA resulted in a loss
of enzyme activity. Treatment with 10% ethanol alone for 2 weeks resulted in only a small,
insignificant increase in NDMA demethylase activity, compared with controls. Levels of
enzyme activity decreased in the mice given 5 or 50 ppm NDMA, significantly so in those
receiving ethanol also. Lack of a similar decrease in those given 20 ppm NDMA may have
been related in part to reduction in water consumption at the higher doses of chemical (an
average of 4.3 ml/day compared with 5.7 ml/day for the mice given 5 ppm NDMA).
DISCUSSIONS
In a previous investigation it was found that exposure of male strain A mice to 500 ppb
NDMA for 16 weeks resulted in lung tumors in 44%, compared with 8% of controls;
increases in lung tumor incidence with lower doses of NDMA were not of statistical
significance. The findings of the present study are in good agreement: 500 ppb NDMA for
16 weeks yielded lung tumors in 48% of the mice. The results further showed that a
significant effect of 500 ppb NDMA can be demonstrated with a different substrain of A
mice and with different diets. Thus, 500 ppb NDMA may be the lowest level that gives an
easily measurable tumorgenic effect in adult strain. A mouse lung, but it does so
reproducibly. It is of considerable interest that this was also a minimally effective
concentration, in our subchronic trials, in giving organs. In mice given 500 ppb NDMA for
2 weeks, about half of the tissue sample assayed had measurable NDMA, in the low ppb
range. Although great caution must be employed in extrapolating from mice to man, thses
results suggest consideration of the possibility that humans with blood levels of 0.1-1 ppb
NDMA may be at risk of tumor initiation in genetically sensitive peripheral target organs.
573

Table 4. Tissue levels of NDMA in mice given low doses of NDMA in drinking water with or without ethanol.
Expt. NDMA
Treatmen
t
1a (males)
1b
(females)
2
(females)
3
(females)
5
5
5
5
5
5
0.5
0.5
Ethanol Average
Water
Consumpti
on a
3.8+0.7
30% 4.7+1.1.
30%
10%
20%
4.2+0.7
4.7+0.4
3.9+0.7
4.5+1.1
4.2+1.5
5.4+1.2
Average
Body
Weight a
25.4+1.9
20.1+2.7
19.2+1.5
17.6+1.7
20.5+2.1
20.5+1.9
20.7+4.3
22.5+2.0
574
NDMA Content
Blood Liver Kidney Lun
g
1
22
4
26
0.6,2 b
4,12 b

At 500 ppb and at all higher doses, inclusion of ethanol in the drinking water led to
consistent increases in the amounts of NDMA in the blood and other organs. Although
there was quantitative variation, as might be expected in an experiment where the
chemical dose was received in the drinking water, the differences in NDMA levels with
and without ethanol were in many cases of considerable magnitude. The obvious
explanation for this effect of ethanol is competitive inhibition of the metabolism of
NDMA, as reported by other workers.This inhibition, causing a reduction in the rate of
formation of cytotoxic metabolites, probably also accounts for the alleviation of
morphologically apparent hepatotoxic effects of NDMA as well. The ethanol did not,
however, prevent the loss of NDMA demethylase activity caused by the higher doses of
NDMA but, in fact, seemed to contribute to this loss. Although ethanol is an inducer of
NDMA demethylase activity in rats after administration in drinking water, at most, only
a small inductive effect was seen in our mice. Inhibition of NDMA metabolism was
clearly the dominant action.
In studies employing ethanol in the drinking water, the argument may be raised
that the observed effects are due to nutritional imbalance secondary to ethanol
consumption, rather than the chemical itself; pair-feeding of liquid diets is thus
recommended. The experiment confirmed that the observed effects of ethanol on blood
and organ content of NDMA were due directly to the ethanol and not to nutritional
differences, since both groups consumed the same amount of nutritionally equivalent
diet. Other workers have similarly found that administration of ethanol in the drinking
water is a sound experimental approach.
In sum, chronic co-treatment of mice with ethanol, along with NDMA, resulted in
a loweing of one hepatotoxic effect of a high dose of NDMA, centrilobular necrosis, but
enhancement of another effect, destruction of NDMA demethylase activity. At all doses
the ethanol resulted in marked increases in circulating levels of NDMA. The
contribution of this effect to tumorgenesis by low doses of NDMA in peripheral target
organs will be an interesting subject for future experiments.
REFERENCES
1. D.H. Fine, “Nistrosamine in the general environment and food”. In “ Nistrosamine
and Human Cancer” (Banbury Report 12) (P.N. Magee, ed.), pp. 199-210. Cold
Spring Harbor Laboratory, New York, 1982
2. L. Lakritz, R.A. Gates, A.M. Gugger and A.E. Wasserman, “Nitrosamine levels in
human blood, urine and gastric aspirate following ingestion of foods containing
potential nistrosamine precursors of preformed nistrosamines”, Food Chem.
Toxicol. 20, 455-459 (1982)
3. T.A. Gough, K.S.Webb and P.F. Swann, “An examination of human blood for the
pressure of volatile nistrosamines”, Food Chem. Toxicol. 21, 151-156.
575

DETERMINATION OF METALLOTHIONEIN, CADMIUM, COPPER AND ZINC
LEVELS IN HUMAN AND RAT TISSUES
Camelia Bara
University of Oradea, Faculty of Environmental Protection, Oradea
26, General Magheru Street, Oradea
ABSTRACT
Metallothionein (MT), zinc (Zn), copper (Cu) and cadmium (Cd) were determined in 10
tissues (brain, heart, kidney cortex, liver, lung, muscle, pancreas, small intestine, spleen and
stomach) from human autopsies (10 male individuals, mean age 43 ± 9 years, all smokers)
and Wistar rats. The mean tissue concentrations of MT in the human samples varied
between 3.8 and 495 µg/g wet weight in spleen and kidney cortex, respectively. In most
tissues human MT levels were high as compared to rats; particularly in liver and kidney
cortex human MT levels exceeded those of rats about 25- and 10-fold, respectively. Positive
linear relationships were observed between Zn or Cu and MT in human liver and between
Cd and MT in human kidney cortex.
INTRODUCTION
Metallothionein (MT), a low molecular weight protein with high cysteine and metal content,
binds both essential (Zn, Cu) and non-essential metals (Cd, Hg). In view of the ability of
MT to be induced by metals, the protein plays a crucial role in the homeostasis and toxicity
of metals. The biological activity of metals thus is determined by both the concentration in
the tissue and the fraction bound to MT. However, there are few reports as to the MT
concentrations in human tissues. In a previous study we found high MT levels in human
kidney cortex, which were even higher in smokers. In these kidney cortices the amount of
cadmium bound to MT directly correlated with the content of renal cadmium but not with
zinc or copper. In a study, Onosaka et al. reported similarly high MT levels in human liver
and a strong positive relationship between hepatic Zn and MT concentrations.
To further elucidate the relationship between tissue levels of MT and metals, we
investigated in this study the levels of MT, zinc, copper and cadmium in 10 different organs
from human autopsies and rats. Some of the rat data have been previously reported.
MATERIALS AND METHODS
Samples of human tissues were obtained from 10 male individuals (20-50 years old, mean
age 43 ± 9, all smokers) necropsied within 2 days after sudden death. None of the
individuals received medication before death. Samples were stored at -80°C until being
processed further. Tissues of humans and rats (Wistar strain, males, 180-200 g) were
homogenized with a Potter-Elvehjem or Ultra-Turrax in 4 or 9 vols, of 30 mM Tris-HCl
buffer pH 7.4, and subsequently centrifuged at 18 000 x g. MT was determined in the
576

supernatants by a modified cadmium saturation method as described previously. Zn, Cu and
Cd were detected in tissue homogenates by atomic emission spectroscopy with inductively
coupled plasma excitation or atomic absorption spectroscopy after wet ashing of the
samples at 140°C with nitric acid.
RESULTS
MT was detected in all human tissues investigated. The highest MT tissue levels were found
in kidney cortex. In all tissues except brain and small intestine, human samples showed
higher MT levels than those of rats. Particularly in liver and kidney, human MT levels
exceeded those of rats about 25- and 10-fold, respectively.
Human tissues showed moderate differences in their Zn or Cu levels whereas regarding Cd
levels pronounced variations were observed. The highest Cd concentrations (29.7 ±13.6
µg/gwet weight) were observed in the kidney cortex.
In most human and rat tissues the levels of Zn and Cu were approximately similar
within a factor of 2. In comparison with the very low levels of Cd in rat tissues, the Cd
levels of all human tissues were remarkably higher. The Cd levels of all rat tissues were
lower than the detection limit of 20 ng Cd/g tissue wet weight. In human liver, positive
relationships between the concentrations of both Zn and Cu and the amount of MT were
observed. The regression coefficients were r = 0.871 (MT (µg) = 9.8 Zn (µg) - 257) and r =
0.955 (MT (µg) = 100 Cu (µg) — 359), respectively.
In human kidney cortex, a positive relationship was observed between Cd and MT with
a regression coefficient of r = 0.779 (MT (µg) = 13.4 Cd (µg) + 98.5). Metallothionein
levels in human kidney and liver were not significantly correlated.
DISCUSSION
MT was detected in all rat and human tissues investigated. The rat data confirm earlier
findings of others, and us although lower levels of brain MT were obtained with a
radioimmunoassay. In most human tissues higher levels of MT were observed than in rats.
Moreover, human MT levels, particularly in liver and kidney showed greater interindividual
variations, e.g., the concentrations of human hepatic MT varied between 11 and 1000 µg/g.
The extraordinarily high levels of MT in human kidney and liver may reflect a different
transcriptional control of the protein, the occurrence of multiple subtypes of human MT
isoforms, or the high exposure of man to environmental metals. In accordance with the
latter, levels of zinc, copper and particularly cadmium in human liver and levels of zinc and
cadmium in human kidney clearly exceeded those of the rat. With a ratio of 7 mol Cd per
mol of MT, MT of human liver and kidney cortex has the capacity to bind total tissue
cadmium. This supports the assumed protective role of MT in the detoxification of Gd.
However, in the renal cortex of both smokers and non-smokers, MT was shown to bind
about 50% of tissue Cd only. The remaining non-MT Cd may represent a significant toxic
potential. Zinc, copper and cadmium are potent inducers of MT in animal tissues. Although
there is no clear evidence for the induction of MT by these metals in human tissues, the
577

present findings on positive correlations between renal MT and cadmium, and hepatic MT
and zinc support this view.
Similarly, a close relationship between renal MT and tissue cadmium of smokers and
non-smokers and a positive correlation between MT and zinc in human liver have been
observed. These data indicate that MT in human kidney is mainly induced by cadmium
whereas in liver it may be preferentially induced by zinc and possibly copper. However,
since induction of MT has also been reported after various stresses, factors like the
hormonal and nutritional status and even the type of death might contribute as well.
There seems to be no correlation between hepatic and renal MT levels which we have
also observed in a recent investigation with 145 postmortem samples of liver and kidneys.
Except in 2 individuals, MT levels in human kidney cortex exceeded those of the liver.
However, in these cases hepatic Zn and renal Cd levels were more than twice and less than
50% of the average values, respectively. This supports the relevance for MT induction of the
high Cd content of the kidney as compared to the liver. In contrast, MT levels of rat liver
and kidney positively correlated with each other either after acute or subchronic exposure to
cadmium and zinc or the acute administration to iodoacetate. Therefore the chronic
exposure of man to low levels of metals, in particular cadmium, and the preferential
distribution of Cd-MT into the kidney may explain the observed lack of correlation.
In conclusion, these results demonstrate that considerable amounts of MT are present
in various human tissues. The particularly high levels of MT in human liver and kidney also
in comparison to rats and the high levels of metals in these tissues suggest that the
expression of MT in human tissues may respond to environmental factors, especially
chronic low-level exposure to metals.
REFERENCES
1. E.C Foulkes (Ed.), Biological Roles of Metallothionein, Elsevier/North-Holland, New
York, 1982.
2. R.J. Cousins, Metallothionein - aspects related to copper and zinc metabolism, J.
Inherit. Metab. Dis., 6 (Suppl. 1) (1983) 15-21.
3. K.H. Summer, G.A. Drasch and H.E. Heilmaier, Metallothionein and cadmium in
human kidney cortex: influence of smoking, Hum. Toxicol., 5 (1986) 27-33.
4. S. Onosaka, K.S. Min, C Fukuhara, K. Tanaka, S.I. Tashiro, I. Shimizu, M. Furuta, T.
Yasutomi, K. Kobashi and K.I. Yamamoto, Concentrations of metallothionein and
metals in malignant and non-malignant tissues in human liver, Toxicology, 38 (1986)
261-268.
5. H.E. Heilmaier and K.H. Summer, Metallothionein content and zinc status in various
tissues of rats treated with iodoacetic acid and zinc, Arch. Toxicol., 56 (1985) 247-251.
6. S. Onosaka and M.G. Cherian, The induced synthesis of metallothionein in various
tissues of rat in response to metals. I. Effect of repeated injection of cadmium salts,
Toxicology, 22 (1981) 91-101.
7. C.V. Nolan and Z.A. Shaikh, Determination of metallothionein in tissues by
578

adioimmunoassay and by cadmium saturation method, Anal. Biochem., 154 (1986)
213-223.
8. C.J. Schmidt and D.H. Hamer, Cell specificity and an effect of ras on human
metallothionein gene expression, Proc. Natl. Acad. Sci. U.S.A., 83 (1986) 3346-3350.
9. J. Chung, N.O. Nartey and M.G. Cherian, Metallothionein levels in liver and kidney of
Canadians - A potential indicator of environmental exposure to cadmium, Arch.
Environ. Health, 41 (1986) 319-323.
579

THE ROLE OF WATER QUALITY IN INFANTILE ALIMENTATION
Andrei Csép, Draghici Sonia, Lenard Ildiko, Mirela Indries, Nicoleta Negrut
University of Oradea, Faculty of Medicine and Farmacy
ABSTRACT
The study makes concrete reference to the risks linked to the water consume, which doesn’t
correspond from a chemical point of view, through the presence and the concentration over
the limits admitted by nitrates and nitrites in the water used at milk powder preparation,
distributed through the national programme and used in the alimentation of babies.
Keywords: water, nitrate content, intoxication, Faecal coliforms, Faecal streptococci
INTRODUCTION
Nitrate intoxication was firstly described and named well water intoxication in 1945
(quoted from Biro O.). It is also known under the name of infantile intoxication, infantile
cyanotic methemoglobinemia, or the newborn’s blue disease, because the dominant
symptom is cyanosis, symptom that can lead to the rapid diagnostication of the disease.
(Chiotan, 1999)
The nitrates enter the organism altogether with the foodstuff of vegetal and animal origin,
but mainly through drinking water. They can have a double origin in water:
a) They can come from soils rich in nitrogen salts, when their origin is considered natural;
b) As a consequence of water pollution, that comes either directly from nitrates (e.g.
industrial and agricultural pollution with nitrogen-based fertilizers), or from organic
substances, which through decomposition release nitrates.
Nitrates are not toxic by themselves. To achieve this quality, they have to undergo a
reduction process and be transformed in nitrites.
Transformation may take place:
a) Exogenously (in water), but only in the warm periods of the year, and in the presence
of a reductive flora;
b) Endogenously (in organism), under the action of the same reductive flora;
This reductive flora either gets up in the distal digestive tract in the proximal region where
the absorption takes place in case of digestive perturbations (coliform germs), or gets down
from nasal-pharynges in the case of some diseases, such as: rhinitis, tonsilitis, otitis etc,
especially if they are caused by streptostaphilococci.
The nitrites that entered the body combine with the haemoglobin (Hb) forming
methemoglobin (MHb), leading to a deficit of oxygen. The seriousness of the disease
(clinical forms) is given by the quantity of bloked haemoglobin (that is the percent of MHb)
(Biro, 1993):
• The superficial form appears between 10 – 25% and is characterized by cyanosis of
face (lips, nose), extremities (nails, finger tips) and mucous membrane
580

• The intermediary form appears between 25 – 45%, and dyspnoea, tachycardia, cardiac
breath, headaches are added to the symptoms characteristic to the superficial form
• The severe form appears over 50%, convulsions, arhytmia, coma and even death (blue
death due to asphyxia) being added to the previously mentioned symptoms
characteristic for superficial and intermediary forms of the disease
The disease is present almost exclusively in little children, from the first year of their life,
who are fed artificially (Ciofu E., Ciofu C, 1999, Aplleton & Lange, 1993), due to the
following special physiological characteristics of the sucklings:
• Gastric acidity is lower in the first months of life;
• 80% of the suckling’s Hb is of fetal type, being more unstable and easier to be oxidized
in comparison with the adult’s Hb.;
• Water quantity (hence of nitrates) ingested by the baby, with respect to the blood
volume, is about 12 times higher than in the case of an adult;
• Some enzymatic deficiencies (deficit of glucoso-6-phosphate-dehydrogenase) favour
the appearance of the disease;
• A high frequency disease rate in athrepsic suckling, with gastro-intestinal perturbations
and infections.
MATERIALS AND METHODS
The study was made on 17 sucklings from some rural places of Bihor county, hospitalized
in 2006. The methodology that was used during the experiment was imposed by Ministry of
Family and Health, by studying t€he medical records sent by the health units with beds to
the Public Health Directorate of Bihor County.
The data collected from these medical records are presented in the following tables:
Table 1. Data collected from the patients’ medical records
Case No. Sex Age till the
debut
(months)
Alimentation Association
with BDA
Clinical
form
1 male 1-2 natural no Sever
2 female 0-1 mixed no Medium
3 female 0-1 Natural no Sever
4 male 0-1 Artificial no Sever
5 female 7-8 Artificial no Medium
6 female 2-3 Natural no Sever
7 female 0-1 Mixed yes Sever
8 male 1-2 Artificial yes Sever
9 female 0-1 natural no Medium
10 male 2-3 artificial no Medium
581

11 female 0-1 artificial yes Sever
12 female 14-15 artificial no Medium
13 male 0-1 mixed no Medium
14 male 2-3 natural no Mediumj
15 male 0-1 mixed no Sever
16 female 0-1 natural no medium
17 male 0-1 artificial no medium
Case no. Sanitary
protection of
well
Table 2. Data regarding well water quality
Chemical pollution Microbiological pollution
% Nitrates
in water
(mg/l)
% Nitrites
in water
(mg/l)
Faecal
coliforms
Faecal
streptococci
1 no 45 0-0,3 ≥10 ≥10
2 no 45 0-0,3 ≥10 ≥10
3 no 46-100 0-0,3 ≥10 ≥10
4 no 101-400 0-0,3 ≥10 ≥10
5 no 101-400 ≥0,3 ≥10 ≥10
6 yes 45 0-0,3 ≥10 ≥10
7 yes 46-100 0-0,3 ≥10 ≥10
8 no 46-100 0-0,3 ≥10 ≥10
9 no 46-100 0-0,3 ≥10 ≥10
10 no 45 0-0,3 ≥10 ≥10
11 no 101-400 0-0,3 ≥10 ≥10
12. no 45 0-0,3 ≥10 ≥10
13 no 46-100 0-0,3 ≥10 ≥10
14 no 45 0-0,3 ≤2 ≤2
15 no 46-100 0-0,3 ≥2 ≤2
16 no 46-100 0-3 ≥10 ≥10
17 no 45 ≥3 ≤2 ≤2
RESULTS AND DISCUSSIONS
1. Out of 192 water probes sampled from public and individual wells for lab analyses in
2006, 57 probes (29.68%) were inadequate as regards the nitrate content, and the rest
of 107 probes (55.72%) were inadequate as regards the nitrite content.
2. Out of the 17 cases, 9 cases (52.94%) involved female sucklings and 8 cases (47.05%)
involved malesucklings, fact that proves the disease doesn’t appear only at a certain
582

sex. In fact, there are no data in the specilized literature regarding a variable sensibility
for one sex or another.
47%
53%
Figure 1. Suckling distribution on sexes.
male
female
3. By having a look at the age of sucklings till the moment of disease appearance, it is
noticed that, out of 17 cases, 15 cases (88.23%) are recorded for sucklings of 0 - 3
months old, one case (5.88%) for sucklings of 6 -12 months old, one case (5.88%) for
sucklings of 12 - 36 months old; out of the 15 cases involving sucklings of 0-3 months
old, 10 cases (58.82%) are with sucklings of 0-1 month old, two cases (11.76%) with
sucklings of 1-2 months old and three cases (17.64%) with sucklings of 2-3 months
old.
4. Out of 17 cases, six sucklings (35.29%) were feeeded naturally, seven sucklings
(41.17%) were feeded artificially (cow milk) and four sucklings were feeded in a
mixed manner (23.52%).
5. In most cases, that is 88.23% (15 cases), the serious form of disease appeared after the
consumption of well water without sanitary protection, even if the distance
recommended by norms, with respect to possible sources of contamination, was
respected (latrines, manure and fertilizer deposits);
6. Out of 17 probes, seven probes (41,17%) have the nitrate concentration within the
limits imposed by the sanitary norms (under 45mg/l), the rest of them (58.82%) exceed
the maximum admitted concentration (MAC), as it follows: seven probes (41.17%)
have the nitrate concentration between 46-100mg/l and three probes (17.64%) have the
concentration between 101-400mg/l. It is noticed that there were superficial and severe
clinical forms regardless the nitrate concentration in the water;
7. Most of the probes, that is, 15 probes (88.23%) had the nitrite concentration within the
limits imposed by the sanitary norms (0-0.3mg/l), only two probes (11.76%) had a
583

concentration higher than 0.3 mg/l. There were also superficial and severe clinical
forms regardless the nitrite concentration in the water.
8. In 14 cases (82.35%), there is recorded an outrunning of the sanitary norms as regards
the microbiological quality of water (faecal coliforms and streptococci, indicators of
faecal pollution over the admitted limit), fact that highlights double water pollution:
chemical and biological, which favor the increase of nitrite-nitrate concentration in
water.
CONCLUSIONS
In Bihor County, the water from individual wells is polluted with nitrites – nitrates in
different rural localities (spread all over the county), without recording predisposed
regions).
An increased morbidity is noticed due to acute intoxication with nitrates at the age
group 0-1 year, which can be avoided by the following measures that can be addressed to all
the factors involved:
• Intensification of the water quality control to maintain MAC of nitrates under toxic
values;
• Mother training regarding the advantages of natural alimentation, at least during the
first 3-4 months, and the prevention of digestive and respiratory infections of the
suckling, as well as the information of the families regarding the above-mentioned
risks;
• Counselling of pregnant women from the rural areas to take a water sample from the
well for control;
• Training the medical-sanitary personnel from the rural health unit to know the situation
of the water supplies from the area, and to recognize the disease, which is often
confused with other diseases associated with cyanosis (heart and pulmonary diseases);
• Controlled usage and storage of the natural and chemical fertilizers, monitoring the
removal of solid and liquid wastes in order to protect the water supplies.
REFERENCES
1. Aplleton & Lange, 1993 – Current Medical Diagnosis and Treatment, Publishing
Division of Prentice Hall, Connecticut, USA
2. Biro G., 1993 – Elelmiszer-higienia, Agroinform Kiado, Budapest
3. Chiotan M., 1999 – Boli infectioase, Editura National, Bucuresti
4. Ciofu E., Ciofu C., 1999 – Esentialul in Pediatrie, Editura Medicala Amaltea, Bucuresti
584

SOME ASPECTS OF SHELTER REGENERATION OF QUERCUS SPP.
FOREST STANDS FROM PRODUCTION UNIT II VALEA LUNGĂ,
FORESTRY DISTRICT, LUGOJ (TIMIŞ COUNTY)
Crsitian C. Crainic
University of Oradea, Romania
ABSTRACTS
The paper presents some characteristics of shelter regeneration of Quercus spp. stands
from Production Unit II Valea Lungă, Forestry District Lugoj (Timiş County) as a
consequence of the treatment with repeated logging. The obtained results permit some
theoretical considerations on shelter wood treatment of Quercus spp.
Stands vegetating in particular site conditions.
Key words: shelter regeneration, stand, treatment, oaks, ecosystem
INTRODUCTION
Choosing and promoting the treatments represent a fundamental problem for the forest
district. The extensive treatments, which are characterised by a possibility of an integral
mechanization and a relatively reduced technicality, offer fewer assortments and
strongly affect the dynamic balance of the ecosystem. The intensive treatments need a
high degree of technicality, a proper equipping with transport machines, and the quality
of the products obtained after their application is superior. Also, when choosing the
treatments we have to take into account the station’s particularities, the physiological
and ecological particularities of the species and not in the end the local experience.
The natural environment, the social-economic requests and the tradition of the
Romanian forestry, plead for preserving the forests’ authenticity within the Carpathian-
Danube-Pontic frame, in all its compositional diversity, for managing this very complex
but efficient bio-cybernetic system optimised by nature, on some criteria compatible
with the social-economic exigency of the future (Giurgiu V, 2005)
MATERIALS AND METHODS
In order to point out the application manner of the regenerating cuttings in the
production unit (U.P.) II Valea Lungă, Forestry District Lugoj, it was analysed the way
of conducting the cuttings, the intensity of the interventions, the periodicity of the
interventions and the dynamic of the natural regeneration, from the valuating documents
and from the records at the forest district.
The distributions of the numbers of trees on diameter category were carried out for
the forest stands implied in the regeneration process on the basis of the centralizing
records from the valuating documents. The stational conditions and the vegetation from
U.P. II Valea Lungă, Forestry District Lugoj were analysed. In order to be able to
present the objective reality from the site, digital photographic recordings were carried
out, on magnetic support using a digital camera brand H.P., with a 3,2 MP resolution.
In the present case study, from objective reasons, it is analysed the dynamic of the
regeneration cuttings at three representative forest stands, in the forest planning units
(u.a.) 3A, 4A şi 11C.
585

RESULTS
Following the results of the data from the site, a series of results were reached at, results
presented in the following lines. If the stational conditions are analysed – especially the
ecologic factors (Doniţă N., et. al., 1977) represented by precipitations and respectively
temperature, it can be noticed that they are premises for establishing the seedlings for
usage in favourable conditions – table 1.
Table 1: Monthly average temperatures and precipitations at Lugoj meteorological
station (*)
Month I II III IV V VI VII VIII IX X XI XII
0 1 2 3 4 5 6 7 8 9 10 11 12
T[°C] -1 1 5 12 17 20 22 22 18 13 6 2
Pp [mm] 40 40 50 60 80 80 60 60 50 60 50 50
Tma = 11[°C]
Pp = 600 [mm], where:
Tma – annual average temperature;
Pp – annual average precipitations;
Pp
I ar = , where: (1)
Tma
+ 10
Iar – de Martonne dryness index
600
I ar = = 28,
5 ≅ 29
11+
10
The value of de Martonne dryness index calculated with formula 1 suggests the fact that
the forest vegetation can be developed in favourable conditions. In this context it can be
noticed the growth dynamic of samples from cerris species (Q. cerris) developed from
seedlings in the forest stand from u.a. 11C, analysing the picture in figure 1.
In the oaks forest stand from U.P. II Valea Lungă non-uniform regeneration cuttings are
applied – progressive cuttings (in gaps), and as a result, the regeneration benefits from
the best bio ecological conditions if the treatment technique is strictly applied and it
adapts correctly to the state and structure of the working forest (Florescu I.I., Nicolescu
N. V., 1998).
During 1995 - 2004, in u.a. 3A, 4A and 11C from U.P. II Valea Lungă interventions in
non-uniform regeneration cuttings were applied – progressive cuttings (in gaps), and
were correlated to the fructification years at the main cvernicee species (Q. robur, Q.
cerris, Q. frainetto, Q. petraea, Stănescu V., et. al.,1997) .
586

In the following lines it is presented the dynamic of the interventions in u.a. 3A, 4A and
11C in U.P. II Valea Lungă, analysing the distribution of the number of trees on
diameter categories and respectively on the intensity of interventions on volume (fig. 2,
3, 4, 5, 6, 7).
Fig. 1 – Growth vigour at cerrus regeneration developed from seedling
Analysing figures 2 and 4, it can be observed a surplus of trees of small diameters, trees
that were not totally extracted at the cultural operations (especially on the occasion of
the first thinning), at the preparatory cutting, but the extraction was done step by step, at
the application of favourable workings, the seedlings establishing, on ecologic grounds,
in order to prevent the establishing of the grassy vegetation, the soil sodding and the
occurrence of the forest vegetation succession, on those areas.
The presence of trees with small diameters in an increased number, that constitutes
an element of a specific forest stand, can be explained by the fact that in the gaps
accidentally opened, at the extraction of the accidental and hygiene products, there were
established seedlings that developed at the shelter of the existent forest stand.
Due to the fact that the interventions have a complex character (character of
preparing, of gaps opening, of gaps enlargement and connection – on some areas), they
will be called P1 respectively progressive 1, P2 – progressive 2, etc.
587

Număr de arbori
700
600
500
400
300
200
100
0
0
-100
20 40 60 80 100
Diametru [cm]
97 98 2001 2004 Arboret
Fig. 2 – Dynamic of the trees number distribution on diameter categories when
applying the regeneration cuttings in u.a. 3 A
R
18%
P3
54%
P1
15%
P2
13%
P1
P2
P3
R
Fig. 3 – Intensity of interventions on volume in u.a. 3 A,
P1 the first intervention (with a preparing character), P2 – the second intervention,
P3 – the third intervention, R – the last intervention of connecting
From the analysis of the data presented in the diagram in figure 3, we can notice that the
intensity of interventions at the regeneration cuttings in the forest stand from u.a. 3A is
almost equal in P1, P2 and R, in P3 extracting almost half of the initial forest stand,
aspect proved by the presence of a very abundant fructification at the oak species in the
years 1999-2000 (Maxim I. C., 2003).
588

Număr de arbori
160
140
120
100
80
60
40
20
0
0
-20
20 40 60 80 100
Diametru [cm]
1998 2000 2001 Arboret
Fig. 4 - Dynamic of the trees number distribution on diameter categories when
applying the regeneration cuttings in u.a. 4 A
From the analysis of the data presented in the diagram in figure 5, we can notice that the
intensity of interventions at the regeneration cuttings is relatively equal in the three
interventions, the forest stand being grown in a period of about 4 – 5 years, due to the
fact that it occurred two very abundant fructifications at the oak species in the years
1993-1994, and respectively 1999-2000, because of the favourable stational conditions
(Florescu I.I., Nicolescu N. V., 1996).
30%
32%
38%
Fig. 5 – The intensity of the interventions on volume in u.a. 4 A
589
P1
P2
R

From the analysis of the experimental distributions presented in figure 6, it can be
noticed the fact that in u.a. 11 C the number of small trees is relatively reduced
compared to u.a. 3A, but there is a surplus, their extraction being carried out step by
step during the interventions, on the reasons mentioned before.
Număr de arbori
100
90
80
70
60
50
40
30
20
10
0
-10 0 20 40 60 80 100
Diametru [cm]
1995 1998 2000 2001 Arboret
Fig. 6 - Dynamic of the trees number distribution on diameter categories when
applying the regeneration cuttings in u.a. 11 C
17%
24%
25%
34%
Fig. 7 - The intensity of the interventions on volume in u.a. 11 C
590
P1
P2
P3
R

Analysing the data presented in the diagram from figure 7, it can be noticed that the first
intervention – respectively P1 has intensity on volume of 34%, P2 an intensity of 25%,
P3 has an intensity of 17% and at the connecting cutting it will raise at 24% from the
forest stand. This dynamic of the intensity of interventions carried out at the
regeneration cuttings is explained by the fact that the fructification at oak species was
abundant and the regeneration is done in the best conditions.
DISCUSSION
Some economical imperatives have imposed the carrying out of interventions at
relatively short periods and with relatively increased intervention intensities. For
reaching a massif state in a relatively short period, it was carried out works for
completing the natural regeneration using rapidly growing species (red oak).
In the regeneration compositions there were promoted some species that regenerated
relatively easy from seeds – field maple, Banat field maple, etc., that favour the
reaching at a massif state in best conditions and ensure the stability of the ecosystems
where they are situated (Enescu V., et. al., 1997); The need of silvotechnical
interventions at the new forest stand is imperative because in some situations the
shrubby vegetation becomes overwhelming, being able to spoil the composition of the
new forest stand.
The need of helping works for a natural regeneration (favouring works of establishing
the natural regeneration) is evident, for a successful valuing of fructification at basic
species figure 8 a and b.
a) b)
Fig. 8 – Favouring works for establishing a natural regeneration in the forest
stand in u.a. 4A
591

The wild boars had a special contribution for the acorn incorporation in the soil, which
in the same time influenced the forest soil’s loosening process in the cvernicee forest
stand that fructificated abundantly.
The local experience must be highly valued, thus, having some benefits from the
abundant fructification of the main oak species, the carrying on of some interventions of
high intensity (punctiform interventions) correlated with a proper system of collecting
establishments allowed a reduction of the regeneration period up to 5-8 years.
REFERRENCES
Doniţă N., Purcelean Şt., Ceianu Ig., Beldie A., 1977, Ecologie forestieră, Editura
Ceres, Bucureşti;
Enescu V., Cherecheş D., Bândiu C., 1997, Conservarea biodiversităţii şi a resurselor
genetice forestiere, Editura Agris - Redacţia revistelor agricole, Bucureşti;
Florescu I.I., Nicolescu N. V., 1996, Silvicultura vol. I Studiul pădurii, Editura Lux-
Libris, Braşov;
Florescu I.I., Nicolescu N. V., 1998, Silvicultura vol. II Silvotehnica, Editura
universităţii Transilvania Braşov;
Giurgiu V., 2005, Principii şi criterii pentru alegerea speciilor, proiectarea şi realizarea
de compoziţii optime ale arboretelor, Editura Ceres, Bucureşti;
Maxim I. C., 2003, Lucrare de diplomă, Universitatea din Oradea, Facultatea de
Protecţia Mediului;
Stănescu V., Şofletea N., Popescu O., 1997, Flora forestieră lemnoasă a României,
Editura Ceres, Bucureşti;
*Amenajamentul Unităţii de Producţie II Valea Lungă, Ocolul Silvic Lugoj, Direcţia
Silvică Timişoara;
**Norme tehnice privind alegerea şi aplicarea tratamentelor, 2000, Ministerul Apelor,
Pădurilor şi Protecţiei Mediului, Bucureşti;
592

WASTEWATER TREATMENT TEHNOLOGIES OF BEER INDUSTRY OF
ROMANIA
Pantea Emilia Valentina*, Mirel I**., Romocea Tamara*, Mitrasca Mihaela*
*University of Oradea, ** University of Timisoara
ABSTRACT
For beer industry, the implementation of the (EMS) ISO 14001 protection managerial
system is primordial, determinant for the study of wastewater treatment optimization.
Beer factories are using a lot of water, so it is very important that they choose
efficient methods for treating the wastewater in different stages of the production with a
minimum impact on the environment.
At first, in our country, an anaerobe treatment method was chosen, either for an
anaerobe treatment of this water, but comparing the efficiency of these two processes as
well as the process combined of anaerobe pre-treatment followed by passing the
anaerobe effluent through an anaerobe treatment stage, it was noticed that this choice is
much more advantageous for used water from beer industry.
This paper is trying to exemplify this technology in the case of a beer factory from
Romania.
Keywords: aerobic, anaerobic, brewery effluent, wastewater treatment
INTRODUCTION
Along time were recorded the effects of waste water coming from breweries on the
environment and especially on the effluents in which they are discharged. Protecting the
environment, especially the water sources, has become one of the primordial objectives
of the government, in this way the adoption of a more efficient water management was
attempted.
Beer breweries from Romania produce different sorts of beer, their number has
been increasing after 1990. The quantity and quality of waste water coming from these
breweries vary between 3-5 and 40-50 hectolitres, being dependant on a series of factors
specific to production process.(Pantea et al., 2005)
The treatment of industrial waste water is considered as a non-productive activity,
so that many industries have tried to minimize the investments in water treatment plants.
But, in the last years, together with our EU adhering the legal framework has become
more restrictive, so it has determined the installation and performing of water treatment
plants of industrial wastewater which should be prerequisite for each industry.
WASTEWATER TREATMENT
Breweries and malt factories are part of the fermentative industries category which use
raw materials like: barley, corn, wheat, hop and which use big quantities of water for the
technological process.
Due to the high content of substances easily fermentable, there are some acid
fermentation phenomena, which give off unpleasant smell and develop fungi and the
content in matters in suspension favour the make up of sediments which affect the
piscicultural environment by using the oxygen. (Negulescu C, 1987)
593

The managerial plan of breweries consist in introducing some measures and methods
meant to assure a minimum consumption and a maximum recirculation, achieved
through integrated systems such as: wastewater treatment by recovering and reusing
both the water and the sub-products resulted after the treatment process.
The main wastewater treatment methods from breweries are based on: mechanical
purification, chemical treatment and biological purification assuring in this way the
removal of CBO5 to maximum 30%, 75% and respectively 95%.
Mechanical treatment is based on physical proprieties aiming at the separation of
suspensions and other materials; it uses decanters.
Chemical purification aims at separating or transforming the settling substances or
in suspension by using chemical reactives, assuring in this way the neutralization, the
separation of colloidal and dissolved substances, etc.
Biological purification is based on the use of dissolved and undissolved organic
substances by the micro organisms for multiplication and conservation. According to
the presence or lack of oxygen, the transformation of organic substances by bacteria can
be achieved in aerobe and/or anaerobe processes.
The residues discharged from breweries represent around 39 kg/hl of produced
beer and are mainly made up of 55% carbon hydrates, 27,8% proteins, 6,7% cellulose
and 6,4% grease. The evacuated wastewater charge is reduced to 70-90% as a result of
fermentation process, the resulted fermented sludge is a very good fertilizer.
Among biological treatment systems one can distinguish between anaerobic and
aerobic processes. Anaerobic treatment is characterized by biological conversion of
organic compounds (COD) into biogas(CH4 70-85% and CO2 15-30% with traces of
hydrogen sulphide). During aerobic treatment oxygen is supplied to oxidize the COD
into CO2 and H2O. Both biological processes produce new biological biomass.
The overall basic reactions are:
Anaerobic: COD → CH4 + CO2 + anaerobic biomass
Aerobic: COD + O2 → CO2 + H2O + aerobic biomass
Table 1 presents a general comparison between anaerobic and conventional aerobic
biological treatment systems.(Driessen W. et al., 2003)
Table 1: Anaerobic treatment as compared to aerobic treatment
AEROBIC SYSTEMS ANAEROBIC
SYSTEMS
COD removal 90 - 98% 70 – 85%
Nutrients (N/P) removal high low
Biosolides production high low
Space requirement high low
Energy consumption high low
Energy production no yes
Anaerobic pre-treatment followed by aerobic post – treatment will result in a positive
energy balance, reduced sludge production and space saving.
594

As a result of the comparisons between the aerobe and anaerobe processes the
advantages of introducing the anaerobe processes for the treatment of wastewater with
medium and high charges were discovered.( Minovsky J.,2002).
CASE STUDY
Brewery’s purification plant was initially designed for the purification of wastewater
using the aerobe biological purification process, but because the results were not
satisfactory, they decided to upgrade it and to introduce an anaerobe pre-purification
phase achieved in a reactor UASB type and then the aerobe treatment of the effluent
corresponding to this phase.
THE DESCRIPTION OF THE TECHNOLOGICAL PROCESS
In table 2 the main characteristics of the process are introduced:
Table 2: Parameters specific to purification process
Parameters Unit of measurement Values
COD mg/l 10500
DEBIT mg/l 4200
COD mg/l 2181
TSS mg/l 750
Temperature
o
C 30
pH - 5-8
The wastewater from the brewery with an 86 m 3 /h output will be pumped to a buffer
tank with a 780 m 3 volume, only after a separation of the solid substances of higher
volume in a static filter, the suspended solids being collected in a container for wastes.
The water is passed from the buffer tank where it is constantly mixed with the help of
two submersible mixers. The period of water retention in the buffer tank is 13 h.
After the evaluation of the costs and for their reduction, it was concluded that the
integration of a neutralization tank inside the buffer tank was a good solution.
The chemical dosage and the recirculation of the anaerobe effluent will be done in
this neutralization tank. The measuring of the pH in this tank is continuous. From the
correction tank the water is pumped for the anaerobe treatment with the help of two
pumps. The influent flow is constantly maintained with the help of an electromagnetic
flow measuring device. The output of the influent is 111 m 3 /h.
The reactors active volume is estimated at about 370 m 3 /reactor, so there is a total
active volume of 740 m 3 . This volume releases a volumetric COD charging of 6,1
kg/m 3 . In the anaerobe reactors a 80 –85% purification productivity should be achieved,
the methane production is 0,3 m 3 CH4/COD removed for a concentration of 80% CH4
out of the biogas production.
The characteristics of the anaerobe effluent are:
- COD: 436 mg/l
- BOD: 260 mg/l
595

The effluent from the anaerobe reactors flows due to gravity towards the tank where the
aerobe purification of the anaerobe pre-treated water takes place. A smaller flow will be
recirculated towards the correction tank through a recirculation pipe.
The percolating filters existent before the plant was extended will be used as
aeration tanks. When the biofilter is filled up to a height of 5,1 m (the tank’s volume is
770 m 3 ), o decrease of BOD is produced for an anaerobe productivity of 80% of: 0,71
kg BOD / m 3 . The four aeration tanks are used in a series configuration.
The anaerobe effluent will flow together with the recirculation course of the sludge
decanters in the first aeration compartment, passing then through all the four
compartments. The dissolved oxygen will be measured with the help of a measuring
device placed in the third and forth aeration compartment. The aeration is achieved with
the help of a bubble aeration system, supplied by two bellows, the retention type being
2,9 hours. The tank is aerated with a capacity of 334 Nm 3 /h.
At the forth compartment outlet of aerobe treatment, a sludge/effluent separation is
achieved with the help of sludge decanters, with a total area of 226 m 2 .
The sludge resulted after the purification should be processed by thickening, as a
result the thickened sludge is separated and the excess water from the sludge thickener
will flow in the pH correcting tank.
The decanted effluent has the following parameters: Flow: 86 m 3 /h
BOD: 82 mg/l
TSS: 158 mg/l
pH : 6,5 – 8,5
temperature: < 30 o C
Figure 1. The scheme of the technological process
596

CONCLUSIONS
Anaerobe-aerobe treatment is the most efficient alternative of wastewater treatment for
breweries, the effluents’ qualities which leave the water treatment station are greatly
improved.
Combined anaerobic/aerobic treatment of brewery effluent has important
advantages over complete aerobic treatment especially regardind: reduced sludge
production, significant low space requirements and a pozitive energy balance.
REFERENCES
1. Dima M., Meglei V., Dima B., Badea C., - 2002- Bazele epurării biologice ale
apelor uzate, ETP Tehnopress, Iaşi
2. Driessen, W., Habets, L. Verejiken, T. – 1997- Novel anaerobic – aerobic process
to meet strict effluent plant design requirements, Ferment. Vol.10. No.4, U.K.,
pp.243 – 250
3. Driessen W., Vereijken T.,- 1972 - Recent developments in biological treatment of
brewery effluent – The Institute and Guild of Brewering Convention, Livingstone,
Zambia, March 2-7, 2003, Nashville, Tennessee
4. Ianculescu, O., Ionescu GH., Racoviţeanu Raluca,- 2001 -Epurarea apelor uzate,
Editura Matrix Rom, Bucureşti
5. Minovsky J., Svatopluk C.,- 2002 - Compararea tratării anaerobe şi aerobe a apei
uzate provenită de la industria alimentară, Sinaia
6. Negulescu M. Antoniu R., Negulescu M.,- 1987 - Epurarea apelor uzate
industriale, Editura Tehnică, Bucureşti
7. Pantea E., Mirel I., Romocea T., - 2005 - The anaeroba-aerobe purge system, an
efficient alternative for the purge of warn-out waters from the beer industry,
Conferinţa internaţională „Tehnologii moderne pentru mileniul III”, Oradea,
pp.115-120
597

Figure 2. Wastewater treatment in a brewery
598

REGIONAL DEVELOPMENT USING CONSTRUCTED WETLANDS FOR
WASTEWATER TREATMENT IN VALEA IERULUI (ÉRMELLÉK) REGION
Botond L. Pete
City Hall of Oradea
P-ţa Unirii nr. 1, Oradea, Romania
ABSTRACT
Valea Ierului (Érmellék) lays one the Romania - Hungary border region, and joins the
settlements of the two country sides. As the level of development is not too high in the
region, new solutions are needed to be found, especially in the domain of the
wastewater treatment. The most suitable solution for communal wastewater treatment
should be the epuration with constructed wetlands, because this technology is less
expensive then the classic methods, and it makes possible to save funds by making
smaller investments. Using this technology, the saved funds can be returned and used in
other projects in the concerning of regional development. The most suitable wetland
plants that can be used in the region are the Phragmites australis (communis),
Schoenoplectus lacustris, Typha latifolia.
Keywords: regional development, Valea Ierului, wastewater treatment, constructed
wetlands
INTRODUCTION
One of the major problems of Europe became the issue of wastewater treatment in the
local communities. The irreversible growth of the cities results in a higher water
consumption and the increase of the wastewater quantity. Due to the national increase of
the Romanian life standards more wastewater treatment systems will be needed and this
fact will raise more problems for the local authorities. Starting with the year of 2007
every village with more than 2000 citizens, must have an adequate wastewater treatment
system. In the same time, the increasing use of chemical substances, in special those
from the domestic use, make necessary the modernization of the purification plants. The
building of a modern infrastructure is very expensive, and the major part of the small
communities does not have the necessary financial resources.
A short presentation of Valea Ierului (Érmellék) Region
Valea Ierului (Érmellék) area can be defined as a region between the foothills of the
Occidental Carpathians, the Bükk, Meszes and the Réz-hegység Mountains, and the
Tisza’s lowland. The bigger part of the region is represented by the Sălaj (Szilágy) and
Bihor (Bihar) counties from Romania and just a smaller part belongs to Hungary. On
the north side it is bordered by the Crasna’s basin, on the east side by the hills of Sălaj
county, on south by the valley of Barcău (Berettyó) river. The town of Săcuieni
(Székelyhíd) can be considered the geographical center of the region.
Valea Ierului (Érmellék) as a region was described in more research studies, but
the geographical limits were not established exactly until now. Some of the specialists
consider that Valea Ierului (Érmellék) is such a region that includes Tasnad (Tasnad)
and the others claim that Carei (Nagykaroly) is also included. According to this theory
599

Valea Ierului (Érmellék) has 150.000 residents. In other specialists’ opinion the region
of Valea Ierului (Érmellék) does not include Carei, Tasnad and other localities, as the
number of the residents did not exceed 77.000 inhabitants. As there are more concepts
related, we have to choose one of them. Therefore, we consider that Valea Ierului
(Érmellék) is a region that has 80 localities and 77.000 inhabitants. The largest localities
are the following: Carei (with 45.000 inhabitants), Marghita (Margita – 29.000
inhabitants), Valea lui Mihai (Ermihalyfalva – 21.000 inhabitants), Tăsnad (Tasnad –
20.000 inhabitants), Sacuieni (Szekelyhid – 18.000 inhabitants), Diosig (Bihardioszeg –
9.000 inhabitants), and on the Hungarian region border, Letavertes (12.000 inhabitants).
Figure 1: The map of Valea Ierului (Érmellék)
(Veliky J, 2005)
The natural structure of Valea Ierului (Érmellék)
The region is characterized - as it is situated in the middle of the Carpathian basin – by a
continental climate and by some fluctuations of the temperature, though the average
value does not pass 10°C. According to the measurements made at the Sacuieni
Meteorological Station in 1996 the average precipitation wasn’t more than 800mm.
The value of the atmospheric pressure is not more than 1000 milibar, the number
of sunny hours are approximately 2000, and this fact indicates the equality of the
number of sunny and cloudy hours.
The surface water system of Valea Ierului (Érmellék) is composed by three rivers.
The most important river is the Ier (Ér), which crosses the area from the north-east side
towards south-west. The other rivers are Crasna (Kraszna) in the north and Barcău
(Berettyó) in the south-west.
The Valea Ierului (Érmellék) of our days is not the same with the one from the
40’s. In those times the region was a marshy, swampy area with a rich biodiversity,
many floral and fish species. Some specialists considered that in that time the region
could compete with the biodiversity from the Danube’s Delta. The flora was
600

characterized by marshy plants like the common reed, sedge and the rush. These plants
are also present today, but unfortunately in a small number.
Related to the characteristics of the soil we have to mention that in this area we can
find chemozem, but the oxygen poor soils with an important content of clay are
dominating the area (general characteristics of the marshes). The sandy soil can also be
found in some places, but that is not characteristic.
Wastewater treatment with constructed wetlands
The wastewater purification with constructed wetlands is not a very new water cleaning
technology. It has already been used since the 60’, when some German scientists
discovered the purification capacity of the natural wetlands, and after that the idea of
wastewater cleaning with plants was spread all of the word. Nowadays, in Hungary
many systems like that are working and their efficiency is really high. The reduction of
P, NOx, NH3, K, metals, phenols and other elements can compete in many cases with
the classic wastewater treatment systems.
The wetlands are very successful wastewater purification systems all over the
world, and they are very various. They can be characterized by many criteria, as their
genesis, location, functioning.
By their genesis wetlands can be:
• Marshes
• Bugs
• Flood area
• Marginalized wetlands
• Periodic wetlands
From another point of view they can be natural or artificial wetlands, lakes or marshes,
and some of them are complex piped systems, with a surface or a subsurface flow. In
these complex systems the input wastewater generally enters in the pipe lines flowing
through under the root zone of the plants (generally common reed), and after that is
evacuated in the nature. At this time the water has to be cleaned and purified.
The technology that is going to be used depends on the situation of the villages, the
number of residents, the composition and quantity of the wastewater, external factors as
climate, type of the soil and many other factors. The origin of the wastewater is another
criterion that has to be taken into consideration in order to select the suitable methods,
because there is a significant difference between the wastewaters coming from
communal, industrial or agricultural use.
The system of constructed wetlands is based on the absorption process of the
pollutants by the emergent, submersed and floating plants, using the biofilters created
under the water surface, and in that way the toxic elements are going to be eliminated
from the water.
There exist more physical, chemical and biological processes that make the
treatment efficient. These processes generally are the following: sedimentation,
filtration, adsorption and absorption, biological decomposing, and there are many other
processes that assure together a complex purification.
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The advantages of this technology are numerous:
• These are ecological and natural systems, and they can represent a habitat for
other species that do not take part in the purification process (plants, birds and
other animals);
• The system doesn’t need too much maintenance work, big stuff (numerous
employees), and compared to the classic technology it has low operating costs;
• It is not as expensive to be built as the classic systems;
• Constructed wetlands can reach the same high efficiency than the classic
systems;
• They can be used as a zone of recreation, of entertainment, sport and other
events for people;
• Corrects the panorama of the region.
General aspects regarding the plants that can be used in the purification process
The world’s flora is divided in more floral zones that are organized in systems and
subsystems, on each continent. Valea Ierului (Érmellék)’s aquatic flora belongs to a
temperate climatic zone and it is characterized by a vegetation period in winter time. In
this period, when the water loses its flowing capacity because it freezes, the life
processes of the plants are going to slow down. After that some enzymatic processes
start to take place and the result is the development of the vegetative organs.
There are more types of constructed wetlands taking into consideration the plants
that are going to be used: wetlands with emergent, submersed and floating plants; they
can also use the both types of plants. The natural growth is influenced by many factors
as: the water’s depth, the extension of the water surface, debit and change of the water
level. As some secondary factors we can mention the atmospheric factors (temperature,
precipitation, wind speed, evaporation, number of hours with sunlight, etc.), pedological
factors (type of the soil, clay quantity, etc.), biological factors (the presence of certain
floral and fauna species) and characters of the bulked wastewater (qualitative and
quantitative composition, ph, temperature, turbidity, content of the organic material).
These factors are going to be researched in the next period of our scientific activity.
The emergent plants are the most frequent plants in the aquatic systems. They can
be found in the marshes and bogs, where the plants are fixed in the sediments with their
roots, having their stems and leaves above the water surface. In the water they create
some biological associations, named biofilters, and in that way they contribute to the
cleaning process. Such plants are the common reed (Phragmatis australis, Phragmatis
comunis), the rush (Schoenoplectus lacustris ) and the bulrushes (Typha latifolia, T.
augustipfolia, T. minima).
Other types of the macropytes are the floating plants, which – due to their special
cellular constitution and their air filled leaves – are drifting on the water surface. They
have two subcategories, some of them being unattached and others attached floating
plants.
The floating unattached plants are drifting on the water surface without touching
the bottom with their 20-30 cm long roots. The Pistia Stratiotes or the Eichornia
Crassipes belong to this category. The floating attached plants are also floating on the
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watersurface, but their roots are touching the bottom sediments of the water. Such plants
are: the Nymphaea alba, the Nuphar lutea, the Lemna ssp. and the Wolffia arrhiza.
The submersed plants are another category of the aquatic flora, which are fixed in
the underwater ground (in some of the cases the roots are missing), and their leaves and
stems are entirely under the water surface. They play several roles in an aquatic habitat
like: production of the organic material, oxygen liberation, transfer of material
providing of life place for the flora and fauna of the underwater ground. Many plants
belong to this category, but we will mention just some of them like: Potamogeton
pectinátu, Elodea canadensis, Chara ssp., Lemna trisulca and Ceratophyllum
demersum.
Final conclusion, remarks
The wastewater treatment with constructed wetlands is the same, or is similarly efficient
as the classical purification systems, so that they could be used in the localities from the
Valea Ierului (Érmellék) region. Taking into consideration that in this area the most
localities don’t have the infrastructure to perform a wastewater treatment, we think, that
this kind of technology could assure a good solution, or at least an alternative for the
problems. The building of the system is not as expensive as the building of the classic
one and thus the capital that has not been used can be reinvested and used in other area
of regional development.
In conclusion this technology can solve in a direct way the wastewater
treatment issue in the region, and in an indirect way, on long term, can strongly
contribute to the local and regional development.
REFERENCES
• Veliky J. (2005): Érmellék Kalauz. Hatvani István Szakkolégium, Debrecen. 21-27,
79-10 p.
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STUDY OF VERTICAL MOVEMENT OF SOME MICROELEMENTS
IN THE SOIL
László Szegedi (1) – Lajos Szabó (2.) – Mária Takács Hájos (2.)
(1.) Károly Róbert College Department of Environmental Management and Agronomy,
(2.) Tessedik Sámuel College, Institute of Environmental Sciences
ABSTRACT
A long term field experiment was established with different doses of some heavy metals
(As, Cr, Cd, Cu, Hg, Pb, Zn) to study their behaviour in the soil-plant system. The soil
of the experimental site was characterized by pH (H2O) 6.2, pH (KCI) 5.4, soil organic
matter content 3%, and clay mineral content 30-35%. To check the vertical movement
of the studied elements, two years after application soil samples were taken from the 0-
30, 30-60 and 60-90 cm layers of plots treated with the highest heavy metal load (270
kg element/ha). Both total (cc.HNO3+H2O2 extractable) and available (NH4 – acetate +
EDTA extractable) element content was determined using the ICP-MS technique.
Vertical movement of Cr (VI) was detected int he full soil profile. Therefore Cr
contamination of the soil profile suggest a significant risk for the quality of groundwater.
As and Pb remained mostly int he 0-30 cm layer. There was only a slight
contamination int he 30-60 cm layer. Cd, Hg, Cu and Zn were fixed in the plough layer.
Both total content and available fraction of these elements were detected in the upper 0-
30 cm layer. These contaminants seem to be resistant to leaching.
INTRODUCTION
Environmental pollution is placing an ever-increasing load on the various resources of
our environment, including soil. Tests and experiments involving heavy metals and
other toxic elements play a very important role in our environmental research programs.
Soils polluted with heavy metals can be found near industrial areas, metropolises, along
major transportation routes, roads, and areas treated with waste-water sludge, but soils
can also be ”polluted” geologically (1). Soils are able to accumulate heavy metals for
many years without the obvious signs of their acute toxic effect. However, the
filtering/buffering capacity of soils is finite and above a certain level soils are no longer
able to absorb these elements and become sources of pollution themselves. Toxic
elements are released into water, absorbed by cultivated crops and plants; they are
assimilated into vegetative and generative organs, and enter the food-chain where they
cause long-term harm (2,3).
The system of relations between heavy metals-soils-plants can be accurately and
reliably studied in targeted field experiments. The Ministry of Environment and the Soil
Research Institute of the Hungarian Academy of Science launched a research program
in 1991, titled ”Heavy metal loading of the environment”. The goal of the research
program is to study the effect of certain heavy metals and other potentially toxic
elements in the soil –plant system and in the food chain. The study is carried out in long
term field experiments in major soil types (carbonate-loam, carbonate-sand, acidicloam,
acidic-sand) (4). The aim of the present scientific paper is to examine the
following:
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• Which of the heavy metals that are dispresed over the soil accumulate in the upper
(tilled) strata/horizons?
• Are the heavy metals that are dispresed over the soil, leached into deeper horizons
and as a result, do these elements endanger ground water?
MATERIAL AND METHODS
A field experiment was set up at the K.R. College Farm in 1994, to study the effect of 8
heavy metals (Al, As, Cd, Cu, Hg, Pb, Zn) at three dosage levels (0/30, 90 and 270
kg/ha of each element). These were repeated three times on 35 m 2 plots (Table 1).
Element
Al
As
Cd
Cr
Cu
Hg
Pb
Zn
Table 1. Field experiments with heavy metal load, Gyöngyös, 1994
1
Load levels kg of element/ha
2 3
0
90
270
30
90
270
30
90
270
30
90
270
30
90
270
30
90
270
30
90
270
30
90
270
Form of salts
applied
Al(NO3)3 · 9H2O
NaAsO2
3CdSO4 · 8H2O
K2CrO4
CuSO4 · 5H2O
HgCl2
Pb(NO3)2
ZnSO4 · 7H2O
The soil type used for the experiment is mildly acidic chernozem brown forest soil
formed on alkaline sediment (andesite, andesite tuff). Key soil parameters are the
following: pH(H2O)=6.2; pH(KCl)=5.4;y1=9.5; CaCO3%=0; humus%=3; KA=45;
L%=70; hy=4.8. Particle size of the soil is dominated by clay and silt fraction. The
textural type is clay-loam with a bulk density of 1.20 g/cm 3 . Soil surface is slightly
sloping, elevation is 150 m above sea level. Water table depth is 10 m, therefore the
probability of pollution through surface leaching is minimal. The soil has good water
absorption, conduction and storage capability. In the event of heavy rainfall, rill erosion
occurs ont illed surfaces.
The soil in the plots was examined and tested each year. Samples were taken from
the following strata of the plots treated with the maximum dosage (270 kg/ha): 0-30, 30-
60, 60-90 cm. Samples were taken from bored holes in cartridges. In each plot, 5 bores
were made at each depth range to obtain at an average sample. The samples were dried,
ground and homogenised before determining the ”total” element content estimated with
the cc.HNO3+cc.H2O2 extraction and the available element content with the Lakanen-
Erviö’s method (6). The analysis of samples for elements was carried out by the ICP
laboratory at the Soil Research Institute (RISSAC) of the Hungarian Academy of
Science, for 25 elements, using the ICP-AES method.
RESULTS AND DISCUSSION
As was dominantly present in the upper stratum (0-30 cm) int he 2nd year of the
experiment ( Figure 1). The 0-60 cm stratum was also polluted as shown in the figure by
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the ”total” and available element content. Enrichment/accumulation in deeper soil
horizons/strata is not significant, with respect to the tolerance of sampling, it is
practically negligible. Leaching, and the extent and dynamics of leaching cannot be
determined from this data.
Figure 1. The effect of 270 kg/ha arsenic load ont he arsenic content of the soil
profile (Chernozem brown forest soil, Gyöngyös, 1996)
Figure 2. The effect of 270 kg/ha cadmium load on the cadmium content of the soil
profile (Chernozem brown forest soil, Gyöngyös, 1996)
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Figure 2 shows that Cd also is concentrated int he upper, tilled stratum of soil. Both
”total” and available content is present int he place of input, deeper horizons show no
sign of accumulation. It seems that this extremely toxic element is highly resistant to
leaching in loamy soils int he initial years. According to literature, Cd is highly mobile
in the soil-plant system (5, 7).
The mobility of Cr(VI) is unambiguously evident (Figure 3). In comparison with
unpolluted control soil, accumulation in the deeper strata between 30-60 and 60-90 cms
is obvious, both in terms of ”total” and available content. Heavy rainfall in 1995 and
1996 may also have contributed to leaching. It is also true that the available Cr content
of the deeper strata is one magnitude less than the values measured at the input point.
The fact that Cr pollution was, although more moderately, apparent between 60-90 cms
after the second year draws attention to the importance of examining the mobility of
Cr(VI) in the horizons beyond 1-meter depth. We will need to monitor the limits of Cr
leaching in the future with deeper test bores.
Figure 3. The effect of 270 kg/ha chromium load ont he chromium content of the soil
profile (Chernozem brown forest soil, Gyöngyös, 1996)
The majority of Pb remained at the point of input, however it is not realistic to assume
that this element will remain completely immobilised in acidic forest soils. Hence Pb
may pose a moderate danger to soil water and the food chain (8). According to the
results of soil examinations, Hg is not leached. Literature shows that HgCl2 is rapidly
bonded by organic matter and clay fractions or precipitated in the form of insoluble
salts. Solubility may increase under acidic conditions, thereby Hg in soils may become
mobilised. Cu and Zn are bonded in the tilled horizon, mobility was not experienced.
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CONCLUSION
1. The rate of transformation and immobilisation of toxic pollutants in soils varied
according to elements. A clear distinction can be made between elements that are
mobile pollutants in soils (Cd, Zn, Pb, Cu) and those that transform rapidly into
insoluble forms and become bonded (As, Hg, Cr).
2. Heavy metals and other pollutants that are spread over soils accumulate in the tilled
soil horizons where root mass is highest. Cd, Cu, Pb, Hg and Zn are bonded in the
tilled horizons, while As moderately polluted the deeper horizons between 30-60
cms.
3. The leaching of Cr input into soils in the form of chromate is a rapid process in
mildly acidic brown forest soils, the element was detected at depths of 60-90 cms
after 2 years. Subterranean water sources are endangered by this element as a result
of rapid leaching.
REFERENCES
1. Csathó, P. 1994. Heavy metal pollution and agricultural production. Thematic
literature review. Akaprint, Budapest
2. Fodor, L. 1977. Mobility of some toxic elements in soil-plant system on brown
forest soil. Proceedings of <strong>International</strong> <strong>Conference</strong> of PhD Students. UM,
Miskolc, pp. 84-90.
3. Fodor, L. 1977. Mobility and plant uptake of some heavy metals on brown forest
soil. Proceedings of the 11th World Fertilizer Congress. Research Center,
Braunschweg-Völkenrode, pp. 14-20.
4. Fodor, L. 2002. Nehézfémek akkumulációja a talaj-növény rendszerben. Doktori
(PhD) értekezés. VE GMK, Keszthely.
5. Fodor, L. 2004. Study of heavy metal leaching in the soil. Proceedings of the 13th
Int. ISCO <strong>Conference</strong>. Brisbane.
6. Lakanen, E., Erviö, R. 1971. A comparison of eight extractans for determination of
plant available microelements in soil. Acta Agr. Fenn. 123, 223-232.
7. Lehoczky, É., Szabados, I., Marth, P. 1996. Cadmium Content of Plants as
Affected by Soil Cadmium Concentration. Soil Sci. Plant Anal. 27. 1765-1777.
8. Szabó, L., Tamás, J., Tóth, P., Kovács, E., Fodor, L. 2003. Ólomszennyezés
migrációjának vizsgálata talajokon. In: Mikroelemek a táplálékláncban (ed: Simon,
L., Szilány, M.) Bersenyi György Könyvkiadó, Nyíregyháza, pp. 221-228.
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