52 LLU Raksti 12 (308), 2004; 47-57 1-18AS1I II IIIX inocS/RMSS cE/DX inocLMX inocS/RMAA2Fig.5.3S/RM 4Fig. 5. Flow diagram of solid state (compsting included) conversion of lignocellulose-containing (non-solubles) raw materials and waste:I – preparation of inoculum, II – pilot/demonstration stage, III – production;1 – submerged bioreactor for production of inoculum; 2 – combined liquid/solid state bioreactor; 3 – mixed SS bioreactor; 4 – windrows;5 – motionless or mixed sieves in climate chamber; 6 – production-scale mixed SS bioreactor; 7 – bin’s technology: special bins for closed composting; S – substrate;RM – raw material; X inoc – seed material; E/D – experimental/demonstration products; A – air; SS – solid state; LM – liquid medium.X inocE/DX inocE/DX inocE/DX inocX inocS/RMX inocAS/RMX inocS/RM5576E/DAProduct to the marketProduct to the marketU. Viesturs et al. Solid State Fermentation Systems for Bioremediation and Biodegradation
U. Viesturs et al. Solid State Fermentation Systems for Bioremediation and Biodegradationviride and Tr. lignorum (Viesturs et al., 1998), and anitrificator association, regulating the circulation ofnitrogen – the ammonification and nitrification processeswere applied as the inoculum for biodegradationof lignocellulose and other types of organic waste inthe composting process.MethodsChemical analysesThe concentrations of ammonia, nitrite, nitrate andhydrogen sulphide were determined by a FIAstar 5020Analyzer. The total nitrogen was measured by the“Buchi” Kjeldahl Line. Total carbon was determinedby the modified Tjurin’s method (Rinkis et al., 1987).Then NO 3-N/NH 4-N and C: N ratios were calculated.Microbiological analysesThe quantification of the total number of microorganisms,fungi, Escherichia coli and Salmonella sp.in the compost was performed. The plate count methodwas used for estimating the total number of microorganismsand fungi per 1 g of dry compost. The mediumfor quantification of the total number of bacteria was:Bacto nutrient agar (DIFCO LABORATORIES, USA);for fungi – Chapec medium, for E. coli – Endo agar; forSalmonella – agar Mak-Konky (Strikauska et al., 1999).Microbiotests- Rotoxkit F TM , which measures the lethal effect oftoxicants of rotifers freshly, hatched from cysts(with rotifers Brachionus calyciflorus), after 24-hexposure (Rotoxkit F TM , 1992).- Thamnotoxkit F TM with Thamnocephalus platyurus,which measures the lethal effect of toxicantsof crustacean freshly, hatched from cysts, after 24-h exposure (Thamnotoxkit F TM , 1995).- Ostracodtoxkit F with the benthos organismHeterocypris incongruens is a “direct contact”chronically toxicity microbiotest. After 6 days, thecrustacean morbidity and growth intensity arecompared with the control (Ostracodtoxikit F,2000).The results of microbiotesting were evaluated accordingto the regulations of the method’s authors,and the effect in percentage was observed in the nodiluted sample (Persoone et al., 2003). The followinghazard classification system was used for determinationof the degree of toxic contamination in compost:- Class I: no acute hazard = none of the tests showeda toxic effect;- Class II: slight acute hazard = a statistically significantEP was reached in at least one test, whilethe effect’s level was below 50%;- Class III: acute hazard = the EP 50was reached orexceeded in at least one test, while the effect’slevel was below 100%;LLU Raksti 12 (307), 2004; 47-57 1-18- Class IV: high acute hazard = the EP 100was reachedin at least one test;- Class V: very high acute hazard = the EP 100wasreached in all tests.IR spectroscopySamples of the fermentation media or compost forIR-spectroscopy were dried at 60 o C. 50 mg of the samplewas mixed with 1g of KBr and milled to obtain ahomogenous mixture of the sample. 210 mg of the mixturewas mixed with 890 mg of KBr, milled and pelleted.For spectral analysis of Na-humate, 25 mg of substanceand 1g of KBr was used.The prepared KBr pellets were registered on a FT-IR spectrometer Perkin Elmer (Spectrum RXIFT-IR), theabsorption mode between 400 and 4000 cm -1 , resolution4 cm -1 , 16 scans.As a characteristic absorption band for identificationof Na-humate, 1390 cm -1 (COOH group vibrations)was chosen (Haberhauer and Grzabek, 1999).Results and discussionTo perform the composting process, two Trichodermastrains and the nitrificator association for regulatingthe circulation of nitrogen–ammonification and nitrificationprocesses were applied. Trichodermalignorum was a more active splitter of cellulose andlignin substrates, while Trichoderma viride was mainlythe producer of Trichodermin (Apsite et al., 1998;Viesturs and Leite, 1996) and also the splitter of lignocellulose.Both of Trichoderma micromycetes were thebasis of the plant protection preparation Trichodermin,which could limit plant infections (Apsite et al., 1998).After a 2-month biodegradation of organic wasteunder anaerobic conditions at the thermophilic regime,the amount of Escherichia coli and Salmonellatyphimurum was 0. However, after 6 months at the aerobicregime, applying the above-mentioned associations,the ammonium content in composting media averaged410 mg per kg of the fresh weight of the compost, andthe C: N ratio was 35–40. The determination of the percentageeffect (EP) obtained with each of the appliedmicrobiotests (Rotoxkit F TM with the rotifers Brachionuscalyciflorus, Tamnotoxkit F TM with Tamnocephalusplatyurus and Ostracodtoxkit F with the benthicostracod crustacean Heterocypris incongruens)showed that the quality of the compost from thebioreactor was slight acute hazard. A statistically significantEP was reached in Ostracodtoxkit (EP – 26.6%)and Rotoxkit (EP – 21.3%) tests.The composting process proceeded still two weeksuntil the content of ammonia decreased to 280 mg perkg of the fresh weight of the compost, and the C: Nratio was 25–30. The applied microbiotests showed thatthe toxicity of the compost was no acute hazard. EPvalues ranged from 3.3% to 13.3%.53
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