Applicability of random cloning method to analyze microbial community in full-scale anaerobic digesters.

Microbial communities were analyzed in six full-scale anaerobic digesters and a lab-scale digester using the random cloning method. The purposes were to confirm the applicability, reproducibility, and error range of this method; to discuss the difference in the dominant microbes determined by this method in different operational conditions of temperature (mesophilic and thermophilic) and substrate (garbage, sewage sludge, and livestock waste); and to determine key microbes in each digester. Each sample was analyzed in triplicate. In one of the samples, 373 clones were analyzed to study the composition of microbial community in the digester. Time course analysis was conducted from the start-up period for approximately one year in one of the digesters. Similar microbial diversity was obtained corresponding to the type of substrate change (sewage sludge to garbage). Operational taxonomic units (OTUs) closely related to Coprothermobacter sp. and unidentified bacterium clones TUG14 and TUG22 disappeared during the first 40 d, while OTUs closely related to Bacillus sp. and Clostridium sp. increased later. Microbial diversity in digesters is strongly affected by the operational conditions, and similar microbial diversity can be obtained in triplicate analysis and under similar operational conditions. The present study verified the applicability of this method to discuss the overall difference in microbial communities.

Chlorine isotope fractionation during reductive dechlorination of chlorinated ethenes by anaerobic bacteria.

Chlorine isotope fractionation during reductive dechlorination of trichloroethene (TCE) and tetrachloroethene (PCE) to cis-1,2-dichloroethene (cDCE) by anaerobic bacteria was investigated. The changes in the 37Cl/35Cl ratio observed during the one-step reaction (TCE to cDCE) can be explained by the regioselective elimination of chlorine accompanied by the Rayleigh fractionation. The fractionation factors (alpha) of the TCE dechlorination by three kinds of anaerobic cultures were approximately 0.994-0.995 at 30 degrees C. The enrichment of 37Cl in the organic chlorine during the two-step reaction (PCE to cDCE) can be explained by the random elimination of one chlorine atom in the PCE molecule followed by the regioselective elimination of one chlorine atom in the TCE molecule. The fractionation factors for the first step of the PCE dechlorination with three kinds of anaerobic cultures were estimated to be 0.987-0.991 at 30 degrees C using a mathematical model. Isotope fractionation during the first step would be the primary factor for the chlorine isotope fractionation during the PCE dechorination to cDCE. The developed models can be utilized to evaluate the fractionation factors of regioselective and multistep reactions.