An anaerobic microbial community was enriched in a chemostat that was operated for more than 8 years with benzene and nitrate as electron acceptor. The coexistence of multiple species in the chemostat and the presence of a biofilm, led to the hypothesis that benzene-degrading species coexist in a syntrophic interaction, and that benzene can be degraded in syntrophy by consortia with various electron acceptors in the same culture. The benzene-degrading microorganisms were identified by DNA-stable isotope probing with [U-(13) C]-labelled benzene, and the effect of different electron donors and acceptors on benzene degradation was investigated. The degradation rate constant of benzene with nitrate (0.7 day(-1) ) was higher than reported previously. In the absence of nitrate, the microbial community was able to use sulfate, chlorate or ferric iron as electron acceptor. Bacteria belonging to the Peptococcaceae were identified as dominant benzene consumers, but also those related to Rhodocyclaceae and Burkholderiaceae were found to be associated with the anaerobic benzene degradation process. The benzene degradation activity in the chemostat was associated with microbial growth in biofilms. This, together with the inhibiting effect of hydrogen and the ability to degrade benzene with different electron acceptors, suggests that benzene was degraded via a syntrophic process.

A bacterium (strain G5G6) that grows anaerobically with toluene was isolated from a polluted aquifer (Banisveld, the Netherlands). The bacterium uses Fe(III), Mn(IV) and nitrate as terminal electron acceptors for growth on aromatic compounds. The bacterium does not grow on sugars, lactate or acetate. Phylogenetic analysis of the 16S rRNA gene sequence indicated that strain G5G6 belonged to the Betaproteobacteria. Its closest, but only distantly related, cultured relative is Sterolibacterium denitrificans Chol-1S(T) (94.6% similarity of the 16S rRNA genes), a cholesterol-oxidizing, denitrifying bacterium. Strain G5G6 possesses the benzylsuccinate synthase A (bssA) gene encoding the alpha-subunit of Bss, which catalyzes the first step in anaerobic toluene degradation. The deduced BssA amino acid sequence is closely related to those of Azoarcus and Thauera species, which also belong to the Betaproteobacteria. Strain G5G6 is the first toluene-degrading, iron-reducing bacterium that does not belong to the Geobacteraceae within the Deltaproteobacteria. Based on phylogenetic and physiological comparison, strain G5G6 could not be assigned to a described species. Therefore, strain G5G6 (DSMZ 19032(T)=JCM 14632(T)) is a novel taxon of the Betaproteobacteria. We propose the name Georgfuchsia toluolica gen. nov., sp. nov.

The genomes of the Betaproteobacteria Alicycliphilus denitrificans strains BC and K601T have been sequenced to get insight into the physiology of the two strains. Strain BC degrades benzene with chlorate as electron acceptor. The cyclohexanol-degrading denitrifying strain K601T is not able to use chlorate as electron acceptor, while strain BC cannot degrade cyclohexanol. The 16S rRNA sequences of strains BC and K601T are identical and the fatty acid methyl ester patterns of the strains are similar. Basic Local Alignment Search Tool (BLAST) analysis of predicted open reading frames of both strains showed most hits with Acidovorax sp. JS42, a bacterium that degrades nitro-aromatics. The genomes include strain-specific plasmids (pAlide201 in strain K601T and pAlide01 and pAlide02 in strain BC). Key genes of chlorate reduction in strain BC were located on a 120 kb megaplasmid (pAlide01), which was absent in strain K601T. Genes involved in cyclohexanol degradation were only found in strain K601T. Benzene and toluene are degraded via oxygenase-mediated pathways in both strains. Genes involved in the meta-cleavage pathway of catechol are present in the genomes of both strains. Strain BC also contains all genes of the ortho-cleavage pathway. The large number of mono- and dioxygenase genes in the genomes suggests that the two strains have a broader substrate range than known thus far.

A pilot-scale reactor treating domestic sewage was operated to promote anaerobic digestion and denitrification using endogenous electron donors. While 55 % of organic matter was removed, nitrogen and sulfur showed a different dynamics during the operation. Pyrosequencing analysis clarified this behavior revealing that specific microbial communities inhabited the anaerobic (47.05 % of OTUs) and anoxic (31.39 % of OTUs) chambers. Analysis of 16S rRNA gene partial sequences obtained through pyrosequencing revealed a total of 1727 OTUs clustered at a 3 % distance cutoff. In the anaerobic chamber, microbial community was comprised of fermentative, syntrophic and sulfate-reducing bacteria. The majority of sequences were related to Aminobacterium and Syntrophorhabdus. In the anoxic chamber, the majority of sequences were related to mixotrophic and strictly autotrophic denitrifiers Arcobacter and Sulfuricurvum, respectively, both involved in sulfur-driven denitrification. These results show that pyrosequencing was a powerful tool to investigate the microbial panorama of a complex system, providing new insights to the improvement of the system.

This paper discusses the results of pentachlorophenol (PCP) anaerobic biodegradation in a horizontal-flow anaerobic immobilized biomass (HAIB) reactor operated under methanogenic and halophylic conditions. The system was inoculated with autochthonous microorganisms taken from a site in the Santos-São Vicente Estuary (state of São Paulo, Brazil) severely contaminated with PCP, phenolic compounds, polychlorinated biphenyls, polycyclic aromatic hydrocarbons, and heavy metals. The inoculum was previously enriched for methanogenesis activity by changing glucose concentrations and under halophylic condition. PCP was added to the HAIB reactor as sodium salt (NaPCP) at an initial concentration of 5 mg l(-1) and increased to 13, 15, and 21 mg l(-1). Organic matter removal efficiency ranged from 77 to 100%. PCP removal efficiency was 100%. Denaturing gradient gel electrophoresis profile showed changes in the structure of Bacteria domain, which was associated with NaPCP and glucose amendments. The diversity of Archaea remained unaltered during the different phases. Scanning electron microscope examinations showed that cells morphologically resembling Methanosarcina and Methanosaeta predominated in the biofilm. These cells were detected by fluorescence in situ hybridization with the Methanosarcinales (MSMX860) specific probe. The results are of great importance in planning the estuary's restoration by using anaerobic technology and autochthonous microorganisms for bioremediation.

No abstract

No abstract

Rapid and simple method to quantify fipronil and intermediates from soil.