Effect of dechlorination and sulfate reduction on the microbial community structure in denitrifying membrane-biofilm reactors

Recent studies showed that the chlorinated solvents trichloroethene (TCE), 1,1,1-trichloroethane (TCA), and chloroform (CF) were reductively dehalogenated in a H₂-based membrane biofilm reactor (MBfR) under denitrifying conditions. Here, we describe a detailed phylogenetic characterization of MBfR biofilm communities having distinctly different metabolic functions with respect to electron-acceptor reduction. Using massively parallel pyrosequencing of the V6 region of the 16S rRNA gene, we detected 312, 592, and 639 operational taxonomic units (OTU) in biofilms of three MBfRs that reduced nitrate; nitrate and TCE; or nitrate, sulfate, and all three chlorinated solvents. Comparative community analysis revealed that 13% of the OTUs were shared by all MBfRs, regardless of the feed, but 65% were unique to one MBfR. Pyrosequencing and real-time quantitative PCR showed that Dehalococcoides were markedly enriched in the TCE+nitrate biofilm. The input of a mixture of three chlorinated compounds, which coincided with the onset of sulfate reduction, led to a more diverse community that included sulfate-reducing bacteria (Desulfovibrio) and nitrate-reducing bacteria (Geothrix and Pseudomonas). Our results suggest that chlorinated solvents, as additional electron acceptors to nitrate and sulfate, increased microbial diversity by allowing bacteria with special metabolic capabilities to grow in the biofilm.