Metagenomics and qPCR analysis of aerobic metabolic TCE degrading bacteria

Abstract

Chloroethenes are common groundwater contaminants. While they were initially considered non-biodegradable, different pathways for both, anaerobic and aerobic degradation have been discovered over the last 40 years. Anaerobic reductive dechlorination of tetrachloroethene and trichloroethene (TCE) leads to the formation and possible accumulation of the toxic intermediate products, dichloroethene (DCE) and vinyl chloride (VC). In contrast, aerobic metabolic processes result in mineralization of the contaminants without the formation of stable intermediates. Therefore, productive aerobic degradation processes can have a considerable advantage, depending on site conditions and chloroethene molecule. The potential for aerobic degradation processes can be assessed by molecular biological analysis. In our study, aerobic metabolic TCE enrichment cultures, bioaugmented groundwater, and a microcosm with intrinsic aerobic TCE degradation potential were analyzed. Amplicon sequencing and shotgun metagenomics were used to identify the Rhodocyclaceae bacterial family in all samples, as well as functional gene sequences, which were relatives of monooxygenases and dehalogenases. Based on these sequencing data, qPCR detection methods for TCE assimilating Rhodocyclaceae (Rho) as well as for different functional genes that were associated with monooxygenases (moABC) and haloacid dehalogenases like hydrolases (hdlh) have been established. The specificity of the qPCR methods was demonstrated with a variety of environmental samples and different chloroethene degrading bacterial cultures. The new metagenomic insights and qPCR methods facilitate the assessment of biodegradation potential and monitoring of aerobic metabolic TCE degradation at contaminated sites. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=77 SRC="FIGDIR/small/667672v1_ufig1.gif" ALT="Figure 1"> View larger version (19K): org.highwire.dtl.DTLVardef@10b51feorg.highwire.dtl.DTLVardef@3414e0org.highwire.dtl.DTLVardef@14694ccorg.highwire.dtl.DTLVardef@d2e4e9_HPS_FORMAT_FIGEXP M_FIG C_FIG HighlightsO_LIRhodocyclaceae bacteria were identified in all aerobic TCE degrading samples C_LIO_LIFunctional monooxygenase and dehalogenase like genes were detected C_LIO_LIqPCR methods for monitoring of aerobic metabolic TCE degradation were developed C_LIO_LISpecificity of qPCR methods was shown with different aerobic chloroethene degrading cultures and environmental samples C_LI