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Diversity of dechlorination pathways and organohalide respiring bacteria in chlorobenzene dechlorinating enrichment cultures originating from river sludge

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Abstract

Anaerobic reductive dechlorination of hexachlorobenzene (HCB) and three isomers of tetrachlorobenzene (TeCB) (1,2,3,4-, 1,2,3,5- and 1,2,4,5-TeCB) was investigated in microcosms containing chloroaromatic contaminated river sediment. All chlorobenzenes were dechlorinated to dichlorobenzene (DCB) or monochlorobenzene. From the sediment, a methanogenic sediment-free culture was obtained which dechlorinated HCB, pentachlorobenzene, three TeCB isomers, three trichlorobenzene (TCB) isomers (1,2,3-, 1,2,4- and 1,3,5-TCB) and 1,2-DCB. Dechlorination involved multiple pathways including the removal of doubly flanked, singly flanked and isolated chlorine substituents. 454-pyrosequencing of partial bacterial 16S rRNA genes amplified from selected chlorobenzene dechlorinating sediment-free enrichment cultures revealed the presence of a variety of bacterial species, including Dehalobacter and Dehalococcoides mccartyi, that were previously documented as organohalide respiring bacteria. A genus with apparent close relationship to Desulfitobacterium that also has been associated with organohalide respiration, composed the major fraction of the operational taxonomic units (OTUs). Another major OTU was linked with Sedimentibacter sp., a genus that was previously identified in strict co-cultures of consortia reductively dehalogenating chlorinated compounds. Our data point towards the existence of multiple interactions within highly chlorinated benzene dechlorinating communities.

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Acknowledgments

We thank the Flemish Environmental and Technology Innovation Platform for funding this research as part of the MIP project. Furthermore, we thank P.J. Haest for his help with the real-time qPCR and S. Ruyters for assistance with analyzing 454-pyrosequencing data.

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  1. Danuta Cichocka

    Present address: School of Life Sciences, Institute for Ecopreneurship, University of Applied Sciences and Arts Northwestern Switzerland, Muttenz, Switzerland

Authors and Affiliations

  1. Division of Soil and Water Management, KU Leuven, Kasteelpark Arenberg 20, 3001, Leuven, Belgium

    Pieter Vandermeeren, Steffi Herrmann, Danuta Cichocka & Dirk Springael

  2. Department of Isotope Biogeochemistry, UFZ - Helmholtz Centre for Environmental Research, Leipzig, Germany

    Hans-Hermann Richnow

  3. Laboratory for Process Microbial Ecology and Bioinspirational Management (PME&BIM), Thomas More Mechelen, Sint-Katelijne-Waver, Belgium

    Pieter Busschaert & Bart Lievens

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  1. Pieter Vandermeeren
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Correspondence to Dirk Springael.

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Supplementary Fig. 1

Concentration of CBs in killed controls of Kanaal Ieper-Ijzer sediment microcosms. Microcosms were amended with a HCB, b 1,2,3,4-TeCB, c 1,2,3,5-TeCB, and d 1,2,4,5-TeCB. Symbols indicate: HCB (X), 1,2,3,4-TeCB (filled triangle), 1,2,3,5-TeCB (open square) and 1,2,4,5-TeCB (filled triangle). Presented data points are averages of two replicate microcosms. Standard error bars are indicated. Supplementary material 1 (PPT 197 kb)

Supplementary Table 1

Selected representative OTU sequences and their closest match in GenBank. The OTUs presented are the first 26 in number and represent 93 % of the sequences of the dataset. The dataset consisted of 7,376 sequences, organised into 161 OTUs (defined by a 3 % sequence dissimilarity cut-off). The coverage is the percent of the query sequence that overlaps the subject sequence in GenBank and the similarity is the percent similarity between the query and subject sequences over the length of the coverage area. Supplementary material 2 (DOC 67 kb)

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Vandermeeren, P., Herrmann, S., Cichocka, D. et al. Diversity of dechlorination pathways and organohalide respiring bacteria in chlorobenzene dechlorinating enrichment cultures originating from river sludge. Biodegradation 25, 757–776 (2014). https://doi.org/10.1007/s10532-014-9697-y

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