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Occurrence of Dehalococcoides and Reductive Dehalogenase Genes in Microcosms, a Constructed Wetland and Groundwater from a Chlorinated Ethene Contaminated Field Site as Indicators for In Situ Reductive Dehalogenation

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Abstract

Thus far, members of the genus Dehalococcoides are the only microorganisms known to dehalogenate chlorinated ethenes to ethene and thereby detoxify these common groundwater pollutants. Therefore, it is important to characterize the taxonomic and functional diversity of these key microorganisms and their reductive dehalogenase (RDase) genes in contaminated aquifers for assessing the natural attenuation potential. Little is known about the diversity of RDase genes under field conditions or in laboratory systems under selective pressure during dechlorination activities. Here, we evaluate the diversity of Dehalococcoides sp. and three RDase genes in groundwater as well as in water from a constructed wetland and microcosms setup with contaminated groundwater from the same field site in Bitterfeld (Saxony-Anhalt, Germany). The presence and relative abundance of Pinellas and Cornell subgroups of Dehalococcoides was evaluated by a novel direct sequencing method, which revealed that all sequences were identical and affiliated to the Pinellas subgroup. Contrarily, our results showed remarkable differences at the functional gene level between the systems. Of the vinyl chloride reductase genes, vcrA was detected in samples from the groundwater, wetland, and microcosms, whereas bvcA was only found in wetland and microcosm samples. The trichloroethene dehalogenase gene, tceA could not be detected at all, although complete dehalogenation activity of higher chlorinated ethenes was observed. Our study demonstrates that although the Dehalococcoides 16S rRNA gene sequences retrieved from the investigated systems were identical, the RDase gene diversity varied among the systems, according to the spectrum of the chlorinated ethenes present.

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Acknowledgments

É. Mészáros and G. Imfeld were supported by a European Union Marie Curie Early Stage Training Fellowship (AXIOM, contract no. MEST-CT-2004-8332). This work was supported by the Helmholtz Centre for Environmental Research – UFZ. We would like to thank D. Cichocka for generously supplying the microcosm and enrichment culture samples used and F.E. Löffler and K. Fletcher for providing the dehalogenase clones. The Department of Groundwater Remediation and the SAFIRA II Project, in particular, H. Weiss and R. Trabitzsch, as well as J. Drangmeister and J. Grossmann of GICON and the Landesanstalt für Altlastenfreistellung des Landes Sachsen-Anhalt – LAF, are acknowledged for support during sampling. We would also like to thank I. Mäusezahl and K. Ethner for help with laboratory work.

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Authors and Affiliations

  1. Department of Microbiology, Eötvös Loránd University of Science, Budapest, Hungary

    Éva Mészáros

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

    Éva Mészáros, Gwenaël Imfeld & Ivonne Nijenhuis

  3. Laboratory of Surface Hydrology and Geochemistry (LHyGeS – UMR 7517), University of Strasbourg/ENGEES, CNRS, Strasbourg, France

    Gwenaël Imfeld

  4. Department of Environmental Biotechnology, Helmholtz Centre for Environmental Research – UFZ, Leipzig, Germany

    Marcell Nikolausz

  5. Department of Bioenergy, Helmholtz Centre for Environmental Research – UFZ, Leipzig, Germany

    Marcell Nikolausz

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  1. Éva Mészáros
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  2. Gwenaël Imfeld
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  3. Marcell Nikolausz
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  4. Ivonne Nijenhuis
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Correspondence to Ivonne Nijenhuis.

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Mészáros, É., Imfeld, G., Nikolausz, M. et al. Occurrence of Dehalococcoides and Reductive Dehalogenase Genes in Microcosms, a Constructed Wetland and Groundwater from a Chlorinated Ethene Contaminated Field Site as Indicators for In Situ Reductive Dehalogenation. Water Air Soil Pollut 224, 1768 (2013). https://doi.org/10.1007/s11270-013-1768-x

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