Characterization of Groundwater Microbial Communities, Dechlorinating Bacteria, and In Situ Biodegradation of Chloroethenes Along a Vertical Gradient

The variability of hydrogeochemical conditions can affect groundwater microbial communities and the natural attenuation of organic chemicals in contaminated aquifers. It is suspected that in situ biodegradation in anoxic plumes of chloroethenes depends on the spatial location of the contaminants and the electron donors and acceptors, as well as the patchiness of bacterial populations capable of reductive dechlorination. However, knowledge about the spatial variability of bacterial communities and in situ biodegradation of chloroethenes in aquifers is limited. Here, we show that changes of the bacterial communities, the distribution of putative dechlorinating bacteria and in situ biodegradation at the border of a chloroethenes plume (Bitterfeld, Germany) are related to local hydrogeochemical conditions. Biotic reductive dechlorination occurred along a 50 m vertical gradient, although significant changes of the hydrogeochemistry and contaminant concentrations, bacterial communities and distribution of putative dechlorinating bacteria (Dehalobacter spp., Desulfitobacterium spp., Dehalococcoides spp., and Geobacter spp.) were observed. The occurrence and variability of in situ biodegradation of chloroethenes were revealed by shifts in the isotope compositions of the chloroethenes along the vertical gradient (δ13C ranging from −14.4‰ to −4.4‰). Our results indicate that habitat characteristics were compartmentalized along the vertical gradient and in situ biodegradation occurred with specific reaction conditions at discrete depth. The polyphasic approach that combined geochemical and biomolecular methods with compound-specific analysis enabled to characterize the spatial variability of hydrochemistry, bacterial communities and in situ biodegradation of chloroethenes in a heterogeneous aquifer.

[1]  J. Drever,et al.  Stable isotope (C, Cl, and H) fractionation during vaporization of trichloroethylene , 1999 .

[2]  K. Sowers,et al.  A PCR-based specific assay reveals a population of bacteria within the Chloroflexi associated with the reductive dehalogenation of polychlorinated biphenyls. , 2005, Microbiology.

[3]  R. Amann,et al.  Sequence heterogeneities of genes encoding 16S rRNAs in Paenibacillus polymyxa detected by temperature gradient gel electrophoresis , 1996, Journal of bacteriology.

[4]  C. Condee,et al.  Large-scale production of bacterial consortia for remediation of chlorinated solvent-contaminated groundwater , 2009, Journal of Industrial Microbiology & Biotechnology.

[5]  R. M. Lehman,et al.  Comparison of Extracellular Enzyme Activities and Community Composition of Attached and Free-Living Bacteria in Porous Medium Columns , 2002, Applied and Environmental Microbiology.

[6]  M. Brusseau,et al.  Assessment of in situ reductive dechlorination using compound-specific stable isotopes, functional gene PCR, and geochemical data. , 2009, Environmental science & technology.

[7]  M. Nikolausz,et al.  Assessment of the natural attenuation of chlorinated ethenes in an anaerobic contaminated aquifer in the Bitterfeld/Wolfen area using stable isotope techniques, microcosm studies and molecular biomarkers. , 2007, Chemosphere.

[8]  K. Hayes,et al.  Kinetics of the Transformation of Trichloroethylene and Tetrachloroethylene by Iron Sulfide , 1999 .

[9]  F. Gillet,et al.  Statistical Assessment of Variability of Terminal Restriction Fragment Length Polymorphism Analysis Applied to Complex Microbial Communities , 2009, Applied and Environmental Microbiology.

[10]  B. Sleep,et al.  Evaluation of isotopic enrichment factors for the biodegradation of chlorinated ethenes using a parameter estimation model: toward an improved quantification of biodegradation. , 2006, Environmental science & technology.

[11]  K. Scow,et al.  A Shallow BTEX and MTBE Contaminated Aquifer Supports a Diverse Microbial Community , 2004, Microbial Ecology.

[12]  James M. Tiedje,et al.  16S rRNA Gene-Based Detection of Tetrachloroethene-Dechlorinating Desulfuromonas andDehalococcoides Species , 2000, Applied and Environmental Microbiology.

[13]  K. Hayes,et al.  Reductive dechlorination of tetrachloroethylene and trichloroethylene by mackinawite (FeS) in the presence of metals: reaction rates. , 2007, Environmental science & technology.

[14]  M. Nikolausz,et al.  Assessment of in situ degradation of chlorinated ethenes and bacterial community structure in a complex contaminated groundwater system. , 2008, Water research.

[15]  A. Nocker,et al.  Genotypic Microbial Community Profiling: A Critical Technical Review , 2007, Microbial Ecology.

[16]  M. Nikolausz,et al.  Characterization of microbial communities in the aqueous phase of a constructed model wetland treating 1,2-dichloroethene-contaminated groundwater. , 2010, FEMS microbiology ecology.

[17]  J. Zeyer,et al.  Activity and Diversity of Sulfate-Reducing Bacteria in a Petroleum Hydrocarbon-Contaminated Aquifer , 2002, Applied and Environmental Microbiology.

[18]  R. M. Lehman,et al.  Understanding of Aquifer Microbiology is Tightly Linked to Sampling Approaches , 2007 .

[19]  S. J. Flynn,et al.  Characterization of Two Tetrachloroethene-Reducing, Acetate-Oxidizing Anaerobic Bacteria and Their Description as Desulfuromonas michiganensis sp. nov , 2003, Applied and Environmental Microbiology.

[20]  E. Edwards,et al.  Microbial composition of chlorinated ethene-degrading cultures dominated by Dehalococcoides. , 2006, FEMS microbiology ecology.

[21]  J. Tukey,et al.  Multiple-Factor Analysis , 1947 .

[22]  U. Göbel,et al.  Microbial structure of an anaerobic bioreactor population that continuously dechlorinates 1,2-dichloropropane. , 2002, FEMS microbiology ecology.

[23]  K. Sowers,et al.  Microbial Reductive Dechlorination of Aroclor 1260 in Baltimore Harbor Sediment Microcosms Is Catalyzed by Three Phylotypes within the Phylum Chloroflexi , 2007, Applied and Environmental Microbiology.

[24]  Katherine H. Kang,et al.  Genome Sequence of the PCE-Dechlorinating Bacterium Dehalococcoides ethenogenes , 2005, Science.

[25]  J. Lehmann,et al.  Integrative approach to delineate natural attenuation of chlorinated benzenes in anoxic aquifers. , 2009, Environmental pollution.

[26]  B. Sleep,et al.  Stable carbon isotope evidence for intrinsic bioremediation of tetrachloroethene and trichloroethene at area 6, Dover Air Force Base. , 2001, Environmental science & technology.

[27]  R. Sanford,et al.  Geobacter lovleyi sp. nov. Strain SZ, a Novel Metal-Reducing and Tetrachloroethene-Dechlorinating Bacterium , 2006, Applied and Environmental Microbiology.

[28]  T. Vogel Natural bioremediation of chlorinated solvents. , 1994 .

[29]  E. Boyd,et al.  Mineralogy Influences Structure and Diversity of Bacterial Communities Associated with Geological Substrata in a Pristine Aquifer , 2007, Microbial Ecology.

[30]  Jeffrey A Cunningham,et al.  Contaminant degradation in physically and chemically heterogeneous aquifers. , 2007, Journal of contaminant hydrology.

[31]  S. Giovannoni,et al.  Bacterial community composition determined by culture-independent and -dependent methods during propane-stimulated bioremediation in trichloroethene-contaminated groundwater. , 2005, Environmental microbiology.

[32]  D. E. Ellis,et al.  Molecular Analysis of Dehalococcoides 16S Ribosomal DNA from Chloroethene-Contaminated Sites throughout North America and Europe , 2002, Applied and Environmental Microbiology.

[33]  S. Thornton,et al.  Challenges in Monitoring the Natural Attenuation of Spatially Variable Plumes , 2004, Biodegradation.

[34]  C. H. Ward,et al.  Handbook of Bioremediation , 1993 .

[35]  Barbara A. Bekins,et al.  Microbial populations in contaminant plumes , 2000 .

[36]  L. Eyers,et al.  Environmental genomics: exploring the unmined richness of microbes to degrade xenobiotics , 2004, Applied Microbiology and Biotechnology.

[37]  D. Lane 16S/23S rRNA sequencing , 1991 .

[38]  F. Chapelle,et al.  Bacteria in deep coastal plain sediments of Maryland: A possible source of CO2 to groundwater , 1987 .

[39]  E. Stackebrandt,et al.  Nucleic acid techniques in bacterial systematics , 1991 .

[40]  N. Sturchio,et al.  Carbon and chlorine isotope fractionation of chlorinated aliphatic hydrocarbons by evaporation , 1999 .

[41]  J. Suflita,et al.  Sulfur Cycling in the Terrestrial Subsurface: Commensal Interactions, Spatial Scales, and Microbial Heterogeneity , 1998, Microbial Ecology.

[42]  D. L. Song,et al.  Stable carbon isotope fractionation during aerobic biodegradation of chlorinated ethenes. , 2004, Environmental science & technology.

[43]  N. Goldscheider,et al.  Review: Microbial biocenoses in pristine aquifers and an assessment of investigative methods , 2006 .

[44]  Jérôme Pagès,et al.  Multiple factor analysis (AFMULT package) , 1994 .

[45]  P. Bradley,et al.  Anaerobic Mineralization of Vinyl Chloride in Fe(III)-Reducing, Aquifer Sediments , 1996 .

[46]  James R. Cole,et al.  The Ribosomal Database Project (RDP-II): previewing a new autoaligner that allows regular updates and the new prokaryotic taxonomy , 2003, Nucleic Acids Res..

[47]  Eoin L. Brodie,et al.  Environmental Whole-Genome Amplification To Access Microbial Populations in Contaminated Sediments , 2006, Applied and Environmental Microbiology.

[48]  Holger Weiss,et al.  Attenuation reactions in a multiple contaminated aquifer in Bitterfeld (Germany). , 2004, Environmental pollution.

[49]  B. Toman,et al.  New Guidelines for δ13C Measurements , 2006 .

[50]  I. Cozzarelli,et al.  Biodegradation in Contaminated Aquifers: Incorporating Microbial/Molecular Methods , 2008, Ground water.

[51]  C. H. Ward,et al.  Ground-Water Treatment for Chlorinated Solvents , 1994 .

[52]  H. Heuer,et al.  Bacterial diversity of soils assessed by DGGE, T-RFLP and SSCP fingerprints of PCR-amplified 16S rRNA gene fragments: do the different methods provide similar results? , 2007, Journal of microbiological methods.

[53]  Daniel Hunkeler,et al.  Monitoring Microbial Dechlorination of Tetrachloroethene (PCE) in Groundwater Using Compound-Specific Stable Carbon Isotope Ratios: Microcosm and Field Studies , 1999 .

[54]  H. Heuer,et al.  Analysis of actinomycete communities by specific amplification of genes encoding 16S rRNA and gel-electrophoretic separation in denaturing gradients , 1997, Applied and environmental microbiology.

[55]  C. Schüth,et al.  Carbon and hydrogen isotope effects during sorption of organic contaminants on carbonaceous materials. , 2003, Journal of contaminant hydrology.

[56]  Alison S. Waller,et al.  Carbon isotopic fractionation during aerobic vinyl chloride degradation. , 2005, Environmental science & technology.

[57]  H. Weiss,et al.  Groundwater pollution and remediation options for multi-source contaminated aquifers (Bitterfeld/Wolfen, Germany). , 2003, Toxicology letters.

[58]  Ashley Eaddy Scale-up and Characterization of an Enrichment Culture for Bioaugmentation of the P-Area Chlorinated Ethene Plume at the Savannah River Site , 2008 .

[59]  Jérôme Pagès,et al.  Multiple factor analysis and clustering of a mixture of quantitative, categorical and frequency data , 2008, Comput. Stat. Data Anal..

[60]  P. Bradley,et al.  Microbial mineralization of VC and DCE under different terminal electron accepting conditions. , 1998, Anaerobe.

[61]  W. Röling,et al.  Relationships between Microbial Community Structure and Hydrochemistry in a Landfill Leachate-Polluted Aquifer , 2001, Applied and Environmental Microbiology.

[62]  J. Gossett,et al.  Isolation of a bacterium that reductively dechlorinates tetrachloroethene to ethene. , 1997, Science.

[63]  D. Rizzo,et al.  A multivariate statistical approach to spatial representation of groundwater contamination using hydrochemistry and microbial community profiles. , 2005, Environmental science & technology.

[64]  R. Meckenstock,et al.  Stable isotope fractionation analysis as a tool to monitor biodegradation in contaminated acquifers. , 2004, Journal of contaminant hydrology.