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Applied and Environmental Microbiology, April 2006, p. 2460-2470, Vol. 72, No. 4
0099-2240/06/$08.00+0     doi:10.1128/AEM.72.4.2460-2470.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Development and Characterization of Stable Sediment-Free Anaerobic Bacterial Enrichment Cultures That Dechlorinate Aroclor 1260

Donna L. Bedard,* Jessica J. Bailey,{dagger} Brandon L. Reiss, and Greta Van Slyke Jerzak{ddagger}

Department of Biology, Rensselaer Polytechnic Institute, Troy, New York 12180

Received 17 November 2005/ Accepted 12 January 2006

We have developed sediment-free anaerobic enrichment cultures that dechlorinate a broad spectrum of highly chlorinated polychlorinated biphenyls (PCBs). The cultures were developed from Aroclor 1260-contaminated sediment from the Housatonic River in Lenox, MA. Sediment slurries were primed with 2,6-dibromobiphenyl to stimulate Process N dechlorination (primarily meta dechlorination), and sediment was gradually removed by successive transfers (10%) to minimal medium. The cultures grow on pyruvate, butyrate, or acetate plus H2. Gas chromatography-electron capture detector analysis demonstrated that the cultures extensively dechlorinate 50 to 500 µg/ml of Aroclor 1260 at 22 to 24°C by Dechlorination Process N. Triplicate cultures of the eighth transfer without sediment dechlorinated 76% of the hexa- through nonachlorobiphenyls in Aroclor 1260 (250 µg/ml) to tri- through pentachlorobiphenyls in 110 days. At least 64 PCB congeners, all of which are chlorinated on both rings and 47 of which have six or more chlorines, were substrates for this dechlorination. To characterize the bacterial diversity in the enrichments, we used eubacterial primers to amplify and clone 16S rRNA genes from DNA extracted from cultures grown on acetate plus H2. Restriction fragment length polymorphism analysis of 107 clones demonstrated the presence of Thauera-like Betaproteobacteria, Geobacter-like Deltaproteobacteria, Pseudomonas species, various Clostridiales, Bacteroidetes, Dehalococcoides of the Chloroflexi group, and unclassified Eubacteria. Our development of highly enriched, robust, stable, sediment-free cultures that extensively dechlorinate a highly chlorinated commercial PCB mixture is a major and unprecedented breakthrough in the field. It will enable intensive study of the organisms and genes responsible for a major PCB dechlorination process that occurs in the environment and could also lead to effective remediation applications.

* Corresponding author. Mailing address: Department of Biology, SC 1W14, Rensselaer Polytechnic Institute, 110 8th St., Troy, NY 12180. E-mail: bedard{at}rpi.edu.

{dagger} Present address: Albany Molecular Research, Inc., 21 Corporate Circle, P.O. Box 15098, Albany, NY 12212.

{ddagger} Present address: The Arbovirus Laboratories, Wadsworth Center, New York State Department of Health, 5668 State Farm Road, Slingerlands, NY 12195.

Applied and Environmental Microbiology, April 2006, p. 2460-2470, Vol. 72, No. 4
0099-2240/06/$08.00+0     doi:10.1128/AEM.72.4.2460-2470.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Adrian, L., Dudkova, V., Demnerova, K., Bedard, D. L. (2009). "Dehalococcoides" sp. Strain CBDB1 Extensively Dechlorinates the Commercial Polychlorinated Biphenyl Mixture Aroclor 1260. Appl. Environ. Microbiol. 75: 4516-4524 [Abstract] [Full Text]  
  • Fagervold, S. K., May, H. D., Sowers, K. R. (2007). Microbial Reductive Dechlorination of Aroclor 1260 in Baltimore Harbor Sediment Microcosms Is Catalyzed by Three Phylotypes within the Phylum Chloroflexi. Appl. Environ. Microbiol. 73: 3009-3018 [Abstract] [Full Text]  
  • Bedard, D. L., Ritalahti, K. M., Loffler, F. E. (2007). The Dehalococcoides Population in Sediment-Free Mixed Cultures Metabolically Dechlorinates the Commercial Polychlorinated Biphenyl Mixture Aroclor 1260. Appl. Environ. Microbiol. 73: 2513-2521 [Abstract] [Full Text]  


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