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Applied and Environmental Microbiology, September 2004, p. 5538-5545, Vol. 70, No. 9
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.9.5538-5545.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Characterization of a Highly Enriched Dehalococcoides-Containing Culture That Grows on Vinyl Chloride and Trichloroethene

Melanie Duhamel, Kaiguo Mo, and Elizabeth A. Edwards*

Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, Ontario, Canada

Received 13 January 2004/ Accepted 12 May 2004

A highly enriched culture that reductively dechlorinates trichloroethene (TCE), cis-1,2-dichloroethene (cDCE), and vinyl chloride (VC) to ethene without methanogenesis is described. The Dehalococcoides strain in this enrichment culture had a yield of (5.6 ± 1.4) x 108 16S rRNA gene copies/µmol of Cl– when grown on VC and hydrogen. Unlike the other VC-degrading cultures described in the literature, strains VS and BAV1, this culture maintained the ability to grow on TCE with a yield of (3.6 ± 1.3) x 108 16S rRNA gene copies/µmol of Cl–. The yields on an electron-equivalent basis measured for the culture grown on TCE and on VC were not significantly different, indicating that both substrates supported growth equally well. PCR followed by denaturing gradient gel electrophoresis, cloning, and phylogenetic analyses revealed that this culture contained one Dehalococcoides 16S rRNA gene sequence, designated KB-1/VC, that was identical (over 1,386 bp) to the sequences of previously described organisms FL2 and CBDB1. A second Dehalococcoides sequence found in separate KB-1 enrichment cultures maintained on cDCE, TCE, and tetrachloroethene was no longer present in the VC-H2 enrichment culture. This second Dehalococcoides sequence was identical to that of BAV1. As neither FL2 nor CBDB1 can dechlorinate VC to ethene in a growth-related fashion, it is clear that current 16S rRNA gene-based analyses do not provide sufficient information to distinguish between metabolically diverse members of the Dehalococcoides group.

* Corresponding author. Mailing address: Department of Chemical Engineering and Applied Chemistry, University of Toronto, 200 College St., Toronto, Ontario M5S 3E5, Canada. Phone: (416) 978-3506. Fax: (416) 978-8605. E-mail: edwards{at}chem-eng.utoronto.ca.

Applied and Environmental Microbiology, September 2004, p. 5538-5545, Vol. 70, No. 9
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.9.5538-5545.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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