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Applied and Environmental Microbiology, February 2003, p. 953-959, Vol. 69, No. 2
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.2.953-959.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Growth of a Dehalococcoides-Like Microorganism on Vinyl Chloride and cis-Dichloroethene as Electron Acceptors as Determined by Competitive PCR

Alison M. Cupples,1 Alfred M. Spormann,1,2,3 and Perry L. McCarty1*

Department of Civil and Environmental Engineering,1 Department of Biological Sciences,2 Department of Geological and Environmental Sciences, Stanford University, Stanford, California 943053

Received 17 June 2002/ Accepted 13 November 2002

A competitive PCR (cPCR) assay targeting 16S ribosomal DNA was developed to enumerate growth of a Dehalococcoides-like microorganism, bacterium VS, from a mixed culture catalyzing the reductive dehalogenation of cis-1,2-dichloroethene (cDCE) and vinyl chloride (VC), with hydrogen being used as an electron donor. The growth of bacterium VS was found to be coupled to the dehalogenation of VC and cDCE, suggesting unique metabolic capabilities. The average growth yield was (5.2 ± 1.5) x 108 copies of the 16S rRNA gene/µmol of Cl- (number of samples, 10), with VC being used as the electron acceptor and hydrogen as the electron donor. The maximum VC utilization rate () was determined to be 7.8 x 10-10 µmol of Cl- (copy-1 day-1), indicating a maximum growth rate of 0.4 day-1. These average growth yield and values agree well with values found previously for dechlorinating cultures. Decay coefficients were determined with growth (0.05 day-1) and no-growth (0.09 day-1) conditions. An important limitation of this cPCR assay was its inability to discriminate between active and inactive cells. This is an essential consideration for kinetic studies.

* Corresponding author. Mailing address: Department of Civil and Environmental Engineering, Terman Engineering Center, Stanford University, Stanford, CA 94305-4020. Phone: (650) 723-4131. Fax: (650) 725-3164. E-mail: pmccarty{at}stanford.edu.

Applied and Environmental Microbiology, February 2003, p. 953-959, Vol. 69, No. 2
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.2.953-959.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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