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Applied and Environmental Microbiology, December 2006, p. 7567-7574, Vol. 72, No. 12
0099-2240/06/$08.00+0     doi:10.1128/AEM.01317-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Detection of Human-Derived Fecal Pollution in Environmental Waters by Use of a PCR-Based Human Polyomavirus Assay{triangledown}

Shannon M. McQuaig,1 Troy M. Scott,2 Valerie J. Harwood,3* Samuel R. Farrah,1 and Jerzy O. Lukasik2

Department of Microbiology and Cell Science, University of Florida, Gainesville, Florida 33611,1 Biological Consulting Services of North Florida, Gainesville, Florida 32609,2 Department of Biology, University of South Florida, Tampa, Florida 336203

Received 8 June 2006/ Accepted 13 September 2006

Regulatory agencies mandate the use of fecal coliforms, Escherichia coli or Enterococcus spp., as microbial indicators of recreational water quality. These indicators of fecal pollution do not identify the specific sources of pollution and at times underestimate health risks associated with recreational water use. This study proposes the use of human polyomaviruses (HPyVs), which are widespread among human populations, as indicators of human fecal pollution. A method was developed to concentrate and extract HPyV DNA from environmental water samples and then to amplify it by nested PCR. HPyVs were detected in as little as 1 µl of sewage and were not amplified from dairy cow or pig wastes. Environmental water samples were screened for the presence of HPyVs and two additional markers of human fecal pollution: the Enterococcus faecium esp gene and the 16S rRNA gene of human-associated Bacteroides. The presence of human-specific indicators of fecal pollution was compared to fecal coliform and Enterococcus concentrations. HPyVs were detected in 19 of 20 (95%) samples containing the E. faecium esp gene and Bacteroides human markers. Weak or no correlation was observed between the presence/absence of human-associated indicators and counts of indicator bacteria. The sensitivity, specificity, and correlation with other human-associated markers suggest that the HPyV assay could be a useful predictor of human fecal pollution in environmental waters and an important component of the microbial-source-tracking "toolbox."

* Corresponding author. Mailing address: Department of Biology, SCA 110, University of South Florida, 4202 E. Fowler Ave., Tampa, FL 33620. Phone: (813) 974-1524. Fax: (813) 974-3263. E-mail: vharwood{at}cas.usf.edu.

{triangledown} Published ahead of print on 22 September 2006.

Applied and Environmental Microbiology, December 2006, p. 7567-7574, Vol. 72, No. 12
0099-2240/06/$08.00+0     doi:10.1128/AEM.01317-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • McQuaig, S. M., Scott, T. M., Lukasik, J. O., Paul, J. H., Harwood, V. J. (2009). Quantification of Human Polyomaviruses JC Virus and BK Virus by TaqMan Quantitative PCR and Comparison to Other Water Quality Indicators in Water and Fecal Samples. Appl. Environ. Microbiol. 75: 3379-3388 [Abstract] [Full Text]  
  • Haack, S. K., Duris, J. W., Fogarty, L. R., Kolpin, D. W., Focazio, M. J., Furlong, E. T., Meyer, M. T. (2009). Comparing Wastewater Chemicals, Indicator Bacteria Concentrations, and Bacterial Pathogen Genes as Fecal Pollution Indicators. J. Environ. Qual. 38: 248-258 [Abstract] [Full Text]  
  • Dumonceaux, T. J., Mesa, C., Severini, A. (2008). Internally Controlled Triplex Quantitative PCR Assay for Human Polyomaviruses JC and BK. J. Clin. Microbiol. 46: 2829-2836 [Abstract] [Full Text]  
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