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A biomarker for the identification of swine fecal pollution in water, using the STII toxin gene from enterotoxigenic Escherichia coli

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Abstract

This research developed a PCR method to identify swine fecal pollution in water, using a portion of the STII toxin gene from enterotoxigenic Escherichia coli as the target sequence. This method showed the gene to have a wide-spread geographical distribution and temporal stability; and the primers demonstrated high specificity, sensitivity, and reliability. A total of 110 DNA extracts from different animal fecal and human sewage samples were screened using the primers and no positives resulted. Centrifugation and filtration methods for concentrating E. coli seeded into stream, ocean, secondary effluent, and dairy lagoon waters resulted in detection limits at the femtogram and attogram levels. E. coli with the biomarker seeded into stream, ocean, and secondary effluent waters remained stable for approximately 2 weeks for all water types. Of the farm lagoon and waste samples tested, 94% were positive for the STII trait, regardless of the number of E. coli screened and 100% were positive when ≥35 E. coli isolates were screened. As the PCR product of the target sequence yielded a single band, the method is applicable to dot blot detection methodology, yielding great accuracy in determining the presence of swine fecal sources.

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Acknowledgements

We acknowledge the USEPA (project number CR826204-01) and the USDA (project number 99-25102-8596) for funding this project. We thank Dr. Gerard Stelma for support and advice throughout this project, Robin Oshiro, who collected fecal samples used for cross-reactivity studies, and Dr. Tom Casey, National Animal Disease Center, Ames, Iowa for the E. coli strain, primer sequences, and information on the STII toxin gene. Also, we thank all the academics and swine farmers for their assistance with field samples. All experiments comply with the current laws of the country in which they were performed, the United States of America.

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  1. Department of Environmental Health Science and Policy, University of California at Irvine, 1368 Social Ecology II, Irvine, CA 92697, USA

    L. A. Khatib, Y. L. Tsai & B. H. Olson

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  1. L. A. Khatib
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  2. Y. L. Tsai
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  3. B. H. Olson
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Correspondence to B. H. Olson.

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Khatib, L.A., Tsai, Y.L. & Olson, B.H. A biomarker for the identification of swine fecal pollution in water, using the STII toxin gene from enterotoxigenic Escherichia coli . Appl Microbiol Biotechnol 63, 231–238 (2003). https://doi.org/10.1007/s00253-003-1373-9

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  • DOI: https://doi.org/10.1007/s00253-003-1373-9

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