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WATER RESOURCES RESEARCH GRANT PROPOSAL

Project ID: 2006MN161B

Title:: Development of a DNA Marker Gene System to Determine Sources of Fecal E. coli in Watersheds

Project Type: Research

Start Date: 03/01/2006

End Date: 02/29/2008

Congressional District: 5th

Focus Categories: #1: Water Quality, Surface Water, Methods

Keywords: E. coli, Gene Probes, Source Tracking, Robotic System, Fecal Pollutants

Principal Investigator: Sadowsky, Michael Jay

Federal Funds: $15,000

Non-Federal Matching Funds: $42,908

Abstract: The contamination of waterways with fecal material remains a persistent threat to public health. Identification of sources of fecal contamination is a vital component for abatement strategies and in the determination of total maximum daily loads. While phenotypic and genotypic techniques have been used to determine potential sources of fecal bacteria in surface waters, most methods require the construction of large known-source libraries, and often fail to adequately differentiate among environmental isolates originating from diverse animal sources. In contrast, library-independent, gene probe methods may provide a cost-effective approach to determine sources of environmental isolates originating from different animal sources. In this proposal, we will use the suppression subtractive hybridization technique to enrich for host source-specific DNA markers for E. coli originating from cows and humans. We have previously used this approach to identify DNA markers for E. coli originating from geese and have used these markers as gene probes in a library independent analysis of sources of E. coli in Lake Superior. The second objective of the proposed research is to use the newly identified DNA markers (and the existing goose marker gene) as gene probes to determine sources of E. coli in the Minneopa Creek watershed. Analyses will be done using a recently developed high throughput, automated, macroarray screening procedure. The identified markers will provide a quantitative, cost-effective, and accurate library-independent method to determine sources of genetically diverse E. coli for use in source tracking studies.

Progress/Completion Report, PDF


U.S. Department of the Interior, U.S. Geological Survey
URL: http://water.usgs.gov/wrri/06grants/2006MN161B.html
Maintained by: John Schefter
Last Updated: Thursday, May 21, 2009
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