The Identification of Nonpoint Sources of Escherichia Coli in Freshwater Ecosystems by DNA Fingerprinting

Chowan University, Murfreesboro NC

Investigators

Abstract

ABSTRACT A grant has been awarded to Ronald Stephens at Ferrum College, to purchase equipment for molecular biological techniques in order to support research which determines the sources of fecal bacteria in freshwater ecosystems by DNA fingerprinting of Escherichia coli bacteria. In 1972 the federal government passed the Clean Water Act whose mission is to "restore and maintain the chemical, physical, and biological integrity of the nations waters." In Virginia, many streams are classified as being impaired because of high levels of fecal bacteria. The EPA has mandated that these problems be mediated by the implementation of the TMDL (Total Maximum Daily Load) Program. Successful water remediation cannot occur until reliable methods of identifying the source(s) of the fecal contamination can be developed. Fecal bacteria are found in the intestines of all warm-blooded animals. Therefore, potential sources of E. coli contamination could be from humans, domestic animals, wild life, or livestock. The presence of fecal bacteria indicate the possibilities of the presence of pathogenic bacteria or viruses. The purpose of this project will be to determine nonpoint (unknown) sources of fecal bacteria in the Blackwater and Pigg River watersheds in Franklin County, Virginia. The specific objectives of the project will be to: Collect fecal bacterial strains from known sources (fecal samples) and unknown sources (water samples). Determine the DNA fingerprint profile of the bacterial samples using the equipment provided. Compare DNA fingerprint profiles from known and unknown sources with existing profiles. Determine the sources of the bacteria in the unknown samples. Increase the DNA fingerprint library database. With the establishment of methodologies to determine the sources of fecal bacteria in the nations waterways, this problem can be more directly addressed and remediation more accurately designed. Otherwise, without specific knowledge of the sources of fecal pollution, expensive remediation programs such as TMDL's will be based on guesswork. The establishment of a DNA fingerprint library database will greatly increase the scientist's ability to determine the sources of fecal pollution. This database could not be established without the purchase of the molecular biology equipment.