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

费勒姆学院的罗纳德斯蒂芬斯获得了一笔赠款，用于购买分子生物学技术设备，以支持通过大肠杆菌的DNA指纹图谱确定淡水生态系统中粪便细菌来源的研究。 1972年，联邦政府通过了《清洁水法》，其使命是“恢复和保持国家沃茨的化学、物理和生物完整性。“在弗吉尼亚州，许多溪流被列为受损，因为高水平的粪便细菌。 美国环保署已授权通过实施TMDL（总最大日负荷）计划来调解这些问题。 只有找到了确定粪便污染源的可靠方法，才能成功地进行水污染治理。 粪便细菌存在于所有温血动物的肠道中。因此，E.大肠杆菌污染可能来自人类、家畜、野生动物或牲畜。 粪便细菌的存在表明病原菌或病毒存在的可能性。本项目的目的是确定弗吉尼亚州富兰克林县黑水河和Pigg河流域粪便细菌的非点源（未知）。 该项目的具体目标是：从已知来源（粪便样本）和未知来源（水样）收集粪便细菌菌株。使用提供的设备确定细菌样本的DNA指纹图谱。将已知和未知来源的DNA指纹图谱与现有图谱进行比较。确定未知样品中的细菌来源。增加DNA指纹库数据库。随着确定国家水道中粪便细菌来源的方法的建立，可以更直接地解决这个问题，并更准确地设计补救措施。 否则，如果没有粪便污染源的具体知识，昂贵的补救方案，如TMDL的将是基于猜测。 DNA指纹库数据库的建立将大大提高科学家确定粪便污染源的能力。如果不购买分子生物学设备，就无法建立这一数据库。

ABSTRACTA 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.