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

Ferrum学院的Ronald Stephens获得了一笔资助，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“恢复和维护国家水域的化学、物理和生物完整性”。在弗吉尼亚州，由于粪便细菌含量高，许多河流被列为受损。美国环境保护署要求通过TMDL（总最大日负荷）计划的实施来解决这些问题。只有开发出确定粪便污染源的可靠方法，水的修复才能取得成功。所有温血动物的肠道中都有粪便细菌。因此，大肠杆菌污染的潜在来源可能来自人类、家畜、野生动物或牲畜。粪便细菌的存在表明存在致病菌或病毒的可能性。该项目的目的是确定弗吉尼亚州富兰克林县黑水和猪河流域的非点源（未知）粪便细菌。该项目的具体目标将是：收集已知来源（粪便样本）和未知来源（水样）的粪便细菌菌株。使用所提供的设备确定细菌样品的DNA指纹图谱。将已知和未知来源的DNA指纹档案与现有档案进行比较。确定未知样品中细菌的来源。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，这一问题可以更直接地解决，也可以更准确地设计补救措施。否则，如果没有对粪便污染源的具体了解，像TMDL这样昂贵的修复计划将基于猜测。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，就不可能建立这个数据库。

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.