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目的应用细菌基因组重复序列PCR技术(rep-PCR)微生物源追踪方法对江苏省盱眙县桂五水库中粪便污染来源进行追踪。方法分别采集不同季节水库周边已知来源粪便标本,分离大肠埃希菌作为指示菌,建立已知污染源指示菌rep-PCR特征指纹库,用基因聚类分析软件计算平均正确归类率,进行判别分析和多元方差分析;同期采集水库水样,分离并确认指示菌,进行rep-PCR扩增,与已知污染源数据库进行对比,判断水样中指示菌污染来源。结果将已知源数据库分为2,3,4,5和9类时,平均正确归类率分别为89.19%,77.58%,76.69%,75.25%和70.92%;已知源数据库可区分指示菌的不同来源。对534株水库水样指示菌微生物源追踪结果显示,人、鸡、鸭、鹅、狗、猪、牛、羊和野生动物各占26.49%,14.74%,7.77%,5.78%,3.98%,7.97%,10.96%,8.76%和8.57%。结论桂五水库粪便污染来源种类繁多,其中人、鸡和牛为主要污染源;该方法为查找水体粪便污染来源及污染整改效果评估提供了新技术。
OBJECTIVE: To trace the source of stool contamination in Gui Wu reservoir in Xuyi County, Jiangsu Province, using the method of bacterial genome repeat PCR (rep-PCR). Methods Stool samples from known sources in different seasons were collected, and Escherichia coli was isolated as the indicator bacteria to establish the fingerprint database of rep-PCR of known indicator bacteria. The average correct classification rate was calculated by genetic clustering analysis software Analysis and multivariate analysis of variance (ANOVA); collect samples of reservoirs in the same period, separate and confirm the indicator bacteria, amplify them by rep-PCR and compare them with the database of known sources of pollution to determine the sources of indicator bacteria contamination in water samples. Results When the known source databases were classified into 2, 3, 4, 5 and 9 categories, the average correct classification rates were 89.19%, 77.58%, 76.69%, 75.25% and 70.92%, respectively; Different sources. The traceability of 534 strains of microorganisms in water samples indicated that 26.49%, 14.74%, 7.77%, 5.78%, 3.98%, 7.97% of human, chickens, ducks, goose, dogs, pigs, cattle, sheep and wild animals %, 10.96%, 8.76% and 8.57%. Conclusion There are many sources of stool pollution in Guiwu reservoir, among them, human, chicken and cow are the main sources of pollution. This method provides a new technology for finding sources of polluted water and assessing the rectification effect of pollution.