应用rep-PCR微生物源方法追踪夏秋季桂五水库粪便污染来源

目的：应用重复序列聚合酶链反应（rep-PCR）微生物源追踪方法，追踪夏秋季江苏桂五水库的粪便污染来源追踪。方法：采集已知来源的人和动物粪便标本以及夏秋两季水样标本，分离并确认指示菌大肠杆菌，以BOXAIR为引物进行PCR扩增；用Bionumerics4．0软件对rep-PCR基因指纹图进行判别分析和多元方差分析并计算各源种类的正确归类率。结果：10、5、3和2分类法平均正确归类率分别为68．13％、74．76％、82．36％和86．03％。判别分析和多元方差分析结果显示rep—PCR基因指纹图能将已知源不同来源指示菌区分开。水样中主要粪便污染来源为人、鸡和牛。结论：建立的rep—PCR微生物源追踪方法能较好地区分水体中指示菌的不同来源，水体标本监测显示桂五水库粪便污染来源种类多，其中人、鸡和牛相对较多，提示应对桂五水库地区加强人、家禽和家畜动物粪便的无害化管理。

Application of rep-PCR microbial source tracking to identify fecal sources in Guiwu reseavoir in the seasons of summer and fall

Objective： Objective： The rep--PCR fingerprinting microbial source tracking method was used to analysis a reservoir of the Jiangsu Guiwu reservoir in the seasons of summer and fall. Methods： Fresh fecal materials were collected from human and animals throughout villages around the reservoir. Escherichia coli were identified as indicators and PCR was expanded with the support of BOXAIR premier. The rates of correct assignment, multivariate analysis of variance （MANOVA） and discriminant analysis were conducted by using Bionumerics 4. 0 software. Watering samples were applied in summer and in fall to identify the main fecal pollutions. Results.. The known source fingerprint library was built and when it was classified as 10, 5, 3 and 2 groups, the average rates of correct classification were respective 68. 13%, 74. 76%, 82. 36% and 86. 03%. MANOVA and discriminant analysis showed that the division of different groups was clear. And the reservoir was most possibly polluted by human, chicken and cow. Conclusions.. The rep--PCR DNA fingerprint analysis was a promising method to determine the source groups of indictor isolates in water. Management practices should intend to decrease the human, poultry and domestic animals fecal pollutions in Guiwu reservoir, more actions should be done to make non--hazard treatment of feces.