Department of Natural, Biotic and Social Environment Engineering, University of Yamanashi, Kofu, Yamanashi, Japan.
Institute of Medicine, Tribhuvan University, Maharajgunj, Kathmandu, Nepal.
J Appl Microbiol. 2018 Aug;125(2):609-619. doi: 10.1111/jam.13884. Epub 2018 May 23.
To validate host-specific Bacteroidales assays to identify faecal-source contamination of drinking water sources in the Kathmandu Valley, Nepal.
A total of 54 composite faecal-source samples were collected from human sewage, ruminants, pigs, dogs, chickens and ducks, which were analysed by quantitative polymerase chain reaction using human-specific (BacHum, HF183 SYBR, gyrB and HF183 TaqMan), ruminant-specific (BacCow and BacR), pig-specific (Pig2Bac and PF163) and dog-specific assays (BacCan SYBR). The BacHum, BacR and Pig2Bac assays were judged the best performing human-specific, ruminant-specific and pig-specific assays respectively. The BacCan SYBR assay highly cross-reacted with other species, resulting in poor performance. Furthermore, these validated assays were applied to microbial source tracking (MST) of 74 drinking water samples. Out of these, 20, 12 and 4% samples were judged contaminated by human, ruminant and pig faeces respectively. Detection ratios of human and ruminant faecal markers were relatively higher in built-up and agricultural areas respectively.
BacHum, BacR and Pig2Bac assays were found suitable for MST and both, human and animal faecal contaminations of drinking water sources were common in the valley.
MST could be an effective tool for preparing the faecal pollution strategies as these are site specific.
验证针对生境特异性的拟杆菌门检测方法，以鉴定尼泊尔加德满都谷地饮用水源的粪便污染来源。
共采集了 54 份人粪、反刍动物、猪、狗、鸡和鸭的复合粪便源样本，采用人源特异性（BacHum、HF183 SYBR、gyrB 和 HF183 TaqMan）、反刍动物特异性（BacCow 和 BacR）、猪特异性（Pig2Bac 和 PF163）和犬特异性（BacCan SYBR）定量聚合酶链反应进行分析。BacHum、BacR 和 Pig2Bac 检测方法被判定为最佳的人源特异性、反刍动物特异性和猪特异性检测方法。BacCan SYBR 检测方法与其他物种高度交叉反应，导致性能不佳。此外，这些经过验证的检测方法应用于 74 个饮用水样本的微生物源追踪（MST）。其中，20%、12%和 4%的样本分别被判定为人粪、反刍动物粪和猪粪污染。在建成区和农业区，人粪和反刍动物粪标记物的检出率相对较高。
BacHum、BacR 和 Pig2Bac 检测方法适用于 MST，并且人类和动物粪便对饮用水源的污染在该谷地很常见。
MST 可以作为一种有效的工具，制定针对特定地点的粪便污染策略。
