Evaluation of host-specific Bacteroidales 16S rRNA gene markers as a complementary tool for detecting fecal pollution in a prairie watershed
Evaluation of host-specific Bacteroidales 16S rRNA gene markers as a complementary tool for detecting fecal pollution in a prairie watershed
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DOI：
10.1016/j.watres.2009.06.045
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发表时间：
2009-11-01
期刊：
影响因子：
12.8
通讯作者：
Yost, C. K.
中科院分区：
文献类型：
--
作者：
Fremaux, B.;Gritzfeld, J.;Yost, C. K.
我们现在和将来识别和消除水的粪便污染的能力，对于减少水传播疾病的发病率是至关重要的。细菌源追踪是最近发展起来的一种识别粪便污染源的方法。伯恩哈德和菲尔德设计的聚合酶链式反应引物[Bernhard，A.E.，菲尔德，K.G.，2000a.根据宿主对普氏杆菌16S rRNA编码基因的差异建立区分人和反刍动物粪便的聚合酶链式反应方法。APPL环境。微生物。66(10)，4571-4574]和Dick等人。[Dick，L.K.，Bernhard，A.E.，Brodeur，T.J.，Santo Domingo，J.W.，Simpson，J.M.，Walters，S.P.，field，K.G.，2005.未培养粪便类杆菌的寄主分布揭示了粪便来源鉴定的遗传标记。APPL环境。微生物。71(6)，3184-3191]用于检测人(HF183)、猪(PF163)和反刍动物(CF128)特异性类杆菌16S rRNA遗传标记在加拿大萨斯喀彻温省粪便污染检测中的适用性。通过检测萨斯喀彻温省12个不同物种的8个未经处理的人类污水样本和265个粪便样本，评估了这些引物的敏感性和特异性。每套引物的特异度为94%。HF183和PF163区分不同物种的准确率为100%，而CF128与22%的猪粪发生交叉反应。对采集于加拿大萨斯喀彻温省曲阿佩尔河沿岸不同水样的70份水样进行了寄主特异性类杆菌标志物和常规指示物大肠埃希氏菌与几种肠道病原体的关系的调查。在41%和14%的水样中分别发现了人类和反刍动物粪便标志物，而在河水中从未检测到猪的标志物。随着HF183和CF128的同时检测，大肠埃希氏菌的最大浓度也随之出现。耐热弯曲杆菌、沙门氏菌。志贺毒素基因(stx1和stx2)阳性的大肠埃希菌(STEC)分别有6、7和63%。然而，在STX阳性的水样中，没有一个对大肠杆菌O157：H7基因标记(UidA)呈阳性。优势比分析表明，CF128可能是沙门氏菌存在的预测因子。在河里调查过了。没有一种粪便指标能够很有把握地预测耐热弯曲杆菌的存在。和STEC。(C)2009爱思唯尔有限公司。保留所有权利。
Our ability to identify and eliminate fecal contamination of water, now and in the future, is essential to reduce incidences of waterborne disease. Bacterial source tracking is a recently developed approach for identifying sources of fecal pollution. PCR primers designed by Bernhard and Field [Bernhard, A.E., Field, K.G., 2000a. A PCR assay to discriminate human and ruminant feces on the basis of host differences in Bacteroides-Prevotella genes encoding 16S rRNA. Appl. Environ. Microbiol. 66(10), 4571-4574] and Dick et al. [Dick, L.K., Bernhard, A.E., Brodeur, T.J., Santo Domingo, J.W., Simpson, J.M., Walters, S.P., Field, K.G., 2005. Host distributions of uncultivated fecal Bacteroidales bacteria reveal genetic markers for fecal source identification. Appl. Environ. Microbiol. 71(6), 3184-3191] for the detection of human (HF183), pig (PF163) and ruminant (CF128) specific Bacteroidales 16s rRNA genetic markers were tested for their suitability in detecting fecal pollution in Saskatchewan, Canada. The sensitivity and specificity of these primers were assessed by testing eight raw human sewage samples and 265 feces from 12 different species in Saskatchewan. The specificity of each primer set was >= 94%. The accuracy of HF183 and PF163 to distinguish between the different species was 100%, whereas CF128 cross-reacted with 22% of the pig feces. occurrence of the host-specific Bacteroidales markers and the conventional indicator Escherichia coli in relation to several enteropathogens was investigated in 70 water samples collected from different sites along the Qu'Appelle River (Saskatchewan, Canada). Human and ruminant fecal markers were identified in 41 and 14% of the water samples, respectively, whereas the pig marker was never detected in the river water. The largest concentrations in E. coli counts were concomitant to the simultaneous detection of HF183 and CF128. Thermotolerant Campylobacter spp., Salmonella spp. and Shiga toxin genes (stx1 and stx2)-positive E. coli (STEC) were detected in 6, 7 and 63% of the water samples, respectively. However, none of the stx positive water samples were positive for the E. coli O157:H7 gene marker (uidA). Odds ratios analysis suggests that CF128 may be predictive for the presence of Salmonella spp. in the river investigated. None of the fecal indicators were able to confidently predict the presence of thermotolerant Campylobacter spp. and STEC. (C) 2009 Elsevier Ltd. All rights reserved.