Identification of nonpoint sources of fecal pollution in coastal waters by using host-specific 16S ribosomal DNA genetic markers from fecal anaerobes
Identification of nonpoint sources of fecal pollution in coastal waters by using host-specific 16S ribosomal DNA genetic markers from fecal anaerobes
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DOI：
10.1128/aem.66.4.1587-1594.2000
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发表时间：
2000-04-01
影响因子：
4.4
通讯作者：
Field, KG
中科院分区：
文献类型：
--
作者：
Bernhard, AE;Field, KG
我们描述了一种新的PCR为基础的方法区分人和牛的粪便污染在沿海沃茨没有培养指示生物，我们表明，该方法可用于跟踪细菌标记序列在复杂的环境中。我们确定了两个人类特异性遗传标记和5个奶牛特异性遗传标记的粪便样本中扩增16 S核糖体DNA（rDNA）片段的双歧杆菌属和类杆菌-普雷沃氏菌组的成员，并进行长度异质性PCR和末端限制性片段长度多态性分析。宿主特异性模式表明，人和奶牛粪便中双歧杆菌和类杆菌-普雷沃菌种群的物种组成存在差异。这些模式在属于同一物种的不同宿主之间具有高度可重复性。此外，所有的主机特定的遗传标记，检测水样中收集的地区经常与粪便污染。易于检测和较长的生存时间在水中的拟杆菌普雷沃氏菌指标优于双歧杆菌指标。粪便16 S rDNA序列对应于我们的拟杆菌-普雷沃氏菌标志物包括密切相关的基因簇，其中没有一个完全匹配先前公布的拟杆菌或普雷沃氏菌序列。我们的方法在污染物浓度为2.8 x 10（-5）至2.8 x 10（-7）g（干重）粪便/升和6.8 x 10（-7）g（干重）污水/升的水中检测到宿主特异性标记。虽然我们的目的是确定粪便污染的非点源，这里描述的方法应广泛适用于监测自然环境中特定细菌群的时空波动。
We describe a new PCR-based method for distinguishing human and cow fecal contamination in coastal waters without culturing indicator organisms, and we show that the method can be used to track bacterial marker sequences in complex environments. We identified two human-specific genetic markers and five cow-specific genetic markers in fecal samples by amplifying 16S ribosomal DNA (rDNA) fragments from members of the genus Bifidobacterium and the Bacteroides-Prevotella group and performing length heterogeneity PCR and terminal restriction fragment length polymorphism analyses. Host-specific patterns suggested that there are species composition differences in the Bifidobacterium and Bacteroides-Prevotella populations of human and cow feces. The patterns were highly reproducible among different hosts belonging to the same species. Additionally, all host-specific genetic markers were detected in water samples collected from areas frequently contaminated with fecal pollution. Ease of detection and longer survival in water made Bacteroides-Prevotella indicators better than Bifidobacterium indicators. Fecal 16S rDNA sequences corresponding to our Bacteroides-Prevotella markers comprised closely related gene clusters, none of which exactly matched previously published Bacteroides or Prevotella sequences. Our method detected host-specific markers in water at pollutant concentrations of 2.8 x 10(-5) to 2.8 x 10(-7) g (dry weight) of feces/liter and 6.8 x 10(-7) g (dry weight) of sewage/liter. Although our aim was to identify nonpoint sources of fecal contamination, the method described here should be widely applicable for monitoring spatial and temporal fluctuations in specific bacterial groups in natural environments.