Lu Jingrang, Santo Domingo Jorge W, Hill Stephen, Edge Thomas A
U.S. Environmental Protection Agency, Office of Research and Development, National Risk Management Research Laboratory, 26 W. Martin Luther King Dr., MS-387, Cincinnati, OH 45268, USA.
Appl Environ Microbiol. 2009 Sep;75(18):5919-26. doi: 10.1128/AEM.00462-09. Epub 2009 Jul 24.
Methods to assess the impact of goose fecal contamination are needed as the result of the increasing number of Canada geese (Branta canadensis) near North American inland waters. However, there is little information on goose fecal microbial communities, and such data are important for the development of host-specific source-tracking methods. To address this issue, 16S rRNA gene clone libraries for Canada goose fecal samples from Ontario, Canada, and Ohio were analyzed. Analyses of fecal clones from Ontario (447) and Ohio (302) showed that goose fecal communities are dominated by the classes "Clostridia" (represented by 33.7% of clones) and "Bacilli" (38.1% of clones) and the phylum "Bacteroidetes" (10.1% of clones). Sequences not previously found in other avian fecal communities were used to develop host-specific assays. Fecal DNA extracts from sewage plants (10 samples) and different species of birds (11 samples) and mammals (18 samples) were used to test for host specificity. Of all the assays tested, one assay showed specificity for Canada goose fecal DNA. The PCR assay was positive for Canada goose fecal DNA extracts collected from three locations in North America (Ohio, Oregon, and Ontario, Canada). Additionally, of 48 DNA extracts from Lake Ontario waters presumed to be impacted by waterfowl feces, 19 tested positive by the assay, although 10 were positive only after a nested PCR approach was used. Due to the level of host specificity and the presence of signals in environmental waters, the assay is proposed as a part of the toolbox to detect Canada goose contamination in waterfowl-contaminated waters.
由于北美内陆水域附近加拿大鹅（Branta canadensis）数量不断增加，因此需要评估鹅粪便污染影响的方法。然而，关于鹅粪便微生物群落的信息很少，而这些数据对于开发宿主特异性溯源方法很重要。为了解决这个问题，对来自加拿大安大略省和俄亥俄州的加拿大鹅粪便样本的16S rRNA基因克隆文库进行了分析。对来自安大略省（447个）和俄亥俄州（302个）的粪便克隆的分析表明，鹅粪便群落以“梭菌纲”（占克隆的33.7%）、“芽孢杆菌纲”（占克隆的38.1%）和“拟杆菌门”（占克隆的10.1%）为主。利用以前在其他鸟类粪便群落中未发现的序列开发宿主特异性检测方法。使用污水处理厂（10个样本）、不同鸟类（11个样本）和哺乳动物（18个样本）的粪便DNA提取物来测试宿主特异性。在所有测试的检测方法中，有一种检测方法对加拿大鹅粪便DNA具有特异性。该PCR检测方法对从北美三个地点（俄亥俄州、俄勒冈州和加拿大安大略省）采集的加拿大鹅粪便DNA提取物呈阳性。此外，在安大略湖水域假定受水禽粪便影响的48份DNA提取物中，有19份检测呈阳性，不过其中10份仅在使用巢式PCR方法后才呈阳性。由于宿主特异性水平以及环境水体中存在信号，该检测方法被提议作为检测水禽污染水体中加拿大鹅污染的工具箱的一部分。
