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鸥粪便中细菌的系统发育多样性及分子检测

Phylogenetic diversity and molecular detection of bacteria in gull feces.

作者信息

Lu Jingrang, Santo Domingo Jorge W, Lamendella Regina, Edge Thomas, Hill Stephen

机构信息

U.S. Environmental Protection Agency, National Risk Management Research Laboratory, Cincinnati, OH 45268, USA.

出版信息

Appl Environ Microbiol. 2008 Jul;74(13):3969-76. doi: 10.1128/AEM.00019-08. Epub 2008 May 9.

Abstract

In spite of increasing public health concerns about the potential risks associated with swimming in waters contaminated with waterfowl feces, little is known about the composition of the gut microbial community of aquatic birds. To address this, a gull 16S rRNA gene clone library was developed and analyzed to determine the identities of fecal bacteria. Analysis of 282 16S rRNA gene clones demonstrated that the gull gut bacterial community is mostly composed of populations closely related to Bacilli (37%), Clostridia (17%), Gammaproteobacteria (11%), and Bacteriodetes (1%). Interestingly, a considerable number of sequences (i.e., 26%) were closely related to Catellicoccus marimammalium, a gram-positive, catalase-negative bacterium. To determine the occurrence of C. marimammalium in waterfowl, species-specific 16S rRNA gene PCR and real-time assays were developed and used to test fecal DNA extracts from different bird (n = 13) and mammal (n = 26) species. The results showed that both assays were specific to gull fecal DNA and that C. marimammalium was present in gull fecal samples collected from the five locations in North America (California, Georgia, Ohio, Wisconsin, and Toronto, Canada) tested. Additionally, 48 DNA extracts from waters collected from six sites in southern California, Great Lakes in Michigan, Lake Erie in Ohio, and Lake Ontario in Canada presumed to be impacted with gull feces were positive by the C. marimammalium assay. Due to the widespread presence of this species in gulls and environmental waters contaminated with gull feces, targeting this bacterial species might be useful for detecting gull fecal contamination in waterfowl-impacted waters.

摘要

尽管公众越来越关注在被水禽粪便污染的水中游泳可能带来的健康风险,但对于水鸟肠道微生物群落的组成却知之甚少。为了解决这个问题,构建并分析了一个海鸥16S rRNA基因克隆文库,以确定粪便细菌的种类。对282个16S rRNA基因克隆的分析表明,海鸥肠道细菌群落主要由与芽孢杆菌(37%)、梭菌(17%)、γ-变形菌(11%)和拟杆菌(1%)密切相关的菌群组成。有趣的是,相当数量的序列(即26%)与海生哺乳动物卡特球菌密切相关,这是一种革兰氏阳性、过氧化氢酶阴性的细菌。为了确定海生哺乳动物卡特球菌在水鸟中的存在情况,开发了物种特异性16S rRNA基因PCR和实时检测方法,并用于检测来自不同鸟类(n = 13)和哺乳动物(n = 26)物种的粪便DNA提取物。结果表明,这两种检测方法对海鸥粪便DNA具有特异性,并且在从北美五个地点(加利福尼亚州、佐治亚州、俄亥俄州、威斯康星州和加拿大安大略省多伦多市)采集的海鸥粪便样本中检测到了海生哺乳动物卡特球菌。此外,对从南加利福尼亚州六个地点、密歇根州五大湖、俄亥俄州伊利湖和加拿大安大略湖采集的水样进行DNA提取,推测这些水样受到海鸥粪便的影响,通过海生哺乳动物卡特球菌检测方法检测,其中48份呈阳性。由于该物种在海鸥和被海鸥粪便污染的环境水域中广泛存在,针对这种细菌物种可能有助于检测受水鸟影响的水域中的海鸥粪便污染。

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