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利用16S rRNA基因测序技术鉴定城市、农业和森林流域中可培养的溪流水细菌。

Identification of culturable stream water bacteria from urban, agricultural, and forested watersheds using 16S rRNA gene sequencing.

作者信息

Belt Kenneth T, Hohn Christina, Gbakima Aiah, Higgins James A

机构信息

USDA Forest Service, UMBC CUERE/BES LTER, Baltimore, MD 21227, USA.

出版信息

J Water Health. 2007 Sep;5(3):395-406. doi: 10.2166/wh.2007.035.

Abstract

Bacteria present in water samples taken on a weekly basis, from June 2004 through June 2005, from three streams, were cultured on Coliscan Easygel agar plates. Colonies representative of a variety of colors and morphologies were subjected to amplification and sequencing of a 1000-1100 nt portion of the 16S rRNA gene. A total of 528 colonies were sequenced; these categorized into 26 genera and 78 species. Of 175 dark blue/purple colonies presumed to be E. coli, sequence analysis indicated that 45 (25%) were actually other genera. For the urban stream Gwynns Falls Gwynns Run, E. coli was the most common genus/species encountered, followed by Klebsiella and Aeromonas. For Pond Branch, a stream located in a forested watershed, it was Serratia, followed by Yersinia and Aeromonas. For McDonogh (MCDN), a stream associated with Zea mays (corn) row crop agriculture, E. coli was the most frequently isolated genus/species, followed by Aeromonas and Enterobacter. ERIC-PCR genotyping of isolates from the most prevalent genera/species, indicated a high degree of diversity within-stream for E. coli and K. pneumoniae. Conversely, genotyping of Y. enterocolitica isolates indicated that some were shared between different streams.

摘要

2004年6月至2005年6月期间,每周从三条溪流采集水样,其中的细菌在科利扫描简易凝胶琼脂平板上培养。对代表各种颜色和形态的菌落进行16S rRNA基因1000 - 1100 nt片段的扩增和测序。总共对528个菌落进行了测序;这些菌落分为26个属和78个种。在175个推测为大肠杆菌的深蓝色/紫色菌落中,序列分析表明有45个(25%)实际上是其他属。对于城市溪流格温斯瀑布 - 格温斯河,大肠杆菌是最常见的属/种,其次是克雷伯菌属和气单胞菌属。对于位于森林流域的池塘支流,最常见的是沙雷氏菌属,其次是耶尔森菌属和气单胞菌属。对于与玉米行作物农业相关的麦克多诺(MCDN)溪流,大肠杆菌是最常分离出的属/种,其次是气单胞菌属和肠杆菌属。对最常见属/种的分离株进行ERIC - PCR基因分型表明,大肠杆菌和肺炎克雷伯菌在溪流内具有高度多样性。相反,小肠结肠炎耶尔森菌分离株的基因分型表明,有些菌株在不同溪流之间是共有的。

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