Weidhaas Jennifer, Mantha Sirisha, Hair Elliott, Nayak Bina, Harwood Valerie J
Civil and Environmental Engineering, West Virginia University, Morgantown, West Virginia, USA
Civil and Environmental Engineering, West Virginia University, Morgantown, West Virginia, USA.
Appl Environ Microbiol. 2015 Jan;81(1):196-202. doi: 10.1128/AEM.02354-14. Epub 2014 Oct 17.
Water quality monitoring techniques that target microorganisms in the order Bacteroidales are potential alternatives to conventional methods for detection of fecal indicator bacteria. Bacteroidales and members of the genus Bacteroides have been the focus of microbial source tracking (MST) investigations for discriminating sources of fecal pollution (e.g., human or cattle feces) in environmental waters. For accurate source apportionment to occur, one needs to understand both the abundance of Bacteroides in host feces and the survival of these host-associated microbial markers after deposition in the environment. Studies were undertaken to evaluate the abundance, persistence, and potential for growth of Bacteroidales originating from poultry litter under oxic and anoxic environmental conditions. Bacteroidales abundance, as determined by quantitative PCR (qPCR) with GenBac primers and probe, increased 2 to 5 log gene copies ml(-1) and 2 log gene copies g litter(-1) under most conditions during incubation of poultry litter in a variety of laboratory microcosm and field mesocosm studies. DNA sequencing of the Bacteroidales organisms in the litter identified taxa with sequences corresponding exactly to the GenBac primer and probe sequences and that were closely related to Bacteroides uniformis, B. ovatus, and B. vulgatus. These results suggest that MST studies using qPCR methods targeting Bacteroidales in watersheds that are affected by poultry litter should be interpreted cautiously. Growth of Bacteroidales originating from poultry litter in environmental waters may occur while Bacteroidales growth from other fecal sources declines, thus confounding the interpretation of MST results.
针对拟杆菌目微生物的水质监测技术是检测粪便指示菌的传统方法的潜在替代方法。拟杆菌目和拟杆菌属成员一直是微生物源追踪(MST)调查的重点,用于区分环境水体中粪便污染的来源(例如人类或牛粪便)。为了进行准确的源分配,需要了解宿主粪便中拟杆菌的丰度以及这些与宿主相关的微生物标记物在环境中沉积后的存活情况。开展了研究以评估在有氧和无氧环境条件下源自家禽粪便的拟杆菌目的丰度、持久性和生长潜力。在各种实验室微观世界和野外中观世界研究中,在家禽粪便培养期间的大多数条件下,用GenBac引物和探针通过定量PCR(qPCR)测定的拟杆菌目丰度增加了2至5个对数基因拷贝毫升⁻¹和2个对数基因拷贝克粪便⁻¹。对粪便中拟杆菌目生物的DNA测序确定了与GenBac引物和探针序列完全对应的分类单元,并且与普氏拟杆菌、卵形拟杆菌和普通拟杆菌密切相关。这些结果表明,在受家禽粪便影响的流域中,使用针对拟杆菌目的qPCR方法进行的MST研究应谨慎解释。当来自其他粪便来源的拟杆菌目生长下降时,源自家禽粪便的拟杆菌目可能在环境水体中生长,从而混淆了MST结果的解释。