Study on quantification of opportunistically infectious microorganisms in sewage and treated sewage and safety assessment of wastewater reclamation.

再生水水质对健康相关微生物提出了新的要求。特别是在经济发达的城市阿里亚斯，处理后的污水回用量不断增加。我们水质科学家和工程师几乎只对病原体感兴趣。然而，在上述现状下，另一个重要的注意力应该放在机会性感染上，因为包括残疾人在内的不确定数量的人最终会暴露于水中的机会性感染微生物。采用聚合酶链反应（PCR）和DNA-DNA杂交方法检测嗜肺军团菌，并对PCR方法进行了改进，使其应用于嗜肺军团菌的定量检测 关于我们 显示嗜肺军团菌通过二级污水处理和消毒过程存活的可能性，其中常规的嗜肺军团菌选择性培养基不可用。PCR-MPN检测嗜肺军团菌的实验结果与荧光染色直接计数法的计数结果具有良好的相关性，并建立了一种检测污水中嗜肺军团菌活菌的分子生物学方法。将在尼龙膜滤器上生长的细菌菌落置于含有四种不抑制嗜肺军团菌生长的抗生素的选择性BCYE-α琼脂平板上，通过先前开发的菌落杂交进行处理。用嗜肺军团菌特异的mip DNA探针进行杂交，结果表明，嗜肺军团菌菌落与其它非军团菌菌落在琼脂培养基上的滤膜上形成的菌落相区别。因此，用新的菌落杂交方法可以选择性地定量检测污水中的活菌（菌落形成）。少

Water quality of reclaimed wastewater is required new standard on health related microorganisms. Quantity of treated wastewater reused has been increased especially in developed urban arias. We water quality scientists and engineers have almost only been interested in the pathogens. Under the resent situation above mentioned, however, another important attention should be taken for opportunistic infection, because an unspecified number of people including handicapped persons in health are eventually exposed to water born opportunistically infectious microorganisms.In this study, the researcher developed new detection methods of Legionellapneumophila, which is an important water born opportunistic-infection bacteria and causes legionnaire disease, in sewage. Polymerase chain reaction (PCR) and DNA-DNA hybridization methods were employed to detect L.pneumophila and PCR method was improved to be applied to quantitative detection of L.pneumophila.The results from PCR-DNA hybridization test … More show the possibility that L.pneumophila survives through secondary sewage treatment and disinfection process in which conventional selective media for L.pneumophila are not available. The experimental data of PCR-MPN for L.pneumophila shows good correlation to the enumeration data obtained by fluorescence stained direct count of this bacteria.The researcher also challenged to establish a molecular biological method for the detection of L.pneumophila viable in sewage. Te bacterial colonies grown on a nylon membrane filter put on a selective BCYE-alpha agar plate that contains four antibiotics which do not restrain the growth of L.pneumophila were processedthrough the colony-hybridization previously developed. This new colony-hyblidization with a L.pneumophila-specific mip DNA probe showed that the L.pneumophila colonies were distinguished from other non-Legionella coloniesdeveloped om the memblane filter on the agar medium. Therefore, viable (colony-forming) L.pneumophila in sewage can be detected selectively and quantativelyby using the new colony hyblidization method. Less