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

再生废水的水质要求对与健康有关的微生物制定新的标准。特别是在发达的城市地区，处理后的废水回用量有所增加。我们水质科学家和工程师几乎只对病原体感兴趣。然而，在上述目前的情况下，应该对机会性感染给予另一个重要的关注，因为包括健康的残疾人在内的数目不详的人最终接触到水中产生的机会性传染性微生物。嗜肺军团菌是一种重要的水源性机会感染细菌，可引起军团病，本研究开发了一种新的检测方法。采用聚合酶链反应（PCR）和DNA-DNA杂交法对嗜肺乳杆菌进行了检测，并对PCR法进行了改进，使其可用于嗜肺乳杆菌的定量检测。PCR- dna杂交试验结果表明，嗜肺乳杆菌有可能通过污水二次处理和消毒过程存活，而传统的嗜肺乳杆菌选择培养基是不可用的。嗜肺乳杆菌的PCR-MPN实验数据与荧光染色直接计数所得的计数数据具有良好的相关性。研究人员还提出了建立一种检测污水中嗜肺杆菌的分子生物学方法的挑战。将生长在尼龙膜过滤器上的细菌菌落置于含有四种不抑制嗜肺杆菌生长的抗生素的选择性bcye - α琼脂板上，通过先前开发的菌落杂交处理。用嗜肺杆菌特异性mip DNA探针进行的新菌落杂交表明，嗜肺杆菌菌落与琼脂培养基上膜过滤器培养的其他非军团菌菌落有明显区别。因此，利用新的菌落杂交方法可以对污水中活菌（形成菌落）进行选择性和定量检测。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