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

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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