Development of purification, culture and detection methods and hydrological characteristics on Salmonella spp.from environmental waters

• Main Authors: Hung-Jen Wang, 王宏仁
• Other Authors: Bing-Mu Hsu
• Format: Others
• Language: zh-TW
• Published: 2010
• Online Access: http://ndltd.ncl.edu.tw/handle/88174347157867965697

碩士 === 國立中正大學 === 地震研究所暨應用地球物理研究所 === 98 === Bacterial pathogens are one of the major health risks associated with aquatic environment. Salmonella, Shigella and the other enteric pathogens have emerged as waterborne pathogens of concern. Salmonella is an important bacterium that causes diseases infecting the human alimentary tract. It can be transmitted via the environmental pollution resulting in continuous exposure to the inhabitants. Salmonella serotypes have been identified based on standard serotyping. Although this method has been widely used, high costs as well as labor and time consumption are still major problems. For effective diagnosis of Salmonella strains in environmental samples, a combination of the following tests were employed in the study: (1) Immunomagnetic Separation (IMS); (2) polymerase chain reaction (PCR); (3) pulsed-field gel electrophoresis (PFGE); (4) serum agglutination tests; (5) Real time PCR for understanding the serotype and the genotyping pattern of Salmonella spp. In this study, we also compared various immunomagnetic separation (IMS) operation conditions, including turbidity, add the amount of beads and bead reaction time, and in the different experimental procedures in order to assess the recovery efficiency of IMS and the detection limit of PCR. In the study, the total of 170 sites of water samples were taken from 5 drinking water treatment plants and Puzih River, and analyzed by direct extraction method, broth extraction method and various culture methods. The occurrence frequency was 25.3% for Salmonella spp. for all collected water samples. The 24 positive water samples by culture method were further identified as S. Typhimurium (3/24), S. Bareilly (1/24), S. Isangi (15/24), S. Paratyphi B var. Java (2/24), S. Potsdam (1/24) and S. Newport (2/24) by biochemical testing, serological identification and PFGE.While Combining IMS in the general purification procedure,, the occurrence of Salmonella was increased. In the study, we analyzed the difference between the present/absent of Salmonella spp. in water samples and water quality parameters. High significant correlation was found between the presence of Salmonella and water temperature and total coliforms.