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THE INFLUENCE OF PH AND THE ELUCIDATION OF CONTAMINATION ROUTE FOR LEGIONELLA IN HOT SPRING WATERS

温泉水中PH值的影响及军团菌污染途径的阐明

基本信息

批准号：
13680634
负责人：
KATO Naoyuki
金额：
$ 1.98万
依托单位：
Toho University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13680634/
关键词：
LEGIONELLA HOT SPRING VIABLE BUT NONCULTURABLE L.PNEUMOPHILA ACANTHAMOEBA CHLORINE STERILIZATION ELCIDATION OF CONTAMINATION ROUTE SOIL PFGE法残留塩素濃度 L. pneumophila PCR法 RAPD法血清群レジオネラ pH 温度電導度

项目摘要

1.Change in the quality due to chloridizing against sampled water collected from indoor or outdoor bathtub of various spa was analyzed and epidemiological study of Legionella spp. in these water was carried out.2.Transition to VNC (viable but not cultivable) stage of L.pneumophila population in microcosms preparated in sampled water was examined under conditions of different pH and temperature(1)Whole cells of L.pneumophila were killed in a short period in microcosm with extreme low pH, while cell numbers were gradually reduced in microcosm with high pH. On the other hand, it was suggested that cell population in microcosm with pH5 shifted to VNC stage. (2)L.pneumophila could survive for a long period in microcosms in a range of pH6.0 to 8.2 or 25 to 42℃ (3)It has previously been thought that L.pneumophila could not survive in high-temperature water of 45℃ however, the present data suggested that L.pneumophila could survive as VNC even under this condition. However, L.pneumophila was q … More uickly killed in microcosms with temperature of 50 or more degrees Celsius.3.Antiseptic effect of chlorine against L.pneumophila and Acanthamoeba was examined.(1)L.pneumophila could survive for one minute or more in microcosm with 0.4 ppm of residual chlorine. In residual chlorine of 1.0 ppm, the organisms were quickly killed. (2)Acanthamoeba cells were added into microcosms, and then L.pneumophila multiplicated in phagosomes of amoeba cells and ate away cells, and repeated these process during incubation. After incubation for 24 hrs, a various concentrations of chlorine were added into microcosms. A small amount of L.pneumonia cells was detected even in microcosm disinfected with 50 ppm of residual chlorine. Forty % of Acanthamoeba cells was living in microcosm with 10 ppm of residual chlorine. These cells might exsist as cyst cell having resistance to chlorine, therefore, L.pneumonia cells were also protected in these cysts from bactericidal effect of chlorine.4.The route that Legionella contaminate spa was studied.(1)Genotype of L.pneumophila isolates with identical serotype isolated from spa and the surrounding soil was determined by randomly amplified polymorphic DNA analysis(RAPD). As so far examined, genetic identity among these isoltes was not recognized. (2)RAPD analysis was done about L.pneumophila isolates of a variety of serotype isolated from bathtub and filtlate tank in spa with circulating system. Genetic identity was shown among isolates of different serotype. Further confirmation by pulse field gell electrophoresis(PFGE) was carried out. Less

1、对各温泉浴场室内外浴缸取样水进行氯化处理后的水质变化进行分析，并进行军团菌流行病学研究。 2.在不同pH和温度条件下,考察了采样水中制备的微环境中嗜肺军团菌种群向VNC(存活但不可培养)阶段的转变(1)在极低pH的微环境中嗜肺军团菌的全细胞在短时间内被杀死,而在高pH的微环境中细胞数量逐渐减少。另一方面，有人认为pH5的微观世界中的细胞群转移到VNC阶段。 (2)嗜肺军团菌可以在pH6.0至8.2或25至42℃的微观世界中长期存活。 (3)以前认为嗜肺军团菌不能在45℃的高温水中存活，但目前的数据表明，即使在这种条件下，嗜肺军团菌也可以作为VNC存活。然而，嗜肺军团菌在50摄氏度以上的微观世界中很快被杀死。3.检测了氯对嗜肺军团菌和棘阿米巴的防腐作用。(1)嗜肺军团菌在余氯为0.4 ppm的微观世界中可存活1分钟或更长时间。余氯在1.0ppm时，微生物很快被杀死。 (2)将棘阿米巴细胞加入到微观世界中，然后嗜肺军团菌在阿米巴细胞的吞噬体中增殖并吞噬细胞，并在培养过程中重复这一过程。孵育24小时后，将不同浓度的氯添加到微观世界中。即使在用50 ppm余氯消毒的微观世界中也检测到少量肺炎李斯特菌细胞。 40% 的棘阿米巴细胞生活在残留氯为 10 ppm 的微观世界中。这些细胞可能以对氯具有抗性的包囊细胞的形式存在，因此包囊内的肺炎杆菌细胞也受到了氯的杀菌作用的保护。4.研究了军团菌污染spa的途径。(1)通过随机扩增多态性DNA测定了从spa及其周围土壤中分离出的血清型相同的嗜肺军团菌基因型。分析（RAPD）。到目前为止，尚未认识到这些孤立株之间的遗传同一性。 (2)对带循环系统的水疗浴缸和滤液池分离的多种血清型嗜肺军团菌进行RAPD分析。不同血清型的分离株之间显示出遗传同一性。通过脉冲场凝胶电泳(PFGE)进一步证实。较少的