军团菌与同一空调水环境中分离的自由生活阿米巴的共培养观察  

Observation of coculture of Legionella and free-living amoeba from the same environmental water sample

作　　者：陈洪友[1] 姜庆五[1] 白云[2] 李勤学[1] 吴红岩[2] 

机构地区：[1]复旦大学公共卫生学院,上海200032 [2]上海市浦东新区疾病预防控制中心,上海200129

出　　处：《中国卫生检验杂志》2006年第7期775-777,共3页Chinese Journal of Health Laboratory Technology

摘　　要：目的:通过从同一环境的同一份水样中分别分离出自由生活阿米巴和军团菌,然后将两者在实验室条件下共培养,观察两者的形态,从而推测两者在自然环境下的相互关系。方法:用微孔滤膜过滤,盐酸处理和抗生素抑制杂菌的方法分别从同一份水样中分离军团菌和自由生活阿米巴,然后将两者在实验室条件下进行共培养;取片后G im enes染色,普通光学显微镜下观察,72 h后收集共培养物做超薄切片电镜下观察,研究两者的形态。结果:分别分离到1株I型嗜肺军团菌和一株棘阿米巴。两者进行共培养发现军团菌可以感染阿米巴,光镜下观察,8 h后阿米巴体内即可发现军团菌,24 h后开始增多,72 h后阿米巴滋养体呈高度寄生状态,玻片上可见被裂解的滋养体遗骸。电镜照片显示被吞食的军团菌被多层膜结构缠绕。结论:阿米巴吞噬细菌是一种缠绕吞噬的方式,水样中分离到的军团菌能在同样本中分离到的自由生活阿米巴内生长繁殖,从侧面证实环境水体中自由生活阿米巴是嗜肺军团菌的自然宿主。Objectlve：To observe the survival and multiplication of Legionella cocultured with amoeba from the same water sample for revealing the relationship between them in environmenL Methods：Legionella was cocultured with amoeba iolated from the same sample by filtration, HCL and antibiotic processing under laboratory condition with the ratio of 1 ： 10. Changes were inves- tigated between Legionella and amoeba in morphological way by using inverted microscopy and Gimenes staining of the cover glass picking up from the flask about every 24 h. The coculture was also be observed under electron microscope after 72 h. Results：One strain of Legionella pneumophila serology type I and one strain of acanthamoeba were isolated. Gimenes staining showed that Legionella was obsevered in the amoebae cocultured after 8 hours, and the number was increased after 24 hours. Trophic amoebae were highly parasited, and remains of amoebae that had been lysed were obsevered. Through electron microscope, the multiple membrane around the bacteria was found. Conclusion ： The way for free - living amoeba to phagocytose bacteria is a coiling phagocyto- sis. Legionella can survive and multiply in free - living amoebae isolated from the same water sample, which suggests that freeliving amoebae are the natural host of Legionella pneumophila in environmental water.

关 键 词：嗜肺军团菌 自由生活阿米巴 共同培养 自然宿主 

分 类 号：R117[医药卫生—公共卫生与预防医学]