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Isolation of enterotoxigenic Pseudomonas fluorescens from Asia region

亚洲地区产肠毒素荧光假单胞菌的分离

基本信息

批准号：
14406008
负责人：
TSUJI Takao
金额：
$ 9.47万
依托单位：
Fujita Health University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-14406008/
关键词：
Pseudomonas fluorescens Enterotoxigenic E coli heat-labile enterotoxin Children diarrhea epidermiology

项目摘要

We examined whether there was Pseudomonas fluorescens (enterotoxigenic P.fluorescens:ETPF), which produced a toxin like heat-labile enterotoxin (LT) of enterotoxigenic Escherichia coli (ETEC), in water at Surabya city in Indoneshia. From many places, we got environmental or house keeping water, isolated P.fluorescens and detected the gene of LT by PCR procedure.Among about 500 strains, we could get one strain of which chromosome carried the gene like that of LT. Then, we determined the full sequence of LT like gene from ETPF. However, it was almost identical to that of LT of ETEC. Therefore, to order to analyze how its gene in ETPF will be transferred from ETEC, we have been determining the full sequence of chromosomal DNA of ETPF.Moreover, we tried to isolate ETPF from diarrheal stools of 150 children in Surabaya city. Among them, although we could not isolate pathogenic bacteria like Shigellas, Vibrios, pathogenic E.coli, Campylobacters and ETPF, Pseudomonas, which caused diarrhea in rabbit ileal loop test, were isolated from 6 diarrheal stools. Then, we tried to purify the toxin, and finished its purification. There are many kinds of diarrheal agents of children in developing country, 20 % of diarrheal agents are not determined. Therefore, Pseudomonas isolated in this project may be one of the agents causing children diarrhea in developing country.

我们检测了印尼苏拉比亚市区的水中是否存在荧光假单胞菌(Enterotoxigen P.flrescens：ETPF)，该菌产生一种类似产肠毒素大肠杆菌(ETEC)的不耐热肠毒素(LT)。从多地采集环境水或保温水，分离荧光假单胞菌，用聚合酶链式反应方法检测LT基因，在约500株菌株中，可获得一株染色体上携带LT基因的菌株。然后，我们从ETPF中测定了LT样基因的全序列。然而，它与ETEC的LT几乎相同。因此，为了分析ETPF中的ETPF基因是如何从ETEC转移过来的，我们一直在测定ETPF的染色体DNA全序列，并试图从泗水地区150例腹泻儿童的粪便中分离到ETPF。其中，虽然未能分离出志贺氏菌、弧菌、致病性大肠杆菌、弯曲杆菌和ETPF等致病菌，但从6份腹泻粪便中分离到引起兔回肠环流试验腹泻的假单胞菌。然后，我们尝试提纯毒素，并完成了它的提纯。发展中国家的儿童腹泻药种类繁多，有20%的腹泻药未检出。因此，本项目分离的假单胞菌可能是引起发展中国家儿童腹泻的病原体之一。