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.

我们检查了印度尼西亚泗水市的水中是否存在荧光假单胞菌（产肠毒素 P. florescens：ETPF），它产生类似于产肠毒素大肠杆菌（ETEC）的不耐热肠毒素（LT）。我们从许多地方获取环境或生活用水，分离荧光假单胞菌，并通过PCR方法检测LT基因。在约500株菌株中，我们找到了一株染色体携带类似LT基因的菌株。然后，我们从ETPF中确定了LT样基因的完整序列。然而，它与ETEC的LT几乎相同。因此，为了分析ETPF中的其基因是如何从ETEC转移的，我们一直在测定ETPF染色体DNA的全序列。此外，我们尝试从泗水市150名儿童的腹泻粪便中分离出ETPF。其中，虽然未能分离到志贺氏菌、弧菌、致病性大肠杆菌、弯曲杆菌、ETPF等致病菌，但在兔回肠袢试验中，从6份腹泻粪便中分离到引起腹泻的假单胞菌。然后，我们尝试净化毒素，并完成了净化。发展中国家儿童腹泻剂种类繁多，20%的腹泻剂尚未确定。因此，本项目分离出的假单胞菌可能是导致发展中国家儿童腹泻的病原之一。