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.

我们检查了是否有假单胞菌（肠毒素p.fluorescens：ETPF），该毒素像肠毒素大肠杆菌（ETEC）一样，产生了毒素，例如热毒素肠毒素（LT），在印度尼亚岛的萨拉比亚市的水中。在许多地方，我们得到了环境或房屋保存水，孤立的p.fluorescens并通过PCR程序检测到LT的基因。在大约500个菌株中，我们可以得到一种染色体的菌株，其基因像LT一样。然后，我们确定了来自ETPF的LT基因的完整序列。但是，它几乎与ETEC的LT相同。因此，为了分析其在ETPF中的基因如何从ETEC转移，我们一直在确定ETPF的染色体DNA的完整序列。此外，我们试图将ETPF与萨巴雅市150名儿童的腹泻粪便分离。在其中，从6个腹泻粪便中分离出我们无法隔离引起兔子回肠循环测试的腹泻的假单胞菌，尽管我们无法分离志贺氏菌，颤音，致病性大肠杆菌，弯曲杆菌和ETPF等致病细菌。然后，我们试图纯化毒素，并完成其净化。发展中国家有许多儿童的腹泻药，尚未确定20％的腹泻药。因此，在该项目中孤立的假单胞菌可能是导致发展中国家儿童腹泻的特工之一。