Studies of development of method for detection of aquatic pathogenic bacteria in viable but non-culturable stage and their ecology.

水生非可培养阶段病原菌检测方法及其生态学研究。

• 批准号: 10557236
• 负责人: SHINODA Sumio
• 金额: $ 6.27万
• 依托单位: OKAYAMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-10557236/
• 关键词: Pathogenic vibrio; VNC; Ecology; Vibrio parahaemolyticus; ビブリオ; 腸炎ビブリオ; 生態学; 水中微生物; VBNC

Some bacterial species such as genera Bacillus and Clostridium adapt against undesirable circumstances by spore formation. Recently it has been believed that a tolerant mechanism other than spore formation against change of environmental condition acts in natural environment, namely viable but non-culturable (VNC) stage of bacterial cells. Some of the aquatic bacteria are pathogenic for human and many of them can be detected by cultivation on/in usual media for clinical diagnostic, but some of them cannot. These undetectable bacteria are supposed to exist as VNC.We tried to develop method for detection of VNC and to apply it to ecological study, special emphasis on ecology of Vibrio parahaemolyticus, a food poisoning bacterium. A polymerase chain reaction method with primers induced from lateral flagellar gene of the vibrio was developed and its usefulness was tested. Although some improvements are necessary, the method is thought to be useful for detection VNC cells. Ecological study of the vibrio was also carried out and some points of problem in use of selective media to ecological study was suggested.

一些细菌物种如芽孢杆菌属和梭菌属通过孢子形成来适应不良环境。近年来，人们认为，在自然环境中，细菌细胞的可存活但不可培养（VNC）阶段，除了孢子形成之外，还有一种耐受环境条件变化的机制。一些水生细菌对人类具有致病性，其中许多细菌可以通过在常规培养基上/中培养来检测用于临床诊断，但其中一些细菌不能。我们尝试建立一种检测VNC的方法，并将其应用于生态学研究，特别是对一种食物中毒菌--副溶血性弧菌的生态学研究。以该弧菌的侧鞭毛基因为引物，建立了一种聚合酶链反应方法，并对该方法的实用性进行了检验。虽然还需要改进，但该方法被认为是有用的，用于检测VNC细胞。并对弧菌的生态学进行了研究，提出了应用选择性培养基进行生态学研究时应注意的问题。

项目成果

Alam, Md.J., Miyoshi, S., Tomochika, K., Shinoda, S.: "Analysis of sea waters for the recovery of culturable Vibrio parahaemolyticus and some other vibrios."Microbiol.Immunol.. 46(in press). (2001)

Alam, Md.J.、Miyoshi, S.、Tomochika, K.、Shinoda, S.：“分析海水以恢复可培养的副溶血性弧菌和其他一些弧菌。”Microbiol.Immunol.. 46（印刷中）

Bi, K., Miyoshi, S., Tomochika, K., Shinoda, S.: "Detection of viable Vibrio mimicus by reverse transcription- polymerase chain reaction."Biocontrol.Sci.. 6(in press). (2001)

Bi, K.、Miyoshi, S.、Tomochika, K.、Shinoda, S.：“通过逆转录聚合酶链反应检测活的拟态弧菌。”Biocontrol.Sci.. 6（出版中）。

Shi, L. et al.: "Presence of hemolysin genes (vmh, tdh and hix) in isolates of Vibrio mimicus determined by polymerase chain reaction"J. Health Sci.. 46. 63-65 (2000)

Shi, L. 等人：“通过聚合酶链反应测定拟态弧菌分离株中溶血素基因（vmh、tdh 和 hix）的存在”J.

Bi,K.,Miyoshi,S.,Tomochika,K.,Shinoda,S.: "Detection of viable Vibrio mimicus by reverse transcription-polymerase chain reaction."Biocontrol.Sci.. 6(in press). (2001)

Bi,K.,Miyoshi,S.,Tomochika,K.,Shinoda,S.：“通过逆转录聚合酶链式反应检测活的拟态弧菌。”Biocontrol.Sci.. 6（印刷中）。

Alam,Md.J.,Miyoshi,S.,Tomochika,K.,Shinoda,S.: "Analysis of sea waters for the recovery of culturable Vibrio parahaemolyticus and some other vibrios."Microbiol.Immunol.. 46(in press). (2001)

Alam，Md.J.，Miyoshi，S.，Tomochika，K.，Shinoda，S.：“分析海水以恢复可培养的副溶血性弧菌和其他一些弧菌。”Microbiol.Immunol.. 46（印刷中）