Alam Munirul, Chowdhury Wasimul B, Bhuiyan N A, Islam Atiqul, Hasan Nur A, Nair G Balakrish, Watanabe H, Siddique A K, Huq Anwar, Sack R Bradley, Akhter M Z, Grim Christopher J, Kam K-M, Luey C K Y, Endtz Hubert P, Cravioto Alejandro, Colwell Rita R
International Center for Diarrhoeal Disease Research, Bangladesh, Mohakhali, Dhaka 1212, Bangladesh.
Appl Environ Microbiol. 2009 Oct;75(19):6268-74. doi: 10.1128/AEM.00266-09. Epub 2009 Aug 14.
Forty-two strains of Vibrio parahaemolyticus were isolated from Bay of Bengal estuaries and, with two clinical strains, analyzed for virulence, phenotypic, and molecular traits. Serological analysis indicated O8, O3, O1, and K21 to be the major O and K serogroups, respectively, and O8:K21, O1:KUT, and O3:KUT to be predominant. The K antigen(s) was untypeable, and pandemic serogroup O3:K6 was not detected. The presence of genes toxR and tlh were confirmed by PCR in all but two strains, which also lacked toxR. A total of 18 (41%) strains possessed the virulence gene encoding thermostable direct hemolysin (TDH), and one had the TDH-related hemolysin (trh) gene, but not tdh. Ten (23%) strains exhibited Kanagawa phenomenon that surrogates virulence, of which six, including the two clinical strains, possessed tdh. Of the 18 tdh-positive strains, 17 (94%), including the two clinical strains, had the seromarker O8:K21, one was O9:KUT, and the single trh-positive strain was O1:KUT. None had the group-specific or ORF8 pandemic marker gene. DNA fingerprinting employing pulsed-field gel electrophoresis (PFGE) of SfiI-digested DNA and cluster analysis showed divergence among the strains. Dendrograms constructed using PFGE (SfiI) images from a soft database, including those of pandemic and nonpandemic strains of diverse geographic origin, however, showed that local strains formed a cluster, i.e., "clonal cluster," as did pandemic strains of diverse origin. The demonstrated prevalence of tdh-positive and diarrheagenic serogroup O8:K21 strains in coastal villages of Bangladesh indicates a significant human health risk for inhabitants.
从孟加拉湾河口分离出42株副溶血性弧菌,并与两株临床菌株一起对其毒力、表型和分子特征进行了分析。血清学分析表明,O8、O3、O1和K21分别是主要的O和K血清群,O8:K21、O1:KUT和O3:KUT占主导地位。K抗原无法分型,未检测到大流行血清群O3:K6。除两株缺乏toxR的菌株外,其余所有菌株均通过PCR确认存在toxR和tlh基因。共有18株(41%)菌株拥有编码耐热直接溶血素(TDH)的毒力基因,一株拥有TDH相关溶血素(trh)基因,但没有tdh。10株(23%)菌株表现出作为毒力替代指标的神奈川现象,其中6株,包括两株临床菌株,拥有tdh。在18株tdh阳性菌株中,17株(94%),包括两株临床菌株,具有血清标记O8:K21,一株为O9:KUT,唯一一株trh阳性菌株为O1:KUT。没有一株具有群体特异性或ORF8大流行标记基因。采用脉冲场凝胶电泳(PFGE)对SfiI酶切DNA进行DNA指纹图谱分析和聚类分析,结果显示菌株之间存在差异。然而,使用来自软数据库的PFGE(SfiI)图像构建的系统发育树,包括不同地理来源的大流行和非大流行菌株的图像,显示本地菌株形成了一个聚类,即“克隆聚类”,不同来源的大流行菌株也是如此。在孟加拉国沿海村庄证实tdh阳性和致泻血清群O8:K21菌株的流行,表明居民面临重大的人类健康风险。