Rattanama Pimonsri, Srinitiwarawong Kanchana, Thompson Janelle R, Pomwised Rattanaruji, Supamattaya Kidchakarn, Vuddhakul Varaporn
Department of Microbiology, Faculty of Science, Prince of Songkla University, Hat Yai, 90112 Songkhla, Thailand.
Dis Aquat Organ. 2009 Sep 23;86(2):113-22. doi: 10.3354/dao02119.
The virulence factors of Vibrio harveyi, the causative agent of luminous vibriosis, are not completely understood. We investigated the correlations between shrimp mortality, hemolysis, the presence of a hemolysin gene (vhh), and a gene involved in the type III secretion system (the Vibrio calcium response gene vcrD). V harveyi HY01 was isolated from a shrimp that died from vibriosis, and 36 other V. harveyi isolates were obtained from fish and shellfish in Hat Yai city, Thailand. An ocean isolate of V. harveyi BAA-1116 was also included. Thirteen isolates including V harveyi HYO1 caused shrimp death 12 h after injection. Most V harveyi isolates in this group (designated as Group A) caused hemolysis on prawn blood agar. None of the shrimp died after injection with V harveyi BAA-1116. Molecular analysis of all V harveyi isolates revealed the presence of vcrD in both pathogenic and non-pathogenic strains. Although vhh was detected in all V harveyi isolates, some isolates did not cause hemolysis, indicating that vhh gene expression might be regulated. Analysis of the V harveyi HYO1 genome revealed a V cholerae like-hemolysin gene, hlyA (designated as hhl). Specific primers designed for hhl detected this gene in 3 additional V harveyi isolates but the presence of this gene was not correlated with pathogenicity. Random amplified polymorphic DNA (RAPD) analysis revealed a high degree of genetic diversity in all V harveyi isolates, and there were no correlations among the hhl-positive isolates or the pathogenic strains.
哈维氏弧菌是发光弧菌病的病原体,其毒力因子尚未完全明确。我们研究了虾死亡率、溶血现象、溶血素基因(vhh)的存在以及与III型分泌系统相关的基因(弧菌钙反应基因vcrD)之间的相关性。哈维氏弧菌HY01是从死于弧菌病的虾中分离得到的,另外36株哈维氏弧菌分离株则取自泰国合艾市的鱼类和贝类。还纳入了一株海洋来源的哈维氏弧菌BAA - 1116。包括哈维氏弧菌HY01在内的13株分离株在注射后12小时导致虾死亡。该组中的大多数哈维氏弧菌分离株(指定为A组)在对虾血琼脂上引起溶血。注射哈维氏弧菌BAA - 1116后没有虾死亡。对所有哈维氏弧菌分离株的分子分析显示,致病菌株和非致病菌株中均存在vcrD。虽然在所有哈维氏弧菌分离株中都检测到了vhh,但一些分离株并未引起溶血,这表明vhh基因的表达可能受到调控。对哈维氏弧菌HY01基因组的分析揭示了一个类似霍乱弧菌的溶血素基因hlyA(指定为hhl)。针对hhl设计的特异性引物在另外3株哈维氏弧菌分离株中检测到了该基因,但该基因的存在与致病性并无关联。随机扩增多态性DNA(RAPD)分析显示,所有哈维氏弧菌分离株具有高度的遗传多样性,hhl阳性分离株或致病菌株之间没有相关性。