AQUAVET, Laboratory of Aquatic Animal Diseases, Department of Veterinary Medicine, Federal University of Lavras, Lavras MG 37200-000, Brazil.
Vet Microbiol. 2010 Aug 26;144(3-4):371-6. doi: 10.1016/j.vetmic.2010.01.021. Epub 2010 Feb 1.
The goals of this study were to develop a PCR technique to detect ascV and aopB genes from the type III secretion system (T3SS), to evaluate the frequency of these genes in Aeromonas hydrophila strains isolated from diseased fish and from aquaculture environments, and to determine the relationship between the presence of these genes and virulence of A. hydrophila in Nile tilapia. The PCR assay developed here successfully detected the target genes, showing three different profiles for the strains ascV+/aopB+, ascV+/aopB-, and ascV-/aopB-. A higher frequency of ascV+/aopB+ was verified in isolates from diseased fish compared to those from aquaculture environments (P<0.05). Among 64 isolates from diseased fish, ascV+/aopB+ (62.5%) was the most frequent profile (P<0.05) and caused more intensive mortality rates. Environmental strains containing the ascV+/aopB+ profile were less virulent than isolates from clinical cases. These results suggest that the presence of a functional T3SS probably increases the virulence of A. hydrophila. The PCR technique was shown to be a specific and efficient tool for detection of T3SS, and this technique can be used for virulence typing of A. hydrophila isolates.
本研究的目的是开发一种 PCR 技术来检测 III 型分泌系统(T3SS)中的 ascV 和 aopB 基因,评估这些基因在来自患病鱼类和水产养殖环境的嗜水气单胞菌菌株中的频率,并确定这些基因的存在与嗜水气单胞菌对尼罗罗非鱼的毒力之间的关系。本研究中开发的 PCR 检测方法成功地检测到了靶基因,显示出菌株 ascV+/aopB+、ascV+/aopB-和 ascV-/aopB-三种不同的图谱。与水产养殖环境中的分离株相比,从患病鱼类中分离出的菌株中 ascV+/aopB+的频率更高(P<0.05)。在来自患病鱼类的 64 个分离株中,ascV+/aopB+(62.5%)是最常见的图谱(P<0.05),并导致更高的死亡率。含有 ascV+/aopB+图谱的环境菌株的毒力低于临床病例的分离株。这些结果表明,功能性 T3SS 的存在可能会增加嗜水气单胞菌的毒力。PCR 技术被证明是一种用于检测 T3SS 的特异性和有效工具,该技术可用于嗜水气单胞菌分离株的毒力分型。