Área de Microbiología, Departamento de Biología Funcional, Facultad de Medicina, IUBA, Universidad de Oviedo, 33006 Oviedo, Asturias, Spain.
Environ Microbiol Rep. 2013 Feb;5(1):179-85. doi: 10.1111/1758-2229.12030. Epub 2013 Jan 15.
In this study, the utilization of bioluminescence imaging (BLI) allowed us to define the progression of Yersinia ruckeri during the infection of rainbow trout. A luminescent Y. ruckeri 150 strain was engineered using the pCS26-Pac plasmid containing the lux operon from Photorhabdus luminescens. Two different models of infection of rainbow trout were defined depending on the route in which bacteria were administered, being the gut the major organ affected following bath immersion. This indicates that this organ is important for bacterial dissemination inside the fish and the establishment of the infection. Moreover, the expression of three previously selected operons by in vivo expression technology (IVET) was analysed, the yhlBA involved in the production of a haemolysin, the cdsAB related to the uptake of cysteine and the yctCBA implicated in citrate uptake. Apart from these factors, the expression of yrp1 encoding a serralysin metalloprotease involved in pathogenesis was also analysed. The results indicated that all of the assayed promoters were expressed during infection of rainbow trout. In addition to these findings, the methodology described in this work constitutes a useful model for studying the infection process in other fish pathogenic bacteria.
在这项研究中,我们利用生物发光成像(BLI)来定义弗氏柠檬酸杆菌在感染彩虹鳟鱼过程中的进展。使用含有发光杆菌 lux 操纵子的 pCS26-Pac 质粒,对发光弗氏柠檬酸杆菌 150 株进行了工程改造。根据细菌给药途径的不同,确定了两种不同的彩虹鳟鱼感染模型,肠道是浴浸后受影响的主要器官。这表明该器官对于细菌在鱼体内的传播和感染的建立很重要。此外,通过体内表达技术(IVET)分析了三个先前选择的操纵子的表达,参与产生溶血素的 yhlBA、与半胱氨酸摄取相关的 cdsAB 以及与柠檬酸摄取相关的 yctCBA。除了这些因素外,还分析了编码丝氨酸蛋白酶金属蛋白酶的 yrp1 的表达,该蛋白酶与发病机制有关。结果表明,在感染彩虹鳟鱼期间,所有检测到的启动子都有表达。除了这些发现外,本工作中描述的方法为研究其他鱼类病原菌的感染过程提供了一个有用的模型。