France Haliotis, Kerazan, Lilia, Plouguerneau, France.
J Invertebr Pathol. 2010 Nov;105(3):289-97. doi: 10.1016/j.jip.2010.08.001. Epub 2010 Aug 6.
Since 1998, episodic mass mortality of the abalone Haliotistuberculata has been observed along the northern Brittany coast of France caused by a complex interaction among the host, pathogen and environmental factors. In the present study, abalone were submitted to two successive infections with the pathogen Vibrioharveyi under controlled conditions. During the first challenge, infection by V.harveyi resulted in 64% mortality of mature abalone. After a second infection of those surviving the first challenge, only 44% mortality was observed. Physiological variability in the host response appears to be a major determinant in susceptibility to V.harveyi. In order to isolate differentially expressed genes in H.tuberculata challenged with this bacterium, suppression subtractive hybridization (SSH) cDNA libraries were constructed from muscle of moribund abalone (susceptibles), surviving individuals (apparently resistant to the bacterium) and control (unexposed) animals. Of the 1152 clones sequenced, 218 different partial cDNA sequences were obtained and represented 69 known genes. Of these, 65 were identified for the first time in H.tuberculata. Using real-time PCR, a time-course study was conducted on 19 of the genes identified by SSH. A majority of differentially expressed transcripts were down-regulated in susceptible individuals as compared to their resistant counterparts. Bacterial challenge of abalone resulted in the up-regulation of three transcripts (encoding ferritin, heat shock protein HSP84 and fatty acid binding protein FABP) in those that survived exposure to V.harveyi. This study has identified potential candidates for further investigation into the functional basis of resistance and susceptibility to summer vibriosis outbreaks in abalone.
自 1998 年以来,法国布列塔尼北部沿海地区一直观察到鲍鱼 Haliotistuberculata 的爆发性大规模死亡,这是宿主、病原体和环境因素复杂相互作用的结果。在本研究中,鲍鱼在受控条件下接受了两次病原体 Vibrio harveyi 的连续感染。在第一次挑战中,V. harveyi 的感染导致成熟鲍鱼的死亡率为 64%。在第一次挑战中幸存下来的个体再次感染后,仅观察到 44%的死亡率。宿主对感染的生理变异性似乎是对 V. harveyi 易感性的主要决定因素。为了分离出受这种细菌挑战的 H. tuberculata 中差异表达的基因,从濒死鲍鱼(易感)、幸存个体(显然对细菌有抗性)和对照(未暴露)动物的肌肉中构建了抑制性消减杂交(SSH) cDNA 文库。在测序的 1152 个克隆中,获得了 218 个不同的部分 cDNA 序列,代表了 69 个已知基因。其中,有 65 个基因是首次在 H. tuberculata 中鉴定到的。使用实时 PCR,对通过 SSH 鉴定的 19 个基因进行了时间过程研究。与具有抗性的个体相比,易感个体中大多数差异表达的转录物均下调。鲍鱼的细菌挑战导致在暴露于 V. harveyi 后幸存下来的个体中,三个转录物(编码铁蛋白、热休克蛋白 HSP84 和脂肪酸结合蛋白 FABP)上调。本研究鉴定了潜在的候选基因,可进一步研究鲍鱼夏季弧菌病暴发的抗性和易感性的功能基础。