哈维弧菌和鲍类疱疹病毒刺激对杂色鲍免疫相关因子的影响

以哈维弧菌(Vibrio harveyi)、鲍类疱疹病毒(Abalone herpesvirus,Ab HV)悬液对杂色鲍(Haliotis diversicolor)注射刺激,研究其血淋巴中可溶性总蛋白浓度,免疫相关酶超氧化物歧化酶(Superoxide Dismutase,SOD)、酸性磷酸酶(Acid Phosphatase,ACP)、碱性磷酸酶(Alkaline Phosphatase,AKP)活性的变化,杂色鲍外套膜、鳃、肝胰腺、腹足中血蓝蛋白(Hemocyanin,Hc)基因Hc1、Hc2相对表达量的变化以及感染后杂色鲍无细胞血浆的抑菌性。结果显示:(1)杂色鲍血淋巴中可溶性总蛋白浓度变化显著,呈现出先升高后降低的趋势。(2)无细胞血浆中的SOD、ACP、AKP活性与对照组相比变化显著,哈维弧菌注射组的SOD活性在感染后第6小时显著升高;Ab HV注射组SOD活性与对照组比较在注射后的各个时相均呈下降趋势,从注射后6 h各个时相的SOD活性与对照组相比均差异显著(P0.05)。哈维弧菌注射组ACP活性在注射后的第48小时显著升高;Ab HV注射组ACP活性在注射后的第12小时显著升高。哈维弧菌注射组AKP活性在注射后48 h显著升高;AbHV注射组AKP活性在注射后第12小时显著升高。(3)杂色鲍外套膜、鳃、肝胰腺以及腹足中血蓝蛋白两个基因Hc1、Hc2的相对表达量均在注射后的不同时间点出现升高。(4)杂色鲍无细胞血淋巴对哈维弧菌、创伤弧菌和溶珊瑚弧菌这三种贝类病原均表现出了不同程度的抑菌性增强的作用。这些结果表明细菌和病毒的刺激引起了杂色鲍免疫相关因子的变化,使得血淋巴的抑菌性增强。本研究为今后进一步研究杂色鲍的非特异性免疫以及在生产实践中应用免疫技术防治病害提供参考。

The effect of Vibrio harveyi and Abalone herpesvirus on immune factors of Haliotis diversicolor

The relative expression levels of hemocyanin mRNA in different tissues, the total protein concentration, the activities of superoxide dismutase, acid phosphatase, alkaline phosphatase and the antibacterial ability of the hemolymph without cells of abalones Haliotis diversicolor were studied after the animals were injected by Vibrio harveyi and abalone herpesvirus. The results showed that after injection with V. harveyi and abalone herpesvirus, the total soluble protein concentration in the hemolymph varied significantly, showing an increasing trend first and then descending gradually. The activities of Superoxide Dismutase (SOD), Acid Phosphatase (ACP) and Alkaline Phosphatase (AKP) in hemolymph of abalones H. diversicolor changed significantly compared with the control group during the experiment period. The SOD activity in hemolymph of abalones H. diversicolor injected with V. harveyi reached culmination after 6 h; while the SOD activity in hemolymph of abalones H. diversicolor injected with Abalone herpesvirus (AbHV) descended gradually, and the statistical results showed significant difference (P<0.05) between experimental and control groups. The activities of ACP from abalones injected with V. harveyi ascended gradually, reaching culmination after 48 h, while the activities of ACP from abalones injected with AbHV increased significantly compared with control group and reached culmination at 12 h. The AKP activities in the hemolymph of abalones injected with V. harveyi ascended and reached culmination at 48 h, while the AKP activities ascended and reached culmination at 12 h after injection with AbHV. The relative expression levels of hemocyanin gene in different tissues ascended at different time injected with V. harveyi and abalone herpesvirus. And the antibacterial ability of the hemolymph of abalones H. diversicolor against V. harveyi, V. vulnificus and V. coralliiluyitcus enhanced significantly after injected with V. harveyi and abalone herpesvirus.