杂交鳢舒伯特气单胞菌弱毒疫苗候选株筛选及其免疫效果评价  

Screening and immune efficacy evaluation of attenuated vaccine candidate strains of Aeromanas schubertii from hybrid snakehead

作　　者：胡蕾 刘礼辉[1] 杨圆圆[3] 方伟[3] 林强[1] 付小哲[1] 梁红茹[1] 刘山桂 李宁求[1] HU Lei;LIU Li-hui;YANG Yuan-yuan;FANG Wei;LIN Qiang;FU Xiao-zhe;LIANG Hong-ru;LIU Shan-gui;LI Ning-qiu(Key Lab of Fishery Drug Development,Ministry of Agriculture and Rural Affairs,Key Lab of Aquatic Animal Immune Technology,Guangdong Province,Pearl River Fisheries Research Institute,Chinese Academy of Fishery Sciences,Guangzhou 510380,China;College of Fisheries and Life,Shanghai Ocean University,Shanghai 201306,China;Guangdong Provincial Aquatic Animal Epidemic Disease Prevention and Control Center,Guangzhou 510220,China)

机构地区：[1]中国水产科学研究院珠江水产研究所,农业农村部渔用药物创制重点实验室,广东省水产动物免疫技术重点实验室,广州510380 [2]上海海洋大学水产与生命学院,上海201306 [3]广东省水生动物疫病预防控制中心,广州510220

出　　处：《淡水渔业》2019年第5期74-79,共6页Freshwater Fisheries

基　　金：中国水产科学研究院基本科研业务费专项(2017HY-ZC0405);2017年度广东省渔港建设和渔业发展专项资金(鱼病防治)项目

摘　　要：为获得一种安全、高效的舒伯特气单胞菌(Aeromanas schubertii)弱毒疫苗候选株,通过鱼体攻毒实验筛选天然弱毒株,用筛选的弱毒株制备弱毒疫苗,对杂交鳢进行免疫后,采用荧光定量RT-PCR方法评价免疫相关基因表达情况,同时采用强毒株攻毒免疫的杂交鳢测定相对免疫保护率(relative percentage survival,RPS),综合评价弱毒株作为活疫苗侯选株的潜力。结果显示:筛选到一株舒伯特气单胞菌天然弱毒株OM170701,其对靶动物杂交鳢与非靶动物大口黑鲈均无明显致病性;该弱毒疫苗候选株能诱导杂交鳢肝脏中IL-8、IL-1β、补体C3表达量上调,免疫后72 h相对表达量达到最高,分别是对照组的相对表达量的4.95倍、5.57倍、3.94倍;免疫组IgM的相对表达量显著高于对照组,其中第28天时免疫组IgM相对表达量是对照组的17.26倍;攻毒后免疫组成活率显著高于对照组,其RPS为65%。结果表明,该弱毒株通过注射免疫能引发杂交鳢非特异性和特异性免疫应答,并提供良好的免疫保护。Natural attenuated strains were screened through fish challenge test to obtain a safe and efficient attenuated vaccine candidate of Aeromanas schubertii.The expression of immune-related in the post-immunized hybrid snakehead was evaluated by fluorescence quantitative RT-PCR.The relative percentage survival(RPS)of immunized hybrid snakehead after challenged with virulent strain was used to evaluate the potential of attenuated strains as live vaccine candidate.The results showed that a natural attenuated strain of A.schubertii OM170701 was successfully screened,no pathogenic to the target animal hybrid snakehead and non-target animals largemouth bass.The candidate strain of attenuated vaccine could induce expression level of IL-8、IL-1β、complement C3 genes up-regulated in the liver of hybrid snakehead,and the relative expression was the highest at 72 h after immunization,which was 4.95 times,5.57 times,3.94 times of the control group,respectively.The relative expression of IgM in the immunized group was significantly higher than that in the control group.The relative expression level of IgM in the immunized group was 17.26 times to control group on the 28th day.The immune group survival rate was significantly higher than that of the control group after the virulent challenge,and the RPS was 65%.These results indicated that the attenuated strain could elicit non-specific and specific immune responses of hybrid snakehead by injection immunization,and provide good immune protection in hybrid snakehead.

关 键 词：杂交鳢 舒伯特气单胞菌(Aeromanas schubertii) 弱毒疫苗 免疫基因 相对免疫保护率 

分 类 号：S948[农业科学—水产养殖]