嗜水气单胞菌灭活疫苗免疫施氏鲟后的免疫应答反应与保护效果  

Immune responses and protective efficacy in Amur sturgeon(Acipenser schrenckii) after immunization with formalin-killed Aeromonas hydrophila

作　　者：周勇[1] 史玉恒[1] 赵建青 代建丽[2] 范玉顶[1] 刘文枝[1] 江南[1] 曾令兵[1] 

机构地区：[1]中国水产科学研究院长江水产研究所,湖北武汉430223 [2]武昌理工学院生命科学学院,湖北武汉430223

出　　处：《中国水产科学》2018年第1期195-203,共9页Journal of Fishery Sciences of China

基　　金：国家973计划项目(2015CB150703)

摘　　要：为了研究嗜水气单胞菌灭活疫苗免疫施氏鲟(Acipenser schrenckii Brandt,1869)后的免疫应答反应与保护效果,本研究采用福尔马林灭活法制备嗜水气单胞菌灭活疫苗。通过腹腔注射途径免疫健康施氏鲟,对施氏鲟外周血的红细胞和白细胞数量、白细胞组成、吞噬活性、吞噬指数、血清酸性磷酸酶活性、血清溶菌酶活性、血清中和抗体效价等免疫指标及疫苗保护效果进行测定。结果显示,免疫组红、白细胞数量迅速上升,于免疫后第4天达峰值,分别为(8.50±0.17)×10~8个/mL和(8.96±0.44)×10~6个/mL;单核细胞和嗜中性粒细胞分类百分比于第7天达峰值,分别为10.50%和15.53%,极显著高于对照组(P<0.01),淋巴细胞分类百分比在第21天才达峰值73.51%;吞噬指数和吞噬百分比均于第4天达到最高值,分别为4.53和30%;血清中溶菌酶活性和酸性磷酸酶活性分别在免疫后第7天和第14天达到峰值,极显著高于对照组(P<0.01);血清抗体效价于免疫后第21天达到峰值1:203,极显著高于对照组(P<0.01)。攻毒感染实验结果表明,免疫组的相对免疫保护率为76.84%。由此可见,嗜水气单胞菌灭活疫苗免疫施氏鲟后,能够显著诱导施氏鲟体内的免疫应答反应,增强鱼体的免疫保护能力。本研究为养殖鲟因嗜水气单胞菌感染引起疾病的免疫预防技术研究奠定了前期基础。In order to study the immune responses and protective efficacy in Amur sturgeon （Acipenser schrenckii Brandt, 1869）, we used formalin-killed Aeromonas hydrophila to immunize healthy Amur sturgeon by peritoneal injection. We investigated the following immune parameters： peripheral blood cell count, differential leucocyte count, phagocytic activity and index, acid phosphatase activity, lysozyme activity, neutralizing antibody, and relative percentage survival. The numbers of erythrocytes and leucocytes in the peripheral blood of immunized fish increased rapidly and reached maximum values of （8.50 ± 0.17） × 108/mL and （8.96 ±0.44） ×106/mL, respectively, at 4 d post-immunization. The percentages of monocytes and neutrophils reached maximum values of 10.50% and 15.53%, respectively, at 4 d post-immunization, and were very significantly higher than those of fish in the control groups （P 〈 0.01）. However, lymphocyte percentage reached a maximum value of 73.51% at 21 d post-immunization. The phagocytic index and percentage of neutrophils were 4.53 and 30%, respectively, at 4 d post-immu- nization. The activities of lysozyme and acid phosphatase in serum reached peaks at 7 d and 14 d post-immunization, respectively, Neutralizing antibodies in the immunized fish reached 1 ： 203 at 21 d post-immunization. When the immunized fish were challenged with live A. hydrophila, the relative percentage survival of the immunize.d group was 76.84%. These results showed that the formalin-killed A. hydrophila was able to elicit immune protection in Amur sturgeon, which lays the foundation for the further research on immunoprophylaxis against the diseases caused by A. hydrophila infection in sturgeons.

关 键 词：嗜水气单胞菌 施氏鲟 灭活疫苗 免疫指标 相对免疫保护率 

分 类 号：S942[农业科学—水产养殖]