眼斑拟石首鱼烂尾病病原菌的分离鉴定及灭活疫苗的免疫效果  

Isolation and Identification of Bacterial Pathogens from Sciaenops ocellatus with Tail-rotted Disease and Evaluation of Immune Efficacy of Inactived Vaccine

作　　者：姚欢[1,2,3] 黄郁葱[1,2,3] 蔡双虎[1,2,3] 鲁义善[1,2,3] 蔡小辉[4,5] 简纪常[1,2,3] YAO Huan;HUANG Yu-cong;CAI Shuang-hu;LU Yi-shan;CAI Xiao-hui;JIAN Ji-chang(Fisheries College of Guangdong Ocean University;Provincial Key Laboratory of Pathogenic Biology and Epidemiology for Aquatic Economic Animals of Guangdong;Key Laboratory of Control for Diseases of Aquatic Economic Animals of Guangdong Higher Education Institute,Zhanjiang 524088, China;Guangxi Key Laboratory of Marine Biotechnology;Guangxi Institute of Oceanology, Beihai 536000, China)

机构地区：[1]广东海洋大学水产学院 [2]广东省水产经济动物病原生物学及流行病学重点实验室 [3]广东高等学校水产经济动物病害控制重点实验室,广东湛江524088 [4]广西海洋研究所 [5]广西海洋生物技术重点实验室,广西北海536000

出　　处：《广东海洋大学学报》2016年第4期37-44,共8页Journal of Guangdong Ocean University

基　　金：广东省海洋经济创新发展区域示范专项(GD2012-B01-004);广西海洋生物技术重点实验室开放基金(GLMBT-201404);农业部农业公益项目(201203085);广东省科技计划项目(2012020800006)

摘　　要：从患烂尾病眼斑拟石首鱼(Sciaenops ocellatus)的溃疡处分离到一株优势菌So GZ1401,回归感染试验证实该菌是引起烂尾病的病原菌,对眼斑拟石首鱼的半致死量(LD50)为8.6×103 CFU·g^(-1);结合其形态学和生理生化特征及HSP60测序分析,鉴定为哈维氏弧菌(Vibrio harveyi);利用杯碟法研究其胞外产物性质,结果表明:其胞外产物具有淀粉酶、明胶酶、酪蛋白酶、卵磷脂酶、脂酶活性以及溶血活性,但无脲酶活性。药敏试验表明,该菌株对氟哌酸、恩诺沙星、氟苯尼考和复方新诺明等抗菌药有较高的敏感性。制备哈维氏弧菌灭活疫苗,采用注射和浸泡2种途径免疫眼斑拟石首鱼,测定受免疫鱼的血清抗体效价和疫苗对眼斑拟石首鱼的免疫保护率。结果显示,免疫组血清中的抗体效价水平均高于对照组,差异有统计学意义(P<0.05),注射组和浸泡组的抗体效价最高值分别达718.4和128,哈维氏弧菌攻毒后免疫保护率分别为75.0%~86.7%和46.4%~56.7%。哈维氏弧菌灭活疫苗对眼斑拟石首鱼具有较好的免疫保护性。A dominant bacterial strain designated as SoGZ1401 was isolated from ulcer of the moribundSciaenops ocellatus with rotted-tail disease and was confirmed to be the pathogen by artificial infectiontest. The 50% lethal dose of SoGZ1401 to the S. ocellatus was 8.6×103 CFU g-1. The strain SoGZ1401 was identified as Vibrio harveyi based on morphology, physiological & biochemical tests and HSP60DNA sequence analysis. The extracellular products of V. harveyi strain were studied by cylinde-platemethod. The results indicated that the extracellular products show the activities of amylase, gelatinase,casease, lipase, lecithinase and haemolysin, but without urease activity. Drug sensitivity tests revealedthat V. harveyi strain was sensitive to norfloxacin, Enrofloxacin, Florfenicol and compoundsulfamethoxazole. The vaccine was prepared with formalin-inactivated strain SoGZ1401 to studyantibody titer and immune efficacy on S. ocellatus. The vaccination was administered by intraperitonealinjection and bath immersion route. Antibody titer in the serum of the injected group and immersiongroup were significantly increased compared with the control group and the maximal titers reach 718.4and 128, respectively. The Relative Percentage Survival (RPS) values of the injected group andimmersion group were 75.0% - 86.7% and 46.4% - 56.7%, respectively. It is concluded that S.ocellatus can produced strong immune response immunized with V. harveyi bacterin, and the vaccine provide good protection efficiency against bacterial disease cause V. harveyi.

关 键 词：眼斑拟石首鱼 烂尾病 病原 哈维氏弧菌 免疫效果 

分 类 号：S941.42[农业科学—水产养殖] S942.5[农业科学—水产科学]