四川农业大学学报 ›› 2020, Vol. 38 ›› Issue (03): 353-358.doi: 10.16036/j.issn.1000-2650.2020.03.015

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龟源迟缓爱德华氏菌分离鉴定及灭活疫苗研制

孙敬1,2, 柯浩1,3,4, 于交平1,2, 冯国清1,3,4, 梁志凌1,3,4, 刘振兴1,3,4, 郝乐1,3,4, 马艳平1,3,4, 马江耀1,3,4   

  1. 1.广东省农业科学院动物卫生研究所水产病害研究室,广州 510000;
    2.上海海洋大学水产与生命学院,上海 201306;
    3.农业农村部兽用药物与诊断技术广东科学观测实验站,广州 510000;
    4.广东省畜禽疫病防治研究重点实验室,广州 510000
  • 收稿日期:2020-02-23 出版日期:2020-06-28 发布日期:2021-02-03
  • 作者简介:孙敬,硕士研究生。*责任作者:柯浩,研究员,硕士,主要从事水产病害及相关防治研究,E-mail:keha@tom.com。
  • 基金资助:
    广东特色水产动物绿色药物研制与开发专项补助资金(粤财农〔2018〕277号); 广东省省级现代农业产业技术体系“水产疫病监测与综合防控共性关键技术研发创新团队”(2019KJ141)

Isolation and Identification of Turtle-Sourced Edwardsella laguna and Development of Inactivated Vaccine

SUN Jing1,2, KE Hao1,3,4, YU Jiaoping1,2, FENG Guoqing1,3,4, LIANG Zhiling1,3,4, LIU Zhenxing1,3,4, HAO Le1,3,4, MA Yanping1,3,4, MA Jiangyao1,3,4   

  1. 1. Laboratory of Aquatic Diseases,Institute of Animal Health,Guangdong Academy of Agricultural Sciences,Guangzhou 510000,China;
    2. College of Fisheries and Life Science,Shanghai Ocean University,Shanghai 201306,China;
    3. Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong Province,Ministry of Agriculture and Rural Affairs,Guangzhou 510000,China;
    4. Key Laboratory of Livestock Disease Prevention of Guangdong Province,Guangzhou 510000,China
  • Received:2020-02-23 Online:2020-06-28 Published:2021-02-03

摘要: 【目的】利用从发病死亡的中华草龟(Chinemys reevesii)分离到的病原菌株制备灭活疫苗。【方法】对该菌进行培养特性、生理生化特征、16S rRNA同源性分析、药敏等试验。人工感染中华花龟(Ocadia sinensis),确定半数致死剂量(LD50)。经0.03%甲醛灭活后与铝胶3∶1制成灭活疫苗,对中华花龟进行腹腔注射免疫,测定血清抗体效价,以5LD50活菌浓度进行攻毒试验测定免疫保护率。【结果】分离鉴定该菌为迟缓爱德华氏菌。两次免疫14 d后抗体效价分别为1∶64和1∶256,在5LD50剂量攻毒后相对免疫保护率分别为60%和73%。【结论】本灭活疫苗二免的相对保护率达73%,有保护效果,可用于龟的迟缓爱德华氏菌病的预防。

关键词: 中华草龟, 迟缓爱德华氏菌, 灭活疫苗, 抗体效价, 免疫保护

Abstract: 【Objective】 Inactivated vaccines were prepared using pathogenic strains isolated from Chinemys reevesii. 【Method】 Experiments were carried out on the culture characteristics, physiological and biochemical characteristics, 16S rRNA homology analysis and drug sensitivity of the bacteria. Artificially infected with Chinese Striped-neck Turtle (Ocadia sinensis) to determine the half lethal dose (LD50). After being inactivated with 0.03% formaldehyde, an inactivated vaccine was made with aluminum glue 3:1. The Chinese Striped-neck Turtle was immunized intraperitoneally to determine the serum antibody titer. The challenge test was conducted with 5LD50 live bacteria concentration to determine the immune protection rate. 【Result】 Isolated and identified the bacterium as Edwardsiella tarda. After 14 days of two immunizations, the antibody titers were 1∶64 and 1∶256. and the relative immune protection rates after 5LD50 dose challenge were 60% and 73%. 【Conclusion】 The relative protection rate of the second inactivated vaccine is 73%, which has a protective effect and can be used for the prevention of delayed Edwardsiosis of turtles.

Key words: Chinemys reevesii, Edwardsiella tarda, inactivated vaccine, antibody titer, immune protection

中图分类号: 

  • S943