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刘明珠,余庆,肖贺贺,吴思婷,覃仙玲,黎思巧,陈宪云,陆兰天,聂振平,罗永巨,肖俊,李鹏飞.广西斑点叉尾鮰源嗜水气单胞菌灭活疫苗的制备[J].广西科学院学报,2019,35(3):239-245. [点击复制]
- LIU Mingzhu,YU Qing,XIAO Hehe,WU Siting,QIN Xianling,LI Siqiao,CHEN Xianyun,LU Lantian,NIE Zhenping,LUO Yongju,XIAO Jun,LI Pengfei.Development and Characterization of Aeromonas hydrophila Inactivated Vaccine for Ictalurus punctatus in Guangxi[J].Journal of Guangxi Academy of Sciences,2019,35(3):239-245. [点击复制]
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| 广西斑点叉尾鮰源嗜水气单胞菌灭活疫苗的制备 |
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刘明珠1, 余庆2, 肖贺贺3, 吴思婷1, 覃仙玲2, 黎思巧2, 陈宪云2, 陆兰天2, 聂振平1, 罗永巨4, 肖俊4, 李鹏飞1,5
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| (1.广西科学院, 广西壮族自治区海洋研究所, 广西海洋生物技术重点实验室, 广西北海 536000;2.广西科学院, 广西近海海洋环境科学重点实验室, 广西南宁 530007;3.河南师范大学生命科学学院, 河南新乡 453007;4.广西壮族自治区水产科学研究院, 广西水产遗传育种与健康养殖重点实验室, 广西南宁 530021;5.广西海洋天然产物与组合生物合成化学重点实验室, 广西南宁 530007) |
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| 摘要: |
| 本研究针对广西地区分离到的致病性嗜水气单胞菌，开发区域定制化的灭活疫苗，以实现鱼病的有效预防和控制。首先，对斑点叉尾鮰源嗜水气单胞菌在LB培养基中的生长曲线，培养基的最适pH值，以及培养基中添加蔗糖对嗜水气单胞菌生长影响进行研究；随后探索嗜水气单胞菌灭活疫苗的制备方法，并在细胞水平和斑点叉尾鮰活体水平上研究疫苗的安全性，开展疫苗对斑点叉尾鮰的免疫保护实验。结果表明嗜水气单胞菌浓度与菌液OD值间具有较好的线性关系，其回归方程为y=2E-10x+0.1399，R2=0.975 4。用于培养嗜水气单胞菌的LB培养基最适pH值为7.0~7.5，且在培养基中添加蔗糖能显著促进嗜水气单胞菌的生长。研究制备的嗜水气单胞菌灭活疫苗在细胞水平和鱼活体水平均安全无毒，且对斑点叉尾鮰的免疫保护率可达到75%。据此，本研究初步建立起广西斑点叉尾鮰源嗜水气单胞菌灭活疫苗的制备工艺。 |
| 关键词: 斑点叉尾鮰 嗜水气单胞菌 灭活疫苗 制备条件优化 免疫保护 水产养殖 |
| DOI：10.13657/j.cnki.gxkxyxb.20190903.001 |
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| 基金项目:广西重点研发计划项目（2018AB52003），广西自然科学基金项目（2017GXNSFBA198176），广西科学院基本科研业务费项目（2018YJJ903，2017YJJ23002），现代农业产业技术体系专项资金项目（CARS-46）和广西南海珊瑚礁研究重点实验室开放课题（GXLSCRSCS2018003）资助。 |
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| Development and Characterization of Aeromonas hydrophila Inactivated Vaccine for Ictalurus punctatus in Guangxi |
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LIU Mingzhu1, YU Qing2, XIAO Hehe3, WU Siting1, QIN Xianling2, LI Siqiao2, CHEN Xianyun2, LU Lantian2, NIE Zhenping1, LUO Yongju4, XIAO Jun4, LI Pengfei1,5
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| (1.Guangxi Key Laboratory for Marine Biotechnology, Guangxi Institute of Oceanography, Guangxi Academy of Sciences, Beihai, Guangxi, 536000, China;2.Guangxi Key Laboratory of Marine Environmental Science, Guangxi Academy of Sciences, Nanning, Guangxi, 530007, China;3.College of Life Science, Henan Normal University, Xinxiang, Henan, 453007, China;4.Guangxi Key Laboratory of Aquatic Genetic Breeding and Healthy Breeding, Guangxi Academy of Fishery Science, Nanning, Guangxi, 530021, China;5.Guangxi Key Laboratory of Marine Natural Products and Combinatorial Biosynthesis Chemistry, Nanning, Guangxi, 530007, China) |
| Abstract: |
| In view of the pathogenic Aeromonas hydrophila isolated from Guangxi,we tried to develop regional customized inactivated vaccine to achieve effective prevention and control of fish diseases.First,the growth curve of Aeromonas hydrophila in LB medium,the optimum pH value of the medium,and the effect of adding sucrose on the growth of Aeromonas hydrophila in the medium were studied.Subsequently,the preparation method of Aeromonas hydrophila inactivated vaccine was explored,and the safety of the vaccine was studied at the cellular level and the living level of Ictalurus punctatus to carry out the immune protection experiment of the vaccine against Ictalurus nctatus.The results showed that there was a good linear relationship between the concentration of Aeromonas hydrophila and the OD value of bacterial liquid,whose regression equation was y=2E-10x+0.1399,R2=0.975 4.The optimum pH of LB medium for Aeromonas hydrophila was 7.0-7.5,and the addition of sucrose to the medium could significantly promote the growth of Aeromonas hydrophila.The inactivated vaccine against Aeromonas hydrophila prepared at the study was safe and non-toxic at both the cellular level and the living fish level,and the immunization protection rate against Ictalurus punctatus could reach 75%.Based on this,the preparation method of inactivated vaccine against Aeromonas hydrophila from Ictalurus punctatus in Guangxi was preliminarily established. |
| Key words: Ictalurus punctatus Aeromonas hydrophila inactivated vaccine optimization of preparation conditions immune protection aquaculture |
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