大菱鲆鳗弧菌灭活疫苗生产工艺条件的优化

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英文篇名：Optimization of production procedure for inactivated turbot Vibrio anguillarum vaccine 作者：孙承文 ; 李杰 ; 赖迎迢 ; 江小燕 ; 莫照兰 ; 黄志斌 ; 陶家发 英文作者：SUN Cheng-wen;LI Jie;LAI Ying-tiao;JIANG Xiao-yan;MO Zhao-lan;HUANG Zhi-bin;TAO Jia-fa;River Fisheries Research Institute,Chinese Academy of Fisheries Sciences; 关键词：大菱鲆 ; 鳗弧菌 ; 灭活疫苗 ; 安全性 ; 效力 英文关键词：Turbot;;Vibrio anguillarum;;Inactivated vaccine;;Safety;;Potency 中文刊名：SWZP 英文刊名：Chinese Journal of Biologicals 机构：中国水产科学研究院珠江水产研究所;中国水产科学研究院黄海水产研究所; 出版日期：2019-03-19 出版单位：中国生物制品学杂志 年：2019 期：v.32 基金：中国水产科学研究院中央级公益性科研院所基本科研业务费专项资金(2016HY-ZD0502);; 广东省科技计划项目(2016B02023-4003,2016B090920089) 语种：中文; 页：SWZP201903015 页数：5 CN：03 ISSN：22-1197/Q 分类号：85-89

摘要

目的优化大菱鲆鳗弧菌VAM003株灭活疫苗的生产工艺条件。方法通过对大菱鲆鳗弧菌VAM003株培养基(TSB和LB)、培养基盐度(1. 5%、2. 0%、2. 5%、3. 0%、3. 5%NaCl)、接种量(5%、10%、15%)、发酵时间(6～12 h)进行筛选,采用最适发酵条件制备大菱鲆鳗弧菌VAM003株发酵菌液,分别加入不同终浓度的甲醛(0. 1%、0. 2%、0. 3%、0. 4%)灭活24、48、72 h,确定最适灭活条件。采用最适生产工艺制备2批大菱鲆鳗弧菌VAM003株灭活疫苗,并进行安全性及效力试验。结果大菱鲆鳗弧菌VAM003株最适发酵培养条件为:用含2. 5%NaCl的TSB培养基,在接种量10%条件下发酵培养10～12 h,活菌量达5×109 CFU/m L以上;最适灭活条件为:甲醛终浓度0. 2%灭活24 h。大菱鲆注射灭活疫苗后,全部健活,未出现临床症状,摄食、游姿及体色均正常,注射部位无红肿,内脏器官无病变;疫苗的免疫保护率达90%以上。结论成功优化了大菱鲆鳗弧菌VAM003株灭活疫苗规模化生产工艺条件,制备的灭活疫苗具有良好的安全性及效力,可用于大菱鲆鳗弧菌病的预防。
        Objective To optimize the production procedure for inactivated vaccine against turbot Vibrio anguillarum VAM003 strain.Methods The medium,(TSB and LB),sodium chloride concentration(1.5%,2.0%,2.5%,3.0%and 3.5%),inoculum size(5%,10%and 15%)and fermentation time(6～12 h)for fermentation of V.anguillarum VAM003 strain of turbot were optimized.Fermentation liquid was prepared under the optimized condition,and inactivated with formaldehyde at various final concentrations(0.1%,0.2%,0.3%and 0.4%)for 24,48 and 72 h to optimize the condition for inactivation.Two batches of inactivated vaccines were prepared with VAM003 strain by the optimized procedure and tested for safety and potency.Results The optimal fermentation condition for VAM003 strain of V.anguillarum was as follows:the strain was inoculated at an inoculum size of 10%into TSB medium containing 2.5%sodium chloride and cultured for 10～12 h until the viable cell count reached more than 5×109CFU/mL.However,the optimal final formaldehyde concentration for inactivation of V.anguillarum VAM003 strain was 0.2%,while the optimal inactivation time was 24 h.All the turbots injected with the inactivated vaccine survived without clinical symptoms,of which the food intake,swimming posture and body color were normal,and no redness and swelling in injection site or pathological change in organs were observed.The protective rate of vaccine was more than 90%.Conclusion The production procedure for inactivated vaccine against V.anguillarum VAM003 strain was successfully optimized,and the prepared inactivated vaccine showed high safety and potency and might be used for the prevention of Vibrio disease in turbots.

引文

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