维氏气单胞菌CA07株灭活疫苗生产工艺的优化  

Optimization of production process of Aeromonas veronii CA07 strain inactivated vaccine

作　　者：孙承文[1] 巩华[1,2] 赖迎迢 江小燕[1] 任燕 陈总会[1] 黄志斌[1,2] 陶家发 SUN Cheng-wen;GONG Hua;LAI Ying-tiao;JIANG Xiao-yan;REN Yan;CHEN Zong-hui;HUANG Zhi-bin;TAO Jia-fa(River Fisheries Research Institute,Chinese Academy of Fisheries Sciences,Guangzhou 510380,Guangdong Province,China)

机构地区：[1]中国水产科学研究院珠江水产研究所,广东广州510380 [2]农业部渔药创制重点实验室广东省水产动物免疫技术重点实验室,广东广州510380

出　　处：《中国生物制品学杂志》2023年第1期76-80,84,共6页Chinese Journal of Biologicals

基　　金：国家重点研发计划(2019YFD0900103);中国水产科学研究院基本科研业务费专项资金(2020XT0405);广州市科技计划项目(20190-4020004)。

摘　　要：目的优化维氏气单胞菌(Aeromonas veronii,AV)CA07株灭活疫苗的生产工艺。方法通过对AV CA07株菌种培养时间(2~16 h)、发酵培养基(优化发酵培养基、LB培养基、NB培养基)、发酵条件[接种量(1%、5%、10%、15%)、通气量(2、4、6、8 L/min)、发酵时间(6、8、10、12 h)]的优化,确定AV CA07株菌液的发酵工艺。通过对灭活剂甲醛溶液的终浓度(0.10%、0.20%、0.30%、0.40%)、灭活温度(28和37℃)、灭活时间(24、48、72 h)的比较,确定最佳灭活工艺。建立AV CA07株灭活疫苗500 L发酵罐规模化生产工艺,并对制备的疫苗进行安全性及免疫原性试验。结果AV CA07株发酵工艺的最佳接种量为5%,通气量为4 L/min,培养时间为10~12 h;最佳灭活条件为:用终浓度0.30%的甲醛溶液于37℃灭活24 h。500 L发酵罐制备的AV CA07株菌液活菌数达8×10^(9)CFU/mL以上,灭活后经腹部免疫鲫鱼,鲫鱼全部存活;攻毒后,相对免疫保护率达90%以上。结论采用优化生产工艺制备的AV CA07株灭活疫苗具有良好的安全性及免疫原性。Objective To optimize the production process of inactivated vaccine of Aeromonas veronii(AV)CA07 strain.Methods The fermentation culture process of AV CA07 strain liquid was determined through the optimization of the culture time(2~16 h),medium(optimized fermentation medium,LB medium and NB medium)and fermentation conditions(inoculation amount of 1%,5%,10%and 15%;ventilation rate of 2,4,6 and 8 L/min and fermentation time of 6,8,10 and 12 h).The optimal inactivation process was determined through the comparison of the final concentration of formaldehyde solution(0.10%,0.20%,0.30%and 0.40%),inactivation temperature(28 and 37℃)and inactivation time(24,48 and 72 h).The large-scale production process of inactivated vaccine of AV CA07 strain in 500 L fermentor was established and the prepared vaccines were tested for safety and immunogenicity.Results The optimal inoculation amount of AV CA07 strain was 5%,ventilation rate was 4 L/min and culture time was 10~12 h.The optimal inactivation condition was adding formaldehyde solution with final concentration of 0.30%incubating at 37℃for 24 h.The number of viable bacteria in the fermentation broth of AV CA07 strain prepared in 500 L fermentor was more than 8×10^(9)CFU/mL.All crucian carps immunized with the inactivated vaccine by abdomen survived.After challenge,the relative immune protection rate was more than 90%.Conclusion AV CA07 strain inactivated vaccine prepared by optimized production process showed good safety and immunogenicity.

关 键 词：维氏气单胞菌 灭活疫苗 生产工艺 安全性 免疫原性 

分 类 号：S917.1[农业科学—水产科学]