基于响应面法的维氏气单胞菌灭活疫苗菌液发酵工艺优化及免疫效力比较  

Optimization of fermentation process by response surface methodology and comparison of immune efficacy of inactivated vaccine of Aeromonas veronii

作　　者：孙承文[1] 赖迎迢[1,2] 巩华 任燕[1,2] 江小燕 陈总会[1] 黄志斌 陶家发[1,2] SUN Chengwen;LAI Yingtiao;GONG Hua;REN Yan;JIANG Xiaoyan;CHEN Zonghui;HUANG Zhibin;TAO Jiafa(Pearl River Fisheries Research Institute,Chinese Academy of Fishery Sciences,Guangzhou 510380,China;Key Laboratory of Fishery Drug Development,Ministry of Agriculture and Rural Affairs,Key Laboratory of Aquatic Animal Immune Technology of Guangdong Province,Guangzhou 510380,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]农业农村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：《大连海洋大学学报》2021年第4期546-553,共8页Journal of Dalian Ocean University

基　　金：中国水产科学研究院基本科研业务费项目(2020XT0405);广州市科技计划项目(201904020004);国家重点研发计划“蓝色粮仓科技创新”项目(2019YFD0900103)。

摘　　要：为优化维氏气单胞菌Aeromonas veronii灭活疫苗菌液发酵工艺,通过单因素试验确定温度、培养基初始pH、转速、接种量对菌液活菌数的影响,应用响应面法的Box-Behnken进行优化,对不同发酵条件下发酵菌液制备灭活疫苗的安全性及免疫效力进行了比较。结果表明:当最优发酵条件为温度28℃、培养基初始pH 7.5、摇床转速230 r/min、接种量5%时,维氏气单胞菌CA07株发酵获得最大的活菌数为11.13×10^(9) CFU/mL,较单因素试验确定的最高活菌数值提高20.59%,与预测值基本相符;优化发酵条件能提升发酵菌液活菌数,从而显著提高制备的灭活疫苗的相对免疫保护率,即使制备的灭活疫苗稀释3倍,免疫鲫鱼Carassius auratus的相对保护率仍可达到70%以上,以维氏气单胞菌LY02株攻毒,相对免疫保护率达40%以上。研究表明,通过响应面法优化维氏气单胞菌灭活疫苗发酵工艺,在显著增加菌液产量的同时可提高疫苗的免疫效力。In order to optimize the fermentation process of the inactivated vaccine of Aeromonas veronii,influence of temperature(25,28,31,34,and 37℃),initial pH of medium(6.5,7.0,7.5,8.0 and 8.5),rotation speed(160,180,200,220,and 240 r/min),and amount of inoculation(1%,5%,10%,15%,and 20%)on the counts of the viable bacteria were optimized in single factor test by the response surface method Box-Behnken,and safety and immune efficacy of inactivated vaccine prepared by fermentation broth under different fermentation conditions were compared.The maximum count(11.13×10^(9) CFU/mL)of viable Aeromonas veronii CA07 strain was observed under the optimal fermentation conditions of 28℃,initial medium pH of 7.50,rotation speed 230 r/min,and inoculation amount 5%.The maximal viable bacterium value was increased by 20.59%compared with the single factor test,which was basically in line with the predicted value.The optimized fermentation conditions led to significantly increase the number of viable bacteria in the fermentation broth,thus significantly improving the relative immune protection rate of the prepared inactivated vaccine when the inactivated vaccine was diluted by 3 times,with over 70%of the relative protection rate in crucian carp Carassius auratus.The relative immune protection rate was found to be more than 40%in the crucian carp immuned challenged with Aeromonas veronii LY02 strain.The findings indicated that the optimization of fermentation process of inactivated vaccine of Aeromonas veronii by response surface method significantly increased the production of bacterial fluid and improved the immune efficacy of the vaccine.

关 键 词：维氏气单胞菌 发酵工艺条件 优化 响应面分析 免疫效力 

分 类 号：S948[农业科学—水产养殖]