一株维氏气单胞菌发酵培养基优化及其制备疫苗免疫效力比较  

Optimization of Fermentation Medium and Comparison of Immune Efficacy on Its Vaccine Preparation of a Aeromonas veronii Strain

作　　者：孙承文[1] 江小燕[1] 赖迎迢[1,2] 赵长臣 刘春花[1] 任燕[1,2] 巩华[1,2] 陈总会 陶家发[1,2] SUN Cheng-wen;JIANG Xiao-yan;LAI Ying-tiao;ZHAO Chang-chen;LIU Chun-hua;REN Yan;GONG Hua;CHEN Zong-hui;TAO Jia-fa(River Fishery Res.Inst.,Chinese Acad.of Fishery Sci.,Guangzhou 510380;Key Lab.of Fishery Drug Devel't in Minist.of Agric.,Key Lab.of Aquatic Animal Immune Technol.of Guangdong Prov.,Guangzhou 510380)

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：《微生物学杂志》2022年第4期48-54,共7页Journal of Microbiology

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

摘　　要：采用单因素试验、响应面试验法对维氏气单胞菌(Aeromonas veronii)发酵培养基的氮源、碳源、无机盐和磷酸盐成分及用量进行优化组合,确定优化培养基组成:胰蛋白胨10.8 g/L,葡萄糖5.0 g/L,牛肉膏3.0 g/L,磷酸二氢钾2.0 g/L,硫酸镁0.4 g/L,NaCl 5.0 g/L。并与基础培养基的发酵活菌数、制备的灭活疫苗免疫效力进行比较,经过验证试验绘制维氏气单胞菌在优化培养基条件下的7 L发酵罐生长曲线。在优化发酵培养基条件下,维氏气单胞菌活菌数为5.94×10^(9) cfu/mL,比基础培养基增幅43.13%;制备的灭活疫苗相对保护率为77.78%,比基础培养基提高了14.81%。7 L发酵罐发酵培养10 h,活菌数达到最大8.85×10^(9) cfu/mL。通过对发酵培养基的优化,可以获得低成本、优质高效的维氏气单胞菌发酵菌液,为今后维氏气单胞菌灭活疫苗规模化发酵培养提供参考。In order to cultivate high quality and efficient bacterial liquid of Aeromonas veronii,the ingredients and dosages of nitrogen source,carbon source,inorganic salt and phosphate were optimized and combined culture medium for A.veronii inactivated vaccine adopting single-factor test,and response surface methodology experiment to determine the composition of the optimized medium and compare with the number of viable fermenting bacteria in the basic medium and immune efficacy of the inactivated vaccine,with a verification test of growth curve of the bacterium in 7 L fermentor tank.The most suitable fermentation cultivation formula for A.veronii was obtained as followed:tryptone 10.8 g/L,glucose 5.0 g/L,beef extract 3.0 g/L,KHPO2.0 g/L,MgSO0.4 g/L,and NaCl 5.0 g/L.Under the optimized fermentation medium,the number of viable Aeromonas veronii was 5.94×10^(9) cfu/mL,increased by 43.13% as compared with the basic medium.The relative protection rate of the inactivated vaccine prepared with the optimized medium was 77.78%,14.81% higher than that of the basic medium.The maximal number of viable A.veronii cultured in 7 L fermentor tank for 10 h was 8.85×10^(9) cfu/mL.A high yield of A.veronii fermentation broth was obtained through optimized fermentation medium,with low cost,high quality,and efficient.This study provided references for large-scale fermentation and cultivation of A.veronii inactivated vaccine in the future.

关 键 词：维氏气单胞菌 灭活疫苗 鲫鱼 发酵培养基 响应面分析 免疫效力 

分 类 号：Q939.93[生物学—微生物学]