Optimization of fermentation process by response surface methodology and comparison of immune efficacy of inactivated vaccine of Aeromonas veronii
SUN Chengwen, LAI Yingtiao, GONG Hua, REN Yan, JIANG Xiaoyan, CHEN Zonghui, HUANG Zhibin, TAO Jiafa
1.Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510380, China; 2.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
Abstract: In order to optimize the fermentation process of the inactivated vaccine of Aeromonasveronii, 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(11.13×109 CFU/mL)of viable Aeromonasveronii 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 Carassiusauratus.The relative immune protection rate was found to be more than 40% in the crucian carp immuned challenged with Aeromonasveronii LY02 strain.The findings indicated that the optimization of fermentation process of inactivated vaccine of Aeromonasveronii by response surface method significantly increased the production of bacterial fluid and improved the immune efficacy of the vaccine.
孙承文, 赖迎迢, 巩华, 任燕, 江小燕, 陈总会, 黄志斌, 陶家发. 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[J]. 大连海洋大学学报, 2021, 36(4): 546-553.
SUN Chengwen, LAI Yingtiao, GONG Hua, REN Yan, JIANG Xiaoyan, CHEN Zonghui, HUANG Zhibin, TAO Jiafa. Optimization of fermentation process by response surface methodology and comparison of immune efficacy of inactivated vaccine of Aeromonas veronii. Journal of Dalian Ocean University, 2021, 36(4): 546-553.