仔猪副伤寒耐热保护剂活疫苗C500株生产工艺的优化

Optimization of Production Process of Piglet Paratyphus Thermo-stable Live Vaccine Strain C500

为优化仔猪副伤寒耐热保护剂活疫苗生产工艺,对细菌生长曲线、耐热保护剂和冻干曲线进行了研究,建立了发酵体积1%、2%和3%的二级种子接种量下分别发酵12 h^20 h的细菌生长曲线,检测3个配苗比例、4种耐热保护剂和3条冻干曲线下成品疫苗的冻干菌存率、活菌存活率和安全性,确定了接种3%二级种子液发酵培养17 h为最佳发酵培养条件,发酵菌数达到428×108CFU/m L;最佳配苗比例为菌液︰耐热保护剂1∶1(V∶V),优化保护剂B作为最佳耐热保护剂,设定冻干程序3#作为最佳冻干曲线。基于此,仔猪副伤寒耐热保护剂活疫苗抗原含量达到40头份/瓶,冻干菌存率和活菌存活率均达到95%以上,大幅度降低了猪体免疫副反应,极大提升了疫苗的产品品质。

In order to optimize the production process of piglet paratyphus thermo - stable live vaccine, the bacterial growth curve, heat - resistant protective agent and freeze - drying curve were studied. The bacterial growth curves were established by fermentation between 12 h to 20 h after inoculation of 1%, 2% and 3% secondary seed inoculation respectively. The vaccines＇ freeze - dried bacteria survival rate, survival rate of live bacteria and safety were detected on the condition of three rations of bacteria solution and heat - resistant protective agent, four heat - resistant protective agents and three freeze - drying curves. It was confirmed that 3% secondary seed liquid fermentation for 17 h was the best fermentation condition, the number of fermentation bacteria reached 428 × 10-8 CFU/mL. The best proportion of seedlings was antigen ： protective agent 1 ： 1 （ V： V）. Freeze - drying program 3# was set as the best freeze - drying curve. Based on our research results, the antigen contents of paratyphus thermo - stable live vaccine reach 40 doses/vial, and the freeze - dried bacteria survival rate and survival rate of the vaccine were both more than reduced. The research has provided technical basement for control of paratyphoid. 95%. The vaccine side reactions to pig were greatly providing high quality vaccine for the prevention and