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

关键词:  仔猪副伤寒  耐热保护剂  发酵  配苗  冻干曲线

Key words:  paratyphoid  heat-resistant protective agent  fermentation  ratio of vaccine  freeze-drying curve

摘要:

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17h为最佳发酵培养条件，发酵菌数达到428×108 CFU/mL；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。

Abstract:

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 428×108 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 95 %. The vaccine side reactions to pig were greatly reduced. The research has provided technical basement for providing high quality vaccine for the prevention and control of paratyphoid.