副猪嗜血杆菌4型H25株、5型H45株及12型H31株的发酵工艺研究  

Study on the fermentation process of Haemophilus parasuis type 4 H25,type 5 H45,and type12 H31 strains

作　　者：张小兰 Zhang Xiaolan(Jofunhwa Biotechnology(Fuzhou)Co.Ltd,Fuzhou350014)

机构地区：[1]兆丰华生物科技(福州)有限公司,福州350014

出　　处：《福建畜牧兽医》2024年第4期49-54,共6页Fujian Journal of Animal Husbandry and Veterinary medicine

摘　　要：本研究旨在确定副猪嗜血杆菌4型H25株、5型H45株、12型H31株的最佳发酵培养条件。试验分别在摇瓶和发酵罐条件下进行液体发酵培养初探。摇瓶培养条件下,通过确定生长曲线、接种量、培养温度、血清和NAD含量等参数,为后续发酵罐控制参数的设定奠定基础。在50 L发酵罐上,结合控制参数如温度、pH、溶解氧、搅拌速度、压力及补料策略等进行发酵试验,通过活菌计数和菌体密度(OD_(600nm))的分析,确定其最佳发酵条件。进一步逐级放大至100 L、1000 L发酵罐验证稳定性。结果显示,在摇瓶培养条件下,通过生长曲线的绘制初步确定副猪嗜血杆菌4型H25株、5型H45株和12型H31株的最佳收获时间为菌体密度(OD_(600nm))到达稳定期后1 h,最佳接种量为1%~3%,最佳培养温度为36~38℃,最佳血清浓度为8%~10%,最佳NAD浓度为8~16μg/mL。在发酵罐培养条件下,副猪嗜血杆菌4型H25株、5型H45株、12型H31株的最佳发酵条件为:温度36~38℃,pH7.2~7.4,DO10%~30%,搅拌转速100~300 r/min,罐压0.02 Mpa,补料策略为流加20%葡萄糖溶液,葡萄糖浓度维持在0.8~1.2 g/L。三种菌株均可达到10^(10)CFU/mL以上的活菌数,满足规程及工业化生产要求。The aim of this study was to determine the optimal fermentation culture conditions for Haemophilus parasuis type4 H25 strain,type5 H45 strain,type12 H31 strain.Preliminary study on liquid fermentation culture was performed under shaking bottle and fermentation tank conditions.Under shake flask cultivation conditions,the growth curve,inoculation amount,cultivation temperature,serum and NAD content,were determined to set the foundation for the subsequent control parameters of fermentation tank.On a 50 L fermentation tank,fermentation experiments were conducted by combining control parameters such as temperature,pH,dissolved oxygen,stirring speed,pressure,and feeding strategy.The optimal fermentation conditions were determined through analysis of live bacterial count and bacterial density(OD_(600nm)).Further scale up to 100 L and 1000 L fermentation tanks to verify stability.The optimal shake flask culture conditions were preliminarily determined by drawing growth curves.The results showed that the optimal harvest time of type4 H25 strain,type5 H45 strain,and type12 H31 strain were one hour after the bacterial density(OD_(600nm))reached stationary phase.The optimal inoculation amount was 1%~3%,the optimal incubation temperature was 36~38°C,the optimal serum concentration was 8%~10%,and the optimal NAD concentration was 8~16μg/mL.Under the fermenter culture conditions,the optimal fermentation conditions for Haemophilus parasuis type4 H25 strain,type5 H45 strain,type12 H31 strain were:temperature 36~38℃,pH7.2-7.4,DO10%~20%,stirring speed 100~300 r/min,tank pressure 0.02 Mpa,feeding strategy was to add 20%glucose solution,and maintain glucose concentration at 0.8~1.2 g/L.All three strains can reach a viable bacterial count of over 10^(10)CFU/mL,meeting the requirements of regulations and industrial production.

关 键 词：副猪嗜血杆菌 发酵工艺 

分 类 号：S859.797[农业科学—临床兽医学] TQ920.6[农业科学—兽医学]