牛溶血性曼氏杆菌灭活疫苗的制备与检定  

Preparation and quality control of inactivated bovine Mannheimia haemolytica vaccine

作　　者：高佳滨[1] 陈为宏[1] 尹辉[1] 乔波[1] 陈楠楠[1] 朱战波[1] 

机构地区：[1]黑龙江八一农垦大学动物科技学院,黑龙江大庆163319

出　　处：《中国生物制品学杂志》2014年第8期990-993,共4页Chinese Journal of Biologicals

基　　金："十二五"农村领域国家科技计划课题(2012BAD12B03-3)

摘　　要：目的制备牛溶血性曼氏杆菌灭活疫苗,并进行检定。方法从黑龙江某牛场发病牛病变的肺组织中分离溶血性曼氏杆菌,经各项鉴定合格后,制备种子液。确定甲醛灭活牛溶血性曼氏杆菌的最佳浓度,将最佳浓度甲醛灭活24 h的牛溶血性曼氏杆菌菌液分别与弗氏佐剂和灭菌Al(OH)3佐剂混合乳化,制备弗氏佐剂灭活疫苗和铝佐剂灭活疫苗,对两种佐剂灭活疫苗进行无菌检验、物理性状检查、安全性观察和效力检验。结果甲醛灭活牛溶血性曼氏杆菌的最佳终浓度为0.3%。制备的两种佐剂疫苗在5%绵羊血琼脂平板上培养24 h,均无细菌生长;物理性状良好;疫苗注射昆明小鼠后,弗氏佐剂灭活疫苗组小鼠除在注射部位部分产生结节外,无其他副作用,而铝佐剂灭活疫苗组小鼠注射部位产生了脓肿和结块;弗氏佐剂灭活疫苗对经牛溶血性曼氏杆菌强毒菌液攻击的小鼠的保护效果较明显,保护率为90%;铝佐剂灭活疫苗产生的保护作用较弱,保护率为80%。结论成功制备了弗氏佐剂和铝佐剂的牛溶血性曼氏杆菌灭活疫苗,其中弗氏佐剂灭活疫苗效果较好,可用于临床溶血性曼氏杆菌引起的呼吸道传染病的预防。Objective To prepare and perform control tests on inactivated bovine Mannheimia haemolytica vaccine. Methods M. haemolytica was extracted from the lung tissue of bovines infected in a cattle ranch in Heilongjiang Province, of which that qualified in control tests was prepared into seed liquid. The concentration of formaldehyde for inactivation of M. haemolytica was optimized. The M. haemolytica liquid inactivated with formaldehyde at the optimal concentration for 24 h were emulsified with Freund adjuvant and sterile aluminum hydroxide adjuvant respectively, and the prepared two inactivated vaccines were tested for sterility, physical property, safety and potency. Results The optimal formaldehyde concentration for inactivation of M. haemolytica was 0. 3%. No growth of bacteria was observed on the agar plate containing 5% sheep blood on which the two vaccines were cultured for 24 h. The vaccines showed good physical properties. No adverse reactions other than the nodules in injection site were observed in KM mice immunized with the vaccine containing Freund adjuvant, while abscess and indurations were observed in the injection sites of mice immunized with the vaccine containing aluminum hydroxide adjuvant. The protective rate of vaccine containing Freund adjuvant against challenge with virulent M. haemolytica strain was 90%, while that of vaccine containing aluminum hydroxide adjuvant was 80%. Conclusion The inactivated bovine M. haemolytica vaccines containing Freund adjuvant and containing aluminum hydroxide adjuvant were successfully prepared, while the former showed satisfactory effect and might be used for prevention of respiratory infectious diseases caused by M. haemolytica in clinic.

关 键 词：牛溶血性曼氏杆菌 灭活疫苗 弗氏佐剂 A(lOH)3佐剂 

分 类 号：S855.12[农业科学—临床兽医学] R392-33[农业科学—兽医学]