羊黑疫单价疫苗的研制及免疫家兔血清抗体测定

Research on Sheep black disease monovalent vaccine and detection of the serum antibody titer of immuned rabbit

从青海省海北州海晏县同宝村绵羊急性死亡病例中分离到一株B型诺维梭菌,利用该分离菌株(TB05)和B型诺维制苗菌株(C61-4)所产毒素经甲醛灭活,分离株加入氢氧化铝胶、磷酸铝胶及明矾佐剂,制苗株(C61-4)加入磷酸铝胶佐剂制备成羊黑疫单价疫苗,对该疫苗分别进行了安全性检验、家兔免疫试验及血清抗体效价测定试验。结果表明,B型诺维梭菌菌液用5%甲醛72 h即可灭活;家兔皮下注射5 m L该疫苗,观察期内无不良反应,安全性良好。家兔免疫试验表明,分离株氢氧化铝苗、磷酸铝苗及明矾苗0.1 m L免疫组免疫21d后,保护力分别为200、200、100 MLD(家兔最小致死量)、0.5 m L组免疫组保护力均为200 MLD,(C61-4)菌株磷酸铝苗0.1 m L免疫组免疫21d后,保护力低于100 MLD、0.5 m L组免疫组保护力为100 MLD。免疫家兔血清抗体测定结果表明,分离株氢氧化铝苗、磷酸铝苗及明矾苗0.1 m L免疫组免疫21 d后,0.1 m L血清中和力分别为300、200、100 MLD(小鼠最小致死量)、0.5 m L组免疫组0.1 m L血清中和力分别为400、300、300 MLD;(C61-4)磷酸铝苗0.1 m L和0.5 m L免疫组免疫21 d后,0.1 m L血清中和力均为100 MLD。上述试验结果表明,用该地方分离株制备的磷酸铝单价疫苗免疫效果优于C61-4株羊黑疫磷酸铝苗。

A Clostridium noviy type B strain was isolated on sheep struck by acute deaths from Haiyan County Haibei.Toxin formthis bacteria and standard strains （C61-4）were inactivated by formaldehyde the isolated strain was adding aluminum hydroxide gel, rubberand aluminum phosphate adjuvant alum, standard strains（C61-4）adding alunlinum phosphate gel produced black sheep monovalent vaccine.the vaccine were carried out safety inspection, rabbit immune and serum antibody measurement test. The results showed that the cultureactioned with 5% formaldehyde in37 ＊C for 72 h can be inactivated; there no adverse reactions during the observation period when rabbitsinjected 5 mL of the vaccine, the security is good. Rabbit immune tests showed that 0.1 mL immtmity groups of aluminum hydroxide vaccine,aluminum phosphate and alum vaccine the protection force was 200,200,100 MLD（minimum lethal dose in rabbits）, 0.5 mL protectiveimmunity groups force was 200 MLD, standard strains （ C61-4）0. lml immunity groups protection was 100MLD, 0.5 mL protective immunitygroups was 100 MILD. Immune rabbit serum antibody test showed that 0.1 mL immunity groups of aluminum hydroxide vaccine, aluminumphosphate and alum vaccine which 0.1 mL serum Can neutralize toxin for 300,200,100 MLD（mouse minimum lethal dose）, 0.5 mLimmunity groups that O. 1 mL serum Can neutralize toxin for 400,300,300 MLD ; standard strains （C61-4）aluminum phosphate vaccine0.1 mL and 0.5 mL immunity groups,which 0.1 mL serum only neutralize toxin for 100 MLD. The experimental results show that theimmunization were batter use the local strain aluminum vaccine than the standard strains.