犬A型产气荚膜梭菌灭活疫苗的制备及免疫学评价

Preparation and immunological evaluation of inactivated vaccine against Clostridium perfringens type A

产气荚膜梭菌是引起犬猝死症的主要病原菌，目前临床上还没有预防犬猝死症的疫苗。本试验以分离自犬的A型产气荚膜梭菌为制苗菌株，首先对利用甲醛进行产气荚膜梭菌培养液脱毒的浓度和时间进行了优化，然后利用脱毒后的产气荚膜梭菌培养液对小鼠和犬进行免疫，并对不同佐剂（铝佐剂和油佐剂）和不同免疫途径（皮下注射和腹腔注射）的免疫效果进行评价。结果显示，产气荚膜梭菌灭活疫苗具有良好的免疫保护效果，免疫后小鼠体内可产生高水平的血清抗体，抗体效价最高可达6．25×10^4。产气荚膜梭菌铝胶灭活疫苗对犬具有良好的安全性，单次免疫（4mL／只）或2次免疫（2mL／只）均可产生良好的免疫保护效果（5／5）。结果表明，本试验制备的犬A型产气荚膜梭菌灭活疫苗可用于预防犬产气荚膜梭菌感染。

Clostridium perfringens is the most important pathogenic bacterium causing sudden death in dogs,there is no vaccine against it by now. In this study,the Clostridium perfringens type A isolated from dogs was selected as the vaccine producing strain,and the conditions of formalde- hyde detoxification were optimized. Inactivated type Clostridium perfringens was used as vaccine for immunization of female Kunming rats,the oil adjuvant and aluminium hydroxide adjuvant in- traperitoneal and subcutaneous injection to determine the optimal immune condition. The results showed that inactivated Clostridium perfringens can produce high levels of exotoxin, protective experiments in mice and laboratory dogs showed that the inactivated vaccine had a good protective effect,and the rate of immune protection was 100%. Immune effects of the antibody in serum were evaluated by ELISA, and the titer of the antibody reached by 6.25 × 10^4. Single immunization （4 mL/each） or double immunization（2 mL/each） could produce good immune protection effect. These results indicate that inactivated vaccine has a good preventive effect on canine sudden death caused by Clostridium perfringens and is of great significance for the development of commercial vaccines.