体内繁殖禽巴氏杆菌的提取及其交叉保护特性

Recovery and Cross-protection of Pasteurella multocida Propagated in Chicken Vivo

通过不同攻毒途径、不同攻毒剂量、不同采血时间及不同采血途径的试验表明 ,以翅静脉途径攻击多杀巴氏杆菌 ,剂量 10亿 /只 ,在鸡濒死挣扎时的即刻颈静脉无菌放血 ,可以自鸡体采集到最大量的含菌血 ,每只鸡 37～ 39m L ,再将血液进行差速和蔗糖密度梯度离心 ,可最大限度地提取到血液中的巴氏杆菌 ,每只鸡 30 0～ 4 0 0亿的总菌量。该结果与报道的提取方法相比 ,效率提高了 10 0 0倍以上。提取的细菌灭活后免疫鸡 ,经异型菌交叉攻毒试验表明 ,体内繁殖的巴氏杆菌具有交叉保护特性 ,而体外培养菌则没有。用该方法提取的巴氏杆菌 ,其总蛋白和总糖含量均高于培养基培养菌 。

The comparisons of different methods and doses of inoculations,the different time and ways of collection blood post-infection were performed.The results showed that the maximum yield of chicken blood contained Pasteurella Multocida(P.M.),which was about 37-39 mL each chicken,could be reached by challenge with P.M. through the chicken wing vein,the dose was 10~9 CFU each chicken,and collection of the blood via the neck vein when the chickens were moribund.And then,the maximum yield of P.M. would be purificated by differential and sucrose density gradient centrifugation.The bacteria of 3×10^(10)-4×10^(10) CFU could be gained from every chicken by this method from these blood,and was over (1 000) times than previous reported method.Another results showed that,while the chickens were vaccinated with this killed bacteria and challenged with heterogenerous type of P.M.,the bacteria propagated in vivo could induce a cross-protection in chicken,but in vitro could not.The total proteins and the glucosum of P.M. in vivo purificated by this method were all over than in vitro,suggesting that this difference may be relevant to the cross-protection.