Preparation of Chromobacterium Inactivated Vaccine and Evaluation of Its Immune Effect
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摘要: 为防控小熊猫色杆菌病,按细菌灭活疫苗制备程序将海峡(福州)大熊猫研究交流中心2008年、2020年分离的2株色杆菌CV08毒株和CV20毒株经条件优化后灭活,分别制备成灭活菌液与铝胶佐剂联用制备成灭活疫苗。以BALB/c小鼠为试验动物,建立色杆菌感染模型,用2种方式接种CV20毒株灭活疫苗,比较2种接种途径效果。进一步设计2株色杆菌半数致死量(LD50)梯度对二免后小鼠进行攻毒保护试验,同时进行交叉攻毒保护试验。结果表明:CV20毒株灭活疫苗肌肉注射效果优于皮下注射。CV08毒株和CV20毒株对每只小鼠的LD50分别为3.2×108、5×107CFU。色杆菌CV08毒株灭活疫苗对不同LD50攻毒小鼠的保护率为83%、 70% 、60% ;色杆菌CV20毒株灭活疫苗对不同LD50攻毒小鼠的保护率为75% 、70% 、10%。试验制备的CV08、CV20毒株疫苗含菌量均为4×109 CFU·mL-1,安全检验合格,能对自身菌株感染产生较好的保护效果但无交叉保护效果。Abstract: In order to prevent and control the Chromobacterium infection in red pandas, according to the preparation procedure of bacterial inactivated vaccine, two strains of Chromobacterium CV08 strain and CV20 virulent strain isolated from Straits (Fuzhou) Giant Panda Research and Exchange Center in 2008 and 2020 were inactivated after the optimization of conditions, and then the inactivated bacteria solution was prepared by combining with the aluminum-containing adjuvants to produce the inactivated vaccine. The BALB/c mice were used as the experimental animals to establish a model of Chromobacterium infection. The inactivated vaccine of CV20 virulent strain was inoculated in two ways to compare the effects of the two inoculation routes. The median lethal dose (LD50) gradient of two strains of Chromobacterium was further designed to carry out the challenge protection test on mice after the secondary immunization, and the cross-challenge protection test was also carried out. The results showed that the effect of intramuscular injection with the inactivated vaccine of CV20 virulent strain was better than that of subcutaneous injection. The LD50 of CV08 virulent strain and CV20 virulent strain to each mouse was 3.2×108 and 5×107 CFU, respectively. The protective rates of the inactivated vaccine of Chromobacterium CV08 virulent strain on the mice challenged with different LD50 were 83%, 70% and 60%, respectively, while the protective rates of the inactivated vaccine of Chromobacterium CV20 virulent strain on the mice challenged with different LD50 were 75%, 70% and 10%, respectively. The bacterial contents of CV08 and CV20 virulent strain vaccines prepared in the experiment were both 4×109 CFU·mL-1, which passed the safety test and could produce better protective effect against the infection of their own strains but had no cross-protective effect.
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Keywords:
- Red panda /
- Chromobacterium /
- Inactivated vaccine /
- Immune protection
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