气肿疽梭菌C54-1株的制备与检定

The Preparation and Certification of Clostridium chauvoei Strain C54-1

关键词:  气肿疽梭菌C54-1株  CVCC60001  毒力  免疫原性  培养基筛选  强化梭菌培养基(RCM)  苏木精-伊红染色法

Key words:  Clostridium chauvoei strain C54-1  CVCC60001  virulence  immunogenicity  medium selection  reinforced Clostridium medium(RCM)  HE staining

摘要:

为研究气肿疽梭菌C54-1株(CVCC60001)的菌种特性，制备了1批气肿疽梭菌C54-1株，并对菌种的形态及生化特性、培养特性、血清学特性、真空度、纯粹、剩余水分、毒力及免疫原性、16S rDNA等进行检定，以及对于该菌的适宜培养基促生长能力等方面进行了比较。实验结果表明，该冻干菌种的形态及生化特性、培养特性、血清学特性、真空度、纯粹、剩余水分、毒力及免疫原性均符合《中华人民共和国兽用生物制品规程》二〇〇〇版质量标准的规定。16S rDNA鉴定为气肿疽梭菌，相似度为99.93%。在免疫原性检定中，使用致死剂量攻毒时，免疫组能够4/4被保护，证明该菌明矾苗免疫效果良好。使用商品化梭菌培养基对于该菌培养进行了比对，证实商品化培养基替代厌气肉肝汤的可能性。本研究为气肿疽梭菌的制备和检定提供参考依据，并使用不同培养基进行比对研究，为该菌的稳定培养和疫苗工艺改进提供了研究基础。

Abstract:

In order to study the characteristics of C. chauvoei strain C54-1(CVCC60001), a batch of C. chauvoei strain C54-1 was prepared, and the strain morphology, biochemical characteristics, culture characteristics, serological characteristics, vacuum degree, purity, residual water, virulence and immunogenicity,16S rDNA of the strain were tested, and the growth promotion ability of the suitable medium for this strain was compared. The experimental results showed that the morphology, biochemical characteristics, culture characteristics, serological characteristics, vacuum degree, purity, residual water, virulence and immunogenicity of the lyophilized strain were in accordance with the quality standard of the Code of Veterinary Biological products regulations of the People’s Republic of China(2000).16S rDNA was identified as C. chauvoei and the similarity was 99.93%. In the immunogenicity test, when lethal dose was used to attack the animals, the immune group could be protected by 4/4, which proved that the immune effect of the alum vaccine was good. The commercial clostridium culture medium was compared to confirm the possibility of commercial medium replacing anaerobic meat liver soup. This study provided a reference for the preparation and verification of C. chauvoei , and comparison studies using different media provided a research basis for the stable culture of the bacteria and the improvement of vaccine technology.