摘要
为研究猪丹毒丝菌CVCC43005株作为生产用菌种的最高扩繁代次以及新型佐剂对该菌种免疫原性的影响,在对猪丹毒丝菌CVCC43005株形态、生化特性、培养特性、纯粹性、血清学特性、真空度、剩余水分含量、毒力及免疫原性等研究的基础上,建立了猪丹毒丝菌CVCC43005株种子批。将CVCC43005株(1986年冻干)用培养基进行了20次传代(F1~F20),并对其中的F5、F10、F15、F20代进行冻干并抽检测定。结果显示:抽检的各代次菌种均纯粹,形态、生化特性、培养特性均符合猪丹毒丝菌的特性,血清型为2型,真空度检查均呈现白色、粉色或紫色辉光,剩余水分含量均<3.0%,9~10CFU活菌皮下注射体重18~22g小白鼠,均于7日内5/5死亡。将抽检的各代次菌种制成铝胶佐剂灭活疫苗,免疫16~18g小白鼠后均能产生10/10保护。以F5代菌株制备菌液,甲醛灭活后,分别用铝胶佐剂、矿物质白油佐剂、洛阳赛威公司的水溶性佐剂、水性复合免疫佐剂以及法国赛比克公司的MONTANIDETMIMS1313VGNST、GEL01PR6种佐剂制备灭活疫苗,经无菌及安全检查合格后进行16~18g小白鼠免疫效果对比实验,最终确定猪丹毒丝菌CVCC43005株基础种子代数为F1~F10代,生产用菌种的最高扩繁代次宜控制在5代以内,法国赛比克公司的GEL01PR佐剂为猪丹毒灭活疫苗的最佳佐剂。该菌种作为生产用菌种最高扩繁代次的确定及疫苗佐剂的筛选,为修订菌种检定依据奠定了基础。
The CVCC43005 strain was conducted experiments of morphology, biochemical characteristics, culture characteristics, purity, serological characteristics, vacuum degree, residual moisture content, virulence and immunogenicity. A seed batch of E. rhusiopathiae was established. The CVCC43005 strain (lyophilized in 1986) was subjected to passage for 20 times (F1 to F20), and the F5, F10, F15 and F20 generations were lyophilized. All the results of the four batches freeze-dried strains were satisfied. Each batch freeze-dried strains were pure, and the morphology, biochemical characteristics and culture characteristics were consistent with the characteristics of E. rhusiopathiae . The serotype was type 2, and the vacuum test was white, pink or purple glow, residual water content were below 3.0%, 9~10 CFU live bacteria subcutaneous injection of 18~22 g mice, 5/5 died within 7 days. The samples of each generation were made into inactivated vaccines with aluminum gel adjuvant, which could 10/10 protect 16~18 g mice after challenge. Prepare the bacterial solution with F5 generation strain, after inactivation with formaldehyde, respectively using aluminum glue adjuvant, mineral white oil adjuvant, water-soluble adjuvant (Luoyang Saiwei), water-based compound adjuvant (Luoyang Saiwei), MONTANIDE TM IMS1313VGNST (Seppic) and GEL01PR (Seppic) to prepare inactivated vaccines. After sterility and safety test, the immune effect compared experiments were conducted. It is finally determined that the basic seed number of E. rhusiopathiae strain CVCC43005 is F1~F10 generation, and the highest expansion for production strain is 5 generations;the GEL01PR(Seppic) is the best E. rhusiopathiae inactivated vaccine adjuvant. The determination of the strain as the highest expansion of production strains and the screening of vaccine adjuvants provided the foundation for the revision of the strains.
作者
张媛
魏财文
李建
张一帜
辛凌翔
王秀丽
彭国瑞
刘博
蒋玉文
ZHANG Yuan;WEI Cai-wen;LI Jian;ZHANG Yi-zhi;XIN Ling-xiang;WANG Xiu-li;PENG Guo-rui;LIU Bo;JIANG Yu-wen(China Institute of Veterinary Drug Control, Beijing 100081, China)
出处
《中国兽药杂志》
2019年第10期7-14,共8页
Chinese Journal of Veterinary Drug