基于新型佐剂的驴流产沙门菌病-兽疫链球菌病二联灭活苗的制备及其安全性和免疫原性  

Preparation of a combined inactivated vaccine against salmonellosis and Streptococcus equi subsp.zoonosis in donkeys and evaluating the immunogenicity

作　　者：刘明 孙长江 顾敬敏 董建宝 韩文瑜 LIU Ming;SUN Changjiang;GU Jingmin;DONG Jianbao;HAN Wenyu(College of Veterinary Medicine,Jilin University,Changchun 130062,China;Shandong Vocational Animal Science and Veterinary College,Weifang,Shandong 261061,China)

机构地区：[1]吉林大学动物医学学院,吉林长春130062 [2]山东畜牧兽医职业学院,山东潍坊261061

出　　处：《中国兽医学报》2023年第6期1187-1193,1202,共8页Chinese Journal of Veterinary Science

基　　金：山东省重点研发计划资助项目(2021TZXD012);山东省驴产业技术体系建设资助项目(SDAIT-27);山东省自然科学基金面上资助项目(ZR2020MC182)。

摘　　要：旨在研制预防以马流产沙门菌(S.abortus equi)和马链球菌兽疫亚种(SEZ)感染引起的孕驴流产和新生驴驹腹泻的驴流产沙门菌病-兽疫链球菌病二联灭活疫苗,并进行安全性与免疫原性评价。从山东某驴场分离和鉴定出的S.abortus equi、SEZ分离株为疫苗候选菌株,经甲醛灭活,分别与氢氧化铝佐剂和角鲨烯佐剂制备驴流产沙门菌病-兽疫链球菌病二联灭活苗。首先,检测灭活疫苗对BALB/c小鼠的安全性,比较2种不同佐剂诱导免疫保护的效果;以驴流产沙门菌病-兽疫链球菌病二联角鲨烯佐剂灭活疫苗免疫驴,凝集试验检测免疫血清的总抗体水平及血清杀菌试验检测功能性抗体水平,进一步评价二联灭活疫苗的免疫原性。结果显示,以氢氧化铝佐剂二联灭活苗免疫对5倍LD_(50)S.abortus equi和SEZ攻击BALB/c小鼠的保护率分别达到62.5%(5/8)和75.0%(6/8),而角鲨烯佐剂二联灭活疫苗对2种病原的攻击保护率均达到100%(8/8);以角鲨烯佐剂制备的二联灭活疫苗免疫驴的血清中抗流产沙门菌(10^(8)CFU/mL)的抗体凝集效价几何平均值可达1∶32,功能性抗体对S.abortus equi和SEZ的血清杀菌效价为1∶16(杀菌效率68.99%)和1∶4(杀菌效率65.47%)。S.abortus equi和SEZ 2种抗原免疫不会产生免疫干扰,角鲨烯配方佐剂与抗原结合诱导的抗S.abortus equi和SEZ致死剂量攻毒产生的保护作用强于氢氧化铝佐剂,且该二联灭活疫苗在小鼠和驴体内具有较强的保护效力与免疫原性,且安全性良好。本研究为该基于新型疫苗佐剂二联疫苗的研发奠定了坚实的基础。To prevent of the abortus of pregnant donkey and the diarrhea of newborn donkey caused by the infection of Salmonella abortus equi and Streptococcus equi subsp.zooepidemicus,a combined inactivated vaccine of donkey salmonellosis and Streptococcus equi subsp.zoonosis were prepared,its immunogenicity and safety were evaluated.Salmonella abortus equi and Streptococcus equi subsp.zooepidemicus isolated from donkey farms in Shandong Province were used as vaccine candidate strains.After inactivated by formaldehyde,the inactivated vaccines of salmonellosis-Streptococcus equi subsp.zoonosis was prepared with aluminum hydroxide adjuvant or squalene formula adjuvant respectively.The safety and immune protections of mice induced by inactivated vaccines was tested,and the immune protection effects of two different adjuvants were compared.The donkeys were immunized with the combined salmonellosis-Streptococcus zooepidemicus inactivated vaccine with qualene formula adjuvant.The total antibody levels of the donkey serum were detected by agglutination test and the functional antibody level was detected by serum bactericidal test.Results showed that the protective rates of BALB/c mice immunized with the aluminum hydroxide adjuvant inactivated vaccine against five times of LD_(50)against Salmonella abortus equi and Streptococcus equi subsp.z0epidemicus,were 62.5%and 75.0%,respectively,while the protective rates of the squalene formula adjuvant inactivated vaccine against the two pathogens were 100%.The agglutination titer of the antibody against Salmonella abortus equi(10^(8)CFU/mL)in the sera of donkeys immunized with the combined inactivated vaccine prepared with squalene formula adjuvant was 1:32,and the bactericidal titer of the functional antibody against Salmonella abortus equi and Streptococcus equi subsp.zoonosis was 1:16(bactericidal efficiency 68.99%)and 1:4(bactericidal efficiency 65.47%).Conclusion:This study shows that the simultaneous immunization of inactivated Salmonella abortus equi and Streptococcus equi subsp.zo

关 键 词：马流产沙门菌 马链球菌兽疫亚种 灭活疫苗 角鲨烯佐剂 免疫原性和安全性 

分 类 号：S852.61[农业科学—基础兽医学]