羊源溶血性曼氏杆菌A1型灭活疫苗的制备及免疫效果评价  

Preparation and evaluation of inactivated vaccine against sheep Mannheimia haemolytica type A1

作　　者：夏子涵 李妍 马媛 张焕容[1] 杨发龙[1] XIA Zihan;LI Yan;MA Yuan;ZHANG Huanrong;YANG Falong(College of Animal and Veterinary Sciences,Southwest Minzu University,Chengdu 610041,China)

机构地区：[1]西南民族大学畜牧兽医学院,四川成都610041

出　　处：《中国兽医学报》2024年第11期2356-2362,共7页Chinese Journal of Veterinary Science

基　　金：国家重点研发计划资助项目(2023YFD180250202);四川省科技计划资助项目(2023YFN0043);中央高校基本科研业务费专项优秀学生培养工程项目(ZYN2024167)

摘　　要：溶血性曼氏杆菌是引起羊呼吸道疾病的重要病原,研制羊源溶血性曼氏杆菌灭活疫苗是防控该病原的有效手段。本研究从某绵羊场采集的病料中分离的溶血性曼氏杆菌,通过小鼠致病性试验筛选出最适制苗菌株,并以4种抗原量(10^(6)、10^(7)、10^(8)、10^(9) CFU/mL)与3种佐剂(ISA 201佐剂、ISA 206佐剂、铝佐剂)组合制备灭活疫苗,经免疫和攻毒保护试验,筛选出灭活疫苗制备的最佳抗原量和佐剂。最后,利用筛选出的最佳条件制备疫苗,观察免疫绵羊的抗体水平动态变化。结果显示,共分离得到24株绵羊源溶血性曼氏杆菌,其中血清A1型12株,为该羊场感染的优势血清型;筛选获得1株绵羊源溶血性曼氏杆菌A1型菌株(Mh1)为制苗用菌株,其对小鼠的半数致死量为3.16×10^(7) CFU/mL,具有较强的致病性,可造成小鼠的心脏、肝脏、脾脏、肺脏和肾组织不同程度的炎性细胞浸润等病理损伤;免疫及攻毒保护试验结果显示,灭活疫苗的最佳抗原量为10^(9) CFU/mL,最适佐剂为ISA 201,刺激产生的抗体水平最高,对小鼠的保护率达100%,免疫绵羊后可诱导产生良好的体液免疫。结果表明,本研究制备的羊源溶血性曼氏杆菌A1型灭活疫苗具有良好的免疫保护效果,为进一步研发羊源溶血性曼氏杆菌商品化疫苗奠定了良好的基础。Mannheimia haemolytica is an important pathogen causing respiratory diseases in sheep,and the development of an inactivated vaccine of sheep origin against Mannheimia haemolytica is an effective means of prevention and control of this pathogen.In this siudy,Mannheimia haemolytica isolated from the diseased material collected from a sheep farm was screened for the most suitable vaccine strain by mouse pathogenicity test,and the inactivated vaccine was prepared by combining four antigenic amounts(10^(6),10^(7),10^(8),10^(9) CFU/mL)with three adjuvants(ISA 201 adjuvant,ISA 206 adjuvant,and aluminum adjuvant),which were used in immunization and attack protection tests,and then screened for immunity and attack protection tests.Immunization and attack protection tests were conducted to screen the optimal antigen amount and adjuvants for inactivated vaccine preparation.Finally,the vaccine was prepared under the optimal conditions and the dynamic changes of antibody levels in immunized sheep were observed.The results showed that a total of 24 strains of Mannheimia haemolytica of sheep origin were isolated,among which 12strains of serotype A1 were the dominant serotypes for the infection in this sheep farm;one strain of Mannheimia haemolytica of sheep origin of type A1(Mh1)was screened and obtained as the strain for vaccine preparation.Its LD_(50)for mice was 3.16×10^(7) CFU/mL,with strong pathogenicity,and could cause pathological damage such as inflammatory cell infiltration of varying degrees in heart,liver,spleen,lung and kidney tissues;the results of the immunization and attack protection test showed that the optimal antigenic amount of the inactivated vaccine was 10^(9)CFU/mL,and the most suitable adjuvant was ISA 201,which stimulated the production of the highest antibody level and provided 100%protection for the mice.The protection rate of mice reached 100%,and good humoral immunity could be induced after immunizing sheep.The inactivated Mannheiaia haemolytica type A1 vaccine prepared in this study has good immun

关 键 词：绵羊 溶血性曼氏杆菌 灭活疫苗 免疫研究 

分 类 号：S852.61[农业科学—基础兽医学]