高原牧区羊黑疫野毒分离株的生物学鉴定及免疫原性分析  

Biological identification and immunogenicity analysis of sheep black plague isolates from plateau pastoral areas

作　　者：李生庆[1] 张西云[1] 李淑萍 胡国元[1] 王亭亭[1] 刘怀新[1] 叶明俊 贾苗 LI Sheng-qing;ZHANG Xi-yun;LI Shu-ping;HU Guo-yuan;WANG Ting-ting;LIU Huai-xin;YE Ming-jun;JIA Miao(The Academy of Animal and Veterinary Science,Qinghai University,Xining 810016,China;College of Agriculture and Animal Husbandry,Qinghai University,Xining 810016,China)

机构地区：[1]青海大学畜牧兽医科学院,青海西宁810016 [2]青海大学农牧学院动物医学系,青海西宁810016

出　　处：《中国兽医杂志》2019年第7期18-21,I0001,共5页Chinese Journal of Veterinary Medicine

摘　　要：为提高羊黑疫疫苗的免疫保护力,从青藏高原牧区病死羊体内分离的6株野毒株中筛选具有高免疫原性的B型诺维氏梭菌制苗用菌株,通过16S rRNA分子鉴定、α毒素基因片段的扩增、产毒素能力对比分析,筛选出1株产毒能力较高的TB05野毒株,以标准株为对照,研制了羊黑疫氢氧化铝菌苗,对其免疫原性进行了对比分析。结果显示,6株菌16S rRNA分子系统发育树亲缘关系较近,均扩增出大小为570 bp的α毒素基因片段;该TB05野毒株在含1.0%葡萄糖,pH值8.4~8.6的VF培养基中37℃72 h产毒最高,可达40000 MLD/mL;疫苗对家兔的免疫保护力是对照组的2倍,血清抗体效价是对照组的3倍,免疫羊180 d后仍可抵抗2个致死量的攻击,免疫期远高于对照组的3个月。表明该野毒株具有较强的产毒稳定性和较好的免疫原性,可作为羊黑疫疫苗生产备用菌株。In order to improve the immune protection effect of sheep black vaccine,six wild-type strains isolated from diseased sheep in pastoral areas of Qinghai-Tibet Plateau were screened for high immunogenic strains of Clostridium Novyi type B strains.The methods included 16S rRNA molecules identification,amplification ofα-toxin gene fragment,comparative analysis of toxin-producing ability,screening out a TB05 wild strain with high toxic ability,and developing amniotic aluminum hydroxide inactivated vaccine.Its immunogenicity was studied and compared with other vaccines.The results showed that the 16S rRNA sub-systems of the six strains were closely related to each other,and a 570 bpα-toxin gene fragment grew amplified.The TB05 wild strain grew in VF medium containing 1.0%glucose and pH 8.4-8.6 at 37℃for 72 h,and toxicity reached up to 40000 MLD/mL.The immune protection of the vaccine to rabbits was twice that of the control group,the serum antibody titer was three times that of the control group,and the immune sheep survived two lethal dose challenges after 180 days.The immunization period was much higher than that of the control group for 3 months.The wild strain has strong toxigenic stability and good immunogenicity,and can be used as a spare strain for the production of live vaccine of sheep black vaccination.

关 键 词：B型诺维氏梭菌 分子鉴定 产毒试验 免疫原性 

分 类 号：S858.27[农业科学—临床兽医学]