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| 西藏那曲牦牛源多杀巴氏杆菌荚膜分型及其毒力基因检测 |
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陈建春1, 王一飞1, 周赛赛1, 任晨玮1, 罗润波1, 贡嘎1, 曲久2, 高家登2, 索朗斯珠1
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| (1.西藏农牧学院 动物科学学院, 西藏 林芝 860000;2.西藏那曲市兽医站, 西藏 那曲 852002) |
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| 摘要: |
| 为确定2018年7-11月西藏那曲牦牛发病的死亡原因，以安多县、班戈县、聂荣县和色尼区的26头病死牦牛内脏组织为研究对象，利用常规细菌培养法和特异性基因PCR方法，对分离菌进行鉴定，并对其荚膜血清类型、毒力基因以及致病性进行研究。结果表明：纯化得到13株分离菌，分离菌的菌落特征、菌体形态、生化鉴定结果均与多杀巴氏杆菌相符，并扩增出多杀巴氏杆菌特异性基因Kmt-1和16S rDNA，菌株Pm-1和Pm-2的Kmt-1基因序列与印度、伊朗、巴基斯坦、俄罗斯的牛源多杀巴氏杆菌的同源性均达98%以上；13株菌的荚膜分型鉴定结果均为B型；对分离菌进行已知的23种毒力基因检测结果显示，此次分离到的13株菌毒力基因的数量在16~19种，ptfA、fimA等16种毒力基因检出率为100%，tadD、toxA等4种毒力基因检出率为0，且每个地区分离株毒力基因的分布相同；致病性试验中对照组小鼠全部存活，处理组小鼠死亡时间为24~72 h，死亡率为100%，病变主要表现为脾脏、肺脏、肝脏、心脏出血坏死。导致此次那曲地区牦牛发病死亡的原因为荚膜B型多杀巴氏杆菌病，本研究可为该病的防治提供依据，也为下一步预防疫苗的研究提供了优势菌株。 |
| 关键词: 牦牛 多杀巴氏杆菌 荚膜 毒力基因 |
| DOI：10.11841/j.issn.1007-4333.2019.09.10 |
| 投稿时间：2019-01-16 |
| 基金项目:国家肉牛牦牛产业体系项目（CARS-37）；西藏科技厅重大项目资助 |
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| Capsule type identification and virulence gene detection of Pasteurella multocida derived from yaks in Naqu,Tibet |
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CHEN Jianchun1, WANG Yifei1, ZHOU Saisai1, REN Chenwei1, LUO Runbo1, Gonga1, Qujiu2, GAO Jiadeng2, Suolangsizhu1
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| (1.Department of Animal Science, Tibet Agricultural and Animal Husbandry College, Linzhi 860000, China;2.Veterinary Station of Naqu City, Naqu 852002, China) |
| Abstract: |
| In order to determine the causes of disease and death of yak in Naqu,Tibet,26 visceral tissues of sick and dead calves from Ando,Bange,Nierong and Seni were taken as research object from July to November 2018.The identification of isolated bacteria were conducted by using conventional bacterial culture and specific gene PCR and the capsular serotype,virulence gene and pathogenicity were studied by PCR.The results showed that a total 13 bacteria strains were purified,the colony characteristics,morphology and biochemical identification results of the isolated bacterias were consistent with those of Pasteurella. multocida.The specific gene Kmt-1 and 16S rDNA of P.multocida were amplified.The homology of Kmt-1 gene sequence of strains Pm-1 and Pm-2 with P.multocida from cattle in India,Iran,Pakistan and Russia was over 98%.The results of capsular typing of 13 strains of bacteria showed that they were all B-type.The results of 23 known virulence genes showed that the number of virulence genes of 13 strains was 16-19.The detection rates of 16 virulence genes such as ptfA and fimA were 100%,The detection rates of other four virulence genes such as tadD and toxA were 0.The distribution of virulence genes of the isolates in each region was similar.In the pathogenicity test,all the mice in the control group survived.The death time of the mice in the treatment group was 24 to 72 h,and the mortality rate was 100%.The lesions mainly showed in the spleen,lung,liver and heart hemorrhage of mice.In conclusion,the cause of the death of yaks in Naqu was capsular B-type P. multocida.This study provides a basis for the prevention and treatment of the disease,and also provides a dominant strain for the next step of vaccine prevention research. |
| Key words: yak Pasteurella multocida capsule virulence gene |
