1株藏猪源荚膜血清D型多杀性巴氏杆菌的分离鉴定及其生物学特性

doi:10.3785/j.issn.1008-9209.2020.02.013

浙江大学学报（农业与生命科学版）

2020, Vol. 46

Issue (5): 611-617 DOI: 10.3785/j.issn.1008-9209.2020.02.013

动物科学与动物医学

1株藏猪源荚膜血清D型多杀性巴氏杆菌的分离鉴定及其生物学特性

贡嘎¹(

),王一飞¹,格桑卓玛²,索朗斯珠¹,尼玛央宗²,拉巴次仁²

^1.西藏农牧学院动物科学学院，西藏林芝 860000
^2.西藏自治区动物疫病预防控制中心，拉萨 850000

Identification of capsular serotype D Pasteurella multocida isolated from Tibetan swine and its biological characteristics

Gongga¹(

),Yifei WANG¹, Gesangzhuoma², Suolangsizhu¹, Nimayangzong², Labaciren²

^1.Animal Science College, Tibet Agriculture & Animal Husbandry University, Linzhi 860000, Xizang, China
^2.Animal Epidemic Prevention and Control Center of Tibet Autonomous Region, Lhasa 850000, China

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摘要：

为了解西藏地区病死藏猪源多杀性巴氏杆菌荚膜血清型、毒力基因分布情况，对无菌采集的60份病死猪样品（肺和扁桃体各30份）进行细菌分离纯化后，对菌株进行荚膜分型及常见毒力基因的鉴定。结果表明，从藏猪肺病料中成功获得了1株与伊朗家禽多杀性巴氏杆菌（AY225343）亲缘关系较近的D型猪源多杀性巴氏杆菌，分离率达3.33%（1/30），但没有在扁桃体中分离到该菌株。经鉴定，该菌株携带16种毒力基因，且对四环素、多西环素、氨苄西林、阿莫西林具有耐药性，对卡那霉素、链霉素、新霉素、大观霉素、环丙沙星、诺氟沙星和恩诺沙星较敏感。本研究结果为西藏猪源多杀性巴氏杆菌病原学和流行病学调查提供了参考依据。

关键词： 多杀性巴氏杆菌; 荚膜血清D型; 分离鉴定; 毒力基因; 耐药性; 藏猪

Abstract:

To understand the prevalence of porcine Pasteurella multocida in Tibet, a total of 60 samples (lungs, n=30; tonsil, n=30) were sterilely collected from the diseased and dead Tibetan swine in Tibet to analyze the capsular serotypes and virulence gene distribution of the porcine P. multocida. The results showed that one D-type porcine bacillus strain was obtained from the lung tissue, and the positive rate was 3.33% (1/30). However, there were not bacillus isolates from the tonsil. Moreover, the isolated strain carried 16 virulence genes and was closely related with P. multocida isolated from the poultry in Iran (GenBank No. AY225343). In addition, the strain was resistant to tetracycline, doxycycline, ampicillin and amoxicillin, but was sensitive to kanamycin, streptomycin, neomycin, spectinomycin, ciprofloxacin, norfloxacin and enrofloxacin. In conclusion, a capsular serotype D porcine P. multocida is isolated from the Tibetan swine, which provides references for the etiology and epidemiology investigation of porcine P. multocida in Tibet.

Key words: Pasteurella multocida capsular serotype D isolation and identification virulence gene drug resistance Tibetan swine

收稿日期: 2020-02-01 出版日期: 2020-11-19

CLC:

S 855.12

基金资助: 元亨基金（2018）;西藏自治区科技厅2020年度重点项目(202005)

通讯作者: 贡嘎 E-mail: xzlzgg@163.com

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引用本文:

贡嘎,王一飞,格桑卓玛,索朗斯珠,尼玛央宗,拉巴次仁. 1株藏猪源荚膜血清D型多杀性巴氏杆菌的分离鉴定及其生物学特性[J]. 浙江大学学报（农业与生命科学版）, 2020, 46(5): 611-617.

Gongga,Yifei WANG, Gesangzhuoma, Suolangsizhu, Nimayangzong, Labaciren. Identification of capsular serotype D Pasteurella multocida isolated from Tibetan swine and its biological characteristics. Journal of Zhejiang University (Agriculture and Life Sciences), 2020, 46(5): 611-617.

链接本文:

http://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2020.02.013 或 http://www.zjujournals.com/agr/CN/Y2020/V46/I5/611

表1 多杀性巴氏杆菌鉴定及荚膜血清PCR分型中的寡核苷酸序列

表2 药敏试验判定标准

图1 多杀性巴氏杆菌的菌落形态

图2 多杀性巴氏杆菌的种特异性基因Kmt1与荚膜血清分型的PCR鉴定M：DNA分子质量标准（100～2 000 bp）；1：荚膜血清A型；2：荚膜血清B型；3：荚膜血清D型；4：荚膜血清E型；5：荚膜血清F型； 6～11：Kmt1基因。

图3 多杀性巴氏杆菌毒力基因检测M：DNA分子质量标准（100～2 000 bp）；1：ptfA基因；2：fimA基因；3：hsf-1基因；4：hsf-2基因；5：pfhA基因；6：tadD基因；7：toxA基因； 8：sodA基因； 9：tbpA基因；10：sodC基因；11：nanB基因；12：nanH基因；13：ompA基因；14：ompH基因；15：oma87基因；16：plpB基因； 17：pmHAS基因；18：exbB基因；19：exbD基因；20：tonB基因；21：hgbA基因；22：hgbB基因；23：Fur基因。N：无条带。

图4 藏猪多杀性巴氏杆菌与其他多杀性巴氏杆菌菌株的系统进化树Pm：多杀性巴氏杆菌；Kp：肺炎克雷伯菌。

表3 分离菌的药敏试验结果

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