Identification of capsular serotype D <i>Pasteurella multocida</i> isolated from Tibetan swine and its biological characteristics

doi:10.3785/j.issn.1008-9209.2020.02.013

Journal of Zhejiang University (Agriculture and Life Sciences)

2020, Vol. 46

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

Animal sciences & veterinary medicine

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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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

Received: 01 February 2020 Published: 19 November 2020

CLC:

S 855.12

Corresponding Authors: Gongga E-mail: xzlzgg@163.com

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Cite this article:

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.

URL:

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

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

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

Table 1 Sequences of the oligonucleotides used in P. multocida multiplex capsular serum PCR typing assay

Table 2 Criterion of antibiotics sensitivity tests

Fig. 1 Colony morphology of P. multocida

Fig. 2 Identification of P. multocidaKmt1 gene and capsular serotype genesM: DNA marker (100-2 000 bp)；1: Capsular serotype A; 2: Capsular serotype B; 3: Capsular serotype D; 4: Capsular serotype E; 5: Capsular serotype F; 6-11: Kmt1 gene.

Fig. 3 Detection of P. multocida virulence genesM: DNA marker (100-2 000 bp)； 1： ptfA gene； 2： fimA gene； 3： hsf-1 gene； 4： hsf-2 gene； 5： pfhA gene； 6： tadD gene； 7： toxA gene； 8： sodA gene； 9： tbpA gene； 10： sodC gene； 11： nanB gene； 12： nanH gene； 13： ompA gene； 14： ompH gene； 15： oma87 gene； 16： plpB gene； 17： pmHAS gene； 18： exbB gene； 19： exbD gene； 20： tonB gene； 21： hgbA gene； 22： hgbB gene； 23： Fur gene. N: No band.

Fig. 4 Phylogenetic tree of P. multocida isolated from Tibet swines and othersPm: P. multocida; Kp: Klebsiella pneumoniae.

Table 3 Drug sensitivity test results of isolated strains


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