中国畜牧兽医 ›› 2026, Vol. 53 ›› Issue (1): 489-498.doi: 10.16431/j.cnki.1671-7236.2026.01.044

• 基础兽医 • 上一篇    下一篇

貉源支气管败血波氏杆菌分离鉴定及其生物学特性分析

王奇林1(), 曹润来1, 刘佩雯1, 刘子瑜1, 薛竹青1, 杜海霞2, 张悦2, 张敏静2, 李亚聪1, 王威阳1, 丛雁方2, 王晓旭1(), 刘志杰1()   

  1. 1.中国农业科学院特产研究所,农业农村部经济动物疫病重点实验室,吉林省特种动植物科技创新国际合作重点实验室,长春 130112
    2.青岛蔚蓝动物保健集团有限公司,青岛 266114
  • 收稿日期:2025-04-22 出版日期:2026-01-05 发布日期:2025-12-26
  • 通讯作者: 王晓旭,刘志杰 E-mail:wangqilin@caas.cn;wangxiaoxussdd@126.com;liuzhijie@caas.cn
  • 作者简介:王奇林,E-mail:wangqilin@caas.cn
  • 基金资助:
    国家重点研发计划项目(2023YFD1800701);中国农业科学院科技创新工程(CAAS-ASTIP-2022-ISAPS)

Isolation, Identification and Biological Characteristics Analysis of Bordetella bronchiseptica from Raccoon dog(Nyctereutes procyonoides

WANG Qilin1(), CAO Runlai1, LIU Peiwen1, LIU Ziyu1, XUE Zhuqing1, DU Haixia2, ZHANG Yue2, ZHANG Minjing2, LI Yacong1, WANG Weiyang1, CONG Yanfang2, WANG Xiaoxu1(), LIU Zhijie1()   

  1. 1.Key Laboratory of International Cooperation for Specialized Animal and Plant Science;and Technology Innovation of Jilin Province,Key Laboratory of Economic Animal;Diseases of Ministry of Agriculture and Rural Affairs,Institute of Special Animal and;Plant Sciences,Chinese Academy of Agricultural Sciences,Changchun 130112,China
    2.Qingdao Vland Animal Health Group Co. ,Ltd. ,Qingdao 266114,China
  • Received:2025-04-22 Online:2026-01-05 Published:2025-12-26
  • Contact: WANG Xiaoxu, LIU Zhijie E-mail:wangqilin@caas.cn;wangxiaoxussdd@126.com;liuzhijie@caas.cn

摘要:

目的 明确河北省某貉场貉死亡的病因,为有效防控该病提供科学依据。 方法 无菌采集病死貉病变组织及呼吸道分泌物,经病原分离培养获得目标菌株。通过革兰染色对分离菌株进行形态学鉴定;利用16S rRNA基因序列比对明确分离菌株分类;采用PCR扩增检测分离菌株的毒力基因及耐药基因,进一步开展药敏试验评估其耐药表型;通过小鼠致病性试验检测分离菌株的致病性。 结果 从貉体内分离获得的菌株在TSB琼脂平板上呈圆形、边缘整齐的灰白色菌落,在MAC琼脂平板上呈光滑、湿润的透明菌落;革兰染色可观察到菌株呈球杆状、两极着色的革兰阴性菌。16S rRNA序列比对分析显示,分离株与GenBank中其他支气管败血波氏杆菌的相似性为97.0%~100%,鉴定该分离株为支气管败血波氏杆菌。毒力基因和耐药基因检测显示,分离株检出prn、bvgs、fla、fhaB、cyaA、dnt 6种毒力基因和blaTEM、blaSHV、parC、tetC、gyrA 5种耐药基因。药敏试验结果显示,分离菌株对青霉素类、大环内酯类、糖肽类、磺胺类、硝基呋喃类抗菌药表现耐药,对氨基糖苷类和喹诺酮类表现敏感。致病性检测显示,分离菌株对小鼠的半数致死量为3.67×107 CFU。 结论 本试验从死亡貉体内分离获得1株支气管败血波氏杆菌,其携带多种毒力基因和耐药基因,对青霉素类、大环内酯类、糖肽类、磺胺类、硝基呋喃类抗菌药表现耐药,对氨基糖苷类和喹诺酮类敏感,具有多重耐药性和较强致病性。本研究结果为貉支气管败血波氏杆菌病的诊断、临床用药及疫苗研发提供了科学依据。

关键词: 貉; 支气管败血波氏杆菌; 分离鉴定; 毒力基因; 耐药基因

Abstract:

Objective The purpose of this experiment was to clarify the cause of the death of raccoon dogs in a raccoon dog farm in Hebei province, and provide a scientific basis for the effective prevention and control of the disease. Method Sterile collection of diseased tissues and respiratory secretions from raccoon dogs was collected, and the target strains were obtained through pathogen isolation and culture. The morphology of the isolate was identified by Gram staining. The classification of isolate was clarified by using 16S rRNA gene sequence alignment. The virulence genes and drug resistance genes of the isolate were detected by PCR amplification, and further drug sensitivity tests were carried out to evaluate the drug resistance phenotypes. The pathogenicity of the isolate was detected through the mouse pathogenicity test. Result The strain isolated from raccoon dogs formed round, neatly edged grayish-white colonies on TSB agar plates and smooth, moist, transparent colonies on MAC agar plates. Gram staining revealed the strain as Gram-negative coccobacilli with bipolar staining. 16S rRNA sequence alignment analysis showed that the similarity of the isolate to other Bordetella bronchiseptica in GenBank was 97.0%-100%, and this isolate was Bordetella bronchiseptica.Detection of virulence and resistance genes demonstrated the presence of six virulence genes (prn, bvgs, fla, fhaB, cyaA and dnt) and five resistance genes (blaTEM, blaSHV, parC, tetC and gyrA). The results of the drug sensitivity test showed that the isolate was resistant to penicillins, macrolides, glycopeptides, sulfonamides and nitrofurans, but sensitive to aminoglycosides and quinolones. Pathogenicity tests showed that the median lethal dose of the isolate to mice was 3.67×107 CFU. Conclusion In this experiment, one strain of Bordetella bronchiseptica was isolated from dead raccoon dogs, which carried multiple virulence genes and drug resistance genes. The isolate showed resistance to penicillins, macrolides, glycopeptides, sulfonamides and nitrofurans, and was sensitive to aminoglycosides and quinolones, demonstrating multidrug resistance and strong pathogenicity. The results of this study provided a scientific basis for the diagnosis, clinical medication, and vaccine development of Bordetella bronchiseptica disease in raccoon dogs.

Key words: raccoon dog; Bordetella bronchiseptica; isolation and identification; virulence genes; drug resistance gene

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