福建山羊溶血性曼氏杆菌的分离鉴定及耐药性分析

林裕胜; 江锦秀; 张靖鹏; 游伟; 胡奇林

doi:10.19303/j.issn.1008-0384.2019.03.011

福建山羊溶血性曼氏杆菌的分离鉴定及耐药性分析

doi: 10.19303/j.issn.1008-0384.2019.03.011

福建省农业科学院畜牧兽医研究所, 福建福州 350013

基金项目:

国家重点研发计划项目 2016YFD0500906

福建省科技计划项目——省属公益类科研院所基本科研专项 2018R1101014-17

福建省农业科学院科技创新项目 PC2018-6

福建省农业科学院青年创新团队建设项目 STIT2017-1-5

福建省农业科学院青年创新团队建设项目 STIT2017-3-10

详细信息

作者简介:
林裕胜(1988-), 男, 硕士, 助理研究员, 主要从事动物传染病研究

通讯作者:
胡奇林(1963-), 男, 研究员, 主要从事动物传染病学和免疫学研究(E-mail:hql562713@163.com)

中图分类号: S852.61
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- 被引次数: 0
出版历程
- 收稿日期: 2018-12-27
- 修回日期: 2019-02-26
- 刊出日期: 2019-03-28

Isolation, Identification and Antibiotic Resistance of Mannheimia Haemolytica from Diseased Goats in Fujin

Institute of Animal Husbandry & Veterinary Medicine, Fujian Academy of Agricultural Sciences, Fuzhou, Fujian 350013, China

摘要

摘要: 目的对福清一山羊养殖场发生的山羊肺炎病原进行确诊，为福建山羊溶血性曼式杆菌的分离鉴定及检测提供研究基础。方法对剖检采取的组织进行细菌分离培养，随后对溶血性曼氏杆菌、羊口疮病毒、绵羊肺炎支原体和丝状支原体簇等羊常见呼吸道病原进行PCR检测，对分离菌进行16S rRNA基因克隆测序，并对分离株进行药物敏感性研究。结果细菌分离培养结果显示：从病死羊肺脏中分离到1株细菌；PCR检测结果显示，除溶血性曼氏杆菌有扩增条带外，未检测到其他病原；16S rRNA测序结果显示：分离株16S rRNA基因序列与GenBank上公布的其他溶血性曼氏杆菌的同源性为97.1%~100.0%，与溶血性曼氏杆菌美国D171参考株同源性高达100%；以上结果表明分离菌为溶血性曼氏杆菌，命名为FJ-FQ2018。药物敏感性试验表明分离株对氟苯尼考、恩诺沙星、头孢噻吩、头孢唑啉、环丙沙星等高敏，对氨苄西林中敏，对多粘菌素B、林可霉素低敏。结论福建某养殖场引进的山羊发生肺炎后经病原分离鉴定确诊为溶血性曼氏杆菌感染，分离菌株对8种抗生素的耐药性存在差异。
- 山羊 /
- 溶血性曼氏杆菌 /
- 分离鉴定 /
- 药敏试验
Abstract: Objective To identify and study the pathogen that caused the goat pneumonia in a farm in Fuqing, Fujian, and provide reference for the isolation, identification and detection of goat Mannheimia. Method Bacterial isolation and culture were conducted on the tissues taken by autopsy on the diseased goats. ORF virus, mycoplasma, ovipneumonia and mycoplasma cluster were performed by PCR. A microorganism was isolated and identified by 16S rRNA gene cloning and sequencing combined with PCR. Subsequently, sensitivities of the isolated strain toward various drugs were tested. Result The PCR detection showed an unique amplified band of the isolated strain that matched Mannheimia haemolytica. The 16S rRNA sequencing of the strain shared 97.1%-100.0% similarities with those of other strains of M. haemolytica published in GenBank which included an 100% homology with the D171 strain. Positively confirmed to be of the M. haemolytica family, the isolate was code-named FJ-FQ2018. The drug sensitivity test showed the isolate to be highly sensitive to flufenicol, enoxacin, cephalothiophene, cefazoline, ciprofloxacin, et al, moderately sensitive to ampicillin, and slightly sensitive to polymyxin B and lincomycin. Conclusion The pathogen of goat pneumonia isotaled from Fujian was a strain of M. haemolytica, and the drug sensitivities of the isolate was deemed varions toward 8 antibiotics.
- goat /
- Mannheimia haemolytica /
- isolation and identification /
- antibiotic resistance

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图 1 目的片段的PCR扩增

注：M为2 000 DNA Ladder，1为溶血性曼氏杆菌FJ-FQ2018 DNA，2为阴性对照。图 2同。Note：M=2 000 DNA Ladder，1=DNA of M. haemolytica FJ-FQ2018，2=negative reference.The same as Fig. 2.

Figure 1. PCR results of target DNA fragment

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图 2 16S rRNA PCR扩增

Figure 2. PCR results of 16S rRNA fragment of DNA

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图 3 16S rRNA基因的进化分析

Figure 3. Phylogenetic tree analysis on 16S rRNA gene

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表 1 溶血性曼氏杆菌参考株

Table 1. Reference strains of M. haemolytica

菌株 Bacterial strains	登录号 Locus	分离地 Isolate site	分离时间 Isolate time
USDA-ARS-USMARC-184	CP006957	USA	2008
187	CP023046	USA	2011
191	CP023044	USA	2010
D153	CP005972	USA	1983
NCTC10609	LS483299	UK	2018
PHL213	DQ301920	Denmark	2005
Th1405	AB621551	Japan	2011
XJKLMY-10-NF	JQ975940	China XinJjiang	2012
NIVEDI/MHS-2	KU598685	India	2015
M42548	CP005383	German	2013
GDZJcattle2014	KM576848	China Guangdong	2014
MB1401	KT013278	Belgium	2015
XJNKY-125-YF1	JX984986	China XinJjiang	2012
D171	CP006573	USA	2013

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