Isolation, Identification and Antibiotic Resistance of Mannheimia Haemolytica from Diseased Goats in Fujin
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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。 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. -
Key words:
- goat /
- Mannheimia haemolytica /
- isolation and identification /
- antibiotic resistance
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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 timeUSDA-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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