广东某种鸡场鸡白痢沙门菌的分离鉴定及耐药性、毒力基因检测  

Isolation,Identification and Detection of Drug Resistance and Virulence Genes of Salmonella Pullorum from a Breeding Chicken Farm of Guangdong

作　　者：周迪 魏科 段千禧 曹伟胜[1,2,3,4] ZHOU Di;WEI Ke;DUAN Qianxi;CAO Weisheng(College of Veterinary Medicine,South China Agricultural University,Guangzhou 510642,China;Guangdong Provincial Key Laboratory of Prevention and Control for Animal-Originated Zoonotic Diseases,Guangzhou 510642,China;National-Local Joint Engineering Laboratory for Zoonotic Disease Control and Prevention,Guangzhou 510642,China;Key Laboratory of Veterinary Vaccine Innovation of the Ministry of Agriculture and Rural Affairs,Guangzhou 510642,China)

机构地区：[1]华南农业大学兽医学院,广州510642 [2]广东省动物源性人兽共患病预防与控制重点实验室,广州510642 [3]人兽共患病防控制剂国家地方联合工程实验室,广州510642 [4]农业农村部兽用疫苗创制重点实验室,广州510642

出　　处：《中国畜牧兽医》2024年第11期5033-5042,共10页China Animal Husbandry & Veterinary Medicine

基　　金：广东省家禽产业技术体系疾病控制岗位专家(2023KJ128);国家肉鸡产业技术体系(CARS-41)。

摘　　要：【目的】了解某种鸡场鸡白痢沙门菌感染情况,并对其耐药性及毒力基因情况进行分析,旨在为鸡白痢防控提供依据。【方法】试验采集疑似患鸡白痢的病鸡肝脏病料进行沙门菌的分离培养,利用特异性引物通过PCR扩增对分离株进行菌种鉴别。使用多位点序列分型(MLST)技术获得其分子分型,利用琼脂稀释法测定分离株的最小抑菌浓度;运用二代基因测序技术分析分离株的耐药性与毒力基因特征。【结果】本研究共分离获得5株鸡白痢沙门菌分离株,分别命名为SPNH05-1~SPNH05-5。MLST分型结果显示,5株分离株均为ST92型。药敏试验结果显示,分离株均为多重耐药,共存在2种耐药谱:氨苄西林+萘啶酸+磺胺异噁唑和氨苄西林+萘啶酸+磺胺异噁唑+多黏菌素B。对分离株SPNH05-1、SPNH05-2进行二代基因测序,结果显示,2株分离株存在β-内酰胺类、喹诺酮类、氨基糖苷类、磺胺类、多肽类等28种耐药基因,存在151种毒力基因,主要与沙门菌黏附、三型分泌系统、应激、毒素等有关。【结论】本研究结果表明该种鸡场存在多重耐药鸡白痢沙门菌感染,分离株携带多种耐药和毒力基因,试验结果为该种鸡场鸡白痢净化提供参考。【Objective】The purpose of this experiment was to understand the infection of Salmonella Pullorum in chickens of a breeding farm,and analyze the drug resistance and virulence genes,in order to provide evidence for the prevention and control of pullorosis.【Method】The liver tissue of sick chickens suspected to be suffering from pullorosis was collected and Salmonella was isolated and cultured.The isolates were identified by PCR amplification using specific primers.Molecular typing was obtained by multilocus sequence typing(MLST)technique.The minimum inhibitory concentration of the isolates was determined by agar dilution method.Next-generation sequencing(NGS)technology was utilized to analyze the drug resistance and virulence genes.【Result】A total of 5 isolates of Salmonella Pullorum were isolated and named SPNH05-1 to SPNH05-5,respectively.MLST typing results showed that all the 5 isolates were ST92.The results of drug sensitivity test showed that the isolates were multi-drug resistant,and there were 2 drug resistance profiles,ampicillin+nalidixic acid+sulfamisoxazole and ampicillin+nalidixic acid+sulfamisoxazole+polycolistin B.NGS was performed on isolates SPNH05-1 and SPNH05-2,which showed that there were 28 drug resistance genes,includingβ-lactam,quinolones,aminoglycosides,sulfonamides and polypeptides,and 151 virulence genes,which were mainly related to Salmonella adhesion,typeⅢsecretion system,stress and toxin.【Conclusion】The results of this study showed that multi-drug resistant Salmonella Pullorum infection existed in this breeding farm,and the isolates carried multiple drug resistance and virulence genes.The results provided reference for the purification of pullorosis.

关 键 词：鸡白痢沙门菌 分离鉴定 耐药性 毒力基因 

分 类 号：S852.6[农业科学—基础兽医学]