黄鸡源沙门菌分离鉴定及毒力和耐药性分析

doi:10.16431/j.cnki.1671-7236.2023.04.040

摘要/Abstract

摘要： 【目的】探究规模化黄鸡孵化场鸡沙门菌流行株的毒力和耐药情况。【方法】采集弱雏卵黄拭子,接种选择培养基分离细菌,挑取疑似菌落进行革兰氏染色镜检,并对沙门菌属特异性基因invA进行PCR扩增鉴定。采用Kirby-Bauer纸片扩散法进行药敏试验,并通过PCR扩增耐药基因对其进行耐药性分析。利用SPF昆明小鼠对分离株进行致病性试验,并采用PCR法扩增相关毒力基因。【结果】分离菌株中有5株(JSS-Q3、JSS-Q4、JSS-Q5、JSS-Q8和ZJS-C1-11)疑似沙门菌,在伊红-美蓝培养基上形成无色透明的小菌落,在沙门菌显色培养基形成紫色透亮的小菌落,且镜检为短杆状革兰氏阴性菌。沙门菌特异性基因invA PCR扩增阳性,测序比对结果显示与沙门菌相似性达99%;鸡伤寒沙门菌特异性基因(IpaJ) PCR扩增阳性,分离到的5株菌株均为鸡伤寒沙门菌。分离菌株共检出15种毒力基因,其中11种毒力岛基因检出率为100%。耐药性分析表明,5株分离菌株耐药性较强,仅对部分抗菌药敏感,分离菌株对头孢吡肟、氟苯尼考、恩诺沙星表现敏感;另JSS-Q5、JSS-Q8分别对多黏菌素B、新霉素敏感,JSS-Q3对头孢他啶、大观霉素敏感性较好,ZJS-C1-11对头孢克洛、大观霉素以及多黏菌素B敏感性较好。部分菌株携带bla_SHV、bla_TEM、tet (A)、tet (B)等耐药基因。致病性分析发现JSS-Q8和ZJS-C1-11菌株毒性较强,对小鼠具有致死性。【结论】本研究涉及的黄鸡孵化场沙门菌分离株为鸡伤寒沙门菌,均表现多重耐药且部分致病性较强,可能是影响该孵化场孵化率和弱雏率的重要原因之一。研究结果为沙门菌病的检测、诊断及治疗提供一定的试验依据。

关键词: 沙门菌; 鸡; 多重耐药; 毒力基因; 耐药基因

Abstract: 【Objective】 The purpose of this study was to investigate the prevalence virulence and drug resistance of salmonella strains isolated from Yellow chicken in a large-scale hatchery.【Method】 The yolk swabs of weak chicks were collected and inoculated with selective medium for purifying and isolating bacteria.The isolated strains were identificated by Gram staining,microscopic examination and PCR amplification were carried out according to the Salmonella specific gene invA.Kirby-Bauer disk diffusion method was used for drug sensitivity test and amplify drug resistance genes to investigate and analyze the bacterial resistance of clinical isolates.The pathogenicity of the isolates was tested by SPF Kunming mice and the related virulence genes were amplified by PCR. 【Result】 The results showed that five isolated strains (JSS-Q3, JSS-Q4, JSS-Q5, JSS-Q8 and ZJS-C1-11) were suspected to be Salmonella.Colorless transparent colonies were formed in Eosin-Methylene blue medium,and purple transparent colonies were formed in Salmonella chromogenic medium,which were detected as short rod-shaped Gram-negative bacteria.Salmonella specific gene (invA) was positive and 99% similarity with Salmonella and the specific gene (IpaJ) amplification of Salmonella Gallinarum was positive,so the five isolated strains were all Salmonella Gallinarum.The results of virulence gene amplification showed that a total of fifteen virulence genes were detected,of which eleven were 100%.Drug resistance analysis showed that five isolated strains had strong resistance and were only sensitive to some antimicrobial agents.The isolates were sensitive to cefepime,flufenicol and enrofloxacin.In addition,JSS-Q5 and JSS-Q8 were sensitive to polymyxins B and neomycin,respectively,JSS-Q3 was sensitive to ceftazme and macromycin,and ZJS-C1-11 was sensitive to cefaclor,macromycin and polymyxins B.Some strains carried bla_SHV,bla_TEM,tet(A),tet(B) and other drug-resistant genes.Pathogenicity analysis showed that JSS-Q8 and ZJS-C1-11 strains were highly virulent and lethal to mice.【Conclusion】 The Salmonella isolates from Yellow chicken hatchery involved in this study were Salmonella Gallinarum.All of them showed multiple drug resistance and some of them were highly pathogenic,which might be one of the important reasons affecting the hatchery hatching rate and weak chick rate.The results of this study provided certain experimental basis for the detection,diagnosis and treatment of salmonella.

Key words: Salmonella; chicken; multiple drug resistance; virulence gene; resistance gene

中图分类号:

S831.7

吴植, 张聪, 袁慧莎, 董洪燕, 戴利霞, 贾鲁起, 吴双, 朱善元. 黄鸡源沙门菌分离鉴定及毒力和耐药性分析[J]. 中国畜牧兽医, 2023, 50(4): 1695-1706.

WU Zhi, ZHANG Cong, YUAN Huisha, DONG Hongyan, DAI Lixia, JIA Luqi, WU Shuang, ZHU Shanyuan. Isolation,Identification,Virulence and Drug Resistance of Salmonella from Yellow Chicken[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1695-1706.

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