内蒙古地区犊牛腹泻大肠杆菌耐药性分析及毒力基因检测

doi:10.16431/j.cnki.1671-7236.2023.09.037

摘要/Abstract

摘要： 【目的】了解内蒙古地区犊牛腹泻大肠杆菌的耐药性及毒力基因携带情况,筛选敏感抗菌药物,指导临床上治疗犊牛腹泻合理用药。【方法】对采集到的犊牛腹泻病料进行大肠杆菌分离纯化、生化鉴定及分子生物学鉴定,采用药敏试验确定分离菌株的耐药表型及多重耐药情况,并对其进行耐药基因及毒力基因检测。【结果】分离菌株在伊红-美蓝培养基上呈紫黑色带有绿色金属光泽的菌落;与靛基质、甲基红和麦芽糖等反应阳性,柠檬酸盐、H₂S等反应阴性。PCR扩增出大小为262 bp的大肠杆菌特异性基因uida目的条带。经分离鉴定从44份病料中共得到32株犊牛腹泻大肠杆菌,分离率为72.7%。药敏试验结果显示,犊牛腹泻大肠杆菌呈现多重耐药,耐3种及3种以上抗菌药物的占比达到了84.4%;其中,对氨苄西林、头孢噻肟、环丙沙星、卡那霉素和复方新诺明耐药明显,对头孢吡肟、头孢西丁、多西环素、米诺环素和加替沙星相对敏感,尤其对美罗培南和多黏菌素B敏感。耐药基因中检出β-内酰胺类基因bla_TEM、bla_CTX-M,喹诺酮类耐药基因qnrA、qnrS,四环素类耐药基因tetA、tetB、tetD和氨基糖苷类耐药基因aadA、aadB、aacC、aac(3')Ⅱa,其中aadA和bla_CTX-M基因检出率最高,分别为100%和87.5%。在对10种毒力基因检测中,只检测到了Irp2、FyuA、astA、LT-Ⅱ和HlyA 5种毒力基因,而Sta、Stb、HlyE、Stx1和Stx2e 毒力基因未被检测到。【结论】内蒙古地区犊牛腹泻大肠杆菌多重耐药现象较严重,携带bla_CTX-M和aadA等多种耐药基因,携带Irp2和FyuA等毒力基因,建议临床上根据药敏试验结果科学合理地选择抗菌药物,有效做好疫病防控。

关键词: 犊牛腹泻; 大肠杆菌; 分离鉴定; 耐药性; 毒力基因

Abstract: 【Objective】 The purpose of the experiment was to understand the drug resistance and virulence gene carrying of Escherichia coli causing calf diarrhea in Inner Mongolia,screen sensitive antibacterial drugs,and guide the rational use of drugs in clinical treatment of calf diarrhea.【Method】 Isolation,purification,biochemical identification and molecular biological identification of Escherichia coli were carried out on the collected calf diarrhea materials.Drug sensitivity test was used to determine the drug resistance phenotype and multiple drug resistance of the isolated strains,and drug resistance genes and virulence genes were detected.【Result】 The isolated strain exhibited a purple black colony with a green metallic luster on Eosin-Methylene blue medium.They reacted positively with indole,methyl red and maltose,but negatively with citrate and H₂S.A 262 bp target band of Escherichia coli specific gene uida was amplified by PCR.A total of 32 strains of Escherichia coli were isolated from 44 diseased materials,with a separation rate of 72.7%.The results of drug sensitivity test showed that Escherichia coli causing calf diarrhea showed multiple drug resistance,and 84.4% of them were resistant to three or more antimicrobial agents,it was significantly resistant to ampicillin,cefotaxime,ciprofloxacin,kanamycin and compound sulfamethoxazole,and relatively sensitive to cefepime,cefoxitin,doxycycline,minocycline and gatifloxacin,especially sensitive to meropenem and polymyxin B.Among the resistance genes β-lactam genes bla_TEM,bla_CTX-M,quinolone resistance genes qnrA,qnrS,tetracycline resistance genes tetA,tetB,tetD and aminoglycoside resistance genes aadA,aadB,aacC,aac(3')Ⅱa were detected,of which aadA and bla_CTX-M genes had the highest detection rates of 100% and 87.5%,respectively.Among the 10 virulence genes tested,only 5 virulence genes Irp2,FyuA, astA,LT-Ⅱ and HlyA were detected,while Sta,Stb,HlyE,Stx1 and Stx2e virulence genes were not detected.【Conclusion】 The phenomenon of multiple drug resistance of Escherichia coli causing calf diarrhea in Inner Mongolia was relatively serious,carrying multiple drug resistance genes such as bla_CTX-Mand aadA,and carrying virulence genes such as Irp2 and FyuA.It was suggested that antibiotics should be selected scientifically and reasonably according to the drug sensitivity test results in clinical practice to effectively prevent and control epidemic diseases.

Key words: calf diarrhea; Escherichia coli; isolation and identification; drug resistance; virulence gene

中图分类号:

S852.61^＋2

孙月, 王琪, 毛伟, 王博, 董海燕, 樊宏亮, 郝普国, 赵红霞. 内蒙古地区犊牛腹泻大肠杆菌耐药性分析及毒力基因检测[J]. 中国畜牧兽医, 2023, 50(9): 3811-3822.

SUN Yue, WANG Qi, MAO Wei, WANG Bo, DONG Haiyan, FAN Hongliang, HAO Puguo, ZHAO Hongxia. Drug Resistance Analysis and Virulence Gene Detection of Escherichia coli Causing Calf Diarrhea in Inner Mongolia[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(9): 3811-3822.

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