山东省禽源产超广谱β-内酰胺酶大肠杆菌的分子流行病学分析

doi:10.16431/j.cnki.1671-7236.2024.01.041

摘要/Abstract

摘要： 【目的】分析山东省禽源产超广谱β-内酰胺酶（extended-spectrum β-lactamases，ESBLs）大肠杆菌的耐药情况、耐药机制、毒力基因、多位点序列分型（multilocus sequence typing，MLST）、亲缘性和Inc型质粒，为禽临床合理使用抗菌药物防治此类细菌病提供参考。【方法】复苏2021年4月－2021年7月分离自山东省禽泄殖腔拭子样本中的38株产ESBLs大肠杆菌，采用微量肉汤稀释法测定其对16种抗菌药物的敏感性，通过全基因组测序检测其携带的耐药基因、毒力基因、MLST、Inc型质粒，使用Parsnp构建菌株系统发育树。【结果】药敏试验结果显示，所有菌株对氨苄西林、头孢噻呋和头孢他啶耐药，对磺胺异噁唑（97.4%）、甲氧苄啶/磺胺甲噁唑（94.7%）、四环素（89.5%）、氟苯尼考（86.8%）、大观霉素（84.2%）耐药严重。全基因组分析结果显示，38株产ESBLs大肠杆菌的bla_CTX-M型中bla_CTX-M-55基因携带率为34.2%，bla_NDM型中bla_NDM-5和bla_NDM-4基因携带率分别为21.1%和10.5%，其携带大量介导临床中常用抗菌药物的耐药基因和黏附类、侵袭类、血清抗性、铁转运类毒力基因。MLST结果显示，ST10（13.2%）为产ESBLs大肠杆菌最流行的ST型，其次是ST616（10.5%）和ST746（10.5%）。系统发育树分析表明，ST10、ST616和ST746分别属于同一起源。Inc型质粒结果显示，IncFIB （AP001918）携带率最高（68.4%），其次为IncHI2（42.1%）。【结论】山东省禽源产ESBLs大肠杆菌耐药状况严峻，bla_CTX-M基因是导致大肠杆菌对超广谱β-内酰胺类抗菌药物耐药的主要原因，且该类菌株携带大量毒力基因，亟需对其加强监测。

关键词: 超广谱β-内酰胺酶; 碳青霉烯; 禽源大肠杆菌; 全基因组测序; 多位点序列分型

Abstract: 【Objective】 The experiment aims to analyze the drug resistance, the drug resistance mechanism, multilocus sequence typing(MLST), the virulence genes, the affinity and Inc-type plasmids of extended-spectrum β-lactamases (ESBLs) producing Escherichia coli of avian origin in Shandong province, so as to provide reference for the rational use of antibiotics in avian clinical prevention and treatment of such bacterial diseases.【Method】 38 strains of ESBLs producing E.coli, isolated from avian farms in Shandong during April 2021 to July 2021, were recovered.The antimicrobial susceptibility test of these isolates against 16 kinds of antimicrobials was determined by the broth microdilution method.The drug resistance, the virulence genes, MLST and Inc-type plasmids were detected by whole genome sequencing.Parsnp was used to analyze the phylogenetic relationship among these isolates.【Result】 The antimicrobial susceptibility test revealed that all strains were resistant to ampicillin, ceftiofur and ceftazidime.The resistance to sulfisoxazole (97.4%), trimethoprim/sulfamethoxazole (94.7%), tetracycline (89.5%), florfenicol (86.8%), spectinomycin (84.2%) was serious. Analysis results of whole genome sequencing showed that 34.2% of 38 strains of ESBLs producing E.coli carried bla_CTX-M-55 in bla_CTX-M type, 22.1% carried bla_NDM-5 and 10.5% carried bla_NDM-4 in bla_NDM type.ESBLs producing E.coli isolates carried a large number of resistance genes that mediate commonly used antimicrobials in clinical practice, and adhesion, invasion, serum resistance, iron transport virulence genes.MLST results revealed that ST10 (13.2%) was the most prevalent ST among ESBLs producing E.coli isolates, followed by ST616 (10.5%) and ST746 (10.5%).Phylogenetic tree analysis showed that ST10, ST616 and ST746 had the same origin, respectively.Inc-type plasmids results revealed that the carrier rate of IncFIB (AP001918) was the highest (68.4%), followed by IncHI2 (42.1%).【Conclusion】 The situation of ESBLs producing E.coli isolates remains very serious, bla_CTX-M mainly accounted for the resistance to extended-spectrum β-lactams antimicrobials.Moreover, this type of strain carried a large number of virulence genes, and it was urgent to strengthen monitoring.

Key words: extended-spectrum β-lactamases; carbapenem; avian Escherichia coli; whole genome sequencing; multi-locus sequence typing

中图分类号:

S852.61⁺2

宋祥彬, 赵晓雨, 李传溥, 门晓冬, 梁萌, 魏秀丽, 李有志, 杨志昆, 张德琛, 郭玉秋, 汤文利. 山东省禽源产超广谱β-内酰胺酶大肠杆菌的分子流行病学分析[J]. 中国畜牧兽医, 2024, 51(1): 407-416.

SONG Xiangbin, ZHAO Xiaoyu, LI Chuanpu, MEN Xiaodong, LIANG Meng, WEI Xiuli, LI Youzhi, YANG Zhikun, ZHANG Dechen, GUO Yuqiu, TANG Wenli. Molecular Epidemiological Analysis of Avian Extended Spectrum β-lactamases Producing Escherichia coli in Shandong Province[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(1): 407-416.

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