陕西地区羊源致病性大肠杆菌耐药性分析与毒力基因检测

doi:10.11843/j.issn.0366-6964.2022.05.033

摘要/Abstract

摘要： 旨在了解陕西省部分地区腹泻羊源致病性大肠杆菌(E. coli)耐药性及毒力基因携带情况，本研究从10个养殖场采集54份腹泻羊拭子样品，经分离纯化、生化鉴定及16S rRNA基因序列分析，共分离得到50株E. coli，对分离菌进行药敏试验、耐药基因及毒力基因检测。结果显示，分离菌对氨苄西林、氟苯尼考和磺胺异噁唑耐药率达90%以上，且98%(49/50)为多重耐药菌，对8~11种抗生素耐药的菌株占68%(34/50)，仅对美罗培南敏感。所有菌株均携带1~6种不同的耐药基因，其中，Sul1(64%)、TetA(34%)、bla_CTX-M(32%)携带率较高，未检测到bla_S_HV。有5株产ESBLs的E. coli携带mcr-1耐药基因。毒力基因检测结果显示，98%(49/50)的菌株携带毒力基因，其中，etrA检出率最高，为80%(40/50)。综上表明，陕西省羊源E. coli多重耐药情况严峻，β-内酰胺类耐药基因与耐药表型不符，提示可能存在其他耐药机制，同时，分离菌具有复杂的毒力谱。本研究为陕西省羊源致病性E. coli感染的防控提供科学依据。

关键词: 大肠杆菌, 致病性, 耐药性, 耐药基因, 毒力基因

Abstract: The aim of this study was to investigate the antimicrobial resistance and virulence genes of Escherichia coli isolated from diarrhea sheep in Shaanxi Province. In this study, 54 diarrhea swabs were collected from 10 sheep farms. After purification, biochemical identification and 16S rRNA analysis, a total of 50 E. coli strains were isolated. For E. coli isolates, antibiotic resistance, resistance genes, and virulence genes were tested. The results showed that 49 E. coli strains display multiple-drug resistance, more than 90% of the strains were resistant to ampicillin, florfenicol, and sulfisoxazole, and 100% (50/50) of the strains were sensitive to meropenem. In addition, 8-11 resistant accounts for 68% of the total isolated strains (34/50). All strains carry 1-6 different resistance genes. Among them, Sul1 (64%), TetA (34%), bla_CTX-M (32%) had higher carrying rates, and bla_SHV (0%) was not detected. It is noteworthy that five ESBLs-strains carry the mcr-1 gene from the isolated. The results showed that the mortality rate of mice infected with the 50 strains of E. coli was 33%-100%, virulence genes were detected in 49 of the 50 isolates, with etrA showing the highest detection rate, accounting for 80% (40/50). The results indicated that the high proportion of virulence genes and the multidrug-resistance phenomena was prevalent in sheep pathogenic E. coli. The resistance phenotypes of β-lactams were basically not correlated with the resistance genes, suggesting that there might be multiple resistance mechanisms in the E. coli strains. The virulence spectrum of isolates was complex. This study provided a scientific basis for prevention and control of the pathogenic E. coli from sheep in Shaanxi area.

Key words: E. coli, pathogenic, antibiotic resistance, resistance genes, virulence genes

中图分类号:

S852.61

赵学亮, 王斌, 苗永强, 赵浩宇, 谢青芳, 王娟, 杨增岐. 陕西地区羊源致病性大肠杆菌耐药性分析与毒力基因检测[J]. 畜牧兽医学报, 2022, 53(5): 1644-1648.

ZHAO Xueliang, WANG Bin, MIAO Yongqiang, ZHAO Haoyu, XIE Qingfang, WANG Juan, YANG Zengqi. Detection of Virulence Genes and Antimicrobial Resistance Analysis of Escherichia coli Isolated from Diarrhea Sheep in Shaanxi Province[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(5): 1644-1648.

参考文献 18

[1]	SHAHZAD A, ULLAH F, IRSHAD H, et al. Molecular detection of Shiga toxin-producing Escherichia coli (STEC) O157 in sheep, goats, cows and buffaloes[J]. Mol Biol Rep, 2021, 48(10):6113-6121.doi:10. 1007/s11033-021-06631-3.
[2]	HABOURIA H, POKHAREL P, MARIS S, et al. Three new serine-protease autotransporters of Enterobacteriaceae (SPATEs) from extra-intestinal pathogenic Escherichia coli and combined role of SPATEs for cytotoxicity and colonization of the mouse kidney[J]. Virulence, 2019, 10(1):568-587.
[3]	WANG Y, XU C Y, ZHANG R, et al. Changes in colistin resistance and mcr-1 abundance in Escherichia coli of animal and human origins following the ban of colistin-positive additives in China:an epidemiological comparative study[J]. Lancet Infect Dis, 2020, 20(10):1161-1171.
[4]	HERNANDO-AMADO S, COQUE T M, BAQUERO F, et al. Defining and combating antibiotic resistance from One Health and Global Health perspectives[J]. Nat Microbiol, 2019, 4(9):1432-1442.
[5]	ZHAO X L, LV Y Y, ADAM F E A, et al. Comparison of antimicrobial resistance, virulence genes, phylogroups, and biofilm formation of Escherichia coli isolated from intensive farming and free-range sheep[J]. Front Microbiol, 2021, 12:699927.
[6]	MA M G, WANG H N, YU Y, et al. Detection of antimicrobial resistance genes of pathogenic Salmonella from swine with DNA microarray[J]. J Vet Diagn Invest, 2007, 19(2):161-167.
[7]	MATTER D, ROSSANO A, LIMAT S, et al. Antimicrobial resistance profile of Actinobacillus pleuropneumoniae and Actinobacillus porcitonsillarum[J]. Vet Microbiol, 2007, 122(1-2):146-156.
[8]	PEREZ F, HUJER A M, HUJER K M, et al. Global challenge of multidrug-resistant Acinetobacter baumannii[J]. Antimicrob Agents Chemother, 2007, 51(10):3471-3484.
[9]	LIU Y Y, WANG Y, WALSH T R, et al. Emergence of plasmid-mediated colistin resistance mechanism MCR-1 in animals and human beings in China:a microbiological and molecular biological study[J]. Lancet Infect Dis, 2016;16(2):161-168.
[10]	宋晓莉. 江苏部分地区肉羊源大肠杆菌的分子流行病学及其部分特性研究[D]. 扬州:扬州大学, 2019.SONG X L. Molecular epidemiology of Escherichia coli from fleshy sheep in parts of Jiangsu and its characteristics[D]. Yangzhou:Yangzhou University, 2019. (in Chinese)
[11]	刘晨阳. 羊源大肠杆菌的分离鉴定及耐药性与致病性分析[D]. 杨凌:西北农林科技大学, 2019.LIU C Y. Isolation and identification of Escherichia coli from goats and analysis of drug resistance and pathogenicity[D]. Yangling:Northwest A&F University, 2019. (in Chinese)
[12]	SHATALIN K, NUTHANAKANTI A, KAUSHIK A, et al. Inhibitors of bacterial H2S biogenesis targeting antibiotic resistance and tolerance[J]. Science, 2021, 372(6547):1169-1175.
[13]	ZHANG S Q, ABBAS M, REHMAN M U, et al. Updates on the global dissemination of colistin-resistant Escherichia coli:an emerging threat to public health[J]. Sci Total Environ, 2021, 799:149280.
[14]	VILLA L, GARCIA-FERNÁNDEZ A, FORTINI D, et al. Replicon sequence typing of IncF plasmids carrying virulence and resistance determinants[J]. J Antimicrob Chemother, 2010, 65(12):2518-2529.
[15]	刘润春, 张梦洁, 恽佳蕾, 等. 华东华北规模化羊场细菌耐药性的调查研究[J]. 中国兽医科学, 2021, 51(12):1599-1608.LIU R C, ZHANG M J, YUN J L, et al. Investigation of antimicrobial susceptibilities of bacterial on East China and North China sheep farms[J]. Chinese Veterinary Science, 2021,51(12):1599-1608. (in Chinese)
[16]	刘正明, 李金泉, 黄德浩, 等. 内蒙古地区羊源大肠杆菌耐药性研究[J]. 中国畜牧兽医, 2017, 44(3):839-846.LIU Z M, LI J Q, HUANG D H, et al. Study on the drug-resistance of Escherichia coli isolated from sheep in Inner Mongolia[J]. China Animal Husbandry & Veterinary Medicine, 2017, 44(3):839-846. (in Chinese)
[17]	REHMAN M UR, ZHANG H, IQBAL M K, et al. Antibiotic resistance, serogroups, virulence genes, and phylogenetic groups of Escherichia coli isolated from yaks with diarrhea in Qinghai Plateau, China[J]. Gut Pathog, 2017, 9(1):24.
[18]	张金宝,马春芳,余婷, 等. 宁夏地区奶牛乳房炎大肠杆菌毒力基因检测和耐药性分析[J]. 畜牧兽医学报, 2016, 47(1):177-182.ZHANG J B,MA C F,YU T,et al. Detection of virulence genes and antimicrobial resistance analysis of Escherichia coli isolated from dairy cow mastitis in Ningxia[J]. Acta Veterinaria et Zootechnica Sinica, 2016, 47(1):177-182.(in Chinese)