广东地区畜禽源大肠杆菌的耐药性与流行性研究

doi:10.16431/j.cnki.1671-7236.2024.02.040

摘要/Abstract

摘要： 【目的】调查广东地区畜禽源大肠杆菌的耐药特征与流行情况,为动物源大肠杆菌耐药性防控提供参考。【方法】对2019年从广东地区10个不同生产用途养殖场采集饲料、饮水和健康动物的肛/泄殖腔拭子等样品进行大肠杆菌分离鉴定、药敏试验、耐药基因和插入序列共同区(insertion sequence common region,ISCR)检测及种族系统进化分析。【结果】共分离获得416株大肠杆菌,其中种猪源78株、肉猪源77株、蛋鸡源122株、肉鸡源139株。菌株呈现多重耐药特点,不同动物源大肠杆菌中,肉鸡源大肠杆菌耐药率最高。79.6%(331/416)的菌株对3种或3种以上药物产生耐药,最多可对13种药物耐药,共组成156种耐药图谱,耐药谱以复方新诺明/氟苯尼考/多西环素/阿莫西林/氨苄西林居多,占比为11.1%。耐药基因以floR基因检出率最高,为50.0%;其次是sul2和sul3基因,检出率分别为44.2%和42.5%。与替加环素、黏菌素耐药表型相关的耐药基因tet(X3)、tet(X4)和mcr-1检出率均低于10%。插入序列共同区ISCR1、ISCR2和ISCR3的检出率分别为14.4%、10.8%和10.6%。酰胺醇类、磺胺类和β-内酰胺类耐药基因与耐药菌株的符合率在60%以上。耐药基因cmlA、tetA、qnrS、bla_CTX-M-9G与ISCR1的共存有统计学意义(P＜0.05);tet(X4)、sul2基因与ISCR2的共存有统计学意义(P＜0.05);tet(X4)、sul3、qnrS、bla_SHV基因与ISCR3的共存有统计学意义(P＜0.05)。不同动物源大肠杆菌以A群分布为主,ISCR相关移动元件阳性菌主要分布在A群。【结论】广东部分地区动物源大肠杆菌呈现多重耐药现状,携带多种耐药基因及可移动元件ISCR。本研究结果可为掌握广东地区动物源细菌耐药特征及耐药基因的流行情况提供依据。

关键词: 大肠杆菌; 畜禽; 耐药性; 耐药基因

Abstract: 【Objective】 The aim of this experiment was to investigate the prevalence and drug resistance of Escherichia coli (E. coli) from livestock and poultry in some areas of Guangdong, and provide reference for drug resistance prevention and control of E. coli derived from animals.【Method】 Samples of feed, drinking water and anal (cloaca) swabs of healthy animals collected from 10 farms in some areas of Guangdong in 2019 were subjected to E. coli isolation and identification, drug sensitivity test, detection of drug resistance genes and insertion sequence common region (ISCR), as well as ethnic phylogenetic analysis.【Result】 A total of 416 E. coli strains were isolated, including 78 strains from breeding pigs, 77 strains from broilers, 122 strains from laying hens and 139 strains from broilers.These strains showed the characteristics of multiple drug resistance, and the drug resistance rate of E. coli from broilers was the highest among different animal sources.79.6% (331/416) strains were resistant to 3 or more drugs, up to 13 drugs, and a total of 156 drug resistance profiles were formed.The main resistance profiles was sulfamethoxazole-trimethoprim/florfenicol/doxycycline/amoxicillin/ampicillin, accounting for 11.1%. floR gene showed the highest detection rate of 50.0%, followed by sul2 and sul3 genes, which were 44.2% and 42.5%, respectively.The detection rates of tet(X3), tet(X4) and mcr-1 genes associated with tigecycline and colistin resistance phenotypes were all lower than 10%.Insert sequence common region (ISCR) elements for ISCR1, ISCR2 and ISCR3 were present in 14.4%, 10.8% and 10.6% isolates, respectively.The coincidence rate of phenicols, sulfonamides and β-lactam resistance genes with drug-resistant strains was more than 60%, respectively.The coexistence of drug resistance genes cmlA, tetA, qnrS, bla_CTX-M-9G with ISCR1, tet(X4), sul2 with ISCR2, and tet(X4), sul3, qnrS, bla_SHV with ISCR3, were statistically significant (P＜0.05), respectively.E. coli from different animals and ISCR-related mobile element-positive bacteria were both mainly distributed in phylogenetic group A.【Conclusion】 E. coli isolates from animals in some areas of Guangdong presented multiple drug resistance, and carried multiple drug resistance genes, as well as mobile elements ISCR.These results provided reference for understanding the drug resistance characteristics and prevalence of resistant genes in animal-derived bacteria in Guangdong province.

Key words: Escherichia coli; livestock and poultry; drug resistance; resistance genes

中图分类号:

S852.61⁺2

李亚菲, 梁晓云, 吴荔琴, 肖田安, 王富华, 曾振灵. 广东地区畜禽源大肠杆菌的耐药性与流行性研究[J]. 中国畜牧兽医, 2024, 51(2): 837-849.

LI Yafei, LIANG Xiaoyun, WU Liqin, XIAO Tianan, WANG Fuhua, ZENG Zhenling. Drug Resistance and Prevalence of Escherichia coli Isolated from Livestock and Poultry in Areas of Guangdong[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(2): 837-849.

参考文献

[1] MUSHTAQ H,BAKHT J,KHAN I,et al.Antimicrobial efficacy and prevalence of colicinogenic E. coli in faecal matter of human,cow and sheep[J].International Journal of Antimicrobial Agents,2021,57(1):106221.
[2] LAIRD T J,ABRAHAM S,JORDAN D,et al.Porcine enterotoxigenic Escherichia coli:Antimicrobial resistance and development of microbial-based alternative control strategies[J].Veterinary Microbiology,2021,258:109117.
[3] COX G W,AVERY B P,PARMLEY E J,et al.A one health genomic investigation of gentamicin resistance in Escherichia coli from human and chicken sources in Canada,2014 to 2017[J].Antimicrobial Agents and Chemotherapy,2022,66(10):e0067722.
[4] HAMAME A,DAVOUST B,HASNAOUI B,et al.Screening of colistin-resistant bacteria in livestock animals from France[J].Veterinary Research,2022,53(1):96.
[5] PENG Z,HU Z,LI Z,et al.Antimicrobial resistance and population genomics of multidrug-resistant Escherichia coli in pig farms in mainland China[J].Nature Communications,2022,13(1):1116.
[6] SODAGARI H R,VARGA C.Evaluating antimicrobial resistance trends in commensal Escherichia coli isolated from cecal samples of swine at slaughter in the United States,2013-2019[J].Microorganisms,2023,11(4):1033.
[7] NOWAKIEWICZ A,ZIEBA P,GNAT S,et al.A significant number of multi-drug resistant Enterococcus faecalis in wildlife animals;Long-term consequences and new or known reservoirs of resistance?[J].Science of the Total Environment,2020,705:135830.
[8] BRENCIANI A,MORRONI G,SCHWARZ S,et al.Oxazolidinones:Mechanisms of resistance and mobile genetic elements involved[J].Journal of Antimicrobial Chemotherapy,2022,77(10):2596-2621.
[9] BENZ F,HALL A R.Host-specific plasmid evolution explains the variable spread of clinical antibiotic-resistance plasmids[J].Proceedings of the National Academy of Sciences of the United States of America,2023,120(15):e2212147120.
[10] CASTAÑEDA-BARBA S,TOP E M,STALDER T.Plasmids,a molecular cornerstone of antimicrobial resistance in the one health era[J].Nature Reviews Microbiology,2023,10:1038.
[11] 王亮亮.猪源主要肠杆菌科细菌替加环素耐药基因tet (X)变异体的流行与传播特征研究[D].郑州:河南农业大学,2021. WANG L L.Epidemic characteristics and transmission of tigecycline resistance gene tet(X) variants in swine Enterobacteriaceae[D].Zhengzhou:Henan Agricultural University,2021.(in Chinese)
[12] CHEN L,CHEN Z,LIU J,et al.Emergence of RmtB methylase-producing Escherichia coli and Enterobacter cloacae isolates from pigs in China[J].The Journal of Antimicrobial Chemotherapy,2007,59(5):880-885.
[13] BOERLIN P,TRAVIS R,GYLES C L,et al.Antimicrobial resistance and virulence genes of Escherichia coli isolates from swine in Ontario[J].Applied and Environmental Microbiology,2005,71(11):6753-6761.
[14] DOI Y,ARAKAWA Y.16S ribosomal RNA methylation:Emerging resistance mechanism against aminoglycosides[J].Clinical Infectious Diseases,2007,45(1):88-94.
[15] SUN N,LIU J,YANG F,et al.Molecular characterization of the antimicrobial resistance of Riemerella anatipestifer isolated from ducks[J].Veterinary Microbiology,2012,158(3-4):376-383.
[16] CLERMONT O,BONACORSI S,BINGEN E.Rapid and simple determination of the Escherichia coli phylogenetic group[J].Applied and Environmental Microbiology,2000,66(10):4555-4558.
[17] CLINICAL AND LABORATORY STANDARDS INSTITUTE.Performance standards for antimicrobial susceptibility testing[S].30th ed.CLSI supplement M100.Wayne,PA:Clinical and Laboratory Standards Institute,2020.
[18] 高玉斌,赵格,邹明,等.我国不同区域鸡源大肠杆菌的致病性与耐药性状分析[J].中国兽医杂志,2019,55(3):81-84. GAO Y B,ZHAO G,ZOU M,et al.Analysis of virulence gene carrying and drug resistance of Escherichia coli isolated from chickens in different regions of China[J].Chinese Journal of Veterinary Medicine,2019,55(3):81-84.(in Chinese)
[19] 王永亮.吉林省部分地区动物源大肠杆菌耐药性及多重耐药株全基因组序列分析[D].长春:吉林农业大学,2022. WANG Y L.Genome sequence analysis of drug-resistant and multidrug-resistant Escherichia coli strains from animals in some areas of Jilin province[D].Changchun:Jilin Agricultural University,2022.(in Chinese)
[20] 杨守深,曾雪花,林敏,等.猪源大肠杆菌耐药性及超广谱β-内酰胺酶流行性分析[J].中国人兽共患病学报,2019,35(1):45-50. YANG S S,ZENG X H,LIN M,et al.Antimicrobial resistance analysis and extended-spectrum-β-lactamaeses genes detection in Escherichia coli isolated from swine[J].Chinese Journal of Zoonoses,2019,35(1):45-50.(in Chinese)
[21] 朱琪琪,张继瑜,王承业,等.河北地区动物源大肠杆菌多重耐药性与流行性分析[J].中国畜牧兽医,2023,50(3):1081-1092. ZHU Q Q,ZHANG J Y,WANG C Y,et al.Analysis of multidrug resistance and prevalence of animal-derived Escherichia coli in Hebei[J].China Animal Husbandry&Veterinary Medicine,2023,50(3):1081-1092.(in Chinese)
[22] 陈月月,王凯,王舒丰,等.新疆伊犁地区不同动物源大肠杆菌耐药性调查[J].新疆农业科学,2018,55(8):1560-1568. CHEN Y Y,WANG K,WANG S F,et al.Investigation into drug resistance of Escherichia coli from different animal sources in Yili,Xinjiang[J].Xinjiang Agricultural Sciences,2018,55(8):1560-1568.(in Chinese)
[23] MOTTA S S,ALDANA M.Adaptive resistance to antibiotics in bacteria:A systems biology perspective[J].Wiley Interdisciplinary Reviews.Systems Biology and Medicine,2016,8(3):253-267.
[24] 张召兴,李佩国,李蕴玉,等.致蛋鸡卵黄性腹膜炎大肠杆菌分子分群、耐药表型及耐药基因型检测与分析[J].中国预防兽医学报,2019,41(9):885-890. ZHANG Z X,LI P G,LI Y Y,et al.Analyzing phylogenetic groups,drug resistance phenotypes,and drug resistance genotypes of Escherichia coli isolates causing salpingoperitionitis in layer chicken[J].Chinese Journal of Preventive Veterinary Medicine,2019,41(9):885-890.(in Chinese)
[25] 陈霞,车洁,骆鹏杰,等.肉鸡养殖场中耐药相关基因散播情况的调查研究[J].中国畜牧兽医,2020,47(3):931-939. CHEN X,CHE J,LUO P J,et al.Investigation on the spread of resistance-related genes in broiler farms[J].China Animal Husbandry&Veterinary Medicine,2020,47(3):931-939.(in Chinese)
[26] LIU D J,WANG T,SHAO D Y,et al.Structural diversity of the ISCR2-mediated rolling-cycle transferable unit carrying tet (X4)[J].Science of the Total Environment,2022,826:154010.