麻旺鸭源大肠杆菌耐药表型分析及β-内酰胺酶、PMQR基因检测

doi:10.11843/j.issn.0366-6964.2024.04.035

摘要/Abstract

摘要： 本研究旨在了解麻旺鸭源大肠杆菌对β-内酰胺类、氨基糖苷类、四环素类、酰氨醇类、氟喹诺酮类和磺胺类药物的耐药情况以及β-内酰胺酶基因和质粒介导的喹诺酮耐药(plasmid-mediated quinolone resistance，PMQR)基因流行情况。从重庆市酉阳县4个麻旺鸭养殖场采集146份泄殖腔拭子样品，经选择性增菌和PCR扩增大肠杆菌特异性phoA基因分离鉴定大肠杆菌。采用纸片扩散法测定分离菌对25种抗菌药物的耐药性并检测分离菌株中的β-内酰胺酶基因和PMQR基因。结果显示：共分离到大肠杆菌146株。这些菌株对氨苄西林、头孢唑啉耐药严重，耐药率超过50%；对庆大霉素、阿米卡星、氧氟沙星完全敏感。分别有111株(76.0%)和83株(56.8%)菌携带至少1种β-内酰胺酶基因和PMQR基因，其中bla_TEM(74.0%，108/146)和qnrS(54.8%，80/146)基因最为流行。63株菌同时携带β-内酰胺酶基因和PMQR基因。麻旺鸭源大肠杆菌对常用抗菌药物存在耐药，且广泛携带β-内酰胺酶基因和PMQR基因。

关键词: 大肠杆菌, 麻旺鸭, 药物敏感性, PCR, bla_TEM, qnrS

Abstract: This experiment was conducted to study the antimicrobial resistance of Escherichia coli isolated from Mawang ducks to β-lactams, aminoglycosides, tetracyclines, amphenicols, fluoroquinolones and sulfonamides, and dissect the prevalence of β-lactamase genes and plasmid-mediated quinolone resistance (PMQR) genes in E. coli isolates. One hundred and forty six cloacal swab samples were collected from Mawang ducks at four farms in Youyang county, Chongqing municipality. E. coli was isolated and identified by selective enrichment and PCR amplification of the E. coli specific phoA gene. Antimicrobial resistance to 25 antimicrobials was determined using the disk diffusion method on Mueller-Hinton plates. The presence of β-lactamase genes and PMQR genes were detected by PCR. A total of 146 E. coli isolates were obtained from Mawang ducks. These isolates were severely resistant to ampicillin and cefazolin, with resistance rates of over 50%. However, they were completely susceptible to gentamicin, amikacin and ofloxacin. One hundred and eleven isolates (76.0%) and 83 isolates (56.8%) carried at least one type of β-lactamase genes and PMQR genes, respectively. The bla_TEM (74.0%, 108/146) and qnrS (54.8%, 80/146) genes were the most prevalent β-lactamase gene and PMQR gene, respectively. Sixty-three isolates carried β-lactamase gene(s) and PMQR gene(s) simultaneously. Mawang duck-derived E. coli was resistant to commonly used antimicrobials and carried β-lactamase genes and PMQR genes extensively.

Key words: Escherichia coli, Mawang duck, antimicrobial susceptibility, PCR, bla_TEM, qnrS

中图分类号:

S852.61

金睿妍, 张阳, 黄仲舜, 冉维, 丁红雷. 麻旺鸭源大肠杆菌耐药表型分析及β-内酰胺酶、PMQR基因检测[J]. 畜牧兽医学报, 2024, 55(4): 1738-1746.

JIN Ruiyan, ZHANG Yang, HUANG Zhongshun, RAN Wei, DING Honglei. Phenotype Analysis of Antimicrobial Resistance and β-lactamase, PMQR Genes Detection of Escherichia coli Isolated from Mawang Ducks[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(4): 1738-1746.

参考文献

[1] ZHANG C L, WANG Y M, XING Y C, et al. Investigation report on germplasm characteristics of Mawang ducks[J]. Shanghai Journal of Animal Husbandry and Veterinary Medicine, 2005(6):26-27. (in Chinese)
张昌莲, 王阳铭, 邢豫川, 等. 麻旺鸭种质特性调查报告[J]. 上海畜牧兽医通讯, 2005(6):26-27.
[2] LAI J C, MU H, ZHOU B Q, et al. Bla_TEM-positive Salmonella enterica serovars Agona and Derby are prevalent among food-producing animals in Chongqing, China[J]. Front Microbiol, 2023, 14:1011719.
[3] Clinical and Laboratory Standards Institute. Performance standards for antimicrobial susceptibility testing:Thirty-second informational supplement M100-S32[M]. Wayne, PA:CLSI. 2022.
[4] Clinical and Laboratory Standards Institute. Performance standards for antimicrobial disk and dilution susceptibility tests for bacteria isolated from animals, approved standard-fourth edition (VET08-A4)[M]. Wayne, PA:CLSI. 2019.
[5] CHEN S, ZHAO S H, WHITE D G, et al. Characterization of multiple-antimicrobial-resistant Salmonella serovars isolated from retail meats[J]. Appl Environ Microbiol, 2004, 70(1):1-7.
[6] QIAO J, ZHANG Q, ALALI W Q, et al. Characterization of extended-spectrum β-lactamases (ESBLs)-producing Salmonella in retail raw chicken carcasses[J]. Int J Food Microbiol, 2017, 248:72-81.
[7] DALLENNE C, DA COSTA A, DECRÉ D, et al. Development of a set of multiplex PCR assays for the detection of genes encoding important β-lactamases in Enterobacteriaceae[J]. J Antimicrob Chemother, 2010, 65(3):490-495.
[8] ROBICSEK A, STRAHILEVITZ J, SAHM D F, et al. qnr prevalence in ceftazidime-resistant Enterobacteriaceae isolates from the United States[J]. Antimicrob Agents Chemother, 2006, 50(8):2872-2874.
[9] CATTOIR V, POIREL L, ROTIMI V, et al. Multiplex PCR for detection of plasmid-mediated quinolone resistance qnr genes in ESBL-producing enterobacterial isolates[J]. J Antimicrob Chemother, 2007, 60(2):394-397.
[10] KIM H B, WANG M H, PARK C H, et al. oqxAB encoding a multidrug efflux pump in human clinical isolates of Enterobacteriaceae[J]. Antimicrob Agents Chemother, 2009, 53(8):3582-3584.
[11] CHEN C L, FENG G, QIN Y, et al. Determination and analysis of antimicrobial resistance of Escherichia coli and Salmonella isolated from chickens in Chongqing area[J]. Heilongjiang Animal Science and Veterinary Medicine, 2019(12):85-88. (in Chinese)
陈春林, 冯刚, 秦杨, 等. 重庆地区鸡源大肠杆菌、沙门氏杆菌耐药性的测定与分析[J]. 黑龙江畜牧兽医, 2019(12):85-88.
[12] LV H L. Isolation, identification and drug resistance analysis of pathogenic Escherichia coli from some pig and chicken farms in Chongqing[D]. Chongqing:Southwest University, 2020. (in Chinese)
吕红林. 重庆部分养殖场猪鸡致病性大肠杆菌分离鉴定与耐药性分析[D]. 重庆:西南大学, 2020.
[13] XU J J, WEN J, ZHU Z F, et al. Isolation, identification and antimicrobial resistance detection of Escherichia coli and Salmonella from a duck farm in Suining, Sichuan[J]. Heilongjiang Animal Science and Veterinary Medicine, 2017(22):112-114, 119. (in Chinese)
徐俊杰, 文娟, 朱子凤, 等. 四川遂宁某鸭场大肠杆菌和沙门氏菌的分离鉴定及耐药性检测[J]. 黑龙江畜牧兽医, 2017(22):112-114, 119.
[14] ZHAO C J, OU J, LIN F G. Isolation, identification and antimicrobial sensitivity test of pathogenic Escherichia coli from duck farms in Wanzhou district, Chongqing[J]. Heilongjiang Aninal Science and Veterinary Medicine, 2016(22):136-137, 300. (in Chinese)
赵婵娟, 欧婧, 林发根. 重庆市万州区鸭场致病性大肠杆菌的分离鉴定与药敏试验[J]. 黑龙江畜牧兽医, 2016(22):136-137, 300.
[15] KHAN M R, AZAM S, AHMAD S, et al. Molecular characterization and epidemiology of antibiotic resistance genes of β-lactamase producing bacterial pathogens causing septicemia from tertiary care hospitals[J]. Antibiotics (Basel), 2023, 12(3):617.
[16] JIANG T, LI G L, HUANG L Y, et al. Genomic and phylogenetic analysis of a multidrug-resistant bla_NDM-carrying Klebsiella michiganensis in China[J]. Infect Drug Resist, 2023, 16:3109-3116.
[17] HE X H, XU L W, DAI H D, et al. Genomic characteristics of a multidrug-resistant ST648 Escherichia coli isolate co-carrying bla_KPC-2 and bla_CTX-M-15 genes recovered from a respiratory infection in China[J]. Infect Drug Resist, 2023, 16:3535-3540.
[18] ZHANG W H, LIN X Y, XU L, et al. CTX-M-27 producing Salmonella enterica serotypes Typhimurium and Indiana are prevalent among food-producing animals in China[J]. Front Microbiol, 2016, 7:436.
[19] ZHANG C Z, DING X M, LIN X L, et al. The emergence of chromosomally located bla_CTX-M-55 in Salmonella from foodborne animals in China[J]. Front Microbiol, 2019, 10:1268.
[20] LI L, LIAO X P, LIU Z Z, et al. Co-spread of oqxAB and bla_CTX-M-9G in non-Typhi Salmonella enterica isolates mediated by ST2-IncHI2 plasmids[J]. Int J Antimicrob Agents, 2014, 44(3):263-268.
[21] LUO G, ZOU H, REN S K, et al. Detection of O-antigen, virulence gene and drug sensitivity test of pathogenic Escherichia coli from duck[J]. China Animal Husbandry & Veterinary Medicine, 2023, 50(2):713-721. (in Chinese)
罗干, 邹宏, 任绍科, 等. 鸭源致病性大肠杆菌O抗原与毒力基因检测及药敏试验[J]. 中国畜牧兽医, 2023, 50(2):713-721.