鸡肉生产链中沙门氏菌耐药性及脉冲场凝胶电泳分型分析

doi:10.16431/j.cnki.1671-7236.2023.08.032

摘要/Abstract

摘要： 【目的】了解拥有养殖-屠宰-运输-销售环节的现代化农牧食品企业鸡肉生产链各环节中沙门氏菌耐药情况及分子分型特征。【方法】本研究对在2020年10月—2021年09月从苏北某农牧食品企业下属的养殖场、屠宰厂、运输车辆、专营店分季度连续采集样品中分离得到的37株沙门氏菌,采用纸片扩散法进行13种常用抗菌药物敏感试验,PCR进行11种耐药基因检测、脉冲场凝胶电泳(PFGE)进行分子分型检测。【结果】37株沙门氏菌均耐药,对氨苄西林耐药率最高,为67.57%,对多西环素、庆大霉素、四环素耐药率超过50%,对头孢西丁耐药率最低,为8.11%,各环节沙门氏菌分离菌株对氨苄西林的耐药率均超过50%,沿该鸡肉生产链氨苄西林耐药率呈现下降趋势,耐药率同样呈现下降趋势的抗菌药有头孢他啶、头孢曲松、环丙沙星、四环素,耐药率呈现上升趋势的抗菌药有头孢西丁、甲氧苄氨嘧啶、庆大霉素、丁胺卡那霉素;耐3种及以上抗菌药的菌株为34株,多重耐药率为94.59%,养殖环节、运输环节和销售环节沙门氏菌分离株的多重耐药率均为100%;养殖环节沙门氏菌分离株耐药物种类数最多集中在耐7种及以上抗菌药,对8种抗菌药的耐药率高于其他环节;耐药基因中bla_PSE的检出率最高,为56.76%,未检测到tetC耐药基因;PFGE分型共有26种不同的PFGE带型产生,相似度100%的带型有7组。【结论】苏北某农牧食品企业鸡肉生产链中沙门氏菌有较高的耐药性,多重耐药严重,养殖环节耐药性高于其他环节,PFGE分型提示耐药菌株能够沿生产链"养殖-屠宰"的途径传播,且长期存在,养殖环节避免耐药沙门菌的产生是保障鸡肉产品安全生产关键点。

关键词: 鸡肉生产; 沙门氏菌; 耐药; 脉冲场凝胶电泳

Abstract: 【Objective】 To understand the drug resistance and molecular typing characteristics of Salmonellain the chicken production chain of modern agricultural and animal husbandry food enterprises with the links of breeding,slaughtering,transportation and sales.【Method】 In this study,37 strains of Salmonella isolated from the continuous samples collected from the farms,slaughterhouses,transport vehicles and franchised stores of an agricultural and animal husbandry food enterprise in Northern Jiangsu from October 2020 to September 2021 were tested for 13 common antibiotics by paper diffusion method,11 drug resistance genes by PCR,and molecular typing by pulsed field gel electrophoresis (PFGE).【Result】 The 37 strains of Salmonella isolated were all resistant,the highest resistance rate to ampicillin was 67.57%,the resistance rate to doxycycline,gentamicin and tetracycline was more than 50%,and the lowest resistance rate to cefoxitin was 8.11%.The resistance rate of Salmonella isolated from all links to ampicillin was more than 50%.Along the chicken production chain,the resistance rate of ampicillin showed a downward trend,and the drug resistance rate of ceftazidime,ceftriaxone,ciprofloxacin and tetracycline also showed a downward trend.The antibiotic resistance rate of cefoxitin,trimethoprim,gentamicin,amikacin showed an upward trend.34 strains were resistant to three or more antibiotics,and the multidrug resistance rate was 94.59%.The multidrug resistance rate of Salmonella isolated in breeding,transportation and marketing was 100%.The number of drug-resistant strains of Salmonella isolated from the links of breeding was mostly concentrated in 7 or more antibiotics,and the drug resistance rate to 8 antibiotics was higher than that of other links.The detection rate of drug resistance gene bla_PSE was the highest,and its detection rate was 56.76%.The drug resistance gene tetC was not detected.PFGE classification had produced 26 different PFGE belt types,and there were 7 groups of belt types with 100% similarity.【Conclusion】 Salmonella isolated from the chicken production chain of a modern agricultural and animal husbandry food enterprise in Northern Jiangsu had high drug resistance and serious multidrug resistance,and the drug resistance in the breeding link was higher than that in other links.PFGE typing suggested that the drug resistant strains could spread along the "Breeding-slaughtering"route of the chicken production chain,and existed for a long time.Avoiding the production of drug resistant Salmonella in the breeding link was the key point to ensure the safe production of chicken products.

Key words: chicken production; Salmonella; drug resistance; PFGE

中图分类号:

S855.1

王玉燕, 张林吉, 迟兰, 陈永亮, 任士飞, 孙朋. 鸡肉生产链中沙门氏菌耐药性及脉冲场凝胶电泳分型分析[J]. 中国畜牧兽医, 2023, 50(8): 3345-3353.

WANG Yuyan, ZHANG Linji, CHI Lan, CHEN Yongliang, REN Shifei, SUN Peng. Analysis of Salmonella Resistance and PFGE Typing in Chicken Production Chain[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(8): 3345-3353.

参考文献

[1] 孙菀霞,杜建萍,张春艳,等.生鸡肉生产链中沙门氏菌风险评估的研究进展[J].生物加工过程,2017,15(2):72-78. SUN W X,DU J P,ZHANG C Y,et al.Progress on risk assessment of Salmonella spp.during theraw chicken production chain[J].Chinese Journal of Bioprocess Engineerin,2017,15(2):72-78.(in Chinese)
[2] 张庆贺,张丹俊,李槿年,等.沙门氏菌耐药性研究进展[J].安徽农业科,2018,46(17):27-29. ZHANG Q H,ZHANG D J,LI J N,et al.Advances in Salmonella resistance[J]. Journal of Anhui Agricultural Sciences,2018,46(17):27-29.(in Chinese)
[3] 吴云凤,袁宝君,乔昕,等.肉鸡胴体中沙门氏菌的分离鉴定及多重耐药谱研究[J].南京医科大学学报(自然科学版),2012,32(1):125-128. WU Y F,YUAN B J,QIAO X,et al.Separation and identification of multiple-antimicrobial-resistant of Salmonella on chicken carcass[J].Journal of Nanjing Medical University(Natural Science Edition),2012,32(1):125-128.(in Chinese)
[4] WALLIS C V,LOWDEN P,MARSHALL-JONES Z V,et al.Distinct fermentation and antibiotic sensitivity profiles exist in Salmonellae of canine and human origin[J].BMC Microbiology,2018,18(1):15-16.
[5] KUMAR N,MOHAN K,GEORGES K,et al.Occurrence of virulence and resistance genes in Salmonella in cloacae of slaughtered chickens and ducks at pluck shops in Trinidad[J].Journal of Food Protection,2021,84(1):39-46.
[6] PARVIN S M,HASAN M M,ALI Y M,et al.Prevalence and multidrug resistance pattern of Salmonella carrying extended-spectrum β-lactamase in frozen chicken meat in Bangladesh[J].Journal of Food Protection,2020,83(12):2107-2121.
[7] YU X,ZHU H W,BO Y H,et al.Prevalence and antimicrobial resistance of Salmonella enterica subspecies enterica serovar Enteritidis isolated from broiler chickens in Shandong province,China,2013-2018[J].Poultry Science,2021,100(2):1016-1023.
[8] CLINICAL AND LABORATORY STANDARDS INSTITUTE.Performance standards for antimicrobial susceptibility testing 29th information supplement M100[S].Wayne,Pennsylvania:Clinical and Laboratory Standards Institute,2019.
[9] ZELENDOVA M,DOLEJSKA M,MASARIKOVA M,et al.CTX-M-producing Escherichia coli in pigs from a Czech farm during production cycle[J].Letters Applied Microbiology,2020,71(4):369-376.
[10] 宋艳.山东省部分种鸡场死胚中沙门氏菌的鉴定及耐药特性研究[D].泰安:山东农业大学,2020. SONG Y.Identification of Salmonella in dead embryos and study on their drug resistance characteristics of some farms in Shandong province[D].Tai'an:Shandong Agricultural University,2020.(in Chinese)
[11] GUERRA B,SOTO S M,ARGVELLES J M,et al.Multidrug resistance is mediated by large plasmids carrying a class 1 integron in the emergent Salmonella enterica serotype[4,5,12:i:-][J].Antimicrobial Agents and Chemotherapy,2001,45(4):1305.
[12] 沈赟,秦思,郑东宇,等.江苏省2015-2019年沙门氏菌的血清型和耐药状况分析[J].现代预防医学,2021,48(12):2263-2267. SHEN Y,QIN S,ZHENG D Y,et al.Serotype and antimicrobial resistance of Salmonella isolated from 2015 to 2019 in Jiangsu province[J].Modern Preventive Medicine,2021,48(12):2263-2267.(in Chinese)
[13] RIBOT E M,FAIR M A,GAUTOM R,et al.Standardization of pulsed-field gel electrophoresis protocols for the subtyping of Escherichia coli O157:H7, Salmonella, and Shigella for pulsenet[J].Foodborne Pathogens and Disease,2006,3(1):59-67.
[14] 宋强强,轩慧勇,刘雪连,等.新疆焉耆地区4种动物源的沙门氏菌耐药性及耐药基因检测[J].湖南农业大学学报(自然科学版),2021,47(6):705-711. SONG Q Q,XUAN H Y,LIU X L,et al.Detection of drug resistance and drug resistance genes of Salmonella from four animal sources in Yanqi,Xinjiang[J].Journal of Hunan Agricultural University(Natural Sciences),2021,47(6):705-711.(in Chinese)
[15] 傅宏庆,刘莉,姚志兰,等.苏北地区白羽肉鸡父母代种鸡及其商品鸡中沙门氏菌的分离、鉴定与生物特性[J].微生物学通报,2020,47(12):4105-4112. FU H Q,LIU L,YAO Z L,et al.Isolation,identification and characterization of Salmonella from a parental breeder farm of white broiler and its commercial[J].Microbiology China,2020,47(12):4105-4112.(in Chinese)
[16] 杨瑞,何云凤,刘梦雪,等.秦皇岛地区鸡源沙门氏菌的分离鉴定及耐药性检测[J].黑龙江畜牧兽医,2022,4:64-68. YANG R,HE Y F,LIU M X,et al.Isolation,identification and drug resistance test of chicken-derived Salmonella in Qinhuangdao area[J].Heilongjiang Animal Science and Veterinary Medicine,2022,4:64-68.(in Chinese)
[17] 胡鑫,汤继华,商妤瑶,等.重庆地区鸡源产品沙门氏菌的分离鉴定和耐药基因检测[J].中国家禽,2021,43(9):115-120. HU X,TANG J H,SHANG Y Y,et al.Isolation,identification and drug resistance gene detection of Salmonella from chicken products in Chongqing[J].China Poultry,2021,43(9):115-120.(in Chinese)
[18] 王迪轩,高东阳,张天姿,等.华中地区鸡源沙门氏菌分离鉴定及耐药性分析[J].中国畜牧兽医,2019,46(3):931-939. WANG D X,GAO D Y,ZHANG T Z,et al.Isolation,identification and drug resistance analysis of avian Salmonella in Central China[J].China Animal Husbandry&Veterinary Medicine,2019,46(3):931-939.(in Chinese)
[19] 李楠,赵思俊,王娟,等.肉鸡生产链中肠炎沙门氏菌耐药性分析及ERIC-PCR分型[J].食品科学,2016,37(3):120-124. LI N,ZHAO S J,WANG J,et al.Resistance and ERIC-PCR genotyping of Salmonella Enteritidis in broiler production chain[J].Food Science,2016,37(3):120-124.(in Chinese)
[20] 余峰玲,许艳平,祝雯雯,等.徐州市市售生肉中3种食源性致病菌污染及药敏情况分析[J].中国卫生检验杂志,2022,32(19):2354-2357. YU F L,XU Y P,ZHU W W,et al.Analysis of contamination and drug susceptibility of three food-borne pathogenic bacteria in raw meat sold in Xuzhou city[J].Chinese Journal of Health Laboratory Technology,2022,32(19):2354-2357.(in Chinese)
[21] 王瑶,张焕容,罗薇.禽源沙门氏菌鉴定、分子分型及耐药性分析[J].动物医学进展,2020,41(11):53-57. WANG Y,ZHANG H R,LUO W.Identification,molecular typing and drug resistance analysis of avian Salmonella[J].Progress in Veterinary Medicine,2020,41(11):53-57.(in Chinese)
[22] 姚玉芳,张竹慧,杨小双.黄山市43株沙门氏菌耐药分析及PFGE分子分型[J].安徽预防医学杂志,2022,28(3):202-205. YAO Y F,ZHANG Z H,YANG X S.Drug resistance analysis and PFGE molecular typing of 43 Salmonella strains in Huangshan city[J].Anhui Journal of Preventive Medicine,2022,28(3):202-205.(in Chinese)
[23] 沈月华,纪蕾,严伟,等.50株鼠伤寒沙门氏菌耐药谱及分子分型特征研究[J].中国卫生检验杂志,2022,32(16):1958-1960. SHEN Y H,JI L,YAN W,et al.Drug resistance spectrum and molecular typing characteristics of 50 Salmonella Typhimurium strains[J].Chinese Journal of Health Laboratory Technology,2022,32(16):1958-1960.(in Chinese)
[24] ZHAN Z,XU X,GU Z,et al.Molecular epidemiology and antimicrobial resistance of invasive non-typhoidal Salmonella in China,2007-2016[J].Infection and Drug Resistance,2019,12:2885-2897.
[25] 何晶晶.沙门氏菌4,[5] ,12:i:-多重耐药性和耐药基因研究[D].扬州:扬州大学,2019. HE J J.The analysis of multidrug resistance and resistant genes in Salmonellae 4,
[5] ,12:i:-islates[D].Yangzhou:Yangzhou University,2019.(in Chinese)
[26] DAHIYA S,SHARMA P,KUMARI B,et al.Characterisation of antimicrobial resistance in Salmonellae during 2014-2015 from four centres across India:An ICMR antimicrobial resistance surveillance network report[J].Indian Journal Medical Microbiology,2017,35(1):61-68.
[27] LIAO J,ORSI R H,CARROLL L M,et al.Serotype-specific evolutionary patterns of antimicrobial-resistant Salmonella enterica[J].BMC Evolutionary Biology,2019,19(1):132.
[28] 李小婷,周变华,贾刘数,等.宠物犬源沙门氏菌的分离鉴定及药敏试验[J].动物医学进,2020,41(1):125-130. LI X X,ZHOU B H,JIA L S,et al.Isolation,identification and drug susceptibility test of Salmonella from pet dogs[J].Progress in Veterinary Medicine,2020,41(1):125-130.(in Chinese)
[29] MORAN R A,ANANTHAM S,HOLT K E,et al.Prediction of antibiotic resistance from antibiotic resistance genes detected in antibiotic-resistant commensal Escherichia coli using PCR or WGS[J]. Journal of Antimicrobial Chemotherapy,2017,72(3):700-704.
[30] RIPABELLI G,TAMBURRO M,GUERRIZIO G,et al.Tracking multidrug-resistant Klebsiella pneumoniae from an Italian hospital:Molecular epidemiology and surveillance by PFGE,RAPD and PCR-based resistance genes prevalence[J].Current Microbiology,2018,75(8):977-987.