土鸡屠宰过程中大肠杆菌毒力基因检测及耐药性分析

doi:10.16431/j.cnki.1671-7236.2020.08.031

摘要/Abstract

摘要： 为了解土鸡屠宰过程中大肠杆菌污染、毒力基因携带及耐药情况，2018年9月-2019年1月在重庆万州、开州、巫溪、奉节4个区县的12个土鸡屠宰地点采集了319份样品，通过菌落形态及显微镜观察、生化鉴定、PCR、药敏试验等方法鉴定分离的大肠杆菌，检测其毒力基因携带和耐药性情况。结果表明：大肠杆菌分离率为22.57%（72/319），其中餐馆、活禽宰杀铺和定点屠宰点分离率依次为29.73%（22/74）、25.00%（24/96）和17.45%（26/149），污水、地面、羽毛、用具和胴体分离率依次为75.00%（18/24）、21.62%（8/37）、20.54%（23/112）、17.65%（6/34）和15.18%（17/112）；11种毒力基因中除estA、estB、elt外均被检出，检出率为58.33%（42/72），共检出14种组合型，有4株分离株4种毒力基因同时存在；分离株对阿米卡星、头孢噻肟最敏感，多重耐药比为90.28%（65/72），以7~10重耐药居多。土鸡屠宰过程中致病性大肠杆菌污染风险高，多重耐药现象严重，应重视养殖合理用药及屠宰卫生环境。

关键词: 大肠杆菌; 屠宰过程; 毒力基因; 耐药性

Abstract: In order to explore the contamination,virulence genes carrying and antibiotic resistance of Escherichia coli(E.coli) in slaughtering process of free-range chicken.319 samples which came from 12 slaughter places of Wanzhou,Kaizhou,Wuxi,Fengjie district in Northeast Chongqing were collected in September 2018 to January 2019.These samples were analyzed by colony morphology,microscopic observation,biochemical test,PCR and drug sersitivity test.The results indicated that the isolation rate of E.coli was 22.57%(72/319).The isolation rates of restaurant,live poultry shop and slaughterhouse were 29.73%(22/74),25.00%(24/96) and 17.45%(26/149) respectively.The rates of samples of waste water,ground,feather,tools and carcass were 75.00%(18/24),21.62%(8/37),20.54%(23/112),17.65%(6/34) and 15.18%(17/112) respectively.All of the virulence genes were detected expect estA,estB and elt genes.The detection rate was 58.33%(42/72) and there were 14 combinations even 4 strains coexisted 4 virulence genes.The strains were most sensitive to amikacin and cefotaxime,and existed multi-antibiotic resistance seriously which most of them were from 7 to 10 folds.There was a high risk to be contaminated by pathogenic E.coli in slaughtering process of free-range chicken and existing multi-antibiotic resistance seriously.So the rational using of drugs and keeping a clean process of slaughter should pay more attention.

Key words: Escherichia coli(E.coli); slaughtering process; virulence genes; antibiotic resistance

中图分类号:

S859.1

陈亚强, 彭津津, 廖明, 廖勤丰. 土鸡屠宰过程中大肠杆菌毒力基因检测及耐药性分析[J]. 中国畜牧兽医, 2020, 47(8): 2615-2624.

CHEN Yaqiang, PENG Jinjin, LIAO Ming, LIAO Qinfeng. Virulence Genes Detection and Antibiotic Resistance Analysis of Escherichia coli Isolated from Slaughtering Process of Free-range Chicken[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(8): 2615-2624.

参考文献

[1] ALTALHI A D,HASSAN S A.Bacterial quality of raw milk investigated by Escherichia coli and isolates analysis for specific virulence-gene markers[J].Food Control,2009,20(10):913-917.
[2] GONZALES L,JOFFRE E,RIVERA R,et al.Prevalence seasonality and severity of disease caused by pathogenic Escherichia coli in children with diarrhoea in Bolivia[J].Journal of Medical Microbiology,2013,62(11):1697-1706.
[3] MELLATA M.Human and avian extraintestinal pathogenic Escherichia coli:Infections,zoonotic risks,and antibiotic resistance trends[J].Foodborne Pathogens and Disease,2013,10(11):916-932.
[4] KYUNG H O,SOO B K,MI S P,et al.Development of a one-step PCR assay with nine primer pairs for the detection of five diarrheagenic Escherichia coli types[J].Journal of Microbiology and Biotechnology,2014,24(6):862-868.
[5] EWERS C,ANTAO EM,DIEHL I,et al.Intestine and environment of the chicken as reservoirs for extraintestinal pathogenic Escherichia coli strains with zoonotic potential[J].Applied and Environmental Microbiology,2009,75(1):184-192.
[6] MATEUS L,HENRIQUES S,MERINO C,et al.Virulence genotypes of Escherichia coli canine isolates from pyometra,cystitis and fecal origin[J].Veterinary Microbiology,2013,166(3-4):590-594.
[7] 周陆红,张鹏飞,张杰,等.屠宰猪中大肠杆菌毒力基因检测及耐药性分析[J].食品科学,2019,40(2):264-268. ZHOU L H,ZHANG P F,ZHANG J,et al.Virulence genes and antimicrobial resistance of Escherichia coli isolated from slaughtered pigs[J].Food Science,2019,40(2):264-268.(in Chinese)
[8] 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.
[9] MULLER D,GREUNE L,HEUSIPP G,et al.Identification of unconventional intestinal pathogenic Escherichia coli isolates expressing intermediate virulence factor profiles by using a novel single-step multiplex PCR[J].Applied and Environmental Microbiology,2007,73(10):3380-3390.
[10] 刘璨颖,李建宏,梅启定,等.禽源致病性大肠杆菌的耐药性和毒力基因研究[J].中国家禽,2018,40(13):21-25. LIU C Y,LI J H,MEI Q D,et al.Analysis on antibiotic resistance and virulence genes of avian pathogenic Escherichia coli[J].China Poultry,2018,40(13):21-25.(in Chinese)
[11] 曾博,王红宁,邹立扣,等.西南地区猪肠外大肠杆菌毒力基因检测及耐药性研究[J].中国预防兽医学报,2013,35(2):126-129. ZENG B,WANG H N,ZOU L K,et al.Virulence gene detection and antibiotic sensitivity of swine extraintestinal Eschericha coli from Southwest China[J].Chinese Journal of Preventive Veterinary Medicine,2013,35(2):126-129.(in Chinese)
[12] 孟含,陈娟,龙虎,等.牦牛屠宰环境中大肠杆菌分离菌株的病原学特征研究[J].黑龙江畜牧兽医,2020,3:80-84. MENG H,CHEN J,LONG H,et al.Pathogenic characteristics of Escherichia coli isolated strains in yak slaughtering environment[J].Heilongjiang Animal Science and Veterinary Medicine,2020,3:80-84.(in Chinese)
[13] 李晴.污水产ESBL大肠杆菌水平传递耐药性的研究[D].泰安:山东农业大学,2018. LI Q.Transfer of antimicrobial resistant genes of ESBL-producing Escherichia coli recovered from waste water[D].Tai'an:Shandong Agricultural University,2018.(in Chinese)
[14] 史秋梅,张艳英,高桂生,等.河北省部分地区肉、蛋食品大肠杆菌污染状况的检测[J].河北科技师范学院学报,2012,4:7-11. SHI Q M,ZHANG Y Y,GAO G S,et al.Analysis on pathogenic germ E.coli in meat and eggs from Hebei province of China[J].Journal of Hebei Normal University of Science & Technology,2012,4:7-11.(in Chinese)
[15] BOUVET J,MONTET M P,ROSSEL R,et al.Effects of slaughter processes on pig carcass contamination by verotoxin-producing Escherichia coli and E.coli O157:H7[J].International Journal of Food Microbiology,2002,77(1-2):99-108.
[16] 张苗.淄博市食源性疾病中致泻性大肠埃希菌基因分型、溯源研究及耐药性分析[D].青岛:青岛大学,2018. ZHANG M.Genotyping,traceability and drug resistance study of diarrheagenic Escherichia coli as foodborne diseases in Zibo[D].Qingdao:Qingdao University,2018.(in Chinese)
[17] HORNITZKY M A,MERCIECA K,BETTELHEIM K A,et al.Bovine feces from animals with gastrointestinal infections are a source of serologically diverse atypical enteropathogenic Escherichia coli and Shiga toxin-producing E.coli strains that commonly possess intimin[J].Applied & Environmental Microbiology,2005,71(7):3405-3412.
[18] RAKIN A,NOELTING C,SCHUBERT S,et al.Common and specific characteristics of the high-pathogenicity island of Yersinia enterocolitica[J].Infection & Immunity,1999,67(10):5265-5274.
[19] 田硕.济南市鸡大肠杆菌病流行病学调查及防治研究[D].泰安:山东农业大学,2014. TIAN S.Study on epidemiological investigation and control of chicken colibacillosis in Jinan region[D].Tai'an:Shandong Agricultural University,2014.(in Chinese)
[20] FRICKE W F,MCDERMOTT P F,MAMMEL M K,et al.Antimicrobial resistance-conferring plasmids with similarity to virulence plasmids from avian pathogenic Escherichia coli strains in Salmonella enterica serovar Kentucky isolates from poultry[J].Applied & Environmental Microbiology,2009,75(18):5963-5971.
[21] 王影,李欣南,韩镌竹,等.某肉鸡场鸡源大肠杆菌耐药性跟踪检测与分析[J].中国畜牧兽医,2015,42(12):3329-3337. WANG Y,LI X N,HAN J Z,et al.Tracking detection and analysis of drug resistance of chicken-derived Escherichia coli in a broiler farm[J].China Animal Husbandry & Veterinary Medicine,2015,42(12):3329-3337.(in Chinese)
[22] 植婵萍,郭小婷,姚旭,等.有机农场鸡、猪源大肠杆菌耐药性研究[J].中国畜牧兽医,2015,42(12):3351-3357. ZHI C P,GUO X T,YAO X,et al.Study on drug resistance of Escherichia coli isolated from chicken and swine in organic farm[J].China Animal Husbandry & Veterinary Medicine,2015,42(12):3351-3357.(in Chinese)
[23] ENNE V I,CLAIRE C,KATHERINE S,et al.A high prevalence of antimicrobial resistant Escherichia coli isolated from pigs and a low prevalence of antimicrobial resistant E.coli from cattle and sheep in Great Britain at slaughter[J].FEMS Microbiology Letters,2008,278(2):193-199.