笼养绿尾虹雉大肠埃希菌耐药特征与毒力基因分析  

Antibiotic resistance characteristics and virulence gene analysis of Escherichia coli in caged Chinese monals(Lophophorus lhuysii)

作　　者：梁驹 吴邦元[1,2] 肖英平 冯少华[1,2] 徐小钦 王彬 周材权[1,4] 陈黎 李万洪[5] 张龙 LIANG Ju;WU Bang-yuan;XIAO Ying-ping;FENG Shao-hua;XU Xiao-qin;WANG Bin;ZHOU Cai-quan;CHEN Li;LI Wan-hong;ZHANG Long(Key Laboratory of Southwest China Wildlife Resources Conservation,Ministry of Education,Nanchong,Sichuan 637009,China;College of Life Science,China West Normal University,Nanchong,Sichuan 637009,China;Institute of Agro-product Safety and Nutrition,Zhejiang Academy of Agricultural Sciences,Hangzhou,Zhejiang 310021,China;Sichuan Wildlife Rehabilitation and Breeding Research Center/Institute of Ecology,China West Normal University,Nanchong,Sichuan 637009,China;Sichuan Fengtongzhai National Nature Reserve Administration,Ya’an,Sichuan 625700,China)

机构地区：[1]西南野生动植物资源保护教育部重点实验室,四川南充637009 [2]西华师范大学生命科学学院,四川南充637009 [3]浙江省农业科学院农产品质量安全与营养研究所,浙江杭州310021 [4]四川省野生动物救护与繁育研究中心/西华师范大学生态研究院,四川南充637009 [5]四川蜂桶寨国家级自然保护区管护中心,四川雅安625700

出　　处：《南方农业学报》2024年第12期3789-3798,共10页Journal of Southern Agriculture

基　　金：国家自然科学基金青年基金项目(32200339)。

摘　　要：【目的】分析笼养绿尾虹雉大肠埃希菌的耐药特征与毒力基因携带状况,了解笼养绿尾虹雉大肠埃希菌耐药性及潜在风险,为绿尾虹雉驯养繁育过程中抗生素的合理使用及动物源细菌耐药性的遏制提供理论依据。【方法】于2022年1月、3月、4月、7月和10月采集四川蜂桶寨国家级自然保护区管护中心笼养绿尾虹雉新鲜粪便样本116份,进行细菌分离鉴定,通过药敏试验测定分离获得的大肠埃希菌对18种抗生素的耐药表型,筛选鉴定产超广谱β-内酰胺酶(ESBL)的菌株,通过PCR扩增对13种耐药基因和9种毒力基因进行检测,并对代表性耐药表型与相关耐药基因进行关联分析。【结果】116份样本共分离获得84株大肠埃希菌,分离率为72.41%,其中有12株不耐受任何抗生素,耐受3种及以上抗生素的耐药菌株有53株,占63.10%,菌株的耐药数量最高为12种,耐药谱以氨苄青霉素、甲氧苄氨嘧啶和庆大霉素为主,平均耐药率分别为78.57%、75.00%和45.24%。耐药基因检测结果显示,多黏菌素类耐药基因pmrA的平均检出率最高,为80.95%。其他平均检出率达50.00%的耐药基因还有β-内酰胺类耐药基因blaTEM(70.24%)、氯霉素类耐药基因floR(57.14%)和磺胺类耐药基因sulⅠ(50.00%);庆大霉素耐药性与氨基糖苷类耐药基因aadA1具有极显著关联性(P<0.01),而氨苄青霉素耐药性与blaTEM基因、甲氧苄氨嘧啶耐药性与sulI基因、环丙沙星耐药性与喹诺酮类耐药基因qnrS均无显著关联性(P>0.05,下同)。筛选鉴定出产ESBL菌36株,产ESBL菌对氨苄青霉素与甲氧苄氨嘧啶的耐药率均为100.00%,但产ESBL菌与blaTEM基因无显著关联性。对7月和10月粪便样本中分离的39株大肠埃希菌进行毒力基因检测,其中蛋白分泌物调节基因(escV)的检出率最高(17.95%),其次为肠定植因子基因(pic)(15.38%),而侵袭性质粒调节基因(invE)未检出。【结论】笼养绿尾虹雉粪便中的�【Objective】To analyze the antibiotic resistance characteristics and virulence gene carriage status of Escherichia coli in caged Chinese monals(Lophophorus lhuysii),explore the antibiotic resistance and potential risks of E.coli in caged Chinese monals,which could provide theoretical basis for the rational use of antibiotics and containment of drug resistance in animal-derived bacteria in the breeding process of Chinese monals.【Method】A total of 116 fresh fecal samples were collected from the artificial breeding ground at the Sichuan Fengtongzhai National Nature Reserve Administration in January,March,April,July and October 2022.Conducted bacterial isolation and identification,determined the antibiotic resistance patterns of E.coli strains obtained through susceptibility testing against 18 types of antibiotics,screened for strains producing extended-spectrum beta-lactamases(ESBLs),and used PCR amplification to detect 13 types of resistance genes and 9 types of virulence genes.Additionally,performed a correlation analysis between representative resistance phenotypes and associated resistance genes.【Result】A total of 84 strains of E.coli were isolated from 116 samples,with a detection rate of 72.41%.Among them,12 strains were sensitive to all tested antibiotics,while 53 strains were resistant to 3 or more tested antibiotics,accounting for 63.10%.The highest number of antibiotic resistance phenotype in the isolates was 12,and a majority of the observed antibiotic resistance phenotype were ampicillin,trimethoprim and gentamicin,with verage resistance rates of 78.57%,75.00%and 45.24%.The antibiotic resistance gene detection results showed that the average detection rate of the polymyxins resistance gene pmrA was the highest(80.95%).Other antibiotic resistance genes with a detection rate exceeding 50.00%includedβ-lactams resistance gene blaTEM(70.24%),chloramphenicol resistance gene floR(57.14%)and sulfonamides resistance gene sulI(50.00%).The resistance to gentamicin was significantly associated with the a

关 键 词：绿尾虹雉 大肠埃希菌 耐药表型 耐药基因 毒力基因 

分 类 号：S858.9[农业科学—临床兽医学]