犊牛腹泻源大肠杆菌耐药情况及HPI相关基因的检测

doi:10.16431/j.cnki.1671-7236.2020.06.034

中国畜牧兽医 ›› 2020, Vol. 47 ›› Issue (6): 1942-1951.doi: 10.16431/j.cnki.1671-7236.2020.06.034

犊牛腹泻源大肠杆菌耐药情况及HPI相关基因的检测

高海慧¹, 黎玉琼¹, 高小斐², 王培柱³, 吴学青³, 康晓冬¹, 梁小军¹

1. 宁夏农林科学院动物科学研究所, 银川 750002;
2. 陕西省榆林市定边县樊学镇畜牧兽医站, 定边 718699;
3. 宁夏吴忠市利通区扁担沟镇畜牧兽医工作站, 吴忠 751100

收稿日期:2020-01-07 出版日期:2020-06-20 发布日期:2020-06-20
通讯作者: 康晓冬, 梁小军 E-mail:kangxd1971@126.com;lxj0520@163.com
作者简介:高海慧(1990-),女,宁夏吴忠人,硕士,助理研究员,研究方向:牛羊等反刍动物病原微生物,E-mail:690718895@qq.com;黎玉琼(1973-),女,宁夏吴忠人,本科,副研究员,研究方向:临床兽医和动物疾病,E-mail:1463805069@qq.com
基金资助:
宁夏回族自治区自然科学基金——宁夏地区犊牛腹泻源性大肠杆菌HPI相关毒力基因检测与序列分析（2018AAC03185）；宁夏农林科学院农业科技自主创新项目——宁夏地区犊牛隐孢子虫流行病学调查及种群结构研究（NKYQ-18-13）；国家肉牛牦牛产业技术体系专项（CARS-37）；奶牛胎衣不下症中兽药新药创制与示范（2019BEF02003）

Drug Resistance and Detection of HPI Related Genes in E.coli from Calves with Diarrhea

GAO Haihui¹, LI Yuqiong¹, GAO Xiaofei², WANG Peizhu³, WU Xueqing³, KANG Xiaodong¹, LIANG Xiaojun¹

1. Institute of Animal Science, Ningxia Academy of Agriculture and Forestry Sciences, Yinchuan 750002, China;
2. Animal Husbandry and Veterinary Station, Fanxue Town, Dingbian County, Dingbian 718699, China;
3. Animal Husbandry and Veterinary Station, Biandangou Town, Wuzhong County, Wuzhong 751100, China

Received:2020-01-07 Online:2020-06-20 Published:2020-06-20

摘要/Abstract

摘要： 本研究旨在明确犊牛腹泻源性大肠杆菌的耐药情况、强毒力岛（HPI）标志基因及相关基因的携带情况，以及大肠杆菌分离株与HPI携带的关系。采集犊牛病理性腹泻样本158份，采用麦康凯培养基和伊红美蓝培养基进行筛选，镜检符合大肠杆菌形态的进行VITEK 2 Compact生化鉴定和PCR鉴定，采用K-B纸片法对大肠杆菌分离株进行药敏试验，应用PCR方法进行分离株HPI相关基因携带情况的检测。结果显示，共分离得到75株大肠杆菌，分离株对阿莫西林、哌拉西林、氨苄西林、头孢唑啉、氟苯尼考、恩诺沙星、四环素的耐药率均>90%，对头孢氨苄、头孢拉定、头孢曲松、环丙沙星、诺氟沙星和氧氟沙星的耐药率均≥ 60.00%，对阿米卡星较敏感，耐药率为9.33%；75株大肠杆菌全部耐3种以上药物，多重耐药（≥ 10）的菌株占85.33%，耐药谱集中在耐14~17种药物，5株对19种药物全部耐药。HPI标志基因irp2的阳性率为100%，其他相关基因fyuA、irp3、irp5、irp8和ytbA的检出率在66.00%以上。综上所述，宁夏地区犊牛腹泻源性大肠杆菌耐药普遍，多重耐药现象严重。

关键词: 犊牛腹泻; 大肠杆菌; 耐药; 强毒力岛

Abstract: The purpose of this study was to determine the drug resistance of Escherichia coli (E.coli) from calf diarrhea,the carrying of high pathogenicity island (HPI) marker gene and related genes,and the relationship between E.coli isolates and HPI carrying.158 pathological diarrhea samples of calves were collected and screened by McConkey medium and Eosin-Methylene blue medium.VITEK 2 compact biochemical identification and PCR identification were carried out for those with the morphology of E.coli.The results showed that 75 strains of E.coli were isolated.The drug resistance rate of the isolates to amoxicillin,piperacillin,ampicillin,cefazolin,florfenicol,enrofloxacin and tetracycline was >90%.The drug resistance rate to cefalexin,cefradine,ceftriaxone,ciprofloxacin,norfloxacin and ofloxacin was ≥ 60.00%.The drug resistance rate to amikacin was 9.33%.75 strains of E.coli were resistant to more than 3 kinds of drugs,85.33% of them were multi resistant (≥ 10),the resistant spectrum was concentrated in 14 to 17 kinds of drugs,and 5 strains were resistant to all 19 kinds of drugs.The positive rate of HPI marker gene irp2 was 100%,and that of other related genes fyuA,irp3,irp5,irp8 and ytbA was over 66.00%.In conclusion,the drug resistance of E.coli from diarrhea in Ningxia was widespread,and the multiple drug resistance was serious.

Key words: diarrhea calves; Escherichia coli; resistance; high pathogenicity island

中图分类号:

S852.61

高海慧, 黎玉琼, 高小斐, 王培柱, 吴学青, 康晓冬, 梁小军. 犊牛腹泻源大肠杆菌耐药情况及HPI相关基因的检测[J]. 中国畜牧兽医, 2020, 47(6): 1942-1951.

GAO Haihui, LI Yuqiong, GAO Xiaofei, WANG Peizhu, WU Xueqing, KANG Xiaodong, LIANG Xiaojun. Drug Resistance and Detection of HPI Related Genes in E.coli from Calves with Diarrhea[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(6): 1942-1951.

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犊牛腹泻源大肠杆菌耐药情况及HPI相关基因的检测

Drug Resistance and Detection of HPI Related Genes in E.coli from Calves with Diarrhea

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