鸭源致病性大肠杆菌O抗原与毒力基因检测及药敏试验

doi:10.16431/j.cnki.1671-7236.2023.02.029

摘要/Abstract

摘要： 【目的】探究荣昌、大足和隆昌三地鸭大肠杆菌分离株的O抗原、毒力基因及耐药性。【方法】将2014年—2021年鸭病料中分离得到的107株细菌在无菌条件下接种于麦康凯培养基中划线进行培养纯化,通过16S rDNA基因扩增测序和生化试验进行细菌鉴定,采用PCR技术对O抗原和16种毒力基因进行检测,采用Kirby-Bauer纸片扩散法进行药敏试验。【结果】107株分离株鉴定为大肠杆菌;O抗原鉴定试验鉴定出9种O抗原,其中优势抗原为O78(37.00%)、O7(25.00%),O121和O145(均为15.00%),并检测到5株O78+O145和O7+O145融合株;共检测出11种毒力因子,其中强致病性毒力基因有Tsh基因(检出率为25.23%)、fyuA基因(检出率为31.78%)、estB基因(检出率为31.78%)、Vat基因(检出率为2.80%)、iucA基因(检出率为44.56%)。3种毒力基因ompA、yijP和ibeB的携带率最高,分别为100.00%、96.26%和85.98%;药敏试验结果表明分离株对氨基糖苷类药物、米诺环素和多黏菌素最为敏感,对大环内酯类药物和克林霉素耐药,分离株均为多重耐药菌,30.00%的分离株表现为7重耐药。【结论】本研究鉴定到107株鸭源致病性大肠杆菌,其致病性和耐药性较强,毒力基因携带情况和耐药性检测发现大肠杆菌的致病力与其耐药性存在一定关系;O抗原检测发现了5株新型O抗原融合株,优势O抗原不断变化,说明西南地区大肠杆菌遗传结构正发生变化,可能存在新的菌体抗原。本研究结果为中国西南地区鸭大肠杆菌病防控及疫苗制备提供依据。

关键词: 鸭源大肠杆菌; O抗原; 毒力基因; 药敏试验

Abstract: 【Objective】 To investigate the O antigen,virulence gene and drug resistance of Escherichia coli (E.coli) isolates from ducks in Rongchang,Dazu and Longchang regions.【Method】 107 strains of bacteria isolated from duck diseased materials from 2014 to 2021 were inoculated in MacConkey medium under sterile conditions for culture and purification,identified by 16S rDNA gene amplification sequencing and biochemical test,detected O antigen and 16 virulence genes by PCR technology,and conducted drug sensitivity test by Kirby-Bauer disk diffusion method.【Result】 107 isolates identified as E.coli.O antigen identification tests identified nine O antigens,of which the dominant antigens were O78 (37.00%),O7 (25.00%),O121 and O145 (both 15.00%),and five O78+O145 and O7+O145 fusion strains were detected.The virulence gene detection test detected 11 virulence factors,of which the strongly pathogenic virulence genes were Tsh gene with a detection rate of 25.23%, fyuA gene with a detection rate of 31.78%, estB gene with a detection rate of 31.78%, Vat gene with a detection rate of 2.80% and iucA gene with a detection rate of 44.56%.3 virulence genes, ompA, yijP and ibeB,had the highest carriage rates of 100.00%,96.26% and 85.98%,respectively.The drug sensitivity test results showed that the isolates were most sensitive to aminoglycosides,minocycline and polymyxin,and resistant to macrolides and clindamycin.All isolates were multi-drug resistant,with 30.00% of isolates showing 7-fold resistance.【Conclusion】 In this study,107 strains of pathogenic E.coli from ducks were identified,which were highly pathogenic and drug resistant,and virulence gene carrier and drug resistance detection found that the virulence of E.coli had a certain relationship with its drug resistance.O antigen detection found five new O antigen fusion strains,the dominant O antigen was constantly changing,indicating that the genetic structure of E.coli in Southwest China was changing,and there might be new bacterial antigens.The results of this study provided a basis for the prevention and control of duck colibacillosis and vaccine preparation in Southwest China.

Key words: Escherichia coli form duck; O-antigen; virulence gene; drug sensitivity test

中图分类号:

Q939.94

罗干, 邹宏, 任绍科, 吴春霞, 邹瑶, 娄银莹, 周洋, 程方俊, 郭建华. 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.

LUO Gan, ZOU Hong, REN Shaoke, WU Chunxia, ZOU Yao, LOU Yinying, ZHOU Yang, CHENG Fangjun, GUO Jianhua. Detection of O-antigen,Virulence Gene and Drug Sensitivity Test of Pathogenic Escherichia coli from Duck[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 713-721.

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