犊牛源肠外致病性大肠杆菌分离鉴定、致病性及耐药性分析

doi:10.16431/j.cnki.1671-7236.2023.12.024

Abstract

Abstract: 【Objective】 The purpose of this study was to identify the bacterial pathogens and its biological characteristics causing the death of calves in a large-scale cattle farm in Kashi,Xinjiang,so as to provide reference for the treatment of bacterial-associated diseases and rational drug for the area.【Method】 The pathological dissection and pathogen isolation on dead calves were conducted,single colony from liver (KS-1) and anal swab (KS-2) were selected for purification and culture.The isolates were identified by morphological observation,biochemical test,16S rRNA gene amplification together with phylogenetic group.The pathogenicity and drug resistance of the isolates were evaluated by virulence gene detection,pathogenicity test and drug sensitivity test.【Result】 After inoculation,the lung,liver,heart blood,joint fluid and mesenteric lymph nodes of dead calves showed a consistent colony growth pattern,as did anal swab samples from healthy calves.The isolated strains were identified as Escherichia coli(E.coli) by the growth morphology of the colony on selective medium,the morphological features of Gram staining microscopy,the biochemical properties and the verification of E.coli 16S rRNA.Phylogenetic analysis showed that KS-1 belonged to group A and KS-2 to group B1.KS-1 contained 8 virulence-related genes (crl,papC,hlyE,eaeA,ompA,ompC,iucD and iutA),and KS-2 contained 3 virulence-related genes (crl,hlyE and ompF).The pathogenicity test results showed that following intraperitoneal injection of 1×10⁸ CFU of the isolate,all mice in KS-1 group died within 16 h,however no mice in KS-2 group died.The results of drug sensitivity tests showed that KS-1 was sensitive to 17 drugs,including piperacillin/tazobactam,cefazolin,cefuroxime,cefuroxime,etc.,it was moderately sensitive to ampicillin/sulbactam and piperacillin,and resistant to ampicillin and cotrimoxazole.KS-2 was sensitive to piperacillin/tazobactam,cefotetan,imipenem,meropenem,amikacin and nitrofurantoin,moderately sensitive to tobramycin,and resistant to the other 14 drugs.【Conclusion】 In this study,two calf-derived E.coli strains were isolated,of which KS-1 was more virulent and carried abundant virulence genes,it was sensitive to most commonly used antibiotics,and was resistant to ampicillin and sulfamethoxazole,while KS-2 carried few virulence genes and showed multi-drug resistance.

Key words: calves; extraintestinal pathogenic Escherichia coli; isolation and identification; virulence genes; drug resistance

CLC Number:

S852.61

SHI Weidi, KANG Lichao, LIU Caixia, KOU Lijun, LYU Shuangfei, REN Huijie, QIAN Ruixuan, MA Xun, WANG Jing, JIANG Jianjun, GAO Shengjie. Isolation,Identification,Pathogenicity and Drug Resistance Analysis of Extraintestinal Pathogenic Escherichia coli from Calves[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(12): 5022-5031.

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Isolation,Identification,Pathogenicity and Drug Resistance Analysis of Extraintestinal Pathogenic Escherichia coli from Calves

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