Karama Musafiri, Johnson Roger P, Holtslander Robert, McEwen Scott A, Gyles Carlton L
Department of Pathobiology, University of Guelph, Guelph, Ontario N1G 2W1.
Can J Vet Res. 2008 Jul;72(4):297-302.
This study determined the prevalence of verotoxin (VT)-producing Escherichia coli (VTEC) in Ontario beef cattle at slaughter and characterized the isolates by serotype, virulence factors, virulence markers, and antimicrobial resistance. Cultures of rectal feces from 500 animals were screened for VT by an enzyme-linked immunosorbent assay (ELISA) and by polymerase chain reaction (PCR) for genes vt1, vt2, and eae. The VT-ELISA-positive samples were tested by a VT-immunoblot to isolate VTEC colonies. The prevalence rates of VTEC by VT-ELISA and PCR were 10.2% [95% confidence interval (CI), 7.8% to 13.2%] and 6.2% (95% CI, 4.4% to 8.7%), respectively. Colonies of VTEC were isolated from 27 (53%) of the 51 VT-ELISA-positive samples and belonged to 24 serotypes, which did not include O157:H7. Twelve of the serotypes have been implicated in disease in humans. Virulence profiling of the isolates by PCR revealed that 2 (8%) were eae-positive, 5 (21%) had vt1 only, and 19 (79%) had vt2, of which 3 had vt2 only, 7 had vt1 + vt2, 4 had vt2 + vt2c, 2 had vt2 + vt2c + vt2d, 2 had vt1 + vt2 + vt2c, and 1 had vt1 + vt2 + vt2c + vt2d. The distribution of selected plasmid-encoded putative virulence genes was as follows: ehxA, 63%; espP, 46%; saa, 67%; and subA, 54%. Nine of the 24 isolates were resistant to 1 or more antimicrobials. Major conclusions are that the VTEC prevalence of 10.2% was among the lower rates reported for beef cattle, a high proportion of the isolates had vt2 genes, the subA gene was reported for the 1st time in Canadian VTEC, and the absence of O157 VTEC likely reflects the use of a technique that detected all VTEC.
本研究确定了安大略省屠宰肉牛中产志贺毒素(VT)的大肠杆菌(VTEC)的流行情况,并通过血清型、毒力因子、毒力标记物和抗菌药物耐药性对分离株进行了特征分析。采用酶联免疫吸附测定(ELISA)和聚合酶链反应(PCR)对500头动物的直肠粪便培养物进行VT筛查,检测vt1、vt2和eae基因。对VT-ELISA阳性样本进行VT免疫印迹检测以分离VTEC菌落。VT-ELISA和PCR检测VTEC的流行率分别为10.2%[95%置信区间(CI),7.8%至13.2%]和6.2%(95%CI,4.4%至8.7%)。从51份VT-ELISA阳性样本中的27份(53%)分离出VTEC菌落,它们属于24种血清型,其中不包括O157:H7。其中12种血清型与人类疾病有关。通过PCR对分离株进行毒力分析表明,2株(8%)eae呈阳性,5株(21%)仅具有vt1,19株(79%)具有vt2,其中3株仅具有vt2,7株具有vt1 + vt2,4株具有vt2 + vt2c,2株具有vt2 + vt2c + vt2d,2株具有vt1 + vt2 + vt2c,1株具有vt1 + vt2 + vt2c + vt2d。所选质粒编码的假定毒力基因的分布如下:ehxA,63%;espP,46%;saa,67%;subA,54%。24株分离株中有9株对1种或多种抗菌药物耐药。主要结论是,10.2%的VTEC流行率在已报道的肉牛流行率中处于较低水平,高比例的分离株具有vt2基因,subA基因在加拿大VTEC中首次报道,且未检测到O157 VTEC可能反映了所采用技术能检测所有VTEC。