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.
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