Sheep have been proposed as a source of human verocytotoxigenic Escherichia coli infection on a number of occasions but few prevalence studies have focused on identifying carriage rates of these pathogens in this species. The focus of this work was to establish the frequency of excretion of E. coli of serogroups O157, O26, O103, O111 and O145 in sheep presented for slaughter in Scotland and to examine their carriage of known virulence determinants. The study involved microbiological isolation of E. coli from 1082 sheep presented for slaughter in four Scottish abattoirs between July 2005 and June 2006. Using faecal enrichment and immunomagnetic separation the isolation rates from these samples was 3.4% for E. coli serogroup O157, 5.2% for E. coli serogroup O26, 2.3% for E. coli serogroup O103 and 0.1% for E. coli serogroup O145. E. coli O111 was not isolated. In the last month of testing, which coincided with sorbitol fermenting E. coli O157 (SFO157) cases in children in Scotland, all 83 recta received were screened and tested negative for SFO157 strains. The study found no verocytotoxin positive strains amongst the E. coli serogroup O103 or O145 isolates. Verocytotoxin positive strains were identified amongst isolates of E. coli serotypes O157:H7 and O26:H11. E. coli O157:H7 was not isolated from samples collected between January and March, a statistically significant drop (p<0.001) in mean shedding relative to other months. There was evidence (p=0.003) of higher shedding of O157 in adults and hogs than in lambs. E. coli O26:H11 was isolated throughout the year, with a statistically significant peak in shedding in the third quarter (p=0.003). The results show that sheep presented for slaughter in Scotland may carry strains of E. coli, particularly of serogroups O157 and O26, which can be presumed to have potential to cause human infection. They did not support a hypothesis that human cases of E. coli O157:H7 are higher in any particular Scottish region as a direct consequence of a higher rate of faecal carriage in sheep in that region.