Details of sample type and source of Escherichia coli isolates investigated for antimicrobial resistance in a One Health study of the beef production system. Sample location/sites comprised composite fecal samples collected from penned cattle in feedlots A, B, C and D as well as water samples from catch basins. Environmental samples comprised constructed wetlands and a creek (adjacent to feedlot C). Wastewater influent and effluent were collected from two water treatment plants while samples at a beef processing facility were obtained from carcasses after hide removal, after final wash, from ground beef and retail meat. Human samples were obtained from blood, urine and abdomen samples collected from hospital patients in southern Alberta.