To determine the distribution of pathogens on cattle hides at the feedlot, samples were collected from six hide surface locations (back, flank, hock, neck, perineum, and ventrum), the oral cavity, the rectal-anal junction, and the feces of feedlot cattle and subjected to Escherichia coli O157 detection via culture methods and to Salmonella detection via PCR. E. coli O157 was isolated from one or more of the sampling locations from 31 (42.5%) of the 73 animals sampled. Location-specific prevalence of E. coli O157 was 5% for back samples, 5% for flank samples, 12% for hock samples, 7% for neck samples, 12% for perineum samples, 8% for ventrum samples, 1% for oral cavity samples, 4% for rectal-anal junction swabs, and 23% for fecal grab samples. Salmonella was isolated from one or more of these sample locations from 100% (50 of 50 samples) of all animals sampled. Location-specific prevalence of Salmonella was 76% for back samples, 74% for flank samples, 94% for hock samples, 76% for neck samples, 88% for perineum samples, 86% for ventrum samples, 94% for oral cavity samples, 64% for rectal-anal junction swabs, and 50% for fecal grab samples. The sampling locations that maximized the likelihood of finding E. coli O157 and Salmonella (84 and 96%, respectively) if the animal was positive at one sampling location or more were the hock, perineum, and fecal grab. These data suggest that the use of multiple sample locations is useful when isolating these pathogens from feedlot cattle. Focusing on one sampling location may underestimate the prevalence.