Two hundred steers and heifers from a large feedyard (65,000-head capacity) were used to determine the prevalence levels of enterohemorrhagic Escherichia coli O157 (EHEC O157) and Salmonella spp. prior to and after shipping to a commercial packing facility. Two samples, a ventral midline hide swab and a fecal sample, were aseptically collected from each animal 2 weeks prior to the date of transportation and at the packing plant immediately after exsanguination. Samples were collected from all trailers (n = 46) before animals were loaded for transport to the packing facility. The average prevalence levels of EHEC O157 on hides (18%) and in feces (9.5%) at the feedyard decreased (P > 0.05) at the packing plant to 4.5 and 5.5%, respectively. The average prevalence levels of Salmonella spp. on hides (6%) and in feces (18%) at the feedyard increased to 89 and 46%, respectively, upon arrival at the packing plant. Average prevalence levels for EHEC O157 and Salmonella spp. on the trailers were 5.43 and 59%, respectively. The results of this study demonstrate that transportation may be a potential stressor for cattle, as evidenced by the increased shedding of Salmonella spp.