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The prevalence of E. coli O157:H7 in feedlot cattle increases during the warmer months of the year and during transportation. This study was conducted to determine whether stress contributes to this increase. Feedlot heifers of four breeds (hide colors: red, black, tan, and white) were selected on the basis of weight, and assigned to pens on the basis of hide color, weight, and fecal prevalence of E. coli O157:H7. Animals were weighed, condition scored, and temperament scored every 28 days. Fecal grab samples were collected for each animal individually at the same time. The presence and levels of E. coli O157:H7 and levels of generic E. coli in feces were determined. The results of this study indicate there were no clear trends between the heat stress or handling stress and fecal generic E. coli levels or E. coli O157:H7 prevalence in feces.