Dried distillers' grain with solubles (DDGS) is a by-product of ethanol production, and its use as cattle feed has increased as a result of the expansion of the fuel ethanol industry. However, the inclusion of corn DDGS into feedlot diets may increase the shedding of Escherichia coli O157:H7. This study investigated whether corn or wheat DDGS at 2 concentrations (20% or 40% vs. 100% barley grain) affected the survival of E. coli O157:H7 in incubations of ruminal digesta and feces. Neither the type nor the level of DDGS had any effect on fermentation or the survival of E. coli O157:H7 in ruminal digesta. However, there was a time by DDGS interaction (p < 0.05), where the numbers of E. coli O157:H7 in feces did not differ after 4 or 12 h of incubation but were greater after 24 h in both 40% wheat and 40% corn DDGS as compared with other treatments. Additionally, after 24 h, the numbers of E. coli O157:H7 were greater in fecal incubations with corn DDGS than with wheat DDGS (p < 0.05). The differences in the numbers of E. coli O157:H7 were not attributable to changes in pH or in concentrations of volatile fatty acids in the media. These results suggest that the inclusion of high levels of corn or wheat DDGS in feedlot diets of cattle may encourage the survival of E. coli O157:H7 in feces.