The present study was designed to investigate hemorrhagic enteritis virus (HEV) as a predisposing factor influencing the susceptibility of young turkeys to Escherichia coli infections. In addition, the pathologic changes caused by administration of E. coli by various routes were compared. Following oral infection with HEV, groups of turkeys were inoculated with various doses of pathogenic E. coli by intravenous (IV), intra-air sac (IA), or intratracheal (IT) routes. A synergistic effect was observed in birds that were exposed to a combined HEV-E. coli challenge, resulting in higher mortality than that caused by either pathogen alone. This synergy was more evident when the bacteria were administered by the IT route than when it was administered by the two other routes. Turkeys infected with HEV and then inoculated IT with E. coli O78 had higher mortality (61%) and higher occurrence of gross body lesions (74%) than birds given E. coli alone (0% mortality and 16% gross lesions). After E. coli inoculation by the IA and IT routes, lesions observed were mainly pericarditis, perihepatitis, lung and air-sac lesions, splenic enlargement, and occasional arthritis. The incidence of lesions was affected by HEV exposure. In contrast, IV inoculation with E. coli O78 usually resulted in arthritis, and its incidence was independent of previous HEV exposure. The synergistic effect between HEV and E. coli administered IT can be used as a challenge model for testing E. coli vaccines.