Previous studies have shown that viral upper respiratory tract (URT) infection predisposes to the development of bacterial otitis media (OM). The mechanisms underlying this phenomenon have been well defined for influenza A virus in the chinchilla model of experimental OM but have not been for adenovirus because of the heretofore lack of an appropriate model. In this study, chinchillas were inoculated intranasally or transbullarly with type 1 adenovirus to assess whether or not the chinchilla could serve as a suitable model for future investigation of adenovirus predisposition to bacterial OM. Data indicated that the chinchilla readily supported an active infection by and responded serologically to this agent and that route of inoculation markedly influenced otoscopic findings, elicitation of middle ear fluids, onset and location of histopathology, and progression of disease. The time course of disease onset and recovery and an assessment of the effect of this virus isolate on the mucosal epithelial integrity and on both the ciliary activity and transport function of the eustachian tube epithelium for both routes of inoculation are presented.