Several aspects of the epidemiology of rotavirus suggest the possibility that transmission may occur by nonenteral routes. We utilized the mouse model of rotavirus infection to investigate the experimental transmission of rotavirus infection by respiratory droplets. Following exposure to a defined dose of aerosolized rotavirus, the kinetics of viral replication within the lung and gastrointestinal tract was studied using a double antibody enzyme immunoassay and indirect immunofluorescence. These studies documented the efficient transmission of rotavirus infection by means of aerosol in all exposed animals. Rotavirus antigen was detected as early as 12 hours after infection in the pulmonary and gastrointestinal tracts of the infected animals and antigen remained detectable in both sites for at least 8 days following infection. Gastrointestinal illness was clearly demonstrable in the animals while pulmonary pathology was not evident. These studies document that rotavirus infection can be transmitted by aerosol droplets under experimental conditions.