The nonenveloped avian reoviruses represent a distinct antigenic subgroup of orthoreoviruses. Unlike their mammalian counterparts, the avian reoviruses exhibit the unusual property of inducing rapid and extensive syncytium formation in cell cultures, a cytopathic effect more commonly associated with enveloped virus replication. While the syncytium-inducing capability of avian reovirus has been known for quite some time, the relationship between cell fusion and the virus replication cycle has not been determined. The conservation of the syncytial phenotype among all field isolates of avian reovirus suggests that avian reovirus-induced syncytium formation either reflects an essential step in the virus replication cycle involving intracellular membrane interactions or that cell fusion contributes to enhanced virus replication in infected animals. In order to distinguish between these possibilities, we have examined several aspects of virus replication in the presence of inhibitors of syncytium formation. Inhibitors of intracellular vesicle transport and O-linked glycosylation eliminated or markedly reduced syncytium formation with little effect on the rate or extent of virus macromolecular synthesis and infectious progeny virus production. Moreover, syncytium formation was not required for virus-induced cytopathology or virus egress but did significantly enhance the rate of both of these processes. The data indicate that, unlike the syncytium-inducing enveloped viruses, the membrane interactions and protein trafficking required for avian reovirus-induced syncytium formation do not reflect the sequelae of an essential step in the virus replication cycle. These results suggest that the conservation of the avian reovirus syncytial phenotype may reflect a fortuitous aspect of virus replication which confers advantages associated with the rapid spread of the virus within an infected host.