Physical, chemical, and serological characterization of rotavirus isolates from turkeys was done. Cesium chloride (CsCl)-gradient isopycnic centrifugation of infected cell cultures revealed the presence of rotavirus particles of three different densities. They were double-shelled, single-shelled, and core particles. The double-shelled particles had a buoyant density (in CsCl) of 1.34 g/cml3, and that of single-shelled particles in CsCl was 1.36 g/cm3. The buoyant density of core particles in CsCl was greater than 1.40 g/cm3. These rotavirus isolates were not inactivated by chloroform and were relatively stable at pH 3.0. Their replication was not affected by 5-bromo-2'-deoxyuridine. Avian rotaviruses were not completely inactivated by heat treatment of 56 C for 8 hr. All six avian rotavirus isolates examined were antigenically related to each other. However, there was no antigenic relationship between mammalian rotaviruses and the avian rotavirus isolates examined.