SUMMARY Until recently, the chicken has been considered the only natural host and the main host for chicken anemia virus (CAV). However, CAV infection has now been reported in other avian species, including Japanese quail, jackdaws, rooks, and rare avian breeds. In this study, we examined chickens, ostriches, and turkeys for seromolecular detection of CAV in Iran. For the study, 375 serum and thymus samples from chickens, 84 serum and blood samples from ostriches, and 100 serum and blood samples from turkeys were collected from various locations in Iran. Serum samples were examined using ELISA. Deoxyribonucleic acid was extracted from the thymus, and blood samples were analyzed for detection of the VP2 genome of CAV by PCR. According to these results, 69.07 and 58.4% of chickens were positive for the antibody to CAV and the VP2 genome of CAV, respectively. In addition, in ostriches 27.38 and 34.52% were positive for the CAV antibody and the CAV VP2 genome, respectively. In turkey flocks, all turkeys (100%) were negative with respect to detection of the VP2 CAV genome and CAV antibody. It was concluded that CAV is distributed in broiler chickens and ostriches throughout Iran. Furthermore, ostriches may be considered a reservoir of CAV, and turkeys appear not to be susceptible to CAV infection.