The Barn Owl, Tyto alba (Scopoli, 1769), occurs worldwide and shows a considerable amount of morphological and geographical variation, leading to the recognition of many subspecies around the world. Yet comprehensive study on this species needs to be done. Data from mitochondrial gene (CYTB) with 620bp length is analyzed for 30 individuals around the world. Maximum likelihood (ML), maximum parsimony (MP) and bayesian analysis showed considerable genetic variation between alba clade (Old World) and furcata clade (New World). The amounts of genetic variation within each of these clades are in ranges from 0.4%-1.6% but variation between clades is 7.21%. This data may suggest that Barn Owls of the Old World were a separate species from those of the New World. We found high amount of genetic variation between T. a. stertens from Indonesia and alba clade and we didn’t find any support for recognition of T. bargei as a separate species.

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