In this study, the localization of 3 beta-hydroxysteroid dehydrogenase (3beta-HSD) activity in the wall of canine perianal sinus (PS) was determined. The 3 beta-HSD activity was found out both in the cytoplasm of cells, situated in the propria and forming clusters adjacently to apocrine glands and in the cytoplasm of some epithelial cells in apocrine cells' glands. The results obtained about the 3 beta-HSD activity allowed us to propose a role of this enzyme in PS development and possibly, in tumourogenesis.