Melanosis of the urinary bladder is a very rare condition characterized by an abnormal black or brownish-black pigmentation of the organ. The pigmentary disorder can involve both the urothelial cell layers and/or the submucosa. The biologic potential of the melanosis of urinary bladder remains unknown because only a few cases have been reported in medical literature. So far melanosis of the urinary bladder is not known to occur in cattle. Here we describe the first case of melanosis of the urinary bladder in an inbred red-spotted, 7-year-old cow. Light, ultrastructural, and immunohistochemic investigations demonstrated melanin pigment in the submucosa and lamina propria but not the urothelium of the bladder. In addition, biochemical characterization of the pigment-laden cells demonstrated that the pigment of this disorder consisted mainly of eumelanin, thus corroborating the morphologic studies. Finally, virologic examination revealed the presence of bovine papillomavirus type 2.