Two cases of cutaneous Pythiosis insidiosi were diagnosed in cattle from the Pantanal region, Brazil. The lesions were observed in the limbs of two 8-month-old beef calves. Close examination showed local swelling and focal ulceration of the skin. Microscopically, there was multifocal granulomatous dermatitis with intralesional Pythium insidiosum hyphae. The diagnosis was based on the morphological aspects, immunohistochemical findings and culture of the etiologic agent.