A dog was examined with a history of weight loss and lameness of the left hind limb. A painful response to examination of the left hip joint, and lymphadenopathy were noted. Amastigote forms of Leishmania sp. were observed by cytology in samples from the popliteus lymph node, and anti-Leishmania sp. antibodies at a titer of 1:640 were detected in serum by indirect immunofluorescence. Radiological changes included osteolysis and a periosteal proliferative reaction in the left femoral greater trochanter. These changes were histologically characterized as an osteolytic granulomatous osteomyelitis associated with amastigotes within macrophages. Non-decalcified fragments of the periosteum were processed for immunohistochemistry, observed with prominent immunolabelling of amastigotes of Leishmania sp. within macrophages. The diagnosis was further confirmed by positive PCR for Leishmania sp., belonging to the Leishmania donovani complex.