A 5-year-old spayed male ferret showed a subcutaneous mass in the right lateral thoracic wall. Microscopic examination revealed that the neoplasm had proliferated in the subcutis with infiltration into the surrounding tissues. A packed bundle of large polymorphic neoplastic cells, containing abundant eosinophilic cytoplasm and a round to ovoid, occasionally bizarre nucleus, were arranged interwoven. The neoplasm had metastasized to the right axillary lymph node. The neoplastic cells were intensively positive for vimentin, desmin and myoglobin. Skeletal muscle type creatine phosphokinase-positive granules were detected in the cytoplasm. Ultrastructurally, various amounts of disorganized myofibrils with focal density resembling the Z-band were shown in the cytoplasm of the neoplastic cells. The neoplasia was diagnosed as rhabdomyosarcoma.