In May 1991, clinical, pathologic, and virologic investigations were carried out on an 8-yr-old male lion (Panthera leo), with recurrent infections, in captivity with two lionesses in the Zoological Garden of Pistoia, Tuscany, Italy. The lion had severe pneumonia, neutropenia, thrombocytopenia, and' an increase in blood urea nitrogen and creatininemia; in spite of therapy, it died within 3 months. At necropsy, the animal had a lymphoma and other lesions similar to those described in feline immunodeficiency virus-infected cats. We identified FIV gag-sequence using PCR amplification of lymph node tissues.