Brucella ovis was isolated for the first time in Italy in 1994 from the genital organs of two domestic rams. In subsequent years bacteriologic and serologic investigations demonstrated an increasing distribution of this disease in domestic sheep. Mouflon (Ovis musimon) occur in several hilly and mountainous areas of Italy where they can potentially contact domestic sheep. To determine if this species may have a role in the epidemiology of B. ovis, four male and four female mouflon, serologically negative for B. ovis and other Brucella spp., were infected intraconjunctivally with B. ovis strain BG1/94. Physical examinations, including collection of blood samples for serology and bacteriology, were performed weekly. The animals were euthanized 8 mo postinoculation (p.i.). Samples of retropharyngeal, parotid, and iliac lymph nodes; bone marrow; kidneys; spleen; epididymis; testicle; bulbourethral glands; seminal vesicles; uterus; and oviducts were collected from each animal as appropriate for histopathology and bacteriology. At the time of euthanasia none of the animals exhibited obvious clinical signs of brucellosis. The animals seroconverted 2 wk p.i. and became seronegative 24 wk p.i. Bacterial cultures, including hemocultures, were negative. No lesions due to B. ovis infection were revealed by histologic examinations. Brucella ovis probably did not infect mouflon and this wild sheep is not likely to play a role in the epidemiology of contagious epididymitis caused by B. ovis.