Between the years 2000 and 2004, 93,107 sera from 1,997 pig herds in 11 regions of Croatia were tested for the presence of antibodies against brucellosis. Positive results were observed in 67 herds from seven regions (mean individual prevalence: approximately 1%; herd prevalence: 3.4%). The herds from all but two of the infected farms were reared outdoors and thus almost certainly came into contact with wildlife. From 2003 to 2004, 424 sera, which were randomly collected from hunted wild boar (Sus scrofa), were also tested and shown to have a mean seroprevalence of 27.6%. Brucella was isolated from 88 out of 151 serologically positive pigs (58.3%) and 7 of the 93 (7.5%) wild boar which were randomly submitted for bacteriological study. All but three isolates were Brucella suis biovar 2; the others being biovar 3. These results suggest that brucellosis is enzootic in Croatian populations of wild boar. These populations represent a potential disease reservoir for free-range pig farms, as they do in other countries of Central and Western Europe. This is the first report of B. suis biovar 3 in swine and wild boar in Europe, which is an issue of serious concern for public health.