From 1981 to 1991, sera of 42 European brown bears (Ursus arctos) from three areas in Croatia were tested for antibodies against 12 Leptospira interrogans serovars: grippotyphosa, sejroe, australis, pomona, canicola, icterohaemorrhagiae, tarassovi, saxkoebing, ballum, bataviae, poi, and hardjo. Diagnostic levels of antibody were found in 17 (40%) of 42 sera. Evidence of exposure to at least one of the serovars was found in seven of 14 free-ranging bears from the Lika region, four of 12 free-ranging bears from the Gorski Kotar region, zero of six orphaned cubs from the Gorski Kotar region, and six of 10 captive bears from the Zagreb Zoo. Based on the antibody titers, we implicated the following serovars: australis in five bears, sejroe in two bears, canicola in one bear, and icterohaemorrhagiae in one bear. There was a strong correlation between serovars implicated by this survey and serovars previously isolated from small mammals in Croatia.