We collected blood and serum from 155 brown bears (Ursus arctos) inhabiting five locations in Alaska during 2013–16 and tested samples for evidence of prior exposure to a suite of bacterial, viral, and parasitic agents. Antibody seroprevalence among Alaska brown bears was estimated to be 15% for Brucella spp., 10% for Francisella tularensis, 7% for Leptospira spp., 18% for canine adenovirus type 1 (CAV-1), 5% for canine distemper virus (CDV), 5% for canine parvovirus, 5% for influenza A virus (IAV), and 44% for Toxoplasma gondii. No samples were seropositive for antibodies to Trichinella spp. Point estimates of prior exposure to pathogens among brown bears at previously unsampled locations generally fell within the range of estimates for previously or contemporaneously sampled bears in Alaska. Statistical support was found for variation in antibody seroprevalence among bears by location or age cohort for CAV-1, CDV, IAV, and Toxoplasma gondii. There was limited concordance in comparisons between our results and previous serosurveys regarding spatial and age-related trends in antibody seroprevalence among Alaska brown bears suggestive of temporal variation. However, we found evidence that the seroprevalence of CAV-1 antibodies is consistently high in bears inhabiting SW Alaska and the cumulative probability of exposure may increase with age. We found evidence for seroconversion or seroreversion to six different infectious agents in one or more bears. Results of this study increase our collective understanding of disease risk to both Alaska brown bear populations and humans that utilize this resource.