The aims of this study were to assess community knowledge, awareness and practices on zoonoses, to gather baseline data on brucellosis in livestock and wildlife, to establish brucellosis seroprevalence in domestic ruminants and humans and risk factors associated with livestock seropositivity, to assess brucellosis dynamics and impact on livestock production and reproduction and to evaluate the performance of the Rose Bengal Plate Test (RBPT) in Tanzania. The results described in this study were carried out through Participatory Rural Appraisal (PRA), cross-sectional and longitudinal studies. In the PRA and cross-sectional studies, rabies, tuberculosis, anthrax and brucellosis were the zoonoses most frequently identified. Cattle were frequently identified as being associated with tuberculosis, anthrax and brucellosis, whereas dogs were frequently identified as being associated with rabies. Small ruminants, pigs, cats and poultry were either infrequently, or not identified as being associated with zoonoses. Recognition of clinical signs of zoonoses in humans was better than in animals. Ingestion of animal products was a route frequently identified as transmitting zoonoses to humans. During the baseline serosurvey, seroprevalences for brucellosis were 6.2% in cattle, 6.5% in small ruminants and 13% in wildlife, respectively. Seropositivity was significantly higher in the pastoral (13.2%), followed by agro-pastoral (5.3%), and lowest in the small holder dairy system (2.3%) (p