Bovine paratuberculosis or Johne's disease is caused by Mycobacterium avium subsp. paratuberculosis (MAP), which affects domestic and wild ruminants around the world. The clinical presentation of MAP infection is characterized by chronic diarrhea unresponsive to treatment. The objective of the present study was to investigate the seroprevalence and risk factors associated with bovine paratuberculosis in cattle herds of Tuta, Boyacá, Colombia. This descriptive cross-sectional study with simple random sampling was performed on 882 blood samples taken from cattle of different racial and age groups. Blood samples were processed using an indirect enzyme-linked immunosorbent assay (PARACHEK ® 2 Kit, Prionics AG, Switzerland). The obtained data were analyzed by the statistical software Epi Info. In this study, a general seroprevalence of 3.1% was found, and seropositivity in females was 3.6%. The highest prevalence of antibodies against MAP was in individuals > 4 years (5%) and the Jersey breed (4.8%). Therefore, the age of over 4 years was identified as a risk factor associated with MAP. Moreover, a statistical association was found between management and biosafety variables, such as pen management (p = 0.012), feeding with concentrate (p = 0.012), and the presence of diarrhea on the farm (p = 0.048). It could be concluded that the disease is present in Tuta, however, considering factors, such as the chronicity of the disease and the diagnostic method used, it is expected that the number of infected animals is much higher than presented in this research. [ABSTRACT FROM AUTHOR]