The aim of this study was to determine the prevalence of bovine viral diarrhoea (BVD), bovine neosporosis (NEOS), enzootic bovine leukosis (EBL) and bovine paratuberculosis (BP) in dual-purpose cows reared under low-tropic conditions. The study was carried out at La Pintada farm located in the municipality of Puerto Boyacá, Boyacá, Colombia, taking blood samples from 840 2–3-year-old cows collected from various departments of the country. Competitive (BVD, NEOS, EBL) and indirect (BP) ELISA tests were performed. A seropositivity of 25.83% to BVD, 16.29% to NEOS, 14.64% to EBL and 4.17% to BP was found.