The objective of the present study was to detect the presence of Leptospira spp. and Mycobacterium spp. by means of molecular techniques in cattle of Los Palmitos municipality, Sucre-Colombia.. One hundred seventy-two blood samples from individuals from 12 farms were studied. They were analyzed by polymerase chain reaction (PCR) with two pairs of primers amplifying a region of the 16S rRNA gene of Mycobacterium spp. and other primers specific for Leptospira DNA gyrase subunit B (gyrB). The segment of interest of the 16S rRNA gene was amplified from 35 samples (20 %), of these, 12 were sent for sequencing. Only eight sequences were informative, of which six belonged to species of the genus Dietzia and two to the genus Mycobacterium . Of the three positive samples to Leptospira spp. (1.7 %), only one was recovered showing identity with the species Leptospira interrogans . These results allow to affirm that cattle in Los Palmitos, Sucre-Colombia, are carriers of bacterial species that can generate a negative impact on livestock production, being also a risk to human health.