Purpose: Antibiotic misuse is a common practice during animal production by livestock farmers in the Sumapaz region (Colombia). Thus, contribution of antibiotic resistance through agricultural and livestock practices is an underestimated problem in Colombia. The purpose of this study was to screen for foodborne and zoonotic pathogens associated with local livestock practices. Methods & Materials: We included twenty-four farms in the Sumapaz region that were selected by random. Nutritional content of the milk was analyzed using an ultrasonic milk analyzer (LactoScan system). Bacterial identification was performed by using phenotypic methods based in biochemical reactions and bacterial metabolism for Gram positive and negative bacteria. Antibiotic susceptibility in the milk samples was assessed according to the International guidelines (Disk diffusion method) for ten recommended antibiotics in veterinary medicine. Results: We found no difference in the nutritional content of the raw milk samples. However, we found that twelve milk samples were contaminated with E. coli and S. aureus resistant to multiple antibiotics mainly to oxytetracycline and erythromycin (73% of all isolates). Other E. coli strains were resistant to Amoxicillin (n=10, 83%) and multiple resistance to fosfomycin, oxytetracycline and gentamicin (n=8, 67%). None of the bacterial isolates were susceptible (except to enrofloxacin). Samples obtained from 1098 animals (spread over forty six livestock farms) were screened for bovine TB and brucellosis. Twenty-three were positive for Brucella sp., representing a 4.2% prevalence. We did not find any positive cases of bovine tuberculosis. Conclusion: Our findings suggest that milk raw could be a source of exposure to multidrug resistant E. coli and S. aureus strains as a result of poor livestock practices in the Sumapaz region (Colombia). Training in good farming practices is key to improve sustainable food production and avoiding antibiotic-resistant foodborne disease outbreaks. Further studies to characterize the mechanisms of antibiotic resistance are necessary.