Background: Antibiotic stewardship during livestock production is performed empirically and regulations are not readily followed in Colombia. Therefore, the risk of emergence of multidrug resistant strains might occur across the food chain and reach the consumer. When antibiotics are administered using sub-therapeutic doses, fewer days of treatment or simply without considering the withdrawal time; it can trigger contamination in bovine milk intended for human consumption. Our aim was to identify pathogenic bacteria and their level of antimicrobial resistance in milk harvester routes in Fusagasugá and Granada, Cundinamarca. Methods & Materials: We identified two main milk harvester routes from the municipalities of Fusagasugá and Granada in Cundinamarca (Colombia). Milk samples were pooled and were microbiologically evaluated for bacterial contaminations according to the National Laboratory of Veterinary Diagnosis–ICA guidelines. Microbiological culture and identification test were performed by phenotypic methods and susceptibility test were carried out by the disk diffusion methods. We designed a survey to evaluate the knowledge in antibiotic stewardship in livestock farmers who provided raw milk to the milk collectors. Results: Escherichia coli and Streptococcus agalactiae were detected in the milk harvester route from the municipality of Fusagasugá and Streptococcus agalactiae of the route from Granada municipality. E. coli presented resistance to four different types of antibiotics (Fosfomycin, Ampicillin, Amoxicillin and Trimethoprim-sulfamethoxazole). On the other hand, S. agalactiae was resistant to eight different types of antibiotics (Erythromycin, Fosfomycin, Oxacillin, Ampicillin, Ceftazidime, Ciprofloxacin, Lincomycin, Gentamicin, Tetracycline and Trimethoprim-sulfamethoxazole). In the survey, 79% of livestock farmers reported that they only used of antibiotics to treat diseases; however, antibiotics were administered by recommendation in 34% of the cases, 24% due to a previous experience and 20% because of quality. Additionally, in 59% never requested veterinary services due to high cost (53%) and difficult access (26%). Conclusion: Poor livestock practices are contributing to the emergence of multidrug resistant strains in Fusagasugá and Granada municipalities. Therefore, it is urgent to screen for microbial contamination in dairy products to restict the marketing and sales and to educate the livestock farmers in antibiotic stewardship.