Antimicrobial resistance poses a critical challenge to global public health, complicating the treatment of common and life-threatening infections caused by bacteria, viruses, fungi, and parasites. This issue is particularly severe in developing regions, where factors such as the inappropriate use of antibiotics and inequalities in healthcare access contribute to the rise of multidrug-resistant strains. Unfortunately, Colombia is no exception to this growing problem. Nationally, hospital-associated infections caused by Klebsiella pneumoniae and Pseudomonas aeruginosa have become increasingly frequent, with resistance to broad-spectrum antimicrobials such as carbapenems being on the rise. Additionally, a descriptive analysis of bacterial isolates from intensive care units (ICUs) across twenty hospitals in twelve Colombian cities from 2019 to 2021 revealed a significant increase in the prevalence of multidrug-resistant microorganisms. In the most recent report (2019) on laboratory surveillance of antimicrobial resistance in healthcare-associated infections (HAIs) by the National Institute of Health (INS) of Colombia, the department of Magdalena exceeded national resistance rates for Escherichia coli, K. pneumoniae, P. aeruginosa (resistant to at least one carbapenem), and Staphylococcus aureus (oxacillin-resistant). However, no updated epidemiological data on antibacterial resistance in this region are currently available. This study presents an epidemiological analysis of the resistance profiles of pathogenic bacterial strains in the department of Magdalena, Colombia, during 2023–2024. A retrospective descriptive study was conducted using data from the Julio Méndez Barreneche University Hospital (HUJMB) in Santa Marta D.T.C.H. The findings allow us to identify the most prevalent Gram-positive and Gram-negative bacterial strains in the region and explore potential risk factors.