This study aims to investigate trends in bloodstream infections and their antimicrobial susceptibility profiles over 12 years in our hospital. This retrospective study was carried out in the Bursa Uludag University Hospital, Turkey, during 2008-2019. Blood cultures from patients were performed using BACTEC System. Isolates were identified with Phoenix System until 2018 and "matrix-assisted laser desorption ionization time-of-flight mass spectrometry" (MALDI-TOF MS) in 2019. Antibiotic susceptibility testing was performed with Phoenix System. Patient data came from the BD EpiCenter (TM) data management system. Escherichia coli was found to be the most common Gram-negative (11.6%), and coagulase-negative staphylococci were the most common Gram-positive (10.1%) monomicrobial growth. Overall, there was a significant increase in rates of extended-spectrum beta-lactamase positive E. coli (p = 0.014) and Klebsiella pneumonia (p < 0.001), carbapenem-resistant E. coli (p < 0.001), and K. pneumoniae (p < 0.001) and colistin-resistant K. pneumoniae (p < 0.001) and Acinetobacter baumannii (p < 0.001) over 12 years. Carbapenem and colistin resistance has increased dramatically in recent years. We believe that regular monitoring of the distribution of pathogens and antibiotic susceptibility profiles, especially in intensive care units, can contribute to evidence for the increase in resistant microorganisms and help prevent their spread with antimicrobial stewardship and infection control policies.