The aim of this study was to investigate the in vitro antimicrobial susceptibilities of clinically important Gram-negative bacteria from seven intensive care units in Taiwan in 2016. Materials and methods: In total, 300 non-duplicate isolates of Escherichia coli (n=100), Klebsiella pneumoniae (n=100), and Pseudomonas aeruginosa (n=100) collected from 300 patients were studied. The minimum inhibitory concentrations (MICs) of these isolates to antimicrobial agents were determined using the broth microdilution method. Carbapenemaseencoding genes (bla KPC , bla NDM , bla IMP , bla VIM , and bla OXA-48-like) were studied for the isolates that were not susceptible to any carbapenems. Sequencing analysis of the mcr genes (mcr-1-5) was conducted for all isolates with colistin MICs ≥4 mg/L. Results: Ertapenem non-susceptibility was detected in 3% (n=3) E. coli and 12% (n=12) K. pneumoniae isolates. The susceptibility rates of imipenem, ceftazidime-avibactam (CAZ-AVB), and ceftolozane-tazobactam (CLZ-TAZ) were 99%, 99%, and 88%, respectively, for E. coli, 91%, 100%, and 80%, respectively, for K. pneumoniae, and 66%, 91%, and 93%, respectively, for P. aeruginosa. Carbapenemase-encoding genes were not detected in E. coli, were detected in four (33.3%) K. pneumoniae isolates that were not susceptible to ertapenem (three harboring bla KPC and one harboring bla OXA-48-like), and were not detected in P. aeruginosa isolates that were not susceptible to imipenem. One K. pneumoniae isolate was resistant to colistin (MIC 4 mg/L) and negative for mcr genes. Conclusion: CAZ-AVB exhibited excellent activity against carbapenem-resistant Enterobacteriaceae, and CLZ-TAZ exhibited good activity against imipenem-resistant P. aeruginosa.