A collection of 94 Gram-negative bacteria isolates, showing different antimicrobial resistance phenotypes including to the carbapenem classes was investigated. Strains were originated form clinical sources from a single hospital in Tripoli, Libya during 2015 and were identified based on culture and phenotype characteristics, and fully characterized by the VITEK automated system. Forty-eight percent (48%) of the collection was Acinetobacter baumannii, 50% was Klebsiella pneumoniae and 2% was Escherichia coli. Resistance to the carbapenem classes was reported in 96% of the A. baumannii strains and 94% of the K. pneumonia strains. Seventy-eight percent (78%) of the isolates showed different multidrug-resistant (MDR) phenotypes, with K. pneumoniae expressing the highest proportion (91%). Emergence of resistance to carbapenems among the Gram-negative strains is a challenging global problem, particularly for Africa. Surveillance of these pathogens is urgently required in the Libyan healthcare settings.