This study investigated the species distribution and antimicrobial resistance among 99 enterococci isolated from hospitalized patients in 12 hospitals in Cuba from October 2000 to September 2001. Species identification was performed by WIDER Automatic System (Francisco Soria Melguizo, Madrid, Spain), and the susceptibility testing was performed by disk diffusion, agar dilution, and E-test methods. Enterococcus faecalis was the most prevalent (85%) species, followed by E. faecium (10%), E. gallinarum (2%), E. casseliflavus (2%), and E. durans-hirae (1%). A higher percentage of resistance to ampicillin (50%), fosfomycin (40%), ciprofloxacin (30%), norfloxacin (20%), and tetracycline (90%) was detected in E. faecium isolates, whereas E. faecalis strains showed higher rates of resistance to erythromycin (52.4%), chloramphenicol (34.5%), rifampicin (62.5%), moxifloxacin (3%), and nitrofurantoin (2.4%). Resistance to glycopeptide was detected in E. faecalis (1.2%) and E. faecium (10%). Thirty-one E. faecalis (37%) and 3 E. faecium (30%) showed a high-level resistance to gentamicin. The results of this work will be very helpful to guide empirical antimicrobial therapy and the implementation of infection control measures in Cuban hospitals.