A total of 330 enterococci strains isolated from several human (272 strains) and animal (27) clinical specimens and environmental sources (31) in Brazil were identified to species level. Major human sources included urine (48.5%), blood (15.8%), and wounds (9.5%). Human isolates were identified as Enterococcus faecalis (90.0%), E. faecium (6.9%), E. gallinarum (1.1%), E. durans (0.8%), E. casseliflavus (0.4%), E. raffinosus (0.4%), and E. mundtii (0.4%). Strains isolated from animals were composed of E. hirae (40.7%), E. faecalis (33.3%), E. faecium (18.5%), and E. casseliflavus (7.5%). Among environmental isolates, 42.0% were E. faecalis, 35.4% E. faecium, 13.0% E. hirae, 6.4% E. casseliflavus, and 3.2% E. durans. Antimicrobial susceptibility tests were performed for 200 strains. Overall, high-level resistance (HLR) to aminoglycosides was found in 66 (33.0%) of the strains tested. HLR to gentamicin was detected in 11.5% of the strains, whereas 19.0% of the strains showed HLR to streptomycin and 26.0% showed HLR to kanamycin. Five (22.7%) of the E. faecium strains were resistant to ampicillin [minimum inhibitory concentration (MIC) > 32 micrograms/ml]. Vancomycin MIC50 and MIC90 were 2 and 4 micrograms/ml, respectively; only eight strains (identified as E. casseliflavus or E. gallinarum) had MIC of 8 micrograms/ml. No beta-lactamase activity was detected by the nitrocefin test.