The aim of this study is to evaluate the frequency of microorganisms isolated from pleural fluids and their resistance to antimicrobial agents. A total of 272 pleural fluids were studied between July 2004 - July 2005 from the patients hospitalized in ICU/surgery (127) and respiratory diseases wards (145) at Marius Nasta Institute. The laboratory investigations included: direct microscopy, cultures for aerobic and anaerobic bacteria, identification, disk diffusion method according with CLSI recommendations for resistance and Etest for detection of metallo-beta-lactamase producing isolates of Pseudomonas aeruginosa. Microorganisms were isolated from 159 samples (58.4%), 48 pleural fluids were positive only in microscopy (17.6%). The most frequent isolated strain was P. aeruginosa (49.6%), followed by Staphylococcus aureus (12.8%) and Enterobacteriaceae (11.2%) polymicrobial infections were mostly due to combinations of Pseudomonas with Enterobacteriaceae. For P. aeruginosa the resistance rate was higher than 71% for all beta-lactams. For aminoglycosides the lower resistance rate was to amikacin (18.0%). For quinolones, resistance of P. aeruginosa was 67.8% to ciprofloxacin. P. aeruginosa isolated from patients hospitalized in ICU/surgery were more resistant to some antimicrobials than the strains isolated in the respiratory diseases wards: resistance to amikacin was 24.5% versus 10% respectively. From 21 P. aeruginosa imipenem and multidrug resistant strains tested, 3 were probably producing of metallo-beta-lactamase. S. aureus showed 47.1% oxacillin resistance, 38.9% resistance to gentamicin and ciprofloxacin and 27.7% to erythromycin. All S.aureus strains were susceptible to linezolid, teicoplanin and vancomycin. The resistance of Enterobacteriaceae strains was high to ampicillin (80.0%), amoxicillin/clavulanic acid and trimethoprim/sulfamethoxazole e(57.6%); the lowest resistance rate was to cefoperazone/sulbactam (7.7%) and to imipenem and ciprofloxacin (10.8%).