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Introduction: Pseudomonas aeruginosa species can be dangerous opportunistic pathogen because of its tolerance to physical, chemical, and antibacterial compounds. In hospitals, P. aeruginosa is a formidable opportunistic pathogen, and therefore, the medical concern with infection of immunologically compromised patients in burns and neonatal units, is well justified. Material and methods: Total 1583 samples like swab, urine, sputum, pus, pleural fluid, bronchoalveolar lavage (BAL), ascitic fluid and blood samples from different clinical departments were tested at Clinical Microbiology Department of B. J. Medical College and Civil Hospital, Ahmedabad, Gujarat during April 2009 to April 2010. Results: Out of 1583 samples, 807 samples turned culture positive. Out of 807 culture positive samples, 100 were culture positive for P. aeruginosa. The maximum number (68%) of P. aeruginosa isolates were obtained from swab samples. The highest number of such isolates (48%) belonged to surgical ward. P. aeruginosa showed highest sensitivity against Cefepime - Tazobactam (97%). Conclusion: This study showed that P. aeruginosa is acquiring resistance to commonly used antibiotics as well as newer antibiotics. The antimicrobial agents are losing their efficacy because of spread of the resistant organism, indiscriminate use of antibiotics, and unhygienic condition. It is the need of the time that antibiotic policies should be formulated and implemented to resist and overcome this serious problem. [ABSTRACT FROM AUTHOR]