Introduction: Acinetobacter species have become commonest cause of hospital acquired infections over the last two decades, causing serious life threatening infections. Over the time there has been emergence of multi drug resistant strains of Acinetobacter spp. which has led to limited therapeutic options for treatment of patient. Aim: To identify & isolate Acinetobacter species from various clinical samples & to determine the antibiotic susceptibility pattern of these isolates. Materials and Methods: The study was conducted in Microbiology department at a tertiary care hospital, Solapur, Maharashtra, over a period of 6 months. All samples received for culture and sensitivity, were processed according to standard protocols. Results: Total 1291 samples received in Microbiology department for culture & sensitivity. 430 (33.3%) samples were sterile & 861(66%) samples were culture positive, of which 609 (66%) were gram negative isolates. Among gram negative isolates Acinetobacter spp. were 147 (17%) Acinetobacter spp. were most commonly isolated from wound swab (62%) followed by blood (12%), sputum (5.44%), CSF (4.76%), urine (4.08%). Antibiotic susceptibility pattern showed maximum resistance to Cefotaxime (89%), followed by Amoxicillin-sulbactam(87%), Piperacillin-tazobactam (80%), Amikacin(79.7%),Imipenem (77%), Ciprofloxacin (70%). Conclusion: Acinetobacter spp. are commonly present in hospital environment and are MDRO, even to newer antimicrobials. Therefore rational use of antibiotics is important and necessary to prevent microbial resistance.