Background: Peritonitis is inflammation of the lining of the abdominal cavity. The present study was conducted to study of organism profile in relation to site of perforation in peritoneal fluid in patients of peritonitis. Materials & Methods: 60 patients with a diagnosis of peritonitis were assessed for site of perforation and peritoneal fluid culture and sensitivity report of every patient was sent to the department of microbiology. Patient was reviewed till the day of discharge. Peritoneal, aerobic culture and sensitivity report was followed, and antibiotics were changed as per the sensitivity pattern of the organism grown in the culture. Results: The majority of the patients were between the age group of 18-29 years [20(33.33%)], followed by 30-44 years [19 (31.67%)]. Male preponderance was observed among patients [46(76.67%)]. The majority of the patients were delayed by 0-2 days [29(48.33%)], followed by a delay of 3-5 days [27(45.00%)]. The number of smokers [31(51.67%)] were more than non-smokers [29(48.33%)] among the enrolled patients. Statistically, insignificant difference was observed among patients. The majority of the patients had perforated appendix [17(28.33%)], followed by perforated gastric[11(18.33%)], perforated ileum [10(16.67%)]. Most of the patients had E. coli-in their peritoneal fluid [25(41.67%)], followed by Enterococcus Faecalis [6 (10.00%)] and so on. At the same time, no-aerobic growth was observed in [18(30.00%)] patients. The majority of the patients showed no post operative complications [33(55.00%)], while [18(30.00%)] patients had SSI. Respiratory failure was seen inonly[1(1.67%)] patient. A total of 25 patients had E. coli growth in the appendix [n=12], followed by the ileum [n=5]. A total of 6 patients had Enterococcus faecalis growth at the duodenum [n=3], followed by ileum [n=2]. Pseudomonas aeuroginos a growth was observed at the duodenum [n=1]. Conclusion: The most common site of perforation was Appendix, followed by gastric and duodenum. Peritonitis was most commonly due to E. coli, followed by Enterococcus faecalis. E. coli has also emerged as the predominant organism implicated in the pathogenesis, even if we consider it a site-specific culture. E. coli was sensitive to Amikacin, Tigecycline, Gentamycin, Imipenem, Meropenem and Cefoperazone-sulbactam.At the same time E. coli was resistant to Cefipime, Ceftriaxone and Ciprofloxacin. Surgical site infection was the most common postoperative complication.