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Background: Spillage of stones in peritoneal cavity is quite common in patients undergoing laparoscopiccholecystectomy. Lost stones may cause wide range of complications postoperatively which reflects theinfective potential of these stones. Objective: The aim of this study was to find out the frequency of viable bacteria in stones retrieved fromthe gallbladder after laparoscopic cholecystectomy & subjecting the stones for culture. Design: Cross-sectional study. Place &Duration: This study was conducted at Surgical Unit 4 of Civil Hospital Karachi,from July 2013 till March 2014. Materials & Metods: 80 cases of cholelithiasis , selected & operated by laparoscopic cholecystectomy wereincluded. During cholecystectomy , stones were collected , intact stones were sent in two separate containersfrom each subject, for microbiological examination. Results: This study included 80 patients, 58 patients(72.5%) were female & 22 patients (27.5%) were male.Out of these 80 patients, 27 patients (33.75%) had positive stone culture, out of these 27 patients , 16 patients(20%) with positive core culture, 11 patients (13.75%) had positive surface culture.The commonest organismwas E.coli in 13 patients (16.3%) followed by Pseudomonas in 3 patients(3.8%), Klebsiella in 3 patients(3.80%), Staphylococcus aureus in 2 patients (2.5%) & Bacteroides fragilis in 1 patient (1.3%). No salmonellaspecies were isolated. All organisms were tested for sensitivity against quinolones,penicillins, cephalosporins,aminoglycosides & carbapenems. Conclusion: This study proves the infective potential of gallstones, as they contain viable bacteria, whichmay cause post operative complications, therefore all spilled stones during surgery should be removed &peritonial cavity should be meticulously cleared.