Background: Bactibilia, or the presence of bacteria in bile from the gallbladder, may contribute to the development of septic complications. It has been related to increased rates of surgical site infection after cholecystectomy. Laparoscopic cholecystectomy is the definitive treatment for cholelithiasis and cholecystitis. Advanced age, acute cholecystitis, ERCP, and gallstones are important risk factors for the development of bactibilia. This study aims at showing the effect of bactibilia on the course of laparoscopic cholecystectomy. Materials and methods: This are a prospective case-control study to be conducted at Govt. General Hospital, Vijayawada at Dept. of General Surgery over 1 year on 50 patients. Patients with asymptomatic Cholelithiasis of age 18-60 are included in the study and patients with hemoglobin < 10 gm%, hypoalbuminemia < 2.5 gm/dL, patients in acute sepsis, patients with HbA1c >6.5%, hypertension (>139/89 mm of Hg) and BMI > 30 and patients in an emergency setting and patients who have not consented for the study were excluded from the study. None of the patients received pre-operative antibiotics. Intra-operatively, bile is collected from the gall bladder and sent for microbiological culture in aerobic and anaerobic media for 3-5 days. Patients are divided into: Group A - Microbiological culture positive for microbes and Group B - Microbiological culture sterile. Both groups will be analyzed for the duration of hospitalization after surgery, post-operative wound infection, and any other complications encountered. Results: Out of the 50 patients, 32 were males and 18 were females. The mean age of the patients was 43.5 ∓ 9.6 years. The mean BMI was 27.1 ∓ 4.7. Of the total 25 samples positive for bile culture, 11 were E.coli, 7 were Klebsiella, 3 were Enterobacter spp., 2 were Acinetobacter spp., and the remaining were 2 Viridans streptococci. All the bacteria were sensitive to Amikacin. All except Acinetobacter were sensitive to gentamycin and imipenem. All except one case of E.coli were susceptible to ciprofloxacin. The incidence of port site infection, prolonged fever, intra-abdominal abscess, pneumonia, wound healing duration, and hospital stay were significantly higher in group A compared to group b (p-value < 0.05). Conclusion: Bactibilia affects the outcomes of laparoscopic cholecystectomy in terms of prolonged hospital stay, increased incidence of wound infection, duration of wound healing, and fever.