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In healthy individuals, the bile is usually sterile, however, in cases of cholecystolithiasis and/or choledocholithiasis, it could be colonized with bacteria (bactibilia) and may lead to surgical site infection after cholecystectomy. In our hospital, the local regimen is to use antibiotics prophylaxis only for patients with high-risk factors, while in case of low-risk patients, the antibiotics used as postoperative treatment; this local regimen has no demonstrated bacteriological or epidemiological basis. The aim of this study is to determine the nature of bacteria in bile and their antimicrobial susceptibility in low-risk patients and the relationship between bactibilia and the presence of some predisposing factors as well as developing postoperative infectious complications. This study was conducted in Al-shiffa General hospital, Basrah, Iraq from April 2018 to May 2019. Forty-three patients with uncomplicated symptomatic gallstones who were candidates for elective laparoscopic cholecystectomy and have no risk factors for infection were included in the study. Under fully aseptic technique, a sterile laparoscopic needle connected to a sterile 10ml disposable syringe used to aspirate 5–7ml of bile from the fundus of gallbladder for culture and antibiotic sensitivity. In this study culture assessment of bile demonstrate that, 20 patients (46.51%) have infected bile; Klebsiella spp. 7(35%) and Pseudomonas spp. 7 (35%) are the most frequent causative agents. Factors like gender, age, duration of complaint and number of stones were found not significantly increase the risk of infection. Antibiotics sensitivity revealed maximum sensitivity to Meropenem (100%), Amikacin (90%) and less sensitivity to the most commonly used Ceftazidime. No cases of superficial or deep-seated surgical site infections were reported. In conclusion, bile infection reported in a significant rate in low risk patients for infection subjected to elective laparoscopic cholecystectomy. Klebsiella spp. and Pseudomonas spp. are the most common isolate which shows high sensitivity to Meropenem and Amikacin and less sensitivity to third generation Cephalosporine; so we recommend the use of Amikacin as a prophylactic antibiotic instead of third generation Cephalosporine. Keywords: Bile infection, Cholecyctectomy, elective surgery, laparoscopy, low risk patients