Introduction: Abdominal pain is a common presentation at out patient and emergency service in Cauca, Colombia, it is the fifth leading cause of consultation and hospitalization. Infections of the bile duct are a significant cause of bacteremia and high mortality, hence the importance of establishing an early and appropriate antibiotic therapy. We sought to describe the most frequently isolated bacteria in cultures of bile and blood cultures of patients with acute cholecystitis and / or cholangitis University Hospital San Jose from, Popayan, Colombia. December 2013 to December 2014 and know their profile sensitivity or resistance to antibiotics. Materials and methods: This was a descriptive, cross-sectional study in patients with acute cholangitis and acute cholecystitis at University Hospital San Jose, in the period December 2013 to December 2014. Bile cultures and blood cultures were taken and antibiotic sensitivity tests were performed. Results: Of a total of 106 patients, positive bile cultures were positive in 49.1% and blood cultures were studied in 19% of patients. Most isolates were: Escherichia coli (51.42%), Klebsiella pneumoniae (17%) and other Gram-positive organisms. Discussion: The results of this and other studies show that despite sharing the causative agent of infection in the bile duct, the identification of such organisms by laboratory testing and antibiotic susceptibility varies considerably, which may be associated with place of residence, the type of antibiotic therapy used regularly at each hospital and the duration of such treatment.