Objective To realize the isolation and drug resistance of major pathogens from bile of patients with biliary tract infection, so as to provide reference for rational use of antimicrobial agents.MethodsBile specimens collected from 197 operated patients in a hospital from January 2010 to December 2011were performed aerobic culture, bacteria were identified and performed antimicrobial susceptibility testing. ResultsOf 197 bile specimens, 130 (65.99%) were positive culture; 151 aerobic isolates were obtained, 21 specimens isolated mixed bacteria. The isolated pathogens included 114(75.50%) isolates of gramnegative bacilli, 36 (23.84%) of grampositive cocci, and 1 (0.66%) of fungus. The top 6 pathogens were Escherichia coli(48.34%), Enterococcus faecium(10.59%), Enterobacter cloacae(6.62%), Enterococcus faecalis(5.96%), Klebsiella pneumoniae(5.96%), and Pseudomonas aeruginosa(4.64%). Sensitive rates of Escherichia coli, Enterobacter cloacae and Klebsiella pneumoniae to carbapemem (imipenem, meropenem) were 100.00%; Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa to piperacillin/tazobactam and amikacin were 77.78%-100.00%; Enterococcus spp. to tigecycline and linezolid were 100.00%, all Enterococcus faecalis to vancomycin were 100.00%, 1 Enterococcus faecium isolate was resistant to vancomycin. Sensitive rates of gramnegative bacilli to penicillins, cephalosporins, and quinolones decreased.ConclusionBiliary tract infection is mainly caused by Enterobacteriaceae and Enterococcus spp., bile culture and antimicrobial susceptibility testing of pathogens should be performed as early as possible, clinicians should choose antimicrobial agents according to antimicrobial susceptibility testing results.