Objective To understand the distribution and antimicrobial resistance of bacteria in biliary tract infection in China, and provide reference for clinical antimicrobial choice. Methods According to China Antimicrobial Resistance Surveillance System(CARSS) technical program, data of bacteria isolated from bile specimen and antimicrobial susceptibility testing results reported by CARSS member hospitals in 2014-2019 were analyzed with WHONET 5.6 software, the same bacteria from the same patient was only adopted the first strain. Results A total of 268 016 bacterial strains were analyzed, the top 8 were Escherichia coli (30.9%), Klebsiella pneumoniae (12.7%), Enterococcus faecium (10.1%), Enterococcus faecalis (8.6%), Pseudomonas aeruginosa (4.9%), Enterobacter cloacae (4.5%), Acinetobacter baumannii (2.2%), and Citrobacter freundii (1.8%). Antimicrobial susceptibility testing results of 6 years showed that resistance rates of Escherichia coli and Klebsiella pneumoniae to third-generation cephalosporins were 33.7%-65.6% and 23.6%-43.5% respectively, to quinolones were 48.9%-56.6% and 22.0%-28.5% respectively, to carbapenems were 1.1%-3.7% and 3.5%-7.6% respectively; resistant rates of Pseudomonas aeruginosa and Acinetobacter baumannii to cefoperazone/sulbactam were 13.4%-19.0% and 29.3%-42.7% respectively, to carbapenems were 18.0%-28.0% and 44.5%-59.9% respectively; resistance rates of Enterococcus faecium and Enterococcus faecalis to vancomycin were 0.6%-1.7% and 0.3%-0.7% respectively. Conclusion Gram-negative bacilli are predominant pathogens in biliary tract infection in China, the mjor bacteria were Enterobacteriaceae; pathogens are resistant to multiple antimicrobial agents, clinical antimicrobial choice should refer to antimicrobial resistance surveillance results.