ObjectiveTo understand the distribution and drug resistance of pathogens from patients with anastomotic fistula infection after esophageal cancer surgery, and provide basis for clinical diagnosis and treatment. MethodsPatients were admitted to a hospital due to anastomotic fistula after esophageal cancer surgery between January 2012 and December 2015, microbial culture and antimicrobial susceptibility testing results of patients were retrospectively analyzed. Results1 986 patients underwent radical resection of esophageal cancer within 4 years, 148 of whom developed anastomotic fistula, 104 (70.27%) were with positive microbial culture. A total of 197 pathogenic strains were isolated, 52（26.40%）and 145 （73.60%）strains were isolated from intrathoracic anastomotic fistula and cervical anastomotic fistula respectively；127 (64.47%）strains were gramnegative bacteria, the major were Pseudomonas aeruginosa, Klebsiella pneumoniae, and Acinetobacter baumannii, 62(31.47%) strains  were grampositive bacteria, the major were Staphylococcus aureus, Enterococcus spp., and Streptococcus viridans, 8 strains (4.06%) were fungi. 49（47.12%） cases were with mixed pathogenic infection. The resistance rates of gramnegative bacteria to imipenem were 17.86%47.62%, to polymyxin B was 0, resistance rates of Pseudomonas aeruginosa to other antimicrobial agents were all<50%, Klebsiella pneumoniae to piperacillin and aztreonam were both>70%, Acinetobacter baumannii to most antimicrobial agents were all>50.00%; resistance rates of grampositive bacteria to clindamycin and tetracycline were both>50.00%, to linezolid, vancomycin, and teicoplanin were all 0, resistance rates of Staphylococcus aureus to penicillin, oxacillin, and ciprofloxacin were all>60%，resistance rate of Enterococcus spp. to quinupristin/dalfopristin was 100.00%. ConclusionPostoperative anastomotic fistula combined with infection can affect the prognosis of patients after esophageal cancer surgery, regular monitoring on distribution and drug resistance of pathogens can provide the basis for initial empirical treatment, and is conducive to the early treatment and rational use of antimicrobial agents.