ObjectiveTo investigate antimicrobial application in 56 township hospitals in Hunan Wuling mountain area. Methods13 348 outpatient prescriptions，1 288 medical records of nonoperated patients, and 280 medical records of operated patients in 56 township hospitals in Hunan Wuling mountain area were selected randomly, antimicrobial application was surveyed. ResultsAntimicrobial usage rates in outpatients, nonoperated inpatients, and operated inpatients were 53.01%, 87.97%, and 98.57% respectively, and the percentage of combined antimicrobial agents were 19.57%, 46.96%, and 39.86% respectively. The percentage of antimicrobial prophylaxis in patients undergoing type I incision surgery was 96.94%, combined antimicrobial usage rate was 36.84%. Antimicrobial use density in  hospitalized patients was 124.92 DDDs/100 patient days. 8 573 antimicrobial agents were prescribed in outpatient prescriptions of 56 hospitals, the top 5 frequently used antimicrobial agents were amoxicillin, ceftriaxone, amoxicillin/clavulanate potassium, levofloxacin, and clindamycin. ConclusionAntimicrobial agents are overused and irrationally used in township hospitals in Hunan Wuling mountain area. Health care workers should strengthen learning, conscientiously implement the relevant regulations on antimicrobial management; health administrative departments should strengthen the supervision and management on clinical application of antimicrobial agents, and promote the rational use of antimicrobial agents in township hospitals.