Introduction: Over-prescription of antimicrobials for patients is a major driver of bacterial resistance. The aim of the present study was to assess the knowledge, attitude and prescription in relation to antimicrobials among physicians in the Zhejiang province in China, and identify the determining factors. Methods: A total of 600 physicians in public county hospitals and township health institutions were surveyed cross-sectionally using a structured electronic questionnaire. Results: The questionnaire was completed by 580 physicians and accordingly, the response rate was 96.67%. The mean score of 11 terms related to antimicrobial knowledge was 6.81, and that an average of 32.1% of patients with upper respiratory tract infections (URIs) would be prescribed antimicrobials. Multivariate analysis indicated that young general practitioners with less training are more likely to contribute to high antimicrobial prescriptions (P<0.05). Older physicians with more trainings are more willing to provide patients with the correct knowledge regarding antimicrobials and less likely to prescribe antimicrobial s to URIs. The results of the correlation analysis showed that a positive connection was found between the doctor’s knowledge, attitude and prescription practice (P<0.05). Conclusion: Proper prescription of antimicrobials depends on adequate knowledge and regular training programs for physicians.