Objectives: The aim of this study was to assess knowledge, attitudes and practices amongst public health undergraduates in relation to antimicrobial resistance (AMR) in China. Methods: A cross-sectional survey was conducted amongst all final-year public health undergraduates from 18 universities across China. A structured questionnaire was used to collect information on AMR-related knowledge, attitudes and practices, whilst multivariable linear and logistic regressions were employed to detect associations among these three aspects. Results: A total of 1115 participants were included in this study. The mean ± standard deviation AMR knowledge score was 7.68 ± 2.56. Moreover, 75.2% of students had performed incorrect antimicrobial practice. Studying in a key university [slope = 1.49, 95% confidence interval (CI) 0.71–2.27], being male (slope = 0.36, 95% CI 0.02–0.70), having clinical experience (slope = 1.71, 95% CI 1.13–2.30) and having an affirmative attitude towards AMR were positively associated with knowledge score. Knowledge score was also positively associated with AMR practice (odds ratio = 1.07, 95% CI 1.00–1.13). Conclusions: A knowledge gap in relation to AMR was determined among Chinese public health students. This gap is associated with attitudes towards AMR and in turn influences antimicrobial use. It is clear that additional measures are needed in the curriculum, including additional AMR-specific clinical practices.