Introduction: The inappropriate antimicrobial usage (AMU) in chicken production has led to an increase in the prevalence of antimicrobial resistance (AMR). In Zambia, there is little information documented regarding the knowledge, attitude, and practices of poultry farmers on AMU and AMR. Therefore, this study assessed the knowledge, attitude and practices regarding AMU and AMR among poultry farmers in Kitwe, Zambia. Methods: This was a cross-sectional study conducted among 106 poultry farmers from November to December 2021 using a structured questionnaire. Data analysis was done using IBM Statistical Package for Social Sciences (SPSS) version 26. Results: Overall, of the 106 participants, 90.6% knew what antimicrobials were, but only 29.2% were aware of AMR. The study showed that 46.2% of the participants had low knowledge, 71.7% had negative attitudes, and 61.3% had poor practices regarding AMR. The prevalence of antibiotic use in poultry production was 83%. The most used antimicrobials were tetracycline (84%) and gentamicin (35.2%). The commonly reported reason for the use of antimicrobials was for the treatment (93.2%) and prevention (89.8%) of diseases.