Objective: e-Bug is a teaching resource that addresses the UK 5-year National Action Plan on antimicrobial resistance (AMR) that pledges to work with educators and local authorities to ensure young people understand infection prevention and control (IPC) and AMR. This study aimed to evaluate the effectiveness and acceptability of the e-Bug face-to-face train-the-trainer intervention with school and community educators. Design: Service evaluation of an educational intervention via surveys. Setting: Workshops were organised by Public Health England (PHE) and collaborators in seven regions of the UK during 2018-2019. Method: Pre- and post-intervention surveys measured satisfaction with training, knowledge of IPC and AMR, and confidence to teach others. Statistical analyses included multilevel and ordinal logistic regression models to measure change in educator knowledge and confidence. Results: In all, 262 educators participated: primary (46%), secondary (17%), college (2%), healthcare (29%) and community (7%). Educators had high pre-intervention knowledge of topics, with significant improvement (p < 0.05) in confidence to teach all topics and some significant IPC knowledge improvement, post-intervention. There was strong evidence for a difference in confidence change between educator types, with primary educators improving the most. Ninety-five percent of educators rated the train-the-trainer workshop positively, valued the interactive workshops and felt confident to use the resources. Conclusion: Confident and knowledgeable educators, achieved via e-Bug train-the-trainer workshops, will enhance education of IPC and AMR topics in schools and communities, and therefore support the UK 5-year AMR action plan. The intervention will be monitored with long-term follow-up surveys to explore how training has been disseminated and to evaluate long-term benefits.