This study examined stakeholder perceptions of gaps in antimicrobial resistance (AMR) mitigation efforts across different sectors and career stages, with the goal of generating insights to support prioritization of One Health interventions.

We conducted a cross-sectional survey of AMR stakeholders using a structured questionnaire between October and November 2025. Stakeholders were recruited through international networks across 33 countries on 5 continents. Respondents listed the top three areas in AMR mitigation efforts that they perceived as receiving insufficient attention but require greater focus to reduce the emergence and spread of drug-resistant bacterial infections. We performed thematic analysis, calculated weighted priority scores, conducted an Impact-Effort Matrix analysis, and applied Fisher’s exact tests to examine associations between themes, professional sector and career stages.

Among 103 respondents, participants spanned nine professional sectors and four career stages. Antimicrobial stewardship (18.8%, weighted score 119) and Environmental AMR and Waste (16.5%, weighted score 108) were the most frequently cited perceived gaps. Impact-Effort Matrix analysis identified four priority areas as Quick Wins (stewardship, IPC, surveillance, and animal health, accounting for 45.7% of impact), two Major Projects (environmental AMR, R&D), three Fill-ins (awareness, One Health, other/cross-cutting), and three Hard Slogs (access, financing, policy). No significant associations were observed between perceived mitigation gaps across professional sector and career stages.

The emphasis on antimicrobial stewardship among stakeholders highlights a perceived disconnect between stewardship policy adoption and on-the-ground practice. The Impact-Effort Matrix provides a decision-support tool that may assist policymakers and funders in prioritizing One Health interventions.