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Abstract Background: Gender equity and social inclusion are critical, yet still at risk of being neglected during the development and implementation of national One Health action plans. The guidance provided by the Quadripartite international organizations (World Health Organization, Food and Agriculture Organization of the United Nations, United Nations Environment Programme, and World Organization for Animal Health) recognize these principles, but they lack specific guidance on how to effectively integrate them at the national level. This article introduces the Gender-Responsive and Inclusive Implementation Guide for National One Health Action Plans (GeRIGOH), developed by the NEOH Gender Working Group, as a practical tool to support the integration of gender equity and social inclusion into national One Health action plans. It is intended for One Health practitioners seeking to embed inclusive, equity-centered approaches throughout One Health planning and implementation. Methods: GeRIGOH aligns with the five key steps of the Quadripartite international organizations’ Guide for implementation of the One Health Joint Plan of Action at the national level: situation analysis, setting up multi-sectoral One Health coordination, planning for implementation, implementation and reviewing, sharing and iteratively acting on lessons learned. Results: GeRIGOH offers 32 key yes/no questions and considerations to support equitable decision making, inclusive stakeholder engagement, fair access to resources, and comprehensive monitoring and evaluation frameworks. Conclusion: Although GeRIGOH focuses on national-level action planning, it offers principles and questions that may inform inclusive approaches more broadly and serves as a vital tool for advancing more inclusive, sustainable, and resilient One Health outcomes. One Health impact statement Gender equity and social inclusion are recognized as critical to achieving the aims of One Health (OH), yet they remain neglected in OH policies, tools, and frameworks. While the Quadripartite international organizations’ OH Joint Plan of Action (JPA) gives guidance on how to implement national OH action plans, including governance structures and defining roles among different ministries, agencies, and stakeholders, it offers limited direction on how to meaningfully integrate gender and inclusion into these processes. To address this gap, we developed the Gender-Responsive and Inclusive Implementation Guide for National One Health Action Plans (GeRIGOH). Developed by the NEOH Gender Working Group, this guide provides 32 key yes/no questions and considerations at each step of the OH JPA implementation process to support equitable, inclusive stakeholder engagement and foster a transformative gender approach. By operationalizing gender and inclusion within national OH action plans, GeRIGOH strengthens governance, enhances the sustainability of OH initiatives, and contributes to more just and effective health outcomes and processes across human, animal, and environmental health domains. It offers a critical tool for the global OH community to mainstream equity and inclusion as a core pillar of OH practice.