The Analysis Framework was developed to assist the SNRD-AP Biodiversity Working Group to implement the OH approach within biodiversity-related projects in the Southeast Asia region. The framework allows assessing if the project has a sufficient base for One Health and already applies key OH principles. It enables the identification of potential gate entries, where a OH element can be integrated, and assesses the measures that could allow building a OH element into the project. Through the application of the framework, owners and stakeholders are guided in the revision of the project adopting the OH lens, identifying new opportunities to work across disciplines and engaging with new actors. The framework should be considered a starting point for the transformative process of adding One Health into biodiversity. A radical shift in thinking is required to break down the process of implementing the OH approach into manageable and feasible measurements that are context- and project-specific. By doing so, small-scale actions and successes will likely increase trust in the process and pave the way for a broader integration of One Health within the conservation community. The Analysis Framework was tested on six projects to verify its validity. Tthe Biodiversity-One Health Taskforce of the SNRD-AP Biodiversity Working Group was trained on the step-by-step process to ensure its application on a wider scale.