Building capacity to conduct zoonotic pathogen surveillance in regions where it is lacking is critical for the prompt recognition and identification of emerging health threats. Successful capacity development requires the strengthening of global, regional, and local networks, as well as increased communication and collaboration across all health sectors. In order to achieve the goal of building effective capacity for zoonotic disease detection and response in developing countries, attitudes and perspectives from key stakeholders regarding current capacity building efforts and priorities must be adequately understood. The PREDICT project, a component of United States Agency for International Development’s Emerging Pandemic Threats program, has introduced capacity building efforts to increase zoonotic disease surveillance in wildlife in global ‘hot spot’ regions where zoonotic disease emergence is likely to occur. Asurvey was administered to wildlife officials, public health officials and PREDICT-implementing in-country project scientists in participating countries in order to determine regional perceptions regarding the current status of zoonotic pathogen surveillance, to identify capacity needs at high priority human-animal interfaces and to guide future recommendations for zoonotic disease detection and response in each area. Representatives from 19 countries completed the capacity tracking survey and results showed diverse perspectives across stakeholder groups. Findings indicated a truly One Health approach that improves zoonotic pathogen surveillance on a global scale is greatly needed, including a greater awareness of the role of wildlife in human disease emergence, the recognition of high priority human-animal interfaces and the promotion of evidence-based practices in capacity building and tracking efforts.