The interface between humans, domestic animals, and wildlife has been implicated in the emergence of infectious diseases and the persistence of endemic human and animal diseases. For individuals who reside at this interface, particularly those in low-resource settings, the development of disease risk assessment and mitigation skills must be prioritized. Using a community engagement-One Health approach, we implemented a training program aimed at advancing these skills among agro-pastoralists living adjacent to conservation areas in South Africa. The program included professional development of local facilitators who then conducted workshops with community members. Workshops used a series of experiential, inquiry-based activities to teach participants the concepts of pathogen transmission and disease risk assessment and mitigation. The program was implemented over four weeks with 10 facilitators and 78 workshop participants. We conducted a within-subjects experimental study using a mixed methods design to evaluate the program in terms of facilitator and participant One Health knowledge and practices. Quantitative data included pre/post written assessments; qualitative data included focus group discussions, semi-structured interviews, and pre/post photographs. Mean post-test scores of facilitators increased by 17% (p = 0.0078). For workshop participants, improvements in knowledge were more likely for females than males (OR = 7.315, 95% CI = 2.258-23.705, p = 0.0009) and participants with a higher versus lower education level, albeit borderline non-significant (OR = 4.781, 95% CI = 0.942-24.264, p = 0.0590). Qualitative analysis revealed the implementation of risk mitigation strategies by 98% (60/61) of workshop participants during the three-month follow-up and included improved personal and domestic hygiene practices and enhanced animal housing. Although further evaluation is recommended, this program may be appropriate for consideration as a scalable approach by which to mitigate human and animal infectious disease risk in high-risk/low-resource communities.