Antimicrobial resistance (AMR) has the potential to reverse gains made in modern medicine and seriously jeopardize the lives of people and animals from infections that were once readily treatable (FAO/OIE 2015). Adopting a one health approach is critical to better understand and mitigate factors driving the emerging concern of AMR. Although the development and transmission of AMR in wildlife is not well understood, there is growing evidence that it is associated with proximity to humans or domestic animals. Papua New Guinea (PNG) faces some unique challenges for zoonotic spillover disease events and transmission of AMR between animals and humans. These challenges arise from the erosion of wildlife habitats due to deforestation and close contact of wildlife with villagers and their village-based livestock raised under free-range systems. Papua New Guinea’s human health system and the animal health field and laboratory services are resource-limited and are facing many human and animal disease challenges. The Fleming Fund Country Grant, implemented by the Burnet Institute Australia, is working with Papua New Guinea government and industry stakeholders to tackle AMR by encouraging information sharing, raising awareness, and supporting laboratory capacity in human and animal health for effective AMR surveillance. In addition, national legislation is being strengthened to support implementation of the Papua New Guinea Antimicrobial Resistance National Action Plan (Government of Papua New Guinea 2019) to encourage the judicious use and effective monitoring of antimicrobial medicines in humans and animals.