There have been consistent calls to incorporate gender frameworks into One Health research and policy. Men’s and women’s roles in relation to animals in many rural and agricultural communities may influence potential pathogen exposure and transmission patterns of zoonotic infectious diseases. However, men’s and women’s roles related to animals in rural societies are not static but change over time. This article draws on the results of qualitative research about perceptions and practices related to animal species and animal products in five sites located in Eastern Highlands Province and East New Britain, two provinces in Papua New Guinea (PNG). The qualitative research, which aimed to explore animal-human interactions that may pose a risk of zoonotic infections through focus group discussions, formed part of a broader study of zoonotic association and risks for extrapulmonary tuberculosis in PNG. We first introduce participants’ descriptions of symptoms of illness that they attributed to zoonotic disease, as well as the animals that they attributed those diseases to. We then draw on an in-depth case study of practices related to rearing pigs — animals with important economic and social value in PNG — to illustrate the impact of social and ecological change on the interplay of gender and relationships with animals. One Health strategies at the community level can focus on gender as a way to understand changes underway in the type and intensity of interactions with animals in rural communities, including their potential impact on zoonotic diseases.