The integration of biodiversity conservation and public health has gained significant traction, leading to new efforts to identify win–win solutions for the environment and health. At the forefront of these efforts is pin-pointing ways in which biodiversity conservation can reduce risk of zoonotic spillover, especially given the consequences of epidemics and pandemics of wild animal origin. However, there is currently an incomplete understanding of the mechanisms by which biodiversity change influences the spillover process, limiting the application of integrated strategies aimed at achieving positive outcomes for both conservation and disease management. Here, we review the literature, considering a broad scope of biodiversity dimensions, to identify cases where zoonotic pathogen spillover is mechanistically linked to changes in biodiversity. By reframing the discussion of biodiversity and disease using mechanistic evidence while encompassing multiple aspects of biodiversity, including functional diversity, landscape diversity, phenological diversity, and interaction diversity, we work toward general principles that can guide future research and more effectively integrate the related goals of biodiversity conservation and spillover prevention. We conclude by summarizing how these principles could be used to integrate spillover prevention into ongoing biodiversity conservation initiatives.