The risk of zoonotic spillover from reservoir hosts, such as wildlife or domestic livestock, to people is shaped by the spatial and temporal distribution of infection in reservoir populations. Quantifying these distributions is a key challenge in epidemiology and disease ecology that requires researchers to make trade-offs between the extent and intensity of spatial versus temporal sampling. We discuss sampling methods that strengthen the reliability and validity of inferences about the dynamics of zoonotic pathogens in wildlife hosts. This article is part of the theme issue ‘Dynamic and integrative approaches to understanding pathogen spillover’.