Forest vertebrates are critical to the subsistence of many tropical forest dwellers enjoying little or no access to other sources of animal protein. Yet the ecological and socioeconomic value of forest wildlife is being undermined as many large vertebrate populations are driven to local extinction by unsustainable hunting practices. Although large mammals that are preferred by hunters are widely variable in their morphology and ecology, they share a set of life history traits, which make them particularly vulnerable to overhunting. In this paper we compile data on game harvest from 31 tribal and nontribal settlements in Neotropical forests to examine how mammal assemblages are affected by the history of hunting within settlement catchment areas. The structure of hunter-kill profiles is related to settlement age and size in an attempt to understand how changes in hunting pressure may affect prey selectivity and the structure of residual game assemblages. There was a predictable shift from a few large-bodied to several small-bodied species harvested by increasingly older villages. Settlement persistence thus explained a significant proportion of the variation in mean body mass and species richness of mammals harvested. We conclude that differences in prey species profiles obtained by subsistence hunters of different ethnic groups can be largely explained by the local depletion status of game stocks, particularly large mammals, rather than by cultural factors.