Abstract   Food chain modeling is often used to assess the risks of chemical contaminants to wildlife. In modeling efforts, bioaccessibility from different dietary components is assumed to be similar. The present study explored potential differences in the in vitro bioaccessibility of metals from a range of insect orders, which are common components of the diet of insectivorous bats, and assessed the implications of this for environmental exposure assessment. Bioaccessibility of metals was assessed using an in vitro gastric model simulating gastric and intestinal conditions of insectivorous bats. In vitro–derived metal bioaccessibility was found to differ significantly across insect orders. Bioaccessibility was found to be greatest in Coleoptera, followed by Lepidoptera and Diptera. To establish the implications for risk assessment, a spatially explicit risk model was employed that included and excluded in vitro bioaccessibility data; to examine the daily oral exposure of metals to 14 bat species. The results show that when bioaccessibility data are included in the model, metal exposure predictions across species are changed and that the ranking of bat species, in terms of metal exposure, are altered. The authors recommend that in vitro bioaccessibility data begin to be employed when establishing the risks of contaminants to wildlife species. Environ Toxicol Chem 2015;34:898–906. © 2014 SETAC