Mercury (Hg) bioaccumulates in aquatic ecosystems and may pose a risk to humans who consume fish. Selenium (Se) has the ability to reduce Hg toxicity, but the current guidance for human consumption of fish is based on Hg concentration alone. The purpose of the present study was to examine the relationship between Se and Hg in freshwater sportfish, for which there is a paucity of existing data. We collected three species of fish from different trophic positions from two drinking water reservoirs in central North Carolina, USA, to assess Hg and Se concentrations in relation to fish total length and to compare two measures of the protective ability of Se, the Se:Hg molar ratio and Se health benefit value (HBV_Se), to current guidance for Hg. According to the Se:Hg molar ratio, all of the low trophic position fish sampled and the middle trophic position fish sampled from one of the reservoirs were safe for consumption. The same number of fish were considered safe using the HBV_Se. More fish were deemed unsafe when using the Se:Hg molar ratio and HBV_Se than were considered unsafe when using the U.S. Environmental Protection Agency (USEPA) Hg threshold. These findings suggest that the measures of Se protection may be unnecessarily conservative or that the USEPA Hg threshold may not be sufficiently protective of human health, especially the health of sensitive populations like pregnant or nursing mothers and young children. Future examination of the Se:Hg molar ratio and HBV_Se from a variety of fish tissue samples would help refine the accuracy of these measures so that they may be appropriately utilized in ecological and human health risk assessment.