The influence of landscape matrix on functional connectivity has been clearly established. Now methods to assess the effects of different land uses on species' movements are needed because current methods are often biased. The use of physiological parameters as indicators of the level of resistance to animal movement associated with different land uses (i.e., matrix resistance) could provide estimates of energetic costs and risks to animals migrating through the matrix. To assess whether corticosterone levels indicate matrix resistance, we conducted experiments on substrate choice and measured levels of corticosterone before and after exposure of toads (Bufo bufo) to 3 common substrates (ploughed soil, meadow, and forest litter). We expected matrix resistance and hormone levels to increase from forest litter (habitat of the toad) to meadows to ploughed soil. Adult toads had higher corticosterone levels on ploughed soil than on forest litter or meadow substrates. Hormone levels did not differ between forest litter and meadow. Toads avoided moving onto ploughed soil. Corticosterone levels in juvenile toads were not related to substrate type; however, hormone levels decreased as humidity increased. Juveniles, unlike adults, did not avoid moving over ploughed soil. The difference in responses between adult and juvenile toads may have been due to differences in experimental design (for juveniles, entire body used to measure corticosterone concentration; for adults, saliva alone); differences in the scale of sensory perception of the substrate (juveniles are much smaller than adults); or differences in cognitive processes between adult and juvenile toads. Adults probably had experience with different substrate types, whereas juveniles first emerging from the water probably did not. As a consequence, arable lands could act as ecological traps for juvenile toads.