Bat species in the monophyletic family Phyllostomidae feed on blood, insects, small vertebrates, nectar, fruit and complex omnivorous mixtures. We used nitrogen stable isotope ratios to characterize bat diets and adopted a phylogenetically informed approach to investigate the physiological changes that accompany evolutionary diet changes in phyllostomids. We found that nitrogen stable isotopes separated plant-eating from animal-eating species. The blood of the latter was enriched in 15N. A recent phylogenetic hypothesis suggests that with the possible exception of carnivory, which may have evolved twice, all diets evolved only once from insectivory. The shift from insectivory to nectarivory and frugivory was accompanied by increased intestinal sucrase and maltase activity, decreased trehalase activity, and reduced relative medullary thickness of kidneys. The shift from insectivory to sanguinivory and carnivory resulted in reduced trehalase activity. Vampire bats are the only known vertebrates that do not exhibit intestinal maltase activity. We argue that these physiological changes are adaptive responses to evolutionary diet shifts. © by Urban & Fischer Verlag.