The Harpy Eagle (Harpia harpyja) is threatened with extinction throughout its distribution in the neotropical forests. In the Atlantic Forest, deforestation has reduced the number of suitable habitats, with only a few remnant forest fragments hosting active nests; currently, the only known nests in this region are in the Central Atlantic Forest Ecological Corridor (CAFEC), in Brazil. Little is known about Harpy Eagle diets in this region, despite this information being essential for developing effective conservation strategies. We classified the composition, frequency, richness, ecological attributes, and conservation status of the species that make up the Harpy Eagle’s diet in its last refuges in the CAFEC. Between 2017 and 2021, we collected and analyzed 152 prey remains and 285 camera trap photographs from seven active nests. We identified at least 16 mammal species (96.7%), one parrot and other bird remains (3.3%). The Harpy Eagle’s diet consisted mainly of medium-sized arboreal, folivorous, frugivorous, and diurnal mammals. Five prey species are currently threatened with extinction at global, six at national and seven at regional levels. The majority of the diet consists of Sapajus robustus, which is threatened, and Bradypus variegatus, which is not threatened. In addition to the effects of habitat loss and hunting, the Harpy Eagle may also suffer from the decline in the populations of their prey in the Atlantic Forest.