Two new species of Gyrodactylus Nordmann, 1832 (Platyhelminthes, Monogenoidea) are described from fishes collected from southeastern Brazil. Gyrodactylus geophagensis n. sp. was collected from the body surface of the "cará", Geophagus brasiliensis (Quoy and Gaimard, 1824) (Cichlidae), from the Rio da Guarda, state of Rio de Janeiro; its major diagnostic features are the morphology of the anchor with a short, truncate superficial root and the shape of the hooks - with a long, delicate shaft. Gyrodactylus trairae n. sp. parasitizes the body surface of the "traíra", Hoplias aff. malabaricus (Bloch, 1794) (Erythrinidae), from the rio Guandu, state of Rio de Janeiro and can be easily differentiated from other species of the genus by having a thin, dorsal bridge, connecting the superficial bar with the spathulated shield. These are the first species of Gyrodactylus formally reported from Brazil. Presently, 26 species of Gyrodactylidae are known from freshwater fishes in the neotropical region; a list of these species is provided.