This report describes the first occurrence of Polystomoides brasiliensis n. sp. (Monogenea: Polystomatidae), a new monogenean species in the buccal and pharyngeal cavities of the freshwater turtles in Brazil. Live monogeneans were collected from Hydromedusa maximiliani and Phrynops geoffroanus at the Mariano Procópio Museum's lake, in Juiz de Fora, Minas Gerais, Brazil. Polystomoides brasiliensis differs from all other species of this genus in having 8–9 genital spines, except for Polystomoides uruguayensis, which has 8–10 genital spines. However, the new species differs morphometrically from P. uruguayensis in the greater size of the outer and inner hamuli, as well as having a testis that is proportionally greater than the pharynx and oral sucker. The current study is the first report of monogeneans in chelonians of Brazil, and the first record of helminths in H. maximiliani.