Monocotyle guttatae n. sp. is described from the gills of the ray Dasyatis guttata (Bloch and Schneider, 1801) (Dasyatidae) off the coast of Brazil The new species can be readily differentiated from the other species of the genus in having a male copulatory organ with 2 loops and an accessory piece, 5-7 sclerites on the marginal haptoral papillae, and the absence of accessory sclerites on the dorsal surface of the posterior body. The present record confirms the presence of the genus in the subtropical waters of South America.