During a parasitological survey of Patagonian freshwater fishes, specimens of a new species of Monogenea were collected from the gills of Percichthys trucha (Perciformes: Percichthyidae). This species is described as the only member of a newly proposed genus of Ancyrocephalinae. Duplaccessorius n. gen. (Dactylogyridae, Ancyrocephalinae) is characterized by hooks with 2 subunits, gonads in tandem, a coiled male copulatory organ with counterclockwise rings, an accessory piece formed by 2 distinct parts, and a nonsclerotized sinistrolateral vagina with a funnel shape vestibule. Duplaccessorius andinus n. sp. infects the gills of P. trucha (Cuvier and Valenciennes, 1840) from Andean Patagonian lakes, being the first species of Ancyrocephalinae described from a South American percichthyid.