Three monogenean species, Anacanthorus luquei n. sp., A. scholzi n. sp. and A. cohenae n. sp. are described from the gills of the tetra fish Markiana nigripinnis (Perugia) (Characidae), collected in the Pantanal wetlands, State of Mato Grosso do Sul, Brazil. Among other differences, Anacanthorus luquei n. sp. differs from the most morphologically similar species, based on the structure of the accessory piece as follows: branches with smooth margins (vs with irregular margins in A. cuticulovaginus), without pointed projections at distal end (vs with projections in A. dipelecinus) and with 2 branches (vs 3 in A. quinqueramus). Anacanthorus scholzi n. sp. is most morphologically similar to A. luquei n. sp., differing from it because one of the branches of the accessory piece is bifurcated at the distal portion. Anacanthorus cohenae n. sp. can be differentiated from the congeners based on the combination of the following features: MCO cylindrical and robust with sclerotised flanges on the extremities, accessory piece V-shaped, bearing two branches similar in length and with blunt distal ends, and hooks with a proximal bulb. This is the first parasitological study on M. nigripinnis and, currently, Anacanthorus allocates 88 species infesting characiform fishes in the Neotropical region, including the three new species described here.