A new nematode species (Neoascarophis mariae n. sp.) is described based on specimens collected from the Argentine goatfish Mullus argentinae (Hubbs et Marini) from coastal waters off the state of Rio de Janeiro, Brazil. In the genus, the new species belongs to the group of species with females that have the vulva near the posterior end of the body. Only males of Neoascarophis longispicula Moravec et Klimpel, 2009 are known and can be distinguished from those of the new species by their larger body, developed and somewhat dorsoventrally expanded flat inner part of the pseudolabia, bifurcate deirids and larger spicules (the left one with a rounded tip) with a different length ratio. Other species with females that have the vulva near the equatorial region are N. yarihige Machida, 1976 and N. bathygadi Machida, 1976. Both males and females of N. yarihige are longer than those of the new species and have a shorter vestibule; males have shorter spicules with a different length ratio. Neoascarophis bathygadi is the only member of the genus that shares the presence of a cephalic vesicle with the new species, which, however, is shorter and arises at 40 µm from the anterior end instead from the deirids, as in the new species. Both males and females of N. bathygadi are also longer than those of the new species and have a shorter vestibule; males have a larger left spicule, but shorter right spicule and a different length ratio. Ascarophis upeneichthys Johnston et Mawson, 1945, a parasite of a mullid host, is transferred to Neoascarophis Machida, 1976 and is distinguished from the new species by having a shorter vestibule in females and shorter spicules (left spicule with a pointed tip) with a different length ratio in males.