| Summary: | Abstract The species of Hypostomus from the Parnaíba River basin were reviewed through molecular and morphological analysis. Five species were found in the basin, including a new species herein described. The distribution of H. pusarum was expanded to this basin, and a closely related species was recorded (H. aff. pusarum), also the presence of H. johnii and H. vaillanti was confirmed. The new species is distinguished from most congeners by its large number of premaxillary and dentary teeth, a wide dental angle of 115° to 135°, presence of a rounded dark spots on a lighter background and anteromedial region of the abdomen depleted of plaques (vs. anteromedial region of the abdomen covered by platelets and odontodes in H. johnii, H. pusarum, H. aff. pusarum and H. vaillanti). Furthermore, an identification key of the species from the Maranhão-Piauí ecoregion and maps with the geographic distribution of these species are presented. The species of Hypostomus in the Parnaíba River basin have different geographic distributions, suggesting different niches or geographical barriers, providing an opportunity for ecological and evolutionary studies.
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