An integrative approach by the congruence of genetics, morphology and ecological niche modelling (ENM) was used to delimit a new species of Hypanus (Rafinesque, 1818), a recently resurrected genus of marine stingrays comprising eight species, five of which occur in the western Atlantic. The species with the widest distribution, Hypanus americanus (Hildebrand and Schroeder, 1928), from the northeastern coast of the United States to southeastern Brazil, was demonstrated to be paraphyletic based on protein-coding mitochondrial genome analyses. This data set also indicates that the genetic distance between the new species Hypanus berthalutzae sp. nov. and its three closely related species (H. americanus, H. longus and H. rudis) varies from 0.82% to 3.14%. In addition, Bayesian Analysis of Population Similarity using the mitochondrial gene mt-nd2 supports the separation of H. berthalutzae sp. nov. (southwestern Atlantic) from its sister species H. rudis (eastern Atlantic). Similarly, morphological and morphometric analyses corroborated four morphotypes within the H. americanus species group and indicated the ventral caudal fold height and length and interspiracular and interorbital lengths as useful measurements to distinguish among them. Claspers of adult males also exhibit morphological differences among species. The ENM agreed with molecular and morphological analyses and delimits the distribution of H. berthalutzae sp. nov. to shallow areas close to shore along the Brazilian coast, from the mouth of the Amazon River to São Paulo State, including the northeastern oceanic islands, suggesting that the great outflow of fresh water and sediments and the Mid-Atlantic Ridge might act as barriers. The integration of these data to describe a new species provides information relevant to their conservation status, because all species of the H. americanus species group are under the "data-deficient" status.