Using non-lethal methods for data collection of individuals and participatory monitoring by fishermen, we provide the first empirical evidence of the use of a nursery area by neonate and young-of-the-year cownose rays Rhinoptera bonasus and R. brasiliensis in southeastern Brazil. Two methods were used to collect data: (1) information provided by fishermen (reports, pictures, and videos) and (2) field sampling by researchers. A total of 746 cownose rays were captured; 113 have been identified as R. bonasus, 15 as R. brasiliensis, and 618 were reported by fishermen and could not be identified to the species. Records of newborns were made only in late spring and summer in 2015, 2016, and 2017, which suggests an annual reproductive cycle, with birth in late spring, extending to summer. A repeated use of this area by R. bonasus suggests that it is potentially important to the reproduction of this species. However, R. brasiliensis requires more studies. Small increases in mortality, resulting from increased fishing or other anthropogenic stressors, can have a disproportionately large effect on population viability. Thus, management of areas used during critical stages of the life cycle of rays is crucial to their conservation.

The genus Rhinoptera is composed of eight species widely distributed in tropical, subtropical and temperate coastal waters, which inhabit bays, estuaries and river mouths. Cownose ray Rhinoptera bonasus has been reported to inhabit the Western Atlantic including the Gulf of Mexico and the Caribbean, whereas the Brazilian cownose ray R. brasiliensis has been considered endemic to the coast of Brazil. Recent reports of R. brasiliensis in the Gulf of Mexico bring about the question of whether the species has a wider range than previously reported. Here, the mitochondrial genes COI, Cytb, NADH2 and the nuclear gene RAG1 were used to distinguish among species and to confirm the presence of R. brasiliensis in the Gulf of Mexico. R. brasiliensis specimens collected along the southern Gulf of Mexico showed a remarkable genetic and morphological affinity when compared with R. brasiliensis specimens from Brazil, supporting the presence of the species in Mexico (from Veracruz through Campeche) and providing evidence that its distribution ranges from Brazil to the Caribbean Sea and the Gulf of Mexico. Both species overlap geographically to a large degree, leading to a reassessment of their conservation status. Our results also show that R. bonasus distribution in the Gulf of Mexico may be restricted to the northern portion, in US waters.