Nematoda of the genus Echinocephalus Molin, 1858 include species from the Gnathostomatidae family, the adult stages of which parasitize the stomach and spiral intestine of elasmobranchs as their final hosts. In the present study, we describe Echinocephalus spinosus n. sp. found parasitizing the spiral valve of the freshwater stingray Potamotrygon motoro from the Tapajós River, in the Amazon Basin, in the state of Pará, Brazil. In the study we performed morphological (light and scanning electron microscopy) and molecular (small subunit ribosomal ribonucleic acid (SSU rDNA) and mitochondrial cytochrome c oxidase I sequencing) analyses. E. spinosus n. sp. is only the second species of the Echinocephalus genus described from a strictly freshwater environment. The SSU rDNA based phylogenetic analyses showed Echinocephalus clade as a sister lineage of Gnathostoma, and that the new species arises as a sister to Echinocephalus cf. pseudouncinatus. Time-calibrated phylogenetic analysis revealed that the origin of the freshwater Echinocephalus coincides with the recently proposed origin of the freshwater host potamotrygonin stingray, namely the late Oligocene to early Miocene, when the western Amazon was dominated by the Pebas wetlands, an epicontinental marine/freshwater system.
