The rio Ribeira de Iguape basin (Paraná State) is highly relevant for the preservation of remnants of the Atlantic Forest. Its high levels of diversity and fish endemism explain the river dynamics promoted by the Ponta Grossa Arch, a geological structure that promotes headwater capture and isolation between the upper rio Paraná drainage, the rio Iguaçu and the rio Ribeira de Iguape. Here, our objective was to provide an unprecedented inventory for headwater streams of the rio Ribeira de Iguape basin at the boundaries of the Ponta Grossa Arch. We found 29 species of fish representing four orders and nine families. Siluriformes was the richest order followed by Characiformes. Nine species presented high abundance of sampled individuals, contributing with 87.8% of the collections. Four species appeared in less than 25% of the sampled sites (occasional), and six species appeared in more than 50% (constant). We identified three non-described species and one non-native (Coptodon rendalli). Two species had their geographical distribution extended and the presence of Astyanax bifasciatus, endemic to the Iguaçu river basin, ratifies recent events of headwater capture between coastal drainages and those that flow into the continent. Studies of biogeographic divisors are necessary to explain the origin and dispersion processes of species in order to direct studies on diversity and preventive management actions. Coptodon rendalli (Tilapia) is an alarming record on the introduction of species in the region.