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Knowledge of the fish species in river basins is among the minimum requirements for the management of water and fish resources. Therefore, the aim of this study was to update the fish species composition of the Piquiri River basin, upper Paraná River basin. Data were gathered from recent information published in specialized literature and records for ichthyology collections. This update reports the occurrence of 152 fish species distributed in 8 orders, 31 families, and 89 genera. Non-native species accounted for 20% of all species, and the construction of the Itaipu Power Plant and its fish ladder were the main vectors of introduction. Three percent of the species were endangered, and 11% were classified as migratory. The Piquiri River basin harbors a large number of species, some of which are rare, endangered, migratory, endemic, and even unknown by science. Because of this, maintaining the integrity of this river basin will support the persistence of regional biodiversity. [ABSTRACT FROM AUTHOR]