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Herein we report the second record for Leporinus tigrinus in the Upper Paraná River basin, based on two specimens collected during field studies at Sucuriú River, Mato Grosso do Sul State and two specimens from Claro River, Goiás State. These records extend the species’ distribution in approximately 500 km southwards from the first record in Corumbá dam area, Goiás State. Moreover, we discuss whether the species is native in the Upper Paraná River basin.