Herein we describe a new species of turtle blood fluke (TBF) and propose a new genus to accommodate it, Pitiutrema revelae n. gen., n. sp. This blood fluke infects the heart of six-tubercled Amazon River turtles (Podocnemis sextuberculata [Cornalia, 1849] [Pleurodira: Podocnemididae]) in the headwaters of the Amazon River near Iquitos, Peru. It resembles the other 2 described species of South American freshwater TBFs (Atamatam amazoniensis Bullard and Roberts, 2019, and Paratamatam iquitosiensis Bullard and Roberts, 2019) by having a dorsoventrally flattened and ovoid body, an oral sucker with anteroventral spines, 2 inter-cecal testes arranged in a column, inter-gonadal terminal genitalia, an inter-cecal and post-ovarian Laurer's canal pore, and a Y-shaped excretory bladder. It differs from all other nominal TBFs by having the combination of an aspinose body that lacks mammillae, a tapered (not broadly rounded) posterior body end, a ventral sucker, slightly M-shaped or inverse U-shaped ceca, a deeply-lobed (dendritic) ovary, a transverse uterus, and a dispersed vitellarium. The new genus is further unique among TBF genera by having an anterior to posterior sequence of ventral sucker, anterior testis, ovary, cirrus sac (lateral to posterior half of ovary), and posterior testis. The phylogenetic results and placement of the new taxon (1) were both predicted by our morphological diagnosis and comparisons with related taxa, (2) further indicated monophyly of the nominal South American freshwater TBFs, (3) reaffirmed the marine derived lineage identity of the nominal South American freshwater TBFs, and (4) highlighted that the single cercarial sequence (TBF sp. W-810) from an ampullariid in Brazil does not share a recent common ancestor with any of the nominal South American freshwater TBFs. The new species is the eighth TBF reported from a side-necked turtle (Pleurodira), the first TBF from a member of Podocnemididae, and the third freshwater TBF from South America.