RESUMO
Pimelodidae family is one of the most diverse and widely distributed fish groups in South America. Phylogenetic analysis in the family have recently indicated the existence of two main clades: "sorubiminae" and the OCP clade, including Pimelodus ornatus, "calophysines" and "pimelodines." The aim of this study was to investigate the karyotype of three Amazonian Pimelodidae species: Calophysus macropterus, Propimelodus eigenmanni, and Exallodontus aguanai associating them to the literature, seeking to reconstruct probable ancestral characters. C. macropterus has 2n = 50, 20m+20sm+10a (fundamental number [FN] = 90), simple interstitial nucleolar organizing regions (NORs), and four 5S rDNA sites terminals, two in synteny with the 18S rDNA. P. eigenmanni has 2n = 56, 28m+20sm+2st+6a (FN = 106), simple NORs, and two 5S rDNA sites terminals. E. aguanai has 2n = 56, 36m+12sm+2st+6a (FN = 106) and 18S and 5S rDNA sites interstitial syntenic in the chromosome 1. All species exhibited a higher amount of heterochromatin, differing from the pattern of the family, and strong marking associated with NORs. The integration between molecular phylogenetic data and karyotype data indicated a high probability that 2n = 56 and simple terminals NORs in the short arm are ancestral characters in Pimelodidae, evidenced in "sorubiminae." In the OCP clade derived traits were observed resulting from chromosomal changes that played a critical role in the karyotype evolution of the group.