Auchenipteridae is divided in two subfamilies, Centromochlinae and Auchenipterinae. Centromochlinae has 31 valid species, from which 13 are included in the genus Tatia Miranda Ribeiro, 1911. Among these, Tatia jaracatia Pavanelli & Bifi, 2009 and T. neivai (Ihering, 1930) are the only two representative species from the Paraná-Paraguay basins. This study aimed to analyze cytogenetically these two species and thus provide the first chromosomal data for the genus. Although T. jaracatia and T. neivai presented 2n=58 chromosomes, some differences were observed in the karyotypic formula. The heterochromatin was dispersed in the centromeric and terminal regions of most chromosomes of T. jaracatia, and only in the terminal region of most chromosomes of T. neivai. The AgNORs were detected in the subtelocentric pair 28 for both species, which was confirmed by FISH with 18S rDNA probe. The 5S rDNA sites were detected in four chromosome pairs in T. jaracatia and three chromosome pairs in T. neivai. Both species of Tatia presented great chromosomal similarities among themselves; however, when compared to other species of Auchenipteridae, it was possible to identify some differences in the karyotype macrostructure, in the heterochromatin distribution pattern and in the number and position of 5S rDNA sites, which until now seems to be intrinsic to the genus Tatia.