We used conventional cytogenetic techniques (Giemsa, C-banding, Ag-NOR), and fluorescent hybridization (FISH) with 5S and 18S rDNA probes to investigate the karyotype and cytogenetic characteristics of (Günther, 1860) from Ecuador. The specimens studied have a karyotype with 2n=54 biarmed chromosomes (32 M + 22 SM) and C-positive heterochromatin located on the centromeric, pericentromeric, interstitial, and terminal regions of some chromosomes. The nucleolus organizer regions occurred terminally on the long arm of chromosome pair 2. FISH confirmed the presence of only one 18S rDNA cluster with nonsyntenic localization with the 5S rDNA. Cytogenetic data allow us to refute the earlier morphological hypothesis of a sister relationship between Fowler, 1941 and Posada Arango, 1909 and support the molecular proposal that is a sister group to the monophyletic clade containing Agassiz, 1829 and .