Loricariidae is one of the largest fish families of the world, with about 650 species separated into six subfamilies. To date, cytogenetic data on only 56 species of this family are available. In the present study, the karyotypes of three Ancistrinae species and five Loricariinae species were studied. The lowest diploid number, 2n=38, was observed in Ancistrus n.sp. 1 (Ancistrinae) and the highest diploid number, 2n=70, was observed in Rineloricaria n.sp. (Loricariinae). The nucleolar organizer regions (NORs) were seen at a terminal position in six species and at an interstitial position in two. The karyotypic analysis of Loricariinae and Ancistrinae species revealed that these groups exhibit a large diversity of diploid numbers, suggesting the occurrence of intense karyotypic evolution during their evolutionary history.