Resumo. Abstract: Aim Monitoring altered environments is crucial to understand the impacts and the possible changes in the local fish fauna. Therefore, we aimed to compare the ichthyofauna within a 67-year-old small run-of-the-river reservoir with surveys carried out in 1986 and 2019-2021 and update the checklist of this reservoir. Methods We used the data provided by Alves et al. (1998) and sampled fish in two sites inside Itutinga reservoir using gill nets, seine nets and hand nets and at the one tributary of the area using only hand nets. Results We collected a total of 12,978 individuals belonging to 37 fish species, and the species accumulation curve showed a tendency to the asymptote. Five of the sampled species are non-native and five are migratory. Alves et al. (1998) sampled a total of 25 fish species, 17 genera, 9 families and 3 orders. We added 12 fish species to the checklist that were not recorded by Alves et al. (1998), including four non-native and two migratory. On the other hand, two migratory species and one rheophilic were registered only in the preview study. In addition, we observed important changes in the species abundance rank. Conclusions The differences in the fish assemblage structure comparing both studies indicate important changes in fish assemblages over more than 33 years. Our results highlighted the importance of long-term fish monitoring in altered environments to assess temporal variation in fish assemblages and their responses to the multiple possibilities of external stressors, such as the shift in abundance of non-native species in these environments.