“…Despite the increasing popularity of this method in the assessment of aquatic ecosystems using fish eDNA or ichthyoplankton pools (Carvalho, 2022;Cilleros et al, 2019;Sales et al, 2019;Sales et al, 2021, Silva et al, 2023, the vast majority of fish metabarcoding studies are still concentrated in temperate regions, in wellcharacterized and reasonably accessible environments (Lawson Handley et al, 2019;McDevitt et al, 2019). Considering the highly diverse Neotropical ichthyofauna, appropriate implementation of metabarcoding biomonitoring is hindered by a lack of knowledge of the local biodiversity (e.g., undescribed or cryptic species), primer biases, and the incompleteness of reference databases (Jackman et al, 2021;Sales, Mariani, et al, 2018;Sato et al, 2017).…”