Aim
The Neotropics, particularly South America, holds unparalleled high levels of species richness, when compared to other major biomes. Some neotropical areas are hotspots of a fragmentary known diversity of insects and are under manifest danger of biodiversity loss and climate change. Therefore, prompt estimates methods of its diversity are urgently required to complement slower traditional taxonomic approaches. Despite a variety of algorithms for delimiting species through single-locus DNA barcodes having been developed and applied for rapid estimates of species diversity in a wide array of taxa; however, tree-based and distance-based methods may lead to different group assignments, either overestimating or underestimating the number of putative species. Here, we investigate the performance of different DNA-based species delimitation approaches for a rapid biodiversity estimate of the diversity of Polypedilum (Chironomidae, Diptera) in South America.