ENAGenomicsGerman Collection of Microorganisms and Cell Cultures GmbHDSMZhttps://www.ebi.ac.uk/ena/browser/view/PRJEB85100Data on species distribution is fundamental for conservation planning. However, acquisition of high quality data demands great sampling effort and time, especially in biodiverse regions. Furthermore, species detectability is frequently not considered during species inventories based on conventional methods and this could potentially compromise biodiversity assessments when detectability is low. In this context, eDNA metabarcoding emerges as a new promising method to study species distribution and community composition though the detection of DNA left by animals in environmental samples (e.g. water). We aimed to test the efficacy eDNA metabarcoding in comparison to conventional field inventory methods in tropical, species rich frog communities with previously documented low detectability (Serra do Cipó, southeasern Brazil). Among species detected by both field survey and eDNA metabarcoding, eDNA provided higher detectability for one third of species and similar performance for the remaining ones. eDNA also detected species hardly seen in field surveys (e.g. Physalaemus deimaticus). Even with an incomplete sequence database (about 50% of species in our study site are represented), in seven out of ten streams eDNA detected mode species than field inventories assessing either adult individuals or all life stages. Thus, eDNA metabarcoding proved to be an efficient method to complement field inventories of species with low detectability. We strongly recommend that eDNA surveys are performed in combination with sequencing of DNA target sequences to improve reference libraries. It is also important to further investigate variables likely to influence detectability (e.g., inhibitors, DNA transport and degradation, DNA release variation among species with different life histories) to improve the efficacy of this method, which is a very promising tool for faster acquisition of species distribution data in the conservation race against the pace of human impacts on the planet.ENAfalseHide and seek: species detectability in a biodiversity hotspot increased by environmental DNASpecies detectability through eDNA in a biodiverse site2025-08-112025-08-11PRJEB85100