Assessing Pantanal fauna through environmental DNA metabarcoding after the 2020 megafire

The environmental DNA (eDNA) metabarcoding is a methodology that, from environmental samples such as soil, water, air, and others, enables the simultaneous identification of multiple species, thus allowing for large-scale mapping of biological diversity in a specific study area. Due to its non-invasive sampling approach, where species are detected from the traces they leave in the environment, eliminating the need to isolate and capture organisms, eDNA metabarcoding emerges as a valuable tool in conservation strategies. This study aims to explore the use of eDNA methodology for biodiversity monitoring and environmental impact assessment caused by the 2020 megafire in the Pantanal of Brazil, focusing on vertebrates. Environmental samples were collected at two protected areas and their surrounding areas, Taiamã Ecological Station (TES) and Pantanal Matogrossense National Park (PMNP), Mato Grosso, Brazil. We identified in TES, 27 mammals, 56 fishes, 12 birds, 4 amphibians, and 4 reptiles, while in PMNP, 43 mammals, 45 fishes, 126 birds, 19 amphibians, and 11 reptiles. Soil sampling proved to be more efficient compared to water sampling: 26 species were exclusively identified in soil samples, while 9 were attributed to water samples. Here, we demonstrated that the primer 12SV5 only a superior efficacy in identifying mammal and herpetofauna species compared to the other markers used (16Smam and MiBird). Moreover, we confirmed the complementary role of eDNA alongside camera trapping, and its advantage to estimate species richness with a single field expedition. We stress the need to optimize sample collection methods for the target group and to reduce the influence of contamination and water flow. This study highlights the importance of eDNA methodology as a crucial tool for biodiversity monitoring and environmental impact assessment, enabling rapid access to biodiversity and long-term monitoring.