Metazoan Diversity in Chilean Hypersaline Lakes Unveiled by Environmental DNA

Abstract Saline and hypersaline wetlands are biodiversity hotspots for metazoans such as aquatic invertebrates and wading birds. However, the survival of these habitats and their biota is increasingly threatened by a combination of pressures from climate change and extractive processes, jeopardizing the long-term ecological functioning of these ecosystems. With the goal of improving conservation efforts through state-of-the-art survey techniques in hypersaline ecosystems, this study tests the use of environmental DNA (eDNA) methods for metazoan biomonitoring. We employed a multi-assay approach utilizing three genetic markers—12S rRNA, 18S rRNA, and COI —to analyze biodiversity in two types of environmental substrates, sediment and water. These samples were collected from three hypersaline lakes situated at high altitude in Northern Chile: Salar de Atacama (Laguna Puilar), Salar de Pujsa, and Salar de Tara. In addition, we compared the eDNA outputs with results generated from aquatic macroinvertebrate assessments using kick-nets to evaluate the potential for complementary sampling approaches. Our eDNA analyses revealed a total of 21 and 22 taxa across the three hypersaline lakes in sediment and water, respectively. Within both substrates, the highest diversity was found in Salar de Tara (15 taxa within sediment and 13 taxa from water). Our multi-assay design was able to detect a range of resident hypersaline taxa with different conservation status, spanning from rotifers ( Encentrum ) to endangered snails ( Heleobia atacamensis ), to amphipods ( Hyalella ) and flamingos ( Pheonicopterus ). Macroinvertebrate presence/absence data derived from conventional kick-net surveys further validated Salar de Tara as the most biodiverse system. Compared to net-based assessments, eDNA analysis allowed more refined taxonomic assignments for copepods and ostracods, while certain taxa such as Ephydridae or Hirudinea were not detected through molecular tests. Overall, this study provides evidence that eDNA is an effective tool to elucidate fine scale taxa assemblages and can refine conservation efforts in hypersaline lakes. Given the fast pace of research developments in the field of molecular ecology, eDNA hosts great potential to become a central actor in hypersaline bioassessments in the near future.