Surveying Tropical Faunal Diversity via Airborne DNA Analyses

Here, we assess the potential of airborne eDNA (airDNA) metabarcoding to characterize arthropod and vertebrate assemblages in a tropical forest. We deployed 28 high-flow air samplers across a 1.5 km2 area on Barro Colorado Island, Panama, over two consecutive 24- hour periods and sequenced captured DNA using general eukaryotic and vertebrate-specific metabarcoding primers. AirDNA sampling detected 1293 arthropod operational taxonomic units (OTUs) alongside 157 vertebrate OTUs representing birds, mammals, reptiles, amphibians, and fish. Arthropod richness was comparable to that quantified locally with light and Malaise trap surveys, while vertebrate richness exceeded that typically observed with conventional techniques. The level of field and lab effort employed in this study captured approximately 84% of the asymptotic richness for arthropods and 76% for vertebrates, implying that relatively little additional richness would have been quantified with additional effort within the same domain of sampling. Observed and asymptotic richness increased with the number of sites, the number of days a site was sampled, and the number of PCR replicates run per sample, highlighting the need to standardize effort among future applications of airDNA to compare richness across systems. These results demonstrate that airDNA metabarcoding can efficiently and non- invasively profile tropical terrestrial faunal biodiversity.