Simplified, portable and cost‐efficient airborne eDNA metabarcoding to expand tropical biodiversity assessments

Aquatic environmental DNA (eDNA) analysis has been widely established as a sensitive and non-invasive species detection toolfor global biodiversity monitoring. In contrast, the uptake of airborne eDNA has been constrained by the absence of robust, high-throughput samplers suited to humid tropical settings. We present a simplified, portable, and cost- efficient active air sampler thatinjects air into a liquid collection medium, shifting airborne eDNA from stationary, resource-heavy set-ups to field-deployable,scalable workflows. Using a greenhouse vertebrate mock community to test the sensitivity of our approach, airborne eDNA sam-pling recovered most taxa present, with high detection rates for birds, mammals, and reptiles. An optimized collection bufferincreased sequencing yield by about one fourth over sterile water. To subsequently assess its application in biodiversity researchand conservation science, we tested its functionality in the Chocó-Andean montane forests of northern South America, usinga tree-of-life metabarcoding approach as part of ongoing biodiversity surveys to inform the establishment of a protected areanetwork. While aquatic eDNA detected hundreds of vertebrates and thousands of arthropods and embryophytes, airborne eDNArecovered slightly lower OTU richness. Airborne sampling was particularly effective for mammals, whereas aquatic samplingrecovered more actinopterid, amphibian, and avian OTUs. Overlap in species detection between methods was low, highlightingthe complementarity of airborne and aquatic eDNA-based species assessments. By integrating accessibility, portability, and scal-ability, our approach broadens the applicability of airborne eDNA, where combining airborne and aquatic eDNA metabarcodingcan rapidly deliver broad-scale biodiversity data to support conservation planning in megadiverse yet resource-limited regions.

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