Airborne DNA and spider webs outperform other eDNA sources for monitoring terrestrial vertebrates

Understanding the strengths and limitations of different environmental DNA substrates is essential for optimizing terrestrial vertebrate surveys and monitoring. However, the performance of newly explored substrates (airborne eDNA, vegetation swabs, spiderwebs) compared to longstanding eDNA sources (water and soil) is uncertain. Using a metabarcoding approach, we assessed vertebrate eDNA diversity across seven substrates: three airborne DNA collection methods (a powered fan system, and two passive collection methods), spider webs, vegetation swabs, water, and soil at two sites. The highest species richness was detected by powered air samples and spider webs (powered air: 83 at Perth Zoo, 44 at Karakamia; spider webs: 62 and 40, respectively), with no significant difference in the community composition, suggesting they capture eDNA from similar sources however, all substrates contributed unique detections. Passive airborne DNA collection, though less efficient than powered devices, (mean ..., , # Airborne DNA and spider webs outperform other eDNA sources for monitoring terrestrial vertebrates

Dataset DOI: [10.5061/dryad.6wwpzgnb8](10.5061/dryad.6wwpzgnb8)

## Description of the data and file structure

Metabarcoding data generated from two assays and seven sample types collected: three different methods were used to collect eDNA from the air - powered air samplers, wind-powered samplers, and bare filter media as a passive collection method and soil, spider webs, swabbed vegetation surfaces and water samples. All samples were collected throughout both Karakamia Wildlife sanctuary and Perth Zoo.

### Files and variables

#### File: bc1_16S.txt and bc2_16S.txt

**Description:** Text file containing the unique ID barcoded tags required to demultiplex raw 16S Reptile sequences

#### File: bc1_12S.txt, bc2_12S.txt, bc3_12S.txt and bc4_12S.txt

**Description:** Text file containing the unique ID barcoded tags required to demultiplex raw 12S-V5 sequences:

#### File: Substrate_compa...,