Airborne environmental DNA (airborne eDNA) analysis leverages the globally ubiquitous medium of air to deliver broad species distribution data and support ecosystem monitoring across diverse environments. As this emerging technology matures, addressing critical challenges and seizing key opportunities will be essential to fully realize its potentially transformative impact. In June 2024, the Southern eDNA Society convened over 100 researchers, industry leaders, and biodiversity management stakeholders in a landmark workshop to evaluate the current state of airborne eDNA research and chart a course for future development. Participants explored opportunities for integrating airborne eDNA into existing monitoring systems, but they unanimously agreed that research must first be applied to improving understanding of airborne eDNA ecology. The workshop emphasized the importance of collaborative engagement with stakeholders—including government agencies, Indigenous communities, and citizen scientists—to ensure practical and ethical implementation. This summary highlights current challenges and actionable recommendations, including improving our understanding of airborne eDNA ecology, harmonizing sampling methodology (e.g., devices, materials, sampling density, duration), identifying and mitigating sources of error, and fostering early, sustained stakeholder collaboration. By addressing these challenges, airborne eDNA analysis can become a transformative tool for biodiversity, biosecurity, and conservation monitoring on a global scale. Its ability to detect diverse taxonomic groups—including fungi, plants, arthropods, microbes, and vertebrates—positions airborne eDNA as a pivotal technology for holistic terrestrial biodiversity assessments that transcend traditional, species‐focused monitoring approaches.

In this study, we evaluate the use of various two-dimensional liquid chromatographic methods to characterize water-soluble, synthetically grafted bio-polymers, consisting of long poly(acrylic acid) chains and short maltodextrin grafts. The confirmation of the presence of grafting and the estimation of its extent is challenging. It is complicated by the limited solubility of polymers, their structural dispersity and chemical heterogeneity. Moreover, the starting materials (and other reagents, reaction products and additives) may be present in the product. Reversed-phase liquid chromatography (RPLC), hydrophilic-interaction liquid chromatography (HILIC) and size-exclusion chromatography (SEC) were used to characterize the product, as well as the starting materials. Additionally, fractions were collected for off-line characterization by infrared spectroscopy and mass spectrometry. The one-dimensional separation methods were found to be inconclusive regarding the grafting question. Breakthrough (the early elution of polymer fractions due to strong injection solvents) is shown to be a perpetual problem. This issue is not solved by comprehensive two-dimensional liquid chromatography (LC × LC), but information demonstrating the success of the grafting reaction could be obtained. SEC × RPLC and HILIC × RPLC separations are presented and discussed.

Airborne environmental DNA (eDNA) offers a scalable tool for biodiversity monitoring, yet our limited understanding of airborne eDNA transport makes it challenging to define sample origin. We address this by investigating local spatial dynamics of eDNA dispersal using passive samplers and wind data. Over 3 weeks, we deployed passive samplers at Rotterdam Zoo (the Netherlands) and collected airborne eDNA at 5 locations for multiple durations over 96 h, where all samplers were started simultaneously. Vertebrate diversity was sequenced at 12S and 16S and resident zoo species identified ( N = 24). Wind‐directed catchment areas were estimated using two models (Circular Sector model, using simple wind‐directed data from a nearby weather station, and Footprint model using gridded meteorological data). Catchment areas were defined by wind characteristics, with size and orientation determined by speed and directional variability. Results showed 100% species detections fell within predicted catchment areas up to 100 m (not all species in zoo detected). At longer ranges up to 655 m, 62% of the total detected species fell within the wind‐directed catchment areas. Species detections outside of the catchment areas suggest additional mechanisms may influence longer range transport of airborne eDNA. Longer sampling durations significantly increased the probability of species detections inside catchment areas and both models performed comparably well. Together, our results suggest simple wind characteristic models accurately identify vertebrate airborne eDNA catchments at local scales, increasing confidence that species signals originate at < 200 m downwind from sampler placement. Our work thus helps to enable more spatially explicit biomonitoring inferences.

Airborne environmental DNA (eDNA) is emerging as a powerful tool for noninvasive assessment of terrestrial biodiversity. We evaluated the effectiveness of three analytical pipelines for characterizing plant diversity in a botanical greenhouse, with a particular focus on how particle size and sampling height influence detection. Air was sampled for up to 4 days using active Total Suspended Particulate samplers, a 14‐stage impactor, and a vertical filter stack, all equipped with quartz filters. To assess methodological variability and identify optimal approach, three analytical pipelines were applied: (A) Illumina amplicon sequencing with a QIIME‐based workflow, and (B) Illumina and (C) Nanopore shotgun sequencing processed with Kraken2. Of ~2100 plant genera recorded in the greenhouse and adjacent garden, ~86% were considered detectable based on rbcL, trnL, ITS1, and ITS2 barcode coverage. Pipeline A (32 samples) detected 149 genera, 88% of which were known from the garden; 55% of species‐level assignments matched cataloged species, with ~11% attributable to misannotations or contamination. Pipelines B (6 samples) and C (2 samples) detected 76 and 88 genera, respectively, with 81%–82% corresponding to garden taxa and 50%–51% confirmed at species level. The two shotgun pipelines produced highly similar taxonomic profiles despite samples being collected 7 weeks apart, and both were strongly complementary to the amplicon results. Size‐dependent sampling yielded elevated detections in the 2.5 μm impactor fraction, indicate substantial contributions from sub‐pollen particles, while larger fractions (> 40 μm) also contained plant eDNA, suggesting multiple particle carriers or aggregates. Overall, airborne eDNA provided robust detection of local plant biodiversity and yielded new insights into particle‐size distributions relevant to eDNA transport and deposition.