ObjectiveSmall mammals such as the Julia Creek dunnart (Sminthopsis douglasi) may be difficult to detect using traditional trapping methods. Here, we conducted a pilot study to determine whether eDNA collected from soil and/or air could detect the presence of terrestrial vertebrates, including S. douglasi, in a semi-arid, open grassland environment.ResultsAirborne eDNA analysis returned vertebrate DNA from five sample sites (n = 7), whereas soil eDNA analysis returned vertebrate DNA from a single site (n = 7). The Julia Creek dunnart was not detected in any of the experimental samples. However, several airborne eDNA samples did return strong matches to three terrestrial vertebrates, the long-haired rat (Rattus villosissimus), red kangaroo (Osphranter rufus) and brown quail (Synoicus ypsilophorus), all native species known to occur commonly in the study area. Overall, our preliminary findings suggest that the effectiveness of airborne and soil-derived eDNA in detecting terrestrial vertebrates was constrained by high human signal and low sampling intensity. For future studies, we recommend a number of field and lab-based refinements to increase the likelihood of detecting more taxa, particularly those that occur at low density.Clinical trial numberNot applicable