Perched high up in a Eucalyptus tree, swaying from side to side, lies a sleepy koala unaware of the means spent each year trying to obtain accurate baseline information about its presence. We have thrown all we could at it, from wildlife surveys to night spotting, bioacoustics, detection dogs, and drones equipped with thermal cameras. Yet, whilst critical to its conservation and management efforts, finding a koala remains an ambitious, time-consuming, and costly endeavour often producing insufficient results. Little did we know that traces of koalas’ presence and that of its predators along with other native, domesticated, and invasive species, float in the air and can be detected through traces of DNA. This study confirms that koalas alongside species belonging to the wallaby, possum family, invasive species such as foxes, domestic dogs and rabbits can successfully be detected by sampling airborne particles. We detected 16 unique taxonomic assignments through sampling of airborne particles, successfully assigning 11 of these to species level. Further, we develop and employ a novel koala-specific qPCR for the cost-effective detection of koala eDNA in the wild. Together, this study demonstrates the potential of airborne eDNA for the detection of threatened terrestrial wildlife under natural conditions and presents achievable optimisation steps to increase its field applicability and validity.