Apple replant disease (ARD) is common to all major apple-growing regions in the world. It occurs when new apple trees are replanted on sites where previously the same or closely related crop species were grown. Biotic (fungi, bacteria and nematodes) and abiotic soil factors (poor soil structure, nutrition) contribute to the development and severity of ARD. However, the aetiology of ARD and effects on higher trophic levels are still unknown. In that sense, Collembola might play an important role, since they are one of the dominant mesofauna groups in many soils. They act as decomposer, fungivores and predators, representing different trophic levels in soil food webs. Therefore, any effect of ARD on the occurrence of Collembola could have ecological impacts on the soil quality and health. Here, we examined the colonization behaviour of two Collembolan species, Folsomia candida and Sinella curviseta, in choice tests and population growth tests using Apple Replant Diseased soil (ARD) and non-ARD soil samples from different field sites and standardized laboratory bioassays. Additionally, Collembola behaviour was quantified by continuous video observations to investigate short-term behavioural changes. Results showed that both Collembolan species significantly preferred colonization of the non-ARD soils compared with ARD soils, independent of the origin of the soil samples or specific disinfection treatments. Moreover, the detailed video analysis of the foraging behaviour indicates rapid colonization of soil samples and low dispersal rates. Most likely, volatile compounds and to a lesser extent feeding stimulants play a vital role for the colonization process for both Collembolan species. Finally, results showed negative effects of ARD on population growth of both Collembolan species already after an 8-week period, implying strong nutritional deficiencies in ARD affected soils. The hypothesis that ARD causing microorganisms directly affected orientation, colonization and population development of Collembola is discussed.