AbstractDecades of agriculture in a former—type of ecosystem, for example, deciduous forests—result in severe habitat degradation. The planted tree species encounter harsh conditions due to physically and chemically transformed soil environments. In addition, afforestation itself significantly modifies the upper soil horizons. It may impact the soil fauna communities, including mites from the Mesostigmata order (Mesostigmata = Gamasida). Due to their sensitivity, mesostigmatid mites are good bioindicators of changes in the soil. A decrease in the density, species richness, and diversity of mites indicates disturbances in the soil environment and may confirm the impact of agricultural practice on the soil environment. Our research aim was to examine the impact of soil properties and land use history on the mesostigmatid mite communities in stands growing on forest and post‐agricultural lands. These sites were afforested with different tree species (Betula pendula Roth., Fagus sylvatica L., Pinus sylvestris L., and Quercus robur L.). Thus, 21 research plots were established on forest and post‐agricultural land, each on rusty soils considered to be one of the most common in Central European forests. We collected a total of 567 soil samples during three sampling sessions in spring and autumn for soil Mesostigmata investigation. We also described the soil profile on each plot and collected soil and litter samples to measure pH, litter thickness, soil organic matter, bulk density, soil moisture, soil organic carbon, and elements content. Our study revealed that 16 out of 30 environmental parameters differed between habitat types. A total of 1355 mites were classified into 58 taxa (50 species and 8 genera). The most numerous species were Veigaia nemorensis (165 ind.; 12.2% of all mesostigmatid mites), Zercon peltatus (156; 11.5%), and Paragamasus conus (141; 10.4%). The highest abundance was recorded in birch stands on forest land, whereas the lowest abundance for oak stands growing on post‐agricultural land. Interestingly, in oak stands we recorded both the highest diversity of mite communities (forest land) and the lowest on post‐agricultural land. Furthermore, our study showed that post‐agricultural land and Na content in forest litter affected the abundance, species richness, and diversity of mesostigmatid mite communities. Species richness was additionally affected by tree species, that is, pine and oak. Our research indicated that long‐term agricultural practice negatively affected the density, species richness, and diversity of Mesostigmata communities 20 years after afforestation.