Microbiological evaluation of sandy grassland soils from two different stages of secondary succession on abandoned fields (4 and 8 years old fallow) was carried out as a part of research focused on restoration of semi-natural vegetation communities in Kiskunság National Park in Hungary. There was an apparent total N and organic C enrichment, stimulation of microbial growth and microbial community structure change on fields abandoned by agricultural practice (small family farm) in comparison with native undisturbed grassland. A successional trend of the microbial community was found after 4 and 8 years of fallow-lying soil. It consisted in a shift of r-survival strategy to more efficient C economy, in a decrease of specific respiration and metabolic activity, forced accumulation of storage bacterial compounds and increased fungal distribution. The composition of microbial phospholipid fatty acids mixture of soils abandoned at various times was significantly different.