Land use strongly influences soil properties and unsuitable practices lead to degradation of soil and environmental quality. The aim of this study was to assess the impact of different land uses on some chemical properties of soils developed from Pliocene clays, within hilly environments of central and southern Italy. The areas investigated are located in Vicarello di Volterra (Pisa, Tuscany), S. Quirico d'Orcia (Siena, Tuscany) and Soveria Simeri (Catanzaro, Calabria). Within each area different land uses were compared, including a natural ecosystem (Mediterranean bush), a perennial grass or pasture and an intensive crop (wheat, as monoculture or in rotation). The soils were sampled at 0.0-0.1, 0.1-0.2 and 0.2-0.4 m depth and analysed for particle size, pH, bulk density, cation exchange capacity and exchangeable cations, total organic carbon (TOC) and humified carbon (HC) concentrations, organic carbon stock and total N. The stratification ratio of soil organic carbon was calculated to characterize soil organic carbon distribution with depth. At all sites, soil under Mediterranean bush contained the largest amounts of TOC (as both concentration and stock), HC, total N and exchangeable K, together with the highest cation exchange capacity and the lowest pH values. The decrease in soil OC stock with land use change from natural to agricultural ecosystem was 65-85% to 0.1 m depth, 55-82% to 0.2 m depth and 44-76% to 0.4 m depth, with the lowest decrements for perennial grass from S. Quirico and the highest decrement for continuous wheat from Soveria Simeri. Continuous wheat cropping, based on conventional tillage, proved to be the least sustainable land use. At Soveria Simeri, the organic carbon content under pasture was not significantly larger than under wheat cultivation, probably because of grazing mismanagement; however, organic carbon under pasture was more humified. At S. Quirico, the perennial grass resulted in a significant increase in soil organic carbon at the soil surface relative to the wheat cultivation, while at Vicarello no differences were observed between alfalfa/wheat rotation and perennial grass. Our results lead to the questioning of sustainability of intensive cereal farming and uncontrolled grazing in the considered environments, emphasizing the need for greater attention to conservative land managements.