Management strategies like organic matter (OM) amendment and mechanical energy inputs are known to influence the soil system. A long-term (13–14-year) field experiment was used to evaluate the effects of these management strategies on soil structural formation, structural stabilization and soil tilth of a sandy loam soil in Denmark. OM was applied as manure and by retention of plant residues (ORG) to be compared with plots dressed only with mineral fertilizer (MIN). The soils were rotovated (ROT), compacted (PAC) or left undisturbed (REF) as split-plot treatments in the main plots with OM management over two years prior to sampling. In two consecutive years, undisturbed soil samples were collected from the 6 to 13cm soil layer in the field grown with winter wheat to assess soil organic carbon (C) fractions (total organic C, polysaccharide C and microbial biomass C), total organic C and polysaccharide C of 1–2mm macro-aggregates, bulk density, hyphal length, aggregate stability, clay dispersibility, aggregate tensile strength, direct tensile strength and shear strength. The ease of fragmentation and the torsional shear strength of soil were measured in the field as well. OM application increased all soil organic C fractions. Response patterns of organic C fractions in aggregates were the same patterns as for whole-soil. Polysaccharide C appeared to be an important agent in the aggregation process. The effect of microbial C and fungal hyphae on the aggregation process was not clear. Extensive tillage and traffic produced unfavourable tilth conditions in terms of a greater degree of clay dispersion, lower aggregate stability, higher soil tensile strength and poorer soil fragmentation. OM affected soil reaction to compressive and tensile stresses applied at differing initial bulk densities. The results also indicated the profitability of supplementing the classical laboratory analysis with in situ measurements to better evaluate management effects on soil structure.