Adoption of sustainable tillage can protect soils from biological degradation and maintain soil quality, as compared with conventional management. This paper presents findings from a long-term tillage experiment carried out in Endocalcari-Endohypogleyic Cambisols on a sandy loam soil in Lithuania. The tillage systems were: conventional (CT), moderate (MT), and no tillage (NT). Tillage intensity positively affected microbial substrate utilization and urease activity, as well as, negatively, dehydrogenase activity, bacteria and fungi amounts, and Shannon diversity index of microbiological community. Higher total porosity provoked higher enzyme activity; but, microbial activity correlated negatively with bulk density.