Deforestation occurs at an alarming rate in upland watersheds of Bangladesh and has many detrimental effects on the environment. This study reports the effects of deforestation on soil biological properties along with some important physicochemical parameters of a southern upland watershed in Bangladesh. Soils were sampled at 4 paired sites, each pair representing a deforested site and a forested site, and having similar topographical characteristics. Significantly fewer (p≤0.001) fungi and bacteria, and lower microbial respiration, active microbial biomass, metabolic and microbial quotients were found in soils of the deforested sites. Soil physical properties such as moisture content, water holding capacity, and chemical properties such as organic matter, total N, available P and EC were also lower in deforested soils. Bulk density and pH were significantly higher in deforested soils. Available Ca and Mg were inconsistent between the two land uses at all the paired sites. Reduced abundance and biomass of soil mesofauna were recorded in deforested soils. However, soil anecic species were more abundant in deforested soils than epigeic and endogeic species, which were more abundant in forested soils than on deforested sites.