The aim of this research was to investigate the microbial activity in forest soil from different sites under deciduous and coniferous trees in Serbia. One site on Stara planina was under beech trees (Fagus sp.) while another under mixture of spruce (Picea sp.) and Douglas fir (Pseudotsuga sp.). The site on Kopaonik was under mixture of beech (Fagus sp.) and spruce (Picea sp.) trees. The site on Tara was dominantly under fir (Abies sp.), beech (Fagus sp.) and spruce (Picea sp.). The total number of bacteria, the number of actinobacteria, fungi and microorganisms involved in N and C cycles were determined using standard method of agar plates. The activities of dehydrogenase and ß-glucosidase enzymes were measured by spectrophotometric methods. The microbial activity was affected by tree species and sampling time. The highest dehydrogenase activity, total number of bacteria, number of actinobacteria, aminoheterotrophs, amylolytic and cellulolytic microorganisms were determined in soil under beech trees. The highest total number of fungi and number of pectinolytic microorganisms were determined in soil under spruce and Douglas fir trees. The correlation analyses proved the existence of statistically significant interdependency among investigated parameters.