The application of animal manure effluents in agriculture in combination with nitrification inhibitors should be beneficial for nutrient recycling, soil quality, plant productivity, and greenhouse gas emission and offer economic advantages to make them an alternative to conventional fertilizers. The present study aims to estimate the effects of the addition of bovine manure effluent alone or together with a nitrification inhibitor (3,4-dymethylpyrazol-phosphate (3,4 DMPP)) on the microbial community dynamics in a Mediterranean soil in an incubation experiment over 28 days. The application of the bovine manure effluent increased respiration, microbial biomass carbon, fungal and bacterial growth, and enzyme activities and changed the microbial community structure evaluated by the phospholipid fatty acid pattern. Adding the bovine manure effluent together with the nitrification inhibitor, although partly negating the positive effect of the effluent on soil microbial activity, still resulted in higher or similar growth and activity as in the control. Our results indicate that the addition of the nitrification inhibitor 3,4 DMPP together with a bovine manure effluent could be a promising solution to control the animal manure effluent application effects on soil microbiological properties and microbial dynamics, as well as counteracting direct inhibiting effects of 3,4 DMPP on the soil heterotrophic community.