Purpose The application of organic and inorganic fertilizers to soil can result in increased gaseous emissions, such as NH3, N2O, CO2, and CH4, as well as nitrate leaching, contributing to climate warming and ground and surface water pollution, particularly in regions with hot climates, where high temperatures and high soil nitrification rates often occur. The use of nitrification inhibitors (NIs) has been shown to effectively decrease nitrogen (N) losses from the soil-plant system. Materials and methods Non-disruptive laboratory incubation experiments were conducted to assess the extent to which temperature (20 and 30 degrees C) and nutrient source (mineral and organic fertilizers) influence the rate of carbon (C)- and N-related microbial processes in soil in response to the NI 3,4-dimethylpyrazole phosphate (DMPP). Furthermore, short-term changes in the ability of microbes to degrade C substrates were evaluated in disruptive soil microcosms using microbial community-level physiological profiling and the abundance of the bacterial 16S rRNA gene as a measure of total bacterial population size. Results and discussion DMPP reduced net nitrification after 2 and 4 weeks of incubation at 30 and 20 degrees C by an average of 78.3 and 84.5 %, respectively, and with similar dynamics for mineral or organic fertilization. The addition of labile organic matter with cattle effluent led to a rapid increase in C mineralization that was significantly reduced by DMPP at both temperatures, whereas no changes could be detected after the addition of mineral fertilizer. The culturable heterotrophic microorganisms showed metabolic diversification in the oxidation of C sources, with organic fertilizer playing a major role in the substrate utilization patterns during the first week of incubation and the DMPP effects prevailing from day 14 until day 28. Furthermore, the copy number of the bacterial 16S rRNA gene was reduced by the application of DMPP and organic fertilizer after 28 days. Conclusions Our results show the marked efficiency of DMPP as an NI at elevated temperatures of incubation and when associated with both mineral and organic fertilization, providing support for its use as a tool to mitigate N losses in Mediterranean ecosystems. However, we also observed impaired C respiration rates and bacterial abundances, as well as shifts in communitylevel physiological profiles in soil, possibly indicating a short-term effect of DMPP and organic fertilizers on non-target Crelated processes and microorganisms.