Samples from a sandy agricultural soil were treated with increasing amounts of a fungicide (Sportak). The effects on the soil microflora were investigated over several weeks by monitoring basal and substrate-induced respiration and basal and substrate-induced heat output. The microbial biomass, metabolic quotient (qCO2), relative heat output (rqheat), lag phase of substrate use, and calorimetric: respirometric ratio were used as ecophysiological parameters. As structural and community-specific parameters, we recorded tryptophan contents and auxin metabolism, and calculated the ratios of fungal to bacterial respiration by antibiotic inhibition of substrate-induced respiration. Sportak either inhibited or stimulated the microbiota, depending on the length of exposure to the fungicide and the amount applied. Mineralization of dead biomass was reflected in increased soil tryptophan contents after the Sportak application. A shortened lag phase demonstrated inhibition and a prolonged lag phase stimulation of substrate use. This changed with the experimental phase. The rqheat and the calorimetric: respirometric ratio proved to be suitable parameters for the detection of stress metabolism (repair processes) in soil microbiota, because thermodynamic processes and catabolic and anabolic metabolism are taken into account at the same time. Following the application of Sportak, indole 3-acetic acid biosynthesis decreased while indole-3-ethanol biosynthesis increased, probably as a result of a transitional community shift from K-strategists towards r-strategists. It was not the fungicide but the formulation (mainly xylol) that damaged the organisms. A shift in the ratio of fungi to bacteria was also observed, suggesting that the bacteria were probably more sensitive to xylol than the fungi.