The aim of this study was to evaluate chemical and biochemical changes of organic matter in fertilized (ammonium nitrate) and amended (vermicompost and manure) soils using pyrolysis and metabolic indices. The metabolic potential [dehydrogenase (DH-ase)/water soluble organic carbon (WSOC)], the metabolic quotient (qCO2) and the microbial quotient (Cmic:Corg) were calculated as indices of soil organic matter evolution. Pyrolysis-gas chromatography (Py-GC) was used to study structural changes in the organic matter. Carbon forms and microbial biomass have been measured by dichromate oxidation and fumigation-extraction methods, respectively. Dehydrogenase activity has been tested using INT (p-Iodonitrotetrazolium violet) as substrate. The results showed that organic amendment increased soil microbial biomass and its activity which were strictly related to pyrolytic mineralization and humification indices (N/O, B/E3). Mineral fertilization caused a greater alteration of native soil organic matter than the organic amendments, in that a high release of WSOC and relatively large amounts of aliphatic pyrolytic products, were observed. Therefore, the pyrolysis and metabolic indices provided similar and complementary information on soil organic matter changes after mineral and organic fertilization.