Abstract: In the context of an ongoing monitoring study on the impacts of gold-mining activities on critical ecosystem processes, we explored the use of soil Denitrifying Enzyme Activity (DEA) and Substrate Induced Respiration (SIR) as ecosystem indicators in tropical rainforests of French Guiana. We also propose DEA/SIR ratio as ecosystem attribute able to describe the state of an ecosystem and to reflect changes in ecological processes. With this purpose, we measured SIR, DEA and DEA/SIR ratio in five gold-mining areas and five surrounding natural reference rainforests. We also measured indicators in two conditions of spontaneous regeneration of vegetation (stratified or not) and two conditions of soil rehabilitation (prior preparation of soils or not). We showed a high variability of DEA, SIR and DEA/SIR ratio in the natural reference forests. This pointed out the necessity to identify relevant reference systems – i.e. proving a close match in all relevant ecological dimensions – to compare with closed perturbed systems in order to assess the levels of alterations after disturbances. Results showed a high impact of gold mine on microbial processes with a strong decrease of DEA (10-fold lower), SIR (2-fold lower) and DEA/SIR ratio (8-fold lower) in perturbed areas in comparison with natural reference forests. The type of spontaneous vegetation (stratified or not) influenced the values of indicators as well as prior rehabilitation of soils, demonstrating the capacity of DEA, SIR and DEA/SIR ratio to respond in proportion to the perturbation (robustness) and to the different levels of restoration (sensitivity). The systematic decrease of the ratio DEA/SIR observed in the studied perturbed situations demonstrates clearly that the structure of microbial communities has been also modified. The ratio DEA/SIR proved to be robust and sensitive, and able to describe in fairly fine way changes of soil microbial communities in terms of structure and function in gold mine areas and during processes of restoration. We propose to use DEA, SIR and DEA/SIR ratio as bioindicators of both structural and functional aspects of C and N cycling in soils. Together with others bioindicators based on key supporting functions in soils, these indicators should accurately evaluate the ecological potential of natural ecosystems and the levels of degradation in case of land-use changes. © 2012 Elsevier Ltd.