As rehabilitation efforts in Guyana are recent, there is little information on the effect of different ecological rehabilitation protocols for Guyana's mining sites on biogeochemical cycles and mercury mobility. This study was conducted to assess the impact of different ecological restoration protocols on soil quality with the use of soil microbial indicators and by estimating the mercury mobility. We sampled soil from six rehabilitated mining sites in French Guyana with different ecological restoration procedures. We carried out measurements of enzymatic activities and an analysis of mercury environmental speciation to assess its potential toxicity according to a mobility gradient. The results obtained in this study show that the rehabilitation of mining sites has been carried out in a heterogeneous manner and soil quality is very variable, even in nearby sites. Sites that have been rehabilitated with fabaceous species have positive soil quality indicators. In addition, the results highlight a change in mercury mobility that is 82.1% correlated after co-inertia analysis with soil texture properties, which also confirms a direct effect of rehabilitation on mercury mobility. The non-restored sites had a much higher potential of mercury mobility and toxicity than the sites where ecological restoration was successful. These results highlight the positive effect of controlled rehabilitation and ecological restoration on microbiological activities and the potential toxicity of mercury.