A diversified approach involving chemical, microbiological and ecotoxicity assessment of soil polluted by heavy mineral oil was adopted, in order to improve our understanding of the biodegradability of pollutants, microbial community dynamics and ecotoxicological effects of various bioremediation strategies. With the aim of improving hydrocarbon degradation, the following bioremediation treatments were assayed: i) addition of inorganic nutrients; ii) addition of the rhamnolipid-based biosurfactant MAT10; iii) inoculation of an aliphatic hydrocarbon-degrading microbial consortium (TD); and iv) inoculation of a known hydrocarbon-degrading white-rot fungus strain of Trametes versicolor. After 200 days, all the bioremediation assays achieved between 30% and 50% total petroleum hydrocarbon (TPH) biodegradation, with the T. versicolor inoculation degrading it the most. Biostimulation and T. versicolor inoculation promoted the Brevundimonas genus concurrently with other α-proteobacteria, β-proteobacteria and Cytophaga-Flexibacter-Bacteroides (CFB) as well as Actinobacteria groups. However, T. versicolor inoculation, which produced the highest hydrocarbon degradation in soil, also promoted autochthonous Gram-positive bacterial groups, such as Firmicutes and Actinobacteria. An acute toxicity test using Eisenia fetida confirmed the improvement in the quality of the soil after all biostimulation and bioaugmentation strategies.