Using earthworms as bioindicators of heavy metal contamination in soils is a relevant tool for environmental risk monitoring. This study examines the combined effects of four distinct concentrations mixtures (M1, M2, M3 and M4) containing Cd, Cr, Cu, Ni, Fe and Mn on Aporrectodea giardi earthworms after 12 and 24 days (12 D/24 D) of exposure via the monitoring of certain biomarkers of stress including total protein content, glutathione (GSH), metallothionein (MT), catalase and lipoxygenase (LOX) activities. The results show a decrease in the total protein level for the M3 mixture after 24 D, whereas it increases for all other treatments regardless of exposure time. Glutathione and metallothionine levels increased for M2 and M3 and decreased for M1 and M4 after 12 D; they increased after 24 D for all the mixtures. Regarding enzyme activities, catalase activity was decreased for all the treatments unless for M3 (P > 0.05). However, LOX increased for M1, M2 and M4 except for M3 after 12 D, when inhibition of this biomarker was observed. LOX activity was inhibited for all the mixtures at the end of the treatment. All the mixtures generated oxidative stress in Aporrectodea giardi, which is minimized by increasing MT levels to remove the metal ions and triggering the antioxidant system, composed primarily of GSH and LOX to restore cellular homeostasis. These findings suggest that the species Aporrectodea giardi could be an excellent candidate for ecotoxicological risk assessment of soils contaminated by metal mixtures and it can be used in bioremediation for its fitness which allows it to tolerate high concentrations of metal mixtures.