Aluminum is one of the most abundant elements in earth’s crust and soils. Global soil acidification has occurred with industrialization and enhances Al solubility in acidic soils, resulting in Al phytoxicity and reducing crop production. Earthworms play an important role in nutrient recycling in soils and maintaining soil health. Rhizosphere microorganism excretes exopolysaccharides (EPS) into the rhizosphere. The objectives of this study is to examine effects of EPS from Rhizobium Tropici on aluminum uptake and ecotoxicity to earthworms in soils and litters. Overall this study provides the scientific understanding of the EPS in alleviation of Al toxicity to earthworms in soils. Results showed the clear mitigation of EPS in reducing earthworm mortality rate and increasing its reproductivity under Al stress. EPS significantly decreased Al uptake from soil and litters, and decreased Al in subcellular distribution (cell membrane, cytosol and organelle fractions) in earthworms from soils. Al in purges was found to be mainly present in the residual fraction, while EPS slightly increased Al in water soluble and exchangeable fractions in purges. Overall water soluble and organic matter bound Al in soils were correlated well with Al in earthworms, but Al in earthworms were controlled by total Al in purges. The current study concluded that earthworm Lumbricus Terrestre may be a potential bio-indicator of Al ecotoxicity in soils and EPS from Rhizobium Tropici effectively mitigated Al ecotoxicity in soils.