Toluene is a highly flammable and commonly used industrial chemical with severe health consequences on humans upon exposure and ingestion. In this study, multispecies bioassays were conducted using a species sensitivity distribution approach to determine acute and chronic hazardous concentrations of toluene in soil. Acute and chronic toluene toxicity tests were conducted with seven soil species from four taxonomic groups. The results from the toxicity tests were used to estimate the acute and chronic HC₅ (hazardous concentration for 5 % of species) of toluene in the terrestrial environment at 58.9 (5.4–639.6) mg kg⁻¹ and 2.2 (0.2–19.8) mg kg⁻¹, respectively. To the best of our knowledge, this is the first study to estimate the hazardous concentration of toluene in soil by conducting a battery of bioassays. These values can be used as references for the environmental risk assessment of chemical accidents involving toluene and estimating its impact on soil to protect the terrestrial environment.

Surface modified lipopeptide biosurfactant (BS) with enhancement of amino acids was produced using Bacillus Malacitensis. The aromatic hydrocarbons from contaminated soil were removed by BS soil washing process and bioremediation using activated functionalized carbon-BS matrix (AFC-BS). The Central Composite Design (CCD) showed the optimum time100 h; pH 7; temperature 30°C on maximum yield of BS. The amino acid profiling of BS reveals the enhancement of amino acids especially polar amino acids and its importance in the formation of micellar structure for the tight packing of aromatic hydrocarbons from industrial contaminated soil. AFC-BS matrix was implanted directly into the contaminated soil for 28 days and found 61.80 % of Total Petroleum Hydrocarbon (TPH) removal efficiency which is high compared to the AFC treated soil. The compounds were extracted from contaminated soil and AFC-BS matrix, found similar peaks in high performance liquid chromatography, which reveals the ability of BS to remove aromatic contaminants. The soil toxicity was also analyzed by seed germination and found improvement in the growth of seeds. The germination of seeds increased from 60 % to 100 % and the phytotoxicity of root and shoot was reduced from 89.50 %, 88.45 % to12.55 %, 11.87 % respectively.

A long-term case of residual pollution is studied after 20 years since the largest mining accident in Spain (the Aznalcóllar spill) happened. This pollution is manifested through a surface zoning consisting of bare soils (B0), sparsely vegetated soils (B1), and densely vegetated and recovered soils (B2). A biopiles treatment with a mixture of contaminated soils (B0 and B1) with recovered soils (B2) at 50% (w/w), and vermicompost addition (50 tons ha-1) was evaluated. To assess the effectiveness of treatments, total, water-soluble, and bioavailable fractions of the most polluting elements in the zone (Cu, Zn, As, Pb, Cd, and Sb) was analyzed. To evaluate the potential risk of contamination for the ecosystem, a bioassay with earthworm Eisenia andrei was carried out. Twenty years after the accident, there are still soils where total As and Pb exceed the regulatory levels and water-soluble Zn and As exceed the toxicity guidelines. According to toxicity bioassay, weight variation and juvenile production of earthworms showed an improvement after biopiles treatment, with values that trend to be similar to those of recovered soils. The only bioaccumulated element in earthworms was Cd (BAF>1), both in polluted as in treated soils, which indicates the possible existence of exclusion mechanisms of the other pollutants by earthworms. The comparison between biopiles and polluted soils showed no significant differences for the bioaccumulation factor of trace elements, with the exception of Zn and Cu, which slightly increased after treatment. According to our results, biopiles treatment combined with vermicompost addition is a good technique for the recovery of residual contaminated areas, by the improvement of soil properties and the reduction of the potential toxicity; anyway, monitoring of soils and organisms is needed to prevent the increase of bioavailability of some potentially pollutant elements over time.