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.

Propylparaben is widely used as a preservative in pharmaceuticals and personal care products and is ultimately excreted by the human body. Thus, propylparaben reaches sewage and enters the soil environment by sludge fertilization and wastewater irrigation. However, there are few existing studies on the toxicity and risks of such chemicals in terrestrial environments. In this study, a multispecies bioassay for propylparaben was performed and protective concentrations (PCs) were derived based on toxicity values by probabilistic ecological risk assessment. Acute and chronic bioassays were conducted on 11 species in eight taxonomic groups (Magnoliopsida, Liliopsida, Clitellata, Entognatha, Entomobryomorpha, Chromadorea, Chlorophyceae, Trebouxiophyceae). Based on the toxicity values calculated, the PC₉₅ values for acute and chronic SSDs were 13 and 6 mg/kg dry soil, respectively. Toxicity varied among taxa, with soil algae emerging as the most sensitive to propylparaben. This may be attributable to differences in exposure pathways among species. The exposure pathway of propylparaben can be altered by adsorption to soil particles. As parabens are presently under-regulated globally in terms of their environmental effects, our findings can serve as the basis to propose standard values for environmental protection.

In the present study, we evaluate our previously developed non-destructive soil algal toxicity method using species from a different class of algae; Class Trebouxiophyceae (Chlorella vulgaris and Chlorella sorokiniana), and directly measure the photosynthetic activity of these species adsorbed onto the soil as a new toxicity endpoint. This study shows that non-destructive soil algal toxicity method is applicable to non-specific test species, including those of Class Trebouxiophyceae as well as Class Chlorophyceae (Chlorococcum infusionum and Chlamydomonas reinhardtii). Furthermore, by performing photosynthesis image analysis, we verify that it is possible to measure the photosynthetic activity of soil algae Chlorella vulgaris adsorbed onto soils without the need to extract algal cells from the soil. We propose that the non-destructive soil algal toxicity method represents a novel technique for 1) evaluating pollutants in soil using non-specific algae and 2) conveniently and rapidly assessing the photosynthetic activity of soil algae Chlorella vulgaris adsorbed onto soil as a new toxicity endpoint.

Trace elements (TEs) availability, biochemical activity and functional gene diversity was studied in a Cu-contaminated soil, revegetated for six years with a mixed stand of willow, black poplar, and false indigo-bush, and amended or not with compost plus dolomitic limestone (OMDL). The OMDL amendment significantly reduced Cu and As availability and soil toxicity, and increased the biochemical activity and microbial functional diversity assessed with the GEOCHIP technique, as compared to the unamended soil (Unt). The OMDL soil showed significantly higher abundance of 25 functional genes involved in decomposition organic compounds, and 11, 3 and 11 functional genes involved in the N, P and S biogeochemical cycles. Functional gene abundance was positively correlated with nutrient contents but negatively correlated with Cu availability and soil toxicity. The abundance of microbial functional genes encoding for resistance to various TEs also increased, possibly due to the microbial proliferation and lower Cu exposure in the presence of high total soil Cu concentration. Genes encoding for antibiotic resistance due to the co-occurrence of TEs and antibiotic resistant genes on genetic mobile elements. Overall, phytomanagement confirmed its potential to restore the biological fertility and diversity of a severely Cu-contaminated soil, but the increase of TEs and antibiotic resistant gene abundances deserve attention in future studies.

Soil algal bioassays have been limited by their inability to evaluate several toxic endpoints because it is difficult to collect pure soil algae growing on and beneath the soil surface. This study describes the extension of a previously developed paper-disc method for analyzing soil toxicity to algae. The method can be used in conjunction with flow cytometric analysis and facilitates the assessment of previously proposed toxicity endpoints, such as the growth zone, biomass, and photosynthetic activity. We assessed the applicability of this paper-disc soil method using the green algae Chlamydomonas reinhardtii and Pseudokirchneriella subcapitata exposed to nickel-contaminated soil; examined cell sizes, cell granularity, enzyme activity, and oxidative stress as new toxicity endpoints using flow cytometry; and identified morphological changes in green algae assayed. The results showed that, used in conjunction with flow cytometry, the extended paper-disc soil method is sufficiently sensitive to detect decreases in cell granularity in C. reinhardtii and esterase activity in P. subcapitata. The method also revealed decreases in growth zone, biomass, and electron transfer from the reaction center to the quinone pool. Collectively, the results of this study indicate that soil algal bioassays using nonspecific algae can be used to assess soil quality, to derive several toxicity endpoints for individual cells, and to evaluate previously established flow cytometric toxicity endpoints.

The probabilistic ecological risk assessment using terrestrial toxicity data has been mainly based on microfauna or mesofauna. Soil algae, which are food source for microfauna and mesofauna, may be alternatively used for assessing soil toxicity. However, there are no internationally recommended guidelines for soil algal bioassays, and the collection of algae from the test soils has some limitations. In this study, we suggested the paper-disc method as an easy-to-use alternative. This method has been widely used for testing the antibacterial toxicity of various chemicals in agar media by measuring the diameter of the inhibition zone around the disc. We adapted the paper-disc method for screening the toxicity of copper (Cu) and nickel (Ni) to the soil alga Chlorococcum infusionum using various evaluation endpoints, such as growth zone, chlorophyll fluorescence, and photosynthetic activity. Chlorophyll fluorescence and photosynthetic activity decreased with the increasing concentrations of Cu+2 or Ni+2 contaminated soils. Algal growth zone was analyzed visually and showed similar results to those of chlorophyll fluorescence. The direct ethanol extraction method and indirect culture medium extraction method were similarly effective; however, the former was easier to perform, while the latter might facilitate the analysis of additional endpoints in future studies. Overall, the results suggested that the paper-disc method was not only a user-friendly assay for screening soil toxicity, but also effective due to its association with indirect soil quality indicators.

Soils are the largest terrestrial sink for methane (CH4). However, heavy metals may exert toxicity to soil microorganisms, including methanotrophic bacteria. We tested the effect of lead (Pb), zinc (Zn) and nickel (Ni) on CH4 oxidation (1% v/v) and dehydrogenase activity, an index of the activity of the total soil microbial community in Mollic Gleysol soil in oxic and hypoxic conditions (oxia and hypoxia, 20% and 10% v/v O2, respectively). Metals were added in doses corresponding to the amounts permitted of Pb, Zn, Ni in agricultural soils (60, 120, 35 mg kg−1, respectively), and half and double of these doses. Relatively low metal contents and O2 status reflect the conditions of most agricultural soils of temperate regions. Methane consumption showed high tolerance to heavy metals. The effect of O2 status was stronger than that of metals. CH4 consumption was enhanced under hypoxia, where both the start and the completion of the control and contaminated treatment were faster than under oxic conditions. Dehydrogenase activity, showed higher sensitivity to the contamination (except for low Ni dose), with a stronger effect of heavy metals, than that of the O2 status.

A phenanthrene-degrading bacterium, Sphingomonas paucimobilis EPA505 was used to construct two fluorescence-based reporter strains. Strain D harboring gfp gene was constructed to generate green fluorescence when the strain started to biodegrade phenanthrene. Strain S possessing gef gene was designed to die once phenanthrene biodegradation was initiated and thus to lose green fluorescence when visualized by a live/dead cell staining. Confocal laser scanning microscopic observation followed by image analysis demonstrates that the fluorescence intensity generated by strain D increased and the intensity by strain S decreased linearly at the phenanthrene concentration of up to 200 mg/L. Such quantitative increase and decrease of fluorescence intensity in strain D (i.e., from 1 to 11.90 ± 0.72) and strain S (from 1 to 0.40 ± 0.07) were also evident in the presence of Ottawa sand spiked with the phenanthrene up to 1000 mg/kg. The potential use of the reporter strains in quantitatively determining biodegradable or toxic phenanthrene was discussed.