The application of biochar in remediation and recovery of heavy metals and/or organic contaminants in water and soil is increasing. However, the adverse effect of biochar to aquatic organisms has not received enough attention. In this study, we conducted a study on the biotoxicity of biochar pyrolyzed from pine needle under oxygen-limited conditions. The toxicity of biochar was expressed with the following endpoints: cell growth, chlorophyll-a (Chl-a), reactive oxygen species (ROS), superoxide dismutase (SOD) content of Scenedesmus obliquus (S. obliquus) and the luminescence of Photobacterium phosphoreum (P. phosphoreum). Here, the effect of free radicals (FRs) contained in biochar was stressed. Our results show that the toxicity of biochar is significantly correlated with the concentration of FRs in biochar particles. Meanwhile, we found the FRs-containing biochar could induce the production of acellular ROS (such as ·OH) in water, which would also induce the production of interior cellular ROS in aquatic organisms. Our findings provide a new insight into the mechanism of toxicity aroused by biochar applications and aid in understanding its potential ecological risk.

Aryl phosphorus-containing flame retardants (aryl-PFRs) have been frequently detected with increasingly used worldwide as one of alternatives for brominated flame retardants. However, information on their adverse effects on human health and ecosystem is insufficient, with limited study on their molecular mode of action in vitro. In this study, the cytotoxicity, DNA damage, mitochondrial impairment and the involved molecular mechanisms of certain frequently detectable aryl-PFRs, including 2-ethylhexyldiphenyl phosphate (EHDPP), methyl diphenyl phosphate (MDPP), bisphenol-A bis (diphenyl phosphate) (BDP), isodecyl diphenyl phosphate (IDPP), cresyl diphenyl phosphate (CDP) and the structurally similar and widely used organophosphorus pesticide chlorpyrifos (CPF), were evaluated in A549 cells using high-content screening (HCS) system. Aryl-PFRs showed different lethal concentration 50 (LC50) values ranging from 97.94 to 546.85 μM in A549 cells using CCK-8 assay. EHDPP, IDPP, CDP, MDPP and CPF demonstrated an ability to induce DNA damage, evidenced by increased DNA content and S phase-reducing cell cycle arrest effect using fluorophore dye cocktail assay. Additionally, the selected aryl-PFRs induced mitochondrial impairment by the increasing mitochondrial mass and decreasing mitochondrial membrane potential. Moreover, BDP, MDPP, and CDP, which contain short alkyl chains showed their potential oxidative stress with intracellular ROS and mitochondrial superoxide overproduction from an initially relatively low concentration. Additionally, based on the promotion of firefly luminescence in p53-transfected A549 cells, p53 activation was found to be involved in aryl-PFRs-induced DNA damage. Further real-time PCR results showed that all selected aryl-PFRs triggered p53/p21/gadd45β-, and p53/p21/mdm2-mediated cell cycle pathways, and the p53/bax mediated apoptosis pathway to induce DNA damage and cytotoxic effects. These results suggest that aryl-PFRs (e.g., BDP, MDPP, CDP) cause oxidative stress-mediated DNA damage and mitochondrial impairment, and p53-dependent pathway was involved in the aryl-PFRs-induced DNA damage and cell cycle arrest. In conclusion, this study improves the understanding of PFRs-induced adverse outcomes and the involved molecular mechanism.

A previous study has revealed that the parameter k∙ECₓ, characterizing the shape of concentration-response curves (CRCs), could predict the combined toxicity of binary mixtures. This study further explored the predictability of multi-component mixtures. Eleven component mixtures were designed using the uniform design ray, and the acute toxicity of the eleven environmental pollutants and their mixtures to Vibrio fischeri was determined using microplate toxicity analysis. We used independent action (IA) and the effect residual ratio (ERRₓ) models to evaluate the combined toxicity of multi-component mixtures and ascertain the functional relationship between σ²₍ₖ∙ECₓ₎, a parameter characterizing the CRC morphological difference of multi-component mixtures, and combined toxicity. The variance σ²₍ₖ∙ECₓ₎ of each component characteristic parameter of multi-component mixtures gradually increased in the concentration range, and the relationship between σ²₍ₖ∙ECₓ₎ and ERRₓ was consistent with the exponential function. The literature verification showed that this rule is generally applicable to the acute toxicity of multi-component mixtures to luminescent bacteria. The exponential function showed the variation rule of the joint action of multi-component mixtures. In the present study, the joint toxicity of multi-component mixtures can be predicted from single toxicity and small amount of multiple toxicity, circumventing complex multi-component toxicity experiments.

Co-pyrolysis of sewage sludge (SL) with plant biomass gains attention as a way to minimize SL-derived biochar drawbacks, such as high amount of toxic substances, low specific surface area and carbon content. The toxicity of soil amended with SL- (BCSL) or SL/biomass (BCSLW)-derived biochar was evaluated in long-term pot experiment (180 days). The results were compared to SL-amended soil. Biochars produced at 500, 600, or 700 °C were added to the soil (podzolic loamy sand) at a 2% (w/w) dose. Samples were collected at four different time points (at the beginning, after 30, 90 and 180 days) to assess the potential toxicity of SL-, BCSL- or BCSLW-amended soil. The bacteria Aliivibrio fischeri (luminescence inhibition – Microtox), the plant Lepidium sativum (root growth and germination inhibition test – Phytotoxkit F), and the invertebrate Folsomia candida (mortality and reproduction inhibition test – Collembolan test) were used as the test organisms. Depending on the organism tested and the sample collection time point variable results were observed. In general, SL-amended soil was more toxic than soil with biochars. The leachates from BCSLW-amended soil were more toxic to A. fischeri than leachate from BCSL-amended soil. A different tendency was observed in the case of phytotoxicity. Leachate from BCSL-amended soil was more toxic to L. sativum compared to BCSLW-amended soil. The effect of biochars on F. candida was very diversified, which did not allow a clear trend to be observed. The toxic effect of SL-, BCSL- or BCSW-amended soil to particular organisms was observed in different time, point's periods, which may suggest the different factors affecting this toxicity.

The study aimed to evaluate the ecotoxicity of soil (S) amended with biochars (BCKN) produced by the thermal conversion of sewage sludge (SSL) at temperatures of 500 °C, 600 °C, or 700 °C and SSL itself. The ecotoxicological tests were carried out on organisms representing various trophic levels (Lepidium sativum in plant, Folsomia candida in invertebrates, and Aliivibrio fischeri in bacteria). Moreover, the study evaluated the effects of three plants (Lolium perenne, Trifolium repens, and Arabidopsis thaliana) growing on BCKN700-amended soil on its ecotoxicological properties. The experiment was carried out for six months. In most tests, the conversion of sewage sludge into biochar caused a significant decrease in toxicity by adding it to the soil. The pyrolysis temperature directly determined this effect. The soil amended with the biochars produced at higher temperatures (600 °C and 700 °C) generally exhibited lower toxicity to the test organisms than the SSL. Because of aging, all the biochars lost their inhibition properties against the tested organisms in the solid-phase tests and had a stimulating influence on the reproductive ability of F. candida. With time, the fertilizing effect of the BCKN700 amended soil also increased. The aged biochars also did not have an inhibitory effect on A. fischeri luminescence in the leachate tests. The study has also demonstrated that the cultivation of an appropriate plant species may additionally reduce the toxicity of soil fertilized with biochar. The obtained results show that the conversion of sewage sludge to biochar carried out at an appropriate temperature can become a useful method in reducing the toxicity of the waste and while being safe for agricultural purposes.

Environment can be affected by a variety of micropollutants. In this paper, we develop a system to assess the toxicity on an environmental sample, based on the expression of a nanoluciferase under the control of the STB5 promotor in a yeast. The STB5 gene encodes for a transcription factor involved in a pleiotropic drug resistance and in the oxidative stress response. The response of the modified yeast was assessed using 42 micropollutants belonging to different families (antibiotics, pain killers, hormones, plasticizers, pesticides, etc.). Among them, 26 induced an increase of the bioluminescence for concentration ranges from pg.L⁻¹ to ng.L⁻¹. Surprisingly, for concentrations higher than 100 ng.L⁻¹, no response can be observed, suggesting that other mechanisms are involved when the stress increases. Analyzing the different responses obtained, we highlighted six nonmonotonic types of responses. The type of response seems to be independent of the properties of the compounds (polarity, toxicology, molecular weight) and of their family. In conclusion, we highlighted that a cellular response exists for very low exposition to environmental concentration of micropollutants and that it was necessary to explore the cellular mechanisms involved at very low concentration to provide a better risk assessment.

An experimental and in silico quantitative structure-activity relationship (QSAR) approach was applied to assess the environmental fate and effects of the antipsychotic drug Thioridazine (THI). The sunlight-driven attenuation of THI was simulated using a Xenon arc lamp. The photodegradation reached the complete primary elimination, whereas 97% of primary elimination and 11% of mineralization was achieved after 256 min of irradiation for the initial concentrations of 500 μg L−1 and 50 mg L−1, respectively. A non-target approach for the identification and monitoring of transformation products (TPs) was adopted. The structure of the TPs was further elucidated using liquid chromatography–high resolution mass spectrometry (LC–HRMS). The proposed photodegradation pathway included sulfoxidation, hydroxylation, dehydroxylation, and S- and N-dealkylation, taking into account direct and indirect photolysis through a self-sensitizing process in the higher concentration studied. The biodegradability of THI and photolytic samples of THI was tested according to OECD 301D and 301F, showing that THI and the mixture of TPs were not readily biodegradable. Furthermore, THI was shown to be highly toxic to environmental bacteria using a modified luminescent bacteria test with Vibrio fischeri. This bacteriotoxic activity of THI was significantly reduced by phototransformation and individual concentration-response analysis confirmed a lowered bacterial toxicity for the sulfoxidation products Thioridazine-2-sulfoxide and Thioridazine-5-sulfoxide. Additionally, the applied QSAR models predicted statistical and rule-based positive alerts of mutagenic activities for carbazole derivative TPs (TP 355 and TP 339) formed through sulfoxide elimination, which would require further confirmatory in vitro validation tests.

Heavy metal(loid) rich ash (≤10,000 mg kg−1 total As, Cr, Cu and Zn) originating from the combustion of contaminated wood was subjected to several experimental procedures involving its incorporation into an upland pasture soil. Ash was added to soil that had been prior amended with local cattle manure, replicating practices employed at the farm scale. Metal(loid) concentrations were measured in soil pore water and ryegrass grown on soil/manure plus ash mixtures (0.1–3.0% vol. ash) in a pot experiment; toxicity evaluation was performed on the same pore water samples by means of a bacterial luminescence biosensor assay. Thereafter a sequential extraction procedure was carried out on selected soil, manure and ash mixtures to elucidate the geochemical association of ash derived metal(loid)s with soil constituents. Predictive modelling was applied to selected data from the pot experiment to determine the risk of transfer of As to meat and milk products in cattle grazing pasture amended with ash.The inclusion of manure to soils receiving ash reduced phyto-toxicity and increased ryegrass biomass yields, compared to soil with ash, but without manure. Elevated As and Cu concentrations in pore water and ryegrass tissue resulting from ash additions were reduced furthest by the inclusion of manure due to an increase in their geochemical association with organic matter. Zinc was the only measured metal(loid) to remain uniformly soluble and bioavailable regardless of the addition of ash and manure. Risk modelling on pot experimental data highlighted that an ash addition of >1% (vol.) to this pasture soil could result in As concentrations in milk and meat products exceeding acceptable limits.The results of this study therefore suggest that even singular low doses of ash applied to soil increase the risk of leaching of metal(loid)s and intensify the risk of As transfer in the food chain.

The antipsychotic drug quetiapine (QUT) has been frequently detected in sewage treatment plants. However, information on the fate of QUT in aquatic environments and its behavior during UV treatment is limited. In this study, QUT is shown not to be readily biodegradable in the Closed Bottle Test and the Manometric Respirometry Test according to OECD guidelines. The main biotransformation product (BTP) formed in the tests, a carboxylic acid derivative, was identified by means of high-resolution mass spectrometry. This BTP is presumably a human metabolite and showed higher detection rates than QUT in a river sampling campaign conducted in northern Germany. UV elimination kinetics of QUT at different initial concentrations (226.5, 45.3, 11.3, and 2.3 μmol L−1) were faster at lower initial concentrations. All seven phototransformation products (PTPs) could be still identified at initial concentration of 11.3 μmol L−1. The photolytic mixture generated after 128 min of photolysis of QUT was not better biodegradable than QUT. Initial UV treatment of QUT led to the formation of several additional BTPs. Four of them were identified. The bacterial cytotoxicity and genotoxicity before and after phototransformation of QUT in a modified luminescent bacteria test (LBT) and the umu-test (ISO/FDIS 13829) showed cytotoxic effects in the LBT for QUT. Furthermore, PTPs had similar cytotoxic effects on luminescent bacteria. The umu-test did not reveal any genotoxic activity for QUT or PTPs. In conclusion, the release of QUT into sewage treatment plants and aquatic environments could result in the formation of a main BTP. Additional UV treatment of QUT would lead to the formation of additional BTPs. Moreover, treatment did not result in lower toxicity to tested organisms. In conclusion, UV treatment of QUT should be considered critically as a potential treatment for QUT in aquatic systems.

Amending contaminated soils with organic wastes can influence trace element mobility and toxicity. Soluble concentrations of metals and arsenic were measured in pore water and aqueous soil extracts following the amendment of a heavily contaminated mine soil with compost and biochar (10% v:v) in a pot experiment. Speciation modelling and toxicity assays (Vibrio fischeri luminescence inhibition and Lolium perenne germination) were performed to discriminate mechanisms controlling metal mobility and assess toxicity risk thereafter. Biochar reduced free metal concentrations furthest but dissolved organic carbon primarily controlled metal mobility after compost amendment. Individually, both amendments induced considerable solubilisation of arsenic to pore water (>2500 μg l−1) related to pH and soluble phosphate but combining amendments most effectively reduced toxicity due to simultaneous reductions in extractable metals and increases in soluble nutrients (P). Thus the measure–monitor-model approach taken determined that combining the amendments was most effective at mitigating attendant toxicity risk.