To Örebro University

As the volume of plastic waste from electrical and electronic equipment (WEEE) continues to rise, a significant portion is disposed of in the environment, with only a small fraction being recycled. Both disposal and recycling pose unknown health risks that require immediate attention. Existing knowledge of WEEE plastic toxicity is limited and mostly relies on epidemiological data and association studies, with few insights into the underlying toxicity mechanisms. Therefore, this study aimed to perform comprehensive chemical screening and mechanistic toxicological assessment of WEEE plastic-associated chemicals. Chemical analysis, utilizing suspect screening based on high-resolution mass spectrometry, along with quantitative target chemical analysis, unveiled numerous hazardous compounds including polyaromatic compounds, organophosphate flame retardants, phthalates, benzotriazoles, etc. Toxicity endpoints included perturbation of morphological phenotypes using the Cell Painting approach, inflammatory response, oxidative stress, and endocrine disruption. Results demonstrated that WEEE plastic chemicals altered the phenotypes of the cytoskeleton, endoplasmic reticulum, and mitochondria in a dose-dependent manner. In addition, WEEE chemicals induced inflammatory responses in resting macrophages and altered inflammatory responses in lipopolysaccharide-primed macrophages. Furthermore, WEEE chemicals activated the nuclear factor erythroid 2-related factor 2 (Nrf2) pathway, indicating oxidative stress, and the aryl hydrocarbon receptor (AhR). Endocrine disruption was also observed through the activation of estrogenic receptor-α (ER-α) and the induction of anti-androgenic activity. The findings show that WEEE plastic-associated chemicals exert effects in multiple subcellular sites, via different receptors and mechanisms. Thus, an integrated approach employing both chemical and toxicological methods is essential for comprehensive assessment of the toxicity mechanisms and cumulative chemical burden of WEEE plastic-associated chemicals.

Environmental context: The widespread use of the insecticide DDT has left a legacy of pollution that still threatens ecosystems today. This study presents a method to accurately measure the bioavailability of DDT and its breakdown products in contaminated soils. This will improve risk assessments and guide sustainable land management practices, helping to protect both the environment and human health.

Rationale: The insecticide dichlorodiphenyltrichloroethane (DDT) and its degradation products (collectively DDX) are persistent organic pollutants that pose significant environmental risks due to their persistence and bioaccumulation in ecosystems. Accurate quantification of DDX bioavailability in soil systems is crucial for effective land management and risk assessment.

Methodology: This study utilised equilibrium passive sampling with polyoxymethylene (POM) to determine the bioavailability of DDX in soil. The sorption dynamics of 10 DDX compounds were investigated (p,p′-DDT, o,p′-DDT, p,p′-dichlorodiphenyldichloroethane (p,p′-DDD), o,p′-DDD, p,p′-dichlorodiphenyldichloroethene (p,p′-DDE), o,p′-DDE, p,p′-dichlorodiphenylmethane (p,p′-DDM), p,p′-dichlorobenzophenone (p,p′-DBP), 1-chloro-2,2-bis(4-chlorophenyl)ethylene (p,p′-DDMU) and dicofol) and their POM–water partition coefficients (KPOM) were determined. The study involved interlaboratory comparisons, using soils from nine historically contaminated sites and ecotoxicology assessments (mortality, reproduction and bioaccumulation in earthworms, Eisenia fetida) to validate the POM method.

Results: KPOM values for 9 of the 10 DDX compounds were successfully determined, allowing for accurate quantification of freely dissolved pore water concentrations of DDX in historically contaminated soils. The interlaboratory study highlighted important considerations in extraction and gas chromatography–mass spectrometry analysis, and the ecotoxicology study demonstrated the potential of POM passive sampling as a reliable tool for assessing DDX bioavailability (bioaccumulation in Eisenia fetida).

Discussion: The POM method proved to be a robust and reliable approach for quantifying freely dissolved DDX, with implications for improving the accuracy of risk assessments and guiding sustainable land management practices. The study also highlighted the need for careful consideration of analytical challenges, such as the potential degradation of DDX compounds during gas chromatography analysis, to ensure accurate quantification.

Indoor dust contains a complex mixture of chemicals, including endocrine-disrupting chemicals (EDCs), which may pose risks to children's health. As children spend most of their time indoors and have frequent dust contact, their exposure is heightened. This study quantified the endocrine disrupting potential of dust from children's indoor environments in Sweden, and assessed associations with flame retardants and plasticizers in dust, handwipes, and urine.

Fifty dust samples from 18 homes and 11 preschool units were analyzed for estrogen, anti-androgen, and thyroid receptor activities using human osteosarcoma cell-based luciferase reporter assays. Associations were evaluated with 21 legacy and 18 emerging halogenated flame retardants (HFRs) and 11 organophosphate esters (OPEs) in dust and handwipes, as well as nine plasticizers (eight phthalates and di-isononyl cyclohexane 1,2-dicarboxylate (DiNCH)) in dust, and 14 plasticizer metabolites in urine. Samples for biological and chemical analyses were collected from the same designated areas within a limited time frame.

Most dust samples exhibited estrogen receptor agonist (ER) and androgen receptor antagonistic (anti-AR) activity, while thyroid receptor (TR) induction was low. Preschool dust showed significantly higher estrogenic activity than home dust. No seasonal variation was observed. Associations were observed between dust hormonal activities and urinary plasticizer metabolites, as well as HFR and OPE concentrations in dust and handwipes. Relative potency (REP) analyses of 36 HFRs and OPEs revealed notable anti-AR activity for 2,2´,4,4´-tetrabromodiphenyl ether (BDE-47) (REP values 0.85±0.10 (EC25) and 0.93±0.07 (EC50)) and 2,2´,4,4´,6-pentabromo diphenyl ether (BDE-100) (REP values 2.74±0.29 (EC25) and 3.23±0.42 (EC50)). Additionally, BDE-100 showed low ER induction.

Climate change is driving extreme weather patterns, leading to prolonged droughts and more frequent intense precipitation events. These environmental changes will impact aquatic systems by altering essential water quality parameters such as temperature, redox potential, pH, suspended solids and organic matter, which influence pollutant solubility and determine ecosystem health as well as drinking water production. In this context, sediments play a crucial role as they represent both a sink and source of pollutants. Therefore, sediment toxicity testing is essential for accurate environmental risk assessments. However, there remains a gap regarding comprehensive sediment testing ap-proaches that integrate multiple biomarker responses.

The SEASON project uses an interdisciplinary approach combining strategies of environmental toxicology, analytical chemistry, andhydro geochemistry. The aim is to develop conceptual models for evaluating, understanding and predicting the impact of climate change effects on the fate, bioavailability, and toxicity of pollutants in the aquatic environment. This project focuses on risks associated with sediments contaminated by organic and inorganic pollutants, specifically metals and PFAS (per- and polyfluoroalkyl substances). By studying factors such as temperature, pH, microbial communities, and sediment-water interactions, the project seeks to understand how different climate change aspects affect pollutant behavior in aquatic ecosystems.

The project consists of four sub-projects. Three investigate different chemical groups and mixtures under varying water conditions, using zebrafish (Danio rerio) as the main model organism. These studies will include in vitro and in vivo assays, microcosm experiments, microbiome studies and chemical analyses. The fourth subproject will integrate the results to develop a predictive model for sediment risk assessment.

Sediment contact assays will be performed to evaluate the effects of contaminated samples on zebrafish embryos by measuring teratogenicity, developmental toxicity, behavioral changes, and gene expression. In microcosm studies, we will vary pH and mimic increased precipitation events to assess pollutant toxicity in adult zebrafish including sex-related toxicity differences, reproduction, and behavior. Effect-directed analysis (EDA) will be used to identify key toxicants in the samples. Microbiome analysis using metagenomic sequencing will focus on how contaminated sediments alter bacterial communities, which in turn can affect pollutant distribution and bioavailability. Chemical analyses will quantify PFAS, metals, and their speciation throughout the study. Ultimately, the project will integrate these data to develop models that increase our understanding of the impact of climate change on sediment contamination and aquatic ecosystem health.

Polycyclic aromatic compounds (PACs) are prevalent in the environment, typically found in complex mixtures and high concentrations. Our understanding of the effects of PACs, excluding the 16 priority polycyclic aromatic hydrocarbons (16 PAHs), remains limited. Zebrafish embryos and in vitro bioassays were utilized to investigate the embryotoxic, behavioral, and molecular effects of a soil sample from a former gasworks site in Sweden. Additionally, targeted chemical analysis was conducted to analyze 87 PACs in the soil, fish, water, and plate material. CALUX® assays were used to assess the activation of aryl hydrocarbon and estrogen receptors, as well as the inhibition of the androgen receptor. Larval behavior was measured by analyzing activity during light and darkness and in response to mechanical stimulation. Furthermore, qPCR analyses were performed on a subset of 36 genes associated with specific adverse outcomes, and the total lipid content in the larvae was measured. Exposure to the sample resulted in embryotoxic effects (LC50 = 0.480 mg dry matter soil/mL water). The mixture also induced hyperactivity in darkness and hypoactivity in light and in response to the mechanical stimulus. qPCR analysis revealed differential regulation of 15 genes, including downregulation of opn1sw1 (eye pigmentation) and upregulation of fpgs (heart failure). The sample caused significant responses in three bioassays (ERα-, DR-, and PAH-CALUX), and the exposed larvae exhibited elevated lipid levels. Chemical analysis identified benzo[a]pyrene as the predominant compound in the soil and approximately half of the total PAC concentration was attributed to the 16 PAHs. This study highlights the value of combining in vitro and in vivo methods with chemical analysis to assess toxic mechanisms at specific targets and to elucidate the possible interactions between various pathways in an organism. It also enhances our understanding of the risks associated with environmental mixtures of PACs and their distribution during toxicity testing.

Healthy soils provide valuable ecosystem services (ES), but soil contamination can inhibit essential soil functions (SF) and pose risks to human health and the environment. A key advantage of using gentle remediation options (GRO) is the potential for multifunctionality: to both manage risks and improve soil functionality. In this study, an accessible, scientific method for soil health assessment directed towards practitioners and decision-makers in contaminated land management was developed and demonstrated for a field experiment at a DDX-contaminated tree nursery site in Sweden to evaluate the relative effects of GRO on soil health (i.e., the ‘current capacity’ to provide ES). For the set of relevant soil quality indicators (SQI) selected using a simplified logical sieve, GRO treatment was observed to have highly significant effects on many SQI according to statistical analysis due to the strong influence of biochar amendment on the sandy soil and positive effects of nitrogen-fixing leguminous plants. The SQI were grouped within five SF and the relative effects on soil health were evaluated compared to a reference state (experimental control) by calculating quantitative treated-SF indices. Multiple GRO treatments are shown to have statistically significant positive effects on many SF, including pollutant attenuation and degradation, water cycling and storage, nutrient cycling and provisioning, and soil structure and maintenance. The SF were in turn linked to soil-based ES to calculate treated-ES indices and an overall soil health index (SHI), which can provide simplified yet valuable information to decision-makers regarding the effectiveness of GRO. The experimental GRO treatment of the legume mix with biochar amendment and grass mix with biochar amendment are shown to result in statistically significant improvements to soil health, with overall SHI values of 141 % and 128 %, respectively, compared to the reference state of the grass mix without biochar (set to 100 %).

Soil contaminants may restrict soil functions. A promising soil remediation method is amendment with biochar, which has the potential to both adsorb contaminants and improve soil health. However, effects of biochar amendment on soil-plant nitrogen (N) dynamics and N cycling microbial guilds in contaminated soils are still poorly understood. Here, a metal- and polycyclic aromatic hydrocarbon (PAH) contaminated soil was amended with either biochar (0, 3, 6 % w/w) and/or peat (0, 1.5, 3 % w/w) in a full-factorial design and sown with perennial ryegrass in an outdoor field trial. After three months, N and the stable isotopic ratio δ15N was measured in soil, roots and leaves, along with microbial responses. Aboveground grass biomass decreased by 30 % and leaf N content by 20 % with biochar, while peat alone had no effect. Peat in particular, but also biochar, stimulated the abundance of microorganisms (measured as 16S rRNA gene copy number) and basal respiration. Microbial substrate utilization (MicroResp™) was altered differentially, as peat increased respiration of all carbon sources, while for biochar, respiration of carboxylic acids increased, sugars decreased, and was unaffected for amino acids. Biochar increased the abundance of ammonia oxidizing archaea, while peat stimulated ammonia oxidizing bacteria, Nitrobacter-type nitrite oxidizers and comB-type complete ammonia oxidizers. Biochar and peat also increased nitrous oxide reducing communities (nosZI and nosZII), while peat alone or combined with biochar also increased abundance of nirK-type denitrifiers. However, biochar and peat lowered leaf δ15N by 2-4 ‰, indicating that processes causing gaseous N losses, like denitrification and ammonia volatilization, were reduced compared to the untreated contaminated soil, probably an effect of biotic N immobilization. Overall, this study shows that in addition to contaminant stabilization, amendment with biochar and peat can increase N retention while improving microbial capacity to perform important soil functions.

Tire wear particles (TWP) stand out as a major contributor to microplastic pollution, yet their environmental impact remains inadequately understood. This study delves into the cocktail effects of TWP leachates, employing molecular, cellular, and organismal assessments on diverse biological models. Extracted in artificial seawater and analyzed for metals and organic compounds, TWP leachates revealed the presence of polyaromatic hydrocarbons and 4-tert-octylphenol. Exposure to TWP leachates (1.5 to 1000 mg peq L-1) inhibited algae growth and induced zebrafish embryotoxicity, pigment alterations, and behavioral changes. Cell painting uncovered pro-apoptotic changes, while mechanism-specific gene-reporter assays highlighted endocrine-disrupting potential, particularly antiandrogenic effects. Although heavy metals like zinc have been suggested as major players in TWP leachate toxicity, this study emphasizes water-leachable organic compounds as the primary causative agents of observed acute toxicity. The findings underscore the need to reduce TWP pollution in aquatic systems and enhance regulations governing highly toxic tire additives.

This study presents a comprehensive chemical and toxicological screening of chemicals extracted from WEEE (waste from electrical and electronic equipment) plastics. Chemical identification was conducted through suspect and target screening methods, revealing a diverse array of hazardous compounds including polycyclic aromatic compounds (PACs), organophosphate flame retardants (OPFRs), phthalates, benzotriazoles, and others. Toxicological endpoints included cell morphological phenotypes, inflammatory response, aryl hydrocarbon receptor (AhR) activation, activation of estrogenic receptor, and anti-androgenic activity. Results demonstrated that WEEE plastic chemicals significantly altered cell morphological phenotypes, particularly affecting the cytoskeleton, endoplasmic reticulum (ER), and mitochondrial measures. Moreover, WEEE chemicals induced inflammatory responses in resting human macrophages and altered ongoing inflammatory responses in lipopolysaccharide (LPS)-primed macrophages. Furthermore, WEEE chemicals exhibited potent AhR agonistic activity, activated estrogen receptor-α (ERα), and inhibited androgen receptor (AR) activation. The findings suggest that WEEE plastic chemicals exert their effects through multiple modes of action, targeting various subcellular sites. Thus, a combined approach utilizing non-target and target screening tools is essential for comprehensively assessing the toxic effects and health hazards associated with WEEE plastic chemicals.

The Arctic is undergoing rapid changes, and biota are exposed to multiple stressors, including pollution and climate change. Still, little is known about their joint impact. Here, we investigated the cumulative impact of crude oil, warming, and freshening on the copepod species Calanus glacialis and Calanus finmarchicus. Adult females were exposed to ambient conditions (control; 0 °C + 33 psu) and combined warming and freshening: 5 °C + 27 psu (Scenario 1), 5 °C + 20 psu (Scenario 2) for 6 days. All three conditions were tested with and without dispersed crude oil. In Scenario 1, fecal pellet production (FPP) significantly increased by 40-78% and 42-122% for C. glacialis and C. finmarchicus, respectively. In Scenario 2, FPP decreased by 6-57% for C. glacialis, while it fluctuated for C. finmarchicus. For both species, oil had the strongest effect on FPP, leading to a 68-83% reduction. This overshadowed the differences between climatic scenarios. All variables (temperature, salinity, and oil) had significant single effects and several joint effects on FPP. Our results demonstrate that Arctic copepods are sensitive to environmentally realistic concentrations of crude oil and climate change. Strong reductions in feeding can reduce the copepods' energy content with potential large-scale impacts on the Arctic marine food web.