PloS oneSusanne Vogeler, Matthew O Parker, Samuel C Robson, Alex T Ford
Behavioural endpoints offer a sensitive, non-invasive alternative to traditional mortality assays. Yet, they remain underutilized in regulatory frameworks due to a lack of standardised methodologies and baseline behavioural data. This study investigates the baseline locomotion of the marine amphipod Marinogammarus marinus to external light triggers to support standardised laboratory behavioural assays. Using an automated observation chamber, here we tracked swimming behaviour in response to light stimuli, quantifying locomotion as distance travelled. Groups of eight amphipods were tested in large crystalizing dishes over four days in an 8-minute trial involving two light exposures. A total of 60 individuals - males, females, and brooding females - were assessed. Light consistently triggered activity, with males showing strong negative phototaxis and brooding females the weakest. A startle response was observed within 3-5 seconds of initial light exposure, though responses diminished with repetition, suggesting potential overstimulation or habituation. Inter-individual and day-to-day variability highlighted the need for repeated measures. Additional trials with 32 males evaluated the effects of experimental conditions. Wall-hugging (thigmotaxis) appeared to be an escape response, not anxiety-related. Notable decreases in dissolved oxygen and pH within 24 hours were attributed to the amphipods' presence rather than activity, while salinity fluctuations due to evaporation did not visibly affect behaviour. Differences between male groups from the two separate experiments suggest behavioural shifts due to seasonal variation. This study establishes baseline behavioural data for M. marinus and demonstrates the value of automated tracking systems in behavioural assays in laboratory settings. It emphasizes the need to account for sex, size, and temporal variation in assay design and supports integrating behavioural endpoints into regulatory protocols.
Ecotoxicology (London, England)Nágela Gardênia Rodrigues Santos, Késsia Fernanda Brito Pinho, Itala de Cassia Sousa Reis, Ricardo Luvizotto-Santos, Marianna Basso Jorge
As a pervasive form of marine pollution, microplastics (MPs) are widely distributed across coastal and oceanic systems, raising concerns about their ecological impacts. This review provides an evidence-based synthesis of microplastic-microalgae interactions, integrating physiological responses and associated toxicity pathways. Marine microalgae are particularly relevant in this context because they constitute the foundation of marine food webs and play a central role in primary productivity and global biogeochemical cycling. This study synthesizes experimental ecotoxicological evidence on the effects of microplastics on marine microalgae, with the aim of identifying dominant toxicity endpoints and key knowledge gaps relevant to ecotoxicological evaluation. Publications indexed in Web of Science and Scopus from 1970 to August 2025 were systematically screened, resulting in 59 selected publications, of which 43 experimental studies met the criteria for the comparative ecotoxicological synthesis. The analysis revealed a rapid expansion of research on microplastic-microalgae interactions, with experimental studies predominantly investigating polystyrene and polyethylene, across a broad range of particle sizes. The most frequently investigated biological responses included growth/population parameters, cellular stress and damage indicators, and photosynthetic responses. Despite this progress, several ecologically relevant endpoints and processes remain insufficiently investigated, including aggregate formation, particle sinking dynamics, DNA damage, and long-term exposure under environmentally realistic conditions. Furthermore, most experimental studies rely on short-term laboratory assays using simplified exposure scenarios, which may limit the ecological relevance of current knowledge. Taken together, the available evidence highlights key research priorities and outlines directions for improving experimental ecotoxicological studies.
Pharmacoepidemiology and drug safetyHeidi Taipale, Jari Tiihonen, Antti Tanskanen, Jonas Forsman
PURPOSE: To present updated estimates from a nationwide Swedish study comparing dispensed medications with forensic toxicological findings (2006-2013), following re-evaluation of the toxicology dataset to address periods with non-uniform analytical detection and inconsistent testing. METHODS: Observations from time periods with non-uniform detection or inconsistent testing were excluded. Analyses were repeated using the PRE2DUP method to model drug exposure. Agreement between PRE2DUP-modelled exposure and post-mortem toxicology was assessed using sensitivity, specificity, predicted adherence, and Cohen's κ. RESULTS: After exclusion of inconsistent testing periods, the number of individuals for affected substances was reduced (10 060 vs. 18 627). Proportional changes in sensitivity and specificity were negligible. In contrast, predicted adherence and Cohen's κ improved across several substances. Moderate to substantial proportional increases were observed for selected medications but translated into only small group-level effects (approximately 2%-7%). The largest improvements were observed for cardiovascular drugs, beta-blockers, for which Cohen's κ and predicted adherence increased substantially (approximately 40%-55%) at the class level; notably, only for these substances did Cohen's κ shift interpretation (from fair to moderate agreement). Overall, proportional improvements across all substances were moderate (approximately 10%-15%). Despite these changes, overall agreement patterns remained comparable to the original results. CONCLUSIONS: After exclusion of periods with non-uniform or incomplete toxicological testing, agreement between PRE2DUP-modelled exposure and forensic toxicology improved overall. Moderate increases observed for a few individual medications translated into only small effects at the group level, except for cardiovascular drugs, for which improvements were substantial. These updates do not materially alter the original interpretation but strengthen the validity of register-based drug exposure modelling.
Journal of analytical toxicologyFranck Saint-Marcoux, Nathan Campos Lapa, Sylvain Dulaurent, Pauline Griffeuille, Souleiman El Balkhi
While oral fluid (OF) is increasingly used in forensic and roadside toxicology, measured drug concentrations remain highly dependent on pre-analytical conditions. This study investigated the impact of buccal-cell content and sample fractionation on concentrations of delta 9-tetrahydrocannabinol (THC), cocaine, benzoylecgonine (BZE), and ecgonine methyl ester (EME). Oral-fluid specimens were collected using FLOQSwabs®. Following elution, sonication, and vortex mixing, various fractions (homogenate, pellet, and expressed swab) were analyzed via liquid chromatography-tandem mass spectrometry. For THC (n = 102), median buccal epithelial cell counts and THC concentrations were significantly higher in the cell-rich pellet than in the homogenate (P < .001), with median enrichment ratios of 4.40 and 3.93, respectively. The association between cellular and THC enrichment was strong to very strong (r = 0.77-0.88) for concentrations above 5 ng/mL. Notably, four specimens were below the limit of detection in the homogenate but detectable in the pellet. In contrast, cocaine and its metabolites showed comparable concentrations between the homogenate and pellet, though the expressed swab fraction consistently yielded the highest levels. In 52 specimens, median expressed swab-to-homogenate ratios were 1.16 for cocaine, 1.28 for BZE, and 1.14 for EME. These findings demonstrate that pre-analytical effects are analyte-dependent: buccal cells act as a reservoir for THC, whereas cocaine-related analytes are primarily affected by incomplete release from the swab. These results identify analyte-dependent pre-analytical sources of variability that may affect oral-fluid concentration measurements obtained with this collection device.
ToxicologyJose L Domingo, Marilia Cristina Oliveira Souza, Fernando Barbosa
Human populations and ecosystems are continuously exposed to complex mixtures of environmental contaminants rather than to individual chemicals in isolation. These mixtures include pesticides, metals and metalloids, persistent organic pollutants, endocrine-disrupting chemicals, per- and polyfluoroalkyl substances, pharmaceuticals, plastic-associated compounds, air pollutants, nanomaterials, and numerous poorly characterized substances. Their combined effects may be additive, synergistic, or antagonistic, and are strongly influenced by dose, component ratio, timing, exposure sequence, and biological susceptibility. Experimental evaluation of all environmentally relevant mixtures is infeasible because the number of possible combinations increases combinatorially. Artificial intelligence (AI) offers a possible way to address this limitation. Machine learning, deep learning, graph neural networks, Bayesian approaches, and natural-language processing can integrate heterogeneous data, including chemical structures, molecular descriptors, toxicokinetics, high-throughput screening results, omics profiles, adverse outcome pathways, biomonitoring data, and epidemiological findings. However, the evidence base is uneven. Relatively few studies have applied AI directly to experimentally characterized mixtures, and much of the current optimism is extrapolated from single-chemical toxicology. This review distinguishes explicitly between applications demonstrated in mixtures, proof-of-concept mixture applications, and approaches whose mixture use remains prospective. It further examines the methodological requirements for mixture prediction, including dose and ratio representation, additivity reference models, applicability domains, external validation, and mechanistic interpretability, and proposes a framework for regulatory-grade implementation. Current evidence does not support autonomous AI-driven regulation of mixtures. AI should complement, not replace, experimental and expert evaluation, supporting a transition toward more predictive, mechanism-informed assessment of real-world chemical exposures.
Journal of forensic and legal medicineTommaso D'Anna, Antonina Argo, Beatrice Belmonte, Augusto Orlandi, Stefania Zerbo, Ginevra Malta
BACKGROUND: Attribution of death to hydrogen sulfide (H2S) is challenging because autopsy findings are often nonspecific, postmortem toxicology is vulnerable to analytical and decomposition-related uncertainty, and scene investigation varies in completeness. A systematic appraisal of how these domains support or weaken published causal conclusions is needed. METHODS: This protocol specifies a systematic review with structured narrative synthesis of peer-reviewed human fatality reports in which H2S exposure is suspected or attributed, together with relevant human postmortem comparator studies. MEDLINE via PubMed, Embase, Scopus, Web of Science, and the Cochrane Library will be searched for studies published from 1 January 2007 onward. English-language eligibility will be assessed during screening. Two reviewers will independently select studies, extract prespecified case-level data, and undertake design-specific JBI critical appraisal without total scores. A rule-based matrix will distinguish interpretable exposure evidence, nonspecific compatibility, unresolved discordance, and missing information. Original authors' conclusions will be recorded separately from reviewers' assessments. Recurrence will be summarized descriptively; discriminatory claims will require appropriate within-study comparators and an independently established attribution basis. Formal GRADE ratings and diagnostic-accuracy pooling are not planned. DISCUSSION: The completed review will examine the documented basis and limitations of forensic attribution, including cases with absent or inconclusive toxicology. The proposed convergence profiles are descriptive constructs, not validated diagnostic thresholds or probabilities of causation. Study and case counts, the PRISMA 2020 flow diagram, and synthesis findings will be reported in the completed review; this protocol contains no review results. REVIEW REGISTRATION: PROSPERO CRD420261324108.
Weapon detonations generate substantial quantities of post-blast residue (PBR), which contributes iron-rich particles to soils. After deposition, these residues undergo redox transformations that produce bioavailable iron compounds, potentially increasing phytotoxic risks to agricultural crops. In the present study, soil contamination by detonation of military weapons was simulated by adding 1% of < 1 mm PBR to soil, and iron mobilization under waterlogged conditions was subsequently monitored. Scanning electron microscopy revealed two components in PBR: iron-bearing spherules and metallic fragments composed of an iron core with an oxidized surface layer. Magnetic analyses confirmed that waterlogging had a limited impact on PBR, with no significant changes in explosion-derived iron spherules; however, a decrease was observed in hematite-like phases associated with surface-corroded iron fragments. This decrease was linked to the interaction of these phases with metabolically active soil microorganisms. Following the addition of a carbon and energy source (potato) to PBR-contaminated soil, the abiotic reaction of Fe0 oxidation to Fe2+ significantly accelerated due to microbial synthesis of organic acids and protons (H+) and iron mobilization by microbial activity. This markedly affected the soft magnetic material (fine pedogenic magnetite and iron core of PBR fragments) as well as highly coercive phases (oxidized coating of iron fragments and lithogenic hematite). Phytotoxic effects were evaluated by assessing seed germination, seedling, and root growth in representative species planted during the period of elevated soluble iron concentrations. Increased iron accumulation in stressed seedlings was a key indicator of PBR toxicity. These laboratory-simulation results improve our understanding of the potential environmental transformation of war-related iron residues and their implications for soil sustainability, which require validation under field conditions.
Per- and polyfluoroalkyl substances (PFAS) are persistent environmental pollutants with bioaccumulation potential, but their associations with kidney diseases remain incompletely understood. This study integrated Mendelian randomization and computational toxicology to examine associations between genetically predicted circulating PFAS levels and kidney disease outcomes and to prioritize candidate toxicogenomic pathway themes. Genetically predicted higher PFOA levels were inversely associated with IgA nephropathy (OR = 0.21, P = 0.004), but positively associated with hypertensive nephropathy (OR = 1.20, P < 0.001) and calculus of kidney (OR = 1.24, P = 0.016). Genetically predicted higher PFOS levels were inversely associated with IgA nephropathy (OR = 0.27, P = 0.046) and urinary tract infection (OR = 0.94, P = 0.003). A primary association was also observed between PFOA and membranous nephropathy (OR = 1.56, P = 0.028), but this association was not retained after targeted SNP-exclusion analyses and was therefore not interpreted as a robust or established causal association. Computational toxicology analyses prioritized database-derived candidate targets and pathway themes related to immune response, inflammation, oxidative stress, and apoptosis. Network-prioritized candidate nodes included CTNNB1, TP53, and EGFR in the PFOA-calculus of kidney network, IGF1 in the PFOA-hypertensive nephropathy network, and CCL2, TLR4, MMP9, and IFNG in the PFOA/PFOS-IgA nephropathy networks. In contrast, IL1B, TNF, and IL6 were observed as shared inflammatory nodes across multiple nephropathy-related networks. These candidates should be interpreted as database-derived and network-prioritized targets rather than experimentally validated causal mediators of kidney disease. Overall, this study provides a hypothesis-generating framework for exploring associations among genetically predicted PFAS-related traits, kidney disease outcomes, and candidate toxicogenomic pathway themes that require experimental validation.
International journal of molecular sciencesKyu-Shik Lee, Yeong Chae Kim, Hye-Ran Kim, Jongwan Kim
Micro- and nanoplastics (MNPs) are pervasive environmental contaminants that pose significant threats to ecosystems and the health of humans and other organisms. Increasing evidence indicates that MNP exposure can induce various biological disturbances, including cytotoxicity, chronic inflammation, endocrine disruption, oxidative stress, metabolic dysfunction, and cellular impairment. Many of these processes are closely associated with cancer initiation and progression. However, the molecular mechanisms underlying the relationship between MNP exposure and carcinogenesis remain unclear. This review summarizes the current evidence regarding MNP-associated alterations in gene expression and discusses their potential implications in cancer development and progression. We highlight the toxicogenomic insights derived from the Comparative Toxicogenomics Database (CTD), focusing on key microplastics, including polyethylene, polyethylene terephthalate, polystyrene, and polyvinyl chloride. Specifically, we discuss the chemical-gene interactions, disease associations, gene ontology annotations, pathway enrichment profiles, and chemical similarity networks linked to these polymers. Overall, the available toxicogenomic evidence implies that MNP exposure is associated with biological processes involved in oxidative stress responses, inflammatory signaling, immune dysregulation, metabolic alterations, and cell cycle control, all of which are implicated in carcinogenesis. Finally, we discuss the strengths and limitations of CTD-based toxicogenomic approaches and propose research directions to better understand the potential contribution of MNP exposure to cancer risk.
Anais da Academia Brasileira de CienciasVictória Alice D Dias, Arthur A Ochiutto, Thainá M Nehme, João Vítor B Calvelli, Antonio R DA Cunha Neto, Erika S Silva, Raquel S Jacob, Lucilaine V S Santos, …
Improper disposal of such waste represents a serious threat to the environment due to its potential to contaminate soil, rivers, and oceans, thereby endangering biota and public health. This study aimed to evaluate the ecotoxicological effects of raw and treated leachate, as well as surface water and groundwater adjacent to a Class II landfill, on germination and early growth parameters of Allium cepa L. and the viability or survival of Aliivibrio fischeri. The tested sample concentrations were 25, 50, 75, and 100%, each diluted in distilled water. In the phytotoxicity assays, A. cepa seeds were exposed to different sample concentrations, with distilled water serving as a negative control. The analyzed parameters included germination percentage, germination speed index, shoot length, root elongation, mitotic index, and chromosomal abnormalities. In the acute toxicity tests, the effect on bacterial luminescence was evaluated. Surface water and groundwater samples did not induce toxicity in A. cepa seeds, whereas raw and treated leachate samples inhibited germination and consequently negatively affected all other evaluated parameters. In A. fischeri assays, leachate samples exhibited toxicity, while surface water and groundwater samples induced a hormesis effect. The results from adjacent water samples demonstrate the landfill's efficiency in mitigating contamination of water bodies.
Environmental monitoring and assessmentYuying Zhang, Weiqin Xing, James A Ippolito, Yale Wang, Yongqiang Yang, Xinjun Huang, Liping Li
Soil heavy metal contamination poses persistent ecological risks to terrestrial ecosystems. Woodlice (Oniscidea), also known as terrestrial isopods, are important soil decomposers with strong capacities for heavy metal accumulation and detoxification. This review synthesizes current knowledge regarding heavy metal accumulation, distribution, physiological effects and ecological implications in woodlice. Heavy metals are predominantly accumulated in the hepatopancreas in woodlice, the principal organ for metal sequestration and detoxification. Copper generally exhibits higher bioconcentration factors (BCFs) than other metals because of its strong association with thiol-rich ligands in hepatopancreatic cells. Based on existing investigations, in contaminated areas, mean woodlouse Cd, Cu, Pb, and Zn concentrations were 42.7, 295, 150, and 472 mg kg-1, respectively; woodlouse BCFs for soil Cd, Cu, Pb, and Zn were 4.72, 16.4, 2.39, and 2.38, respectively, while its BCFs for plant Cd, Cu, Pb, and Zn were 9.19, 14.1, 1.29, and 4.54, respectively. Heavy metal accumulation is influenced by metal type, concentration, and exposure duration, and body concentrations generally increase with exposure time until a steady-state equilibrium is reached. Elevated heavy metal exposure can impair feeding, reduce growth, induce hepatopancreatic histopathological damage, and increase mortality, potentially leading to reduced population density and simplified community structure under severe contamination. Although woodlice show potential as bioindicators of soil heavy metal contamination, nonlinear relationships between environmental and body metal concentrations limit their quantitative applicability. Current understanding of trophic transfer of heavy metals from woodlice to higher trophic levels remains limited and warrants further investigation.
Ecotoxicology and environmental safetyYujian Fan, Xiaokai Guo, Jiaying Lyu, Xinyu Wang, Yisheng He
Drug safety science has expanded beyond patient-level pharmacovigilance to address the environmental consequences of pharmaceutical use. Pharmaceuticals are now detected across diverse ecosystems, but traditional single-compound, high-dose toxicological frameworks remain inadequate for evaluating chronic, low-dose, multi-compound environmental exposures. This review provides a mechanistic analysis of pharmaceutical ecotoxicity, tracing how molecular interactions with conserved biological targets in non-target organisms translate into population-level ecological effects. Synthetic estrogens activate nuclear hormone receptors at nanogram-per-liter concentrations, driving reproductive failure through receptor-mediated transcriptional reprogramming. Psychoactive drugs disrupt neurotransmitter systems conserved across vertebrates, altering predator avoidance and reproductive behavior at sub-microgram-per-liter levels. Anti-inflammatory drugs cause species-specific toxicity through differential phase II metabolism, as the diclofenac-vulture crisis demonstrates. Anticancer agents produce genotoxic effects in aquatic organisms through the same DNA-damaging mechanisms underlying their therapeutic activity. A unifying theme emerges when these endpoints are considered alongside antimicrobial resistance: sub-inhibitory antibiotic concentrations in environmental hotspots select for resistant bacteria and accelerate horizontal gene transfer, with resistance genes returning to human pathogens through water, food, and occupational exposure, completing a circular threat linking environmental contamination to clinical treatment failure. Mitigation strategies are evaluated across the pharmaceutical lifecycle, including biodegradable molecular design, manufacturing discharge controls, antimicrobial stewardship, and advanced wastewater treatment. The analysis demonstrates that effective intervention requires targeting root causes across the pharmaceutical lifecycle rather than relying on end-of-pipe remediation, and that integrating environmental sustainability into pharmaceutical safety assessment is essential for protecting both ecological and human health.
Molecules (Basel, Switzerland)Gabriella Kanižai Šarić, Marija Paurević, Andrea Dandić, Martina Šrajer Gajdošik, Vesna Rastija
Imidazoles have been proven to be very effective pesticides, especially against phytopathogenic fungi and insects. Due to their negative effects on the environment, only a few imidazoles have been approved for use by the European Commission (EC). There is an urgent need to develop new imidazole derivatives with high efficiency and a wide spectrum of action against numerous pests that are, at the same time, safe for the environment and beneficial for organisms and humans. In order to reduce expensive and time-consuming experiments, an in silico approach based on quantitative structure-activity relationship (QSAR) models is valuable for predicting the toxicity of new or untested chemicals. In this study, we used the Vega and ChemFREE web platforms to evaluate the pesticide similarity, environmental risk properties, and ecotoxicological effects of imidazole derivatives designed for potential synthesis. Adamantane-, alkyl-, and triazole-amide, ester, carbamate, and ketone derivatives were filtered for the evaluated properties, and four alkyl-amides were highlighted as potentially effective and environmentally safe antifungal, herbicidal, and insecticidal agents. Molecular docking studies indicated the possible mechanism of action of the antifungal, herbicidal, insecticidal, and antibacterial activities of the observed compounds and revealed structural features important for binding to specific receptors.
Scientific reportsMadusanka H K S, Dinupamali H A, Jayasuriya J A M S, Aruggoda A G B, Chathurika J A S, Weerakoon S R
Plant-mediated 'green' synthesis of nanoparticles (NPs) is widely reported, but the exact functional role of the retained phytochemical capping layer versus the core metal remains contested. Furthermore, the impact of thermal calcination-a common post-synthesis purification step-on the bio-functional and ecological profile of these NPs is poorly understood. We synthesized four distinct biogenic NPs-Ag and Fe using Salvinia molesta extract, and Cu and Zn using Mimosa pigra extract. While the Cu, Zn, and Fe NPs were evaluated in both non-calcined (as-synthesized) and thermally calcined states, the Ag NPs were evaluated exclusively in their highly active, non-calcined state. We evaluated their physicochemical properties, in vitro antioxidant capacity (with Ag NPs showing 42.08 mg TE/g), and antibacterial efficacy against Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa. Additionally, the ecotoxicological impact was evaluated via a one-month soil microbial respiration assay for the calcined metal oxides (Cu, Zn, Fe) and the non-calcined Ag NPs. Characterization confirmed that calcination successfully formed highly crystalline metal oxides but stripped the Cu, Zn, and Fe NPs of their organic phytochemical corona. Consequently, the non-calcined NPs exhibited significant antioxidant activity, which was substantially abolished in the metal oxides following calcination. Antibacterial assays revealed a strict metal-dependency; Fe, Cu, and Zn NPs showed no significant antibacterial action even at high screening concentrations, regardless of calcination. In contrast, the non-calcined Ag NPs exhibited potent antimicrobial efficacy, with a minimum inhibitory concentration (MIC) of 7.8 ppm. Crucially, 4-week soil respiration assays (at doses up to 1000 ppm) demonstrated that neither the calcined metal-oxides (Fe, Cu, Zn) nor the highly reactive non-calcined Ag NPs exerted long-term toxic effects on the soil microbiome. Our findings demonstrate that the 'green' bioactivity (antioxidant potential) of biogenic NPs is primarily mediated by the uncalcined phytochemical corona, whereas cytotoxicity (antibacterial action) is governed by the core metal identity. The absence of significant suppression of CO₂ respiration suggests minimal acute metabolic disruption in soil microbiomes, providing preliminary evidence for short-term microbial tolerance. These findings warrant further investigation into the potential agricultural applications of these biogenic nanomaterials.
Journal of analytical toxicologyHelen H Chang, Kacey Cliburn, Jesse Kemp, Dani C Mata
Fentanyl prevalence has increased exponentially in all forensic toxicology case types since the start of the current epidemic in 2016. Prior to 2016, fentanyl detected in postmortem cases was typically seen from use or misuse of medicinal administration. However, with the current epidemic, fentanyl use is from the illicit drug market and concentrations are substantially higher complicating interpretation. Additionally, factors contributing to postmortem redistribution of fentanyl can play a large role in interpretation. In combination with peripheral blood (PPH), brain concentrations can aid interpretation due to the isolated and protected location of the brain. A total of 566 cases were evaluated, and after excluding four outliers, the 562 cases had an overall fentanyl PPH mean of 18 ng/mL, median of 10 ng/mL, and range of 0.53-555 ng/mL. Overall mean, median, and range of fentanyl concentrations in brain samples were 61, 34, and 0.73-3100, respectively. Three concentration ranges, low (≤10 ng/mL), medium (>10-50 ng/mL), and high (>50 ng/mL), derived from PPH were established to evaluate cases based on various concentrations. The median blood-to-brain ratio doubled between the low and high bins. There was a greater percentage of females in the high concentration bin than that in the low and medium bins. The younger age group, aged 15 to 24 years, showed a higher prevalence of fentanyl in the medium group than in the others. The blood: brain ratio for all cases had a mean of 0.36, a median of 0.29, and a range of 0.03-2.57. The wide range of blood: brain ratios indicated that using singular information to interpret the case can be misleading. However, it does show the benefits of testing brain samples in suspected overdose cases where the fentanyl blood concentrations are low.
Journal of analytical toxicologySabrina Amundarain, Zachary Currie, Karen Woodall
Driving under the influence of cannabis is a significant public safety issue. Tetrahydrocannabinol (THC), the main psychoactive compound in cannabis, can impair driving ability and when used in combination with alcohol, the level of impairment can increase. Case studies documenting blood concentrations of THC and associated observations of impairment in individuals suspected of impaired driving continue to provide important contributions to the scientific literature. Retrospective analysis was performed using data from impaired driving investigations in which THC alone, and in combination with alcohol, were the only findings in blood samples submitted to the Centre of Forensic Sciences between 2021 and 2023, inclusive. In cases where only THC was present, blood concentrations ranged from 1.1 to 72 ng/mL (mean 12 ng/mL, median 9.5 ng/mL). When both THC and alcohol were present, the mean blood THC concentration was 13 ng/mL, and the mean blood alcohol concentration was 145 mg/100 mL. The most common driving and drug recognition expert (DRE) observations included collisions, erratic driving, eyelid tremors, poor balance and coordination, and bloodshot eyes. Observations of driving impairment were more frequently observed in cases where both THC and alcohol were detected in the blood. These findings provide additional data to inform the interpretation of blood THC concentrations in the context of DRE observations and suspected impaired driving investigations.
Microplastics (MPs) inevitably undergo aging processes in the environment, which alter their physicochemical properties and may modify their ecological effects. To synthesize the current evidence on aged-MP ecotoxicity, we conducted a meta-analysis of 54 studies comprising 1400 observations across animals, microalgae, and plants. Compared with pristine MPs, aged MPs were generally associated with reduced locomotion and neurological performance in aquatic and terrestrial animals, suppressed microalgal growth, and apparent stimulation of some plant growth-related end points. These divergent responses were associated with aging-related descriptors, particularly the carbonyl index (CI) and fouling index (FI), although their relevance varied across taxa, end points, polymers, and exposure contexts. The CI was negatively associated with animal locomotor responses, while the FI was more closely related to physiology-related responses in the primary producers. Targeted validation experiments using polystyrene (PS) and polyvinyl chloride (PVC) further showed that an increased CI was accompanied by reduced zebrafish locomotion and elevated ROS levels, whereas FI-associated algal responses were accompanied by oxidative stress. Overall, the CI and FI can help compare aged-MP effects across studies and support more generalized and more realistic assessments of aged-MP ecotoxicity. However, these descriptors remain simplified indicators and cannot fully capture the complexity of actual environmental systems, including mixed aging processes, leachate chemistry, multistressor exposure, and matrix-specific interactions.
Chemistry & biodiversityAgenor P Luz-Filho, Thais C Silva, Matheus B Silva, Abraão P Sousa, Helivaldo D S Souza, Priscila S V Lima, Alexandre Almeida-Júnior, Fábio C Sampaio, Petrônio…
Motivated by the promising antimicrobial and ecotoxicological effects of 4-alkylamino-7-chloroquinoline and N-alkylphthalimide derivatives, these privileged nuclei were linked to investigate how lipophilicity influences antibacterial activity and ecotoxicity. For this purpose, seven 4-alkylamino-7-chloroquinolines (2a-g) were prepared, of which four were used as precursors (2a-d) and reacted with different anhydrides. Using a clean synthetic approach, 4-alkylamino-7-chloroquinoline-succinimides (3a-d) and -phthalimides (5a-d) were obtained in yields of 60%-99% after only filtration with distilled water. In vitro assays were performed to determine the minimum inhibitory/bactericidal concentration (MIC/MBC) against Klebsiella pneumoniae, Pseudomonas aeruginosa, Staphylococcus aureus, and Streptococcus mutans. Lipophilicity (log P) and toxicity were predicted using the pkCSM program. In vitro toxicity tests on Artemia salina larvae were used as an indicator of ecotoxicity. 4-Alkylamino-7-chloroquinolines (2b, 2f, 2g) revealed a moderate MIC of 156.25 µg·mL-1 against Gram-(-) and Gram-(+) bacteria, whose predicted toxicity does not pose a potential risk to human health. Results indicate a correlation between increased lipophilicity and higher ecotoxicity, where less lipophilic compounds were less toxic to A. salina (LC50 133.01-777.50 µg·mL-1) than 4-alkylamino-7-chloroquinoline-phthalimides, suggesting further investigation of their cytotoxicity.
Environmental science & technologyStephen Short, Amaia Green Etxabe, Elmer Swart, Claudia Rivetti, Bruno Campos, Rama Krishnan, Peter Kille, David J Spurgeon
Predicting species-specific chemical sensitivity using in silico approaches has the potential to transform environmental risk assessment, conservation, and biomonitoring, while reducing, and ultimately replacing, animal testing. Genomic and transcriptomic data capture extensive sensitivity-relevant variation, including differences in molecular targets, xenobiotic metabolism, and damage mitigation pathways. Large-scale sequencing initiatives therefore offer an unprecedented opportunity to address ecotoxicology's "too many species" problem. Although existing omic-based predictive tools provide proof of concept, they have so far been applied to a narrow set of relatively straightforward prediction scenarios. To achieve broader applicability, current and future tools must be firmly grounded in the diverse molecular mechanisms underlying differential chemical responses. Here, we critically evaluate the emerging field of predicting species sensitivity using molecular variation inferred from omic data. We analyze the strengths and limitations of current omic-based approaches and identify major sequence and ecotoxicological data gaps, as well as critical bioinformatic challenges. We then review the current knowledge of how molecular biology underlies differential chemical sensitivity, outlining research paths to allow the next generation of sensitivity prediction tools to exploit ever expanding omic data.
Veterinary medicine and scienceHyunYoung Chae, Jae-Won Byun, So-Jeong Yim, Bok-Kyung Ku, Tae-Wan Kim, JeongWoo Kang
BACKGROUND: Anticoagulant rodenticides (ARs) are important toxicants in veterinary forensic investigations involving suspected poisoning in cats. OBJECTIVES: This study aimed to develop and validate a standardized LC-MS/MS-QTRAP method for the simultaneous identification and quantification of eight ARs in gastric matrices from cats and to assess its applicability to veterinary forensic toxicology. METHODS: Automated sample preparation was implemented to reduce inter-analyst variability. Dual confirmation using multiple reaction monitoring (MRM) and enhanced product ion (EPI) spectra was applied to improve identification reliability and minimize false-positive results caused by structurally related compounds. The method was applied to 747 cat forensic toxicology cases submitted between January 2020 and August 2025, in which gastric tissue or gastric contents were available for analysis. RESULTS: ARs were detected in 2.5% of cases (19/747). Coumatetralyl was the most frequently detected AR, followed by flocoumafen and bromadiolone. Flocoumafen was detected despite not being an officially registered active ingredient in Korea, highlighting the need for broad multi-analyte screening. CONCLUSIONS: The standardized LC-MS/MS-QTRAP method enabled reliable detection and quantification of ARs in gastric matrices from cats. This approach provides useful forensic evidence of recent AR exposure in suspected poisoning cases and may support improved toxicovigilance for registered and unregistered rodenticides.
Ecotoxicology (London, England)Karyne Marriel Moreira, Augusto César Santos Oliveira, Victor Ventura de Souza, Renata Alice Campos, José Marcello Salabert de Campos, Tatiana da Silva Souza
Glyphosate and atrazine are among the most extensively applied herbicides worldwide and are often detected together in aquatic environments due to their concurrent agricultural use. This study investigated their individual and combined effects on Allium cepa at environmentally realistic concentrations: 62.5-1000 µg L⁻¹ for glyphosate and 0.25-4 µg L⁻¹ for atrazine. Five mixtures were also tested: M1 (62.5 + 0.25), M2 (125 + 0.5), M3 (250 + 1), M4 (500 + 2), and M5 (1000 + 4). Multiple endpoints were analyzed, including germination, root elongation, mitotic index, chromosomal and mitotic abnormalities, cell-cycle phase distribution, sub-G₁ fraction, fluorescence intensity, light-scattering parameters (FSC, SSC), membrane integrity (Evans Blue assay), and mitochondrial activity (TTC assay). Both herbicides and mixtures triggered cytotoxic and genotoxic responses, though most mixtures did not exceed the toxicity of the individual compounds, suggesting non-interactive or saturable mechanisms. Mixture M5, containing concentrations comparable to those reported in agricultural surface waters, produced stronger responses for some biomarkers. Flow-cytometric and mitochondrial endpoints were the most responsive following mitotic index and chromosomal aberrations. On the other hand, germination, root elongation, and membrane integrity showed limited sensitivity. Overall, the data indicate that even concentrations within current environmental guidelines can impair cellular function in A. cepa. This study provides a novel integrated assessment of glyphosate, atrazine, and their mixtures at environmentally relevant concentrations by combining classical A. cepa cytogenotoxicity assays with flow cytometry, membrane integrity, and mitochondrial activity biomarkers.
ChiralityIgor E Oliveira, Daniela Pereira, Ana R Carvalho, Renata Vidal, Lara da Silva Cabral, Lucas H Martorano, Fernando M S Junior, Cláudia Ribeiro, Ana R L Ribeiro,…
Emerging contaminants are an increasing global concern, with antibiotics representing a notable case due to their continuous release and possible contribution to antimicrobial resistance. Fluoroquinolones (FQs) combine high stability, low biodegradability, and biological activity at trace levels, which result in persistence, ecotoxicity, and bioaccumulation in aquatic systems. Nadifloxacin (NAD) is a chiral FQ prescribed as a racemate for dermatological infections, but (S)-NAD is more potent than the racemate and 64 to 256 times more active than (R)-NAD against both Staphylococcus aureus and Propionibacterium acnes, as well as other gram-positive and gram-negative pathogens. Though the stereochemistry plays a decisive role in both pharmacological activity and environmental fate, the enantioselective ecotoxicological evaluation of NAD is of high importance. Thus, this work described the enantioseparation of NAD by chiral liquid chromatography (cLC) on a Lux amylose-1 column (4.6 mm I.D. × 250 mm L, particle size 5 μm) under normal-phase conditions. Optimized analytical conditions provided good chromatographic parameters (Rs = 5.92, α = 2.77, retention 6-10 min) and were successfully scaled up to semipreparative enantioseparation on a CHIRALPAK AD-H DAICEL column (10 mm I.D. × 250 mm L, particle size 5 μm), allowing the separation of both enantiomers in > 98% enantiomeric purity. The absolute configuration and the elution order of the enantiomers were determined by the agreement between the experimental and calculated data of electronic circular dichroism. The acute ecotoxicity of the NAD racemate and its enantiomers was evaluated using Daphnia magna as an aquatic model organism.
International journal of molecular sciencesChi-Fen Chang, Wen-Hsin Lin, Chao-Yuan Huang, Chia-Cheng Yu, Victor C Lin, Te-Ling Lu, Shu-Pin Huang, Bo-Ying Bao
Diisononyl cyclohexane-1,2-dicarboxylate (DINCH), a non-phthalate plasticizer adopted as a safer alternative for food-contact and medical-grade materials, is ubiquitously detected in human biomonitoring studies. Despite widespread exposure, its transcriptional effects in prostate cells and the potential prostate cancer relevance of DINCH-responsive genes remain unclear. We integrated transcriptomic profiling of DINCH-exposed human prostate epithelial cells with exploratory genetic association analyses in 630 patients with prostate cancer receiving androgen deprivation therapy (ADT). Haplotype-tagged single-nucleotide polymorphisms (SNPs) in candidate DINCH-responsive genes were evaluated for their association with overall survival (OS) and cancer-specific survival (CSS). The prostate cancer relevance of the prioritized genes was further validated using pooled multi-cohort bioinformatic analyses. DINCH exposure produced an exploratory molecular signature comprising 83 genes across all tested doses, broadly suppressing cell-matrix adhesion pathways and activating chromatin remodeling. Exploratory genetic screening identified nominal associations of LPP rs1040033 with OS (p = 0.0002, q = 0.131) and FAM111B rs7110278 with CSS (p = 0.0010, q = 0.575); neither association remained significant after multiple-testing correction. DINCH exposure significantly downregulated LPP and upregulated FAM111B expression in prostate epithelial cells. Independently, pooled analyses demonstrated reduced LPP and elevated FAM111B expression in prostate cancer tissues compared with normal prostate tissues. Higher LPP expression predicted a favorable prognosis, whereas elevated FAM111B predicted worse survival. Pathway analyses linked low LPP expression to impaired adhesion signaling and metabolic reprogramming, whereas high FAM111B expression was associated with mitotic and cell-cycle activation. DINCH exposure induced exploratory transcriptional alterations involving LPP-related adhesion pathways and FAM111B-related proliferative signaling. The genetic findings are exploratory and require independent validation. Although public datasets support the prognostic relevance of LPP and FAM111B in prostate cancer, they do not link these genes to DINCH exposure.
Fish occupy a central position in aquatic ecosystems and serve as important bioindicators for environmental monitoring, as well as powerful translational models for understanding toxic mechanisms conserved across higher vertebrates. In recent years, omics techniques have proven to be powerful tools to address complex environmental questions that conventional toxicology methods cannot answer. Despite this potential, a critical translational gap remains between molecular findings and their use in ecological risk assessment frameworks. This review critically synthesizes advances across omics techniques including epigenomics, transcriptomics, metabolomics and proteomics and their integration. Special emphasis is placed on methodological considerations and practical aspects of these techniques in fish environmental toxicology and environmental monitoring. Evidence from single-omics studies suggests conserved biomarker signatures across species while characterizing complex phenomena like non-monotonic dose-response relationships, mixture toxicity and transgenerational and stereoselective effects with implications for population level monitoring. Multi-omics studies, especially those involving triple omics, further enhance mechanistic resolution by reconstructing adverse outcome pathways. We further evaluate using case studies when additional molecular layers provide critical insight and when they offer limited advantage, a strategic distinction with direct implications in environmental monitoring programmes. Finally, current limitations and future directions that will ultimately bridge the translational gap and hold promise for advancing mechanistic ecotoxicology and predictive environmental monitoring are discussed.
Ecotoxicology (London, England)Vanessa Silva Granadeiro Garcia, Lenita de Freitas Tallarico, Flávio Kiyoshi Tominaga, Maria Clara Feitosa Guimarães, Eliana Nakano, Sueli Ivone Borrely
The presence of surfactants in natural waters has become a growing environmental concern, as these substances, widely used in detergents, cleaning products, cosmetics, and industrial processes, can cause serious impacts on aquatic ecosystems, including deleterious effects on biota, depletion of dissolved oxygen, and eutrophication. This study presents a battery of ecotoxicological assays with different aquatic organisms to evaluate the acute, chronic, and sublethal toxic effects of anionic surfactants Linear Alkylbenzene Sulfonate (LAS) and Sodium Dodecyl Sulfate (SDS). The assays were performed with Vibrio fischeri bacteria, Daphnia similis crustaceans, Hyalella azteca amphipods, and Biomphalaria glabrata snails. Regarding LAS acute exposure, H. azteca was the most sensitive organism, followed by D. similis, B. glabrata, and V. fischeri, with E(L)C50 between 5.04 and 28.47 mg/L. Among the organisms exposed to SDS, the most sensitive were V. fischeri bacteria, followed by D. similis, H. azteca, and B. glabrata, with E(L)C50 values between 0.62 and 36.87 mg/L. Chronic effects on the reproduction of D. similis exposed to both surfactants were also observed, as well as teratogenic and lethal effects on B. glabrata embryos. The data obtained herein demonstrated the importance of evaluating the toxicity of surfactants to aquatic ecosystems, as these stressors caused different adverse effects in the exposed species.
Proceedings of the National Academy of Sciences of the United States of AmericaTao Sun, Huifeng Wu, Lennart Weltje, Evgenios Agathokleous, Edward J Calabrese, John P Sumpter
A long-standing issue in ecotoxicology is the arbitrarily chosen and ambiguous definition of "environmentally relevant" concentrations, which undermines research comparability and hampers risk characterization. Here, we propose a probabilistic framework that anchors exposure levels to percentiles of environmental concentration distributions, defining low (<5th percentile), typical (5th to 95th percentiles), and high (>95th percentile) concentrations, as well as worst-case scenarios (e.g., the 99th percentile) for specific contexts. This framework transforms test concentration selection from a subjective assertion into a statistically justified practice, where each concentration corresponds to an explicit occurrence probability. Using global monitoring data, we demonstrate that environmental concentrations reliably follow cumulative probability distributions, validating the fundamental assumption. Crucially, the framework offers a statistical solution for designing proof-of-relevance and proof-of-concept studies. Collectively, this work provides an empirically grounded, immediately usable template for designing ecotoxicological experiments that bridge environmental monitoring, laboratory testing, and regulatory decision-making.
Environmental science & technologyGabriel M Moulatlet, Mariana V Capparelli, Daniela M Truchet, Andreu Rico, Fabricio Villalobos
With the spread of contaminants across the globe, ecosystems are increasingly exposed to pollutants at varying levels of biological organization. The effects of a wide range of contaminants on individuals have been extensively studied within the discipline of ecotoxicology, but understanding the generality of species' responses across taxa and ecosystems remains a major challenge. This is because such responses are shaped by ecophysiological, geographic, and evolutionary factors, dimensions that ecotoxicology has not been able to fully capture on its own. While these dimensions are frequently explored in macroecological studies, such research rarely considers environmental contamination as a variable capable of influencing species occurrence and distribution patterns. Here, we explore the potential of macroecotoxicology as a subdiscipline capable of integrating the core principles of both ecotoxicology and macroecology. By benefiting from recent advances in these disciplines, macroecotoxicological research would allow us to scale responses from individual organisms' sensitivity up to broader patterns of species distribution and ecosystem response, contributing to the development of predictive frameworks for biodiversity change in a rapidly transforming world.
Scientific reportsCarl Söderberg, Oleg Sysoev, Rasmus Magnusson, Fredrik C Kugelberg, Henrik Green
Intoxication-induced death, both intentional and unintentional, is a global public health concern, contributing substantially to mortality in many regions. Postmortem toxicological interpretation typically relies on reference concentrations of individual substances, which fails to account for combinatorial effects when multiple drugs are present. More comprehensive approaches that analyse multiple substances simultaneously could improve diagnostic accuracy. This study applied computational anomaly detection methods to identify toxicological profiles deviating from known non-intoxication patterns and assessed whether such deviations correspond to fatal intoxications or could serve as a screening tool for cases requiring further review. Postmortem femoral blood drug concentrations covering 191 substances from 16,710 forensic autopsies were analyzed. Cases were divided into a training set of non-intoxications (n = 8320) and separate validation (n = 2674) and test sets (n = 3342) containing both intoxications and non-intoxications. A machine learning-based anomaly detection model was trained and optimized before being applied to the test set. The model distinguished intoxications from normal patterns with 83.9% accuracy. False negatives often involved opioids and ethanol, while false positives were associated with rare substances, numerous detections, or high concentrations. Overall, this approach demonstrates strong potential as a screening tool for identifying toxicological patterns of forensic interest.
Environmental geochemistry and healthCem Tokatlı, Said Muhammad, Bayram Yüksel, Mehmet Metin Yazman, Fikret Ustaoğlu
Rivers discharging into the Marmara Sea integrate contaminant inputs from industrial, urban, agricultural, mining, and geogenic sources. Dissolved potentially toxic elements (PTEs) and phosphorus were measured in spring 2024 at 60 freshwater river channels immediately upstream of their discharge into the Marmara Sea. Concentrations showed substantial spatial heterogeneity. The WHO drinking-water guideline values for boron (2400 µg/L) and arsenic (10.00 µg/L) were exceeded at 2/60 and 10/60 stations, respectively. Dilovası Creek in the İzmit Gulf represented the principal industrial-metal hotspot, with the basin-wide maxima for Ni (119.2 µg/L), Al (368.4 µg/L), Cr (19.2 µg/L), Zn (142.3 µg/L), and Co (1.99 µg/L). Çapraz Stream in the Susurluk Basin had the highest B (3522.8 µg/L) and As (36.8 µg/L) concentrations and the poorest index-based water-quality status (WQI = 93.826; HPI = 232.391), whereas Şerefli Creek in the Tekirdağ Region showed the maximum dissolved-P concentration (6656.3 µg/L). Multivariate analysis and positive matrix factorization distinguished industrial-metal, B-As-enriched, and nutrient-related chemical profiles. Arsenic was the principal toxicological concern: Monte Carlo 95th-percentile HI values were 1.28 for adults and 1.60 for children, while corresponding CR values were 5.81E-04 and 7.29E-04. These screening results identify B-As-enriched southwestern sub-basins and industrialized eastern sites as priorities for targeted monitoring and source-specific mitigation.
Sharp-force fatalities represent a major proportion of homicidal deaths in South Africa, yet the contribution of alcohol and drugs of misuse and abuse to these incidents remains poorly characterised. This retrospective, cross-sectional study reviewed 248 sharp-force fatalities admitted to a large urban forensic service in Cape Town during 2024. Demographic, injury, and toxicological data were extracted from forensic records, and post-mortem blood and urine specimens were analysed using validated headspace gas chromatography for alcohol and liquid chromatography-tandem mass spectrometry targeting 29 drugs. Blood alcohol was detected (>0.01 g/100 mL) in 66.1% of tested cases, while 140 cases (56.5%) were positive for at least one drug. The most frequently identified analytes were 11-nor-9-carboxy-tetrahydrocannabinol (29.0%), methaqualone (27.0%), and methamphetamine (25.8%). Polydrug use occurred in 57.6% of drug-positive cases, most commonly involving methaqualone and methamphetamine, demonstrating a recurrent stimulant-sedative co-use pattern. Of the 140 drug-positive cases, 72 (51.4%) had blood alcohol concentrations below 0.05 g/100 mL, including those with no detectable alcohol, indicating that alcohol was unlikely to have been a major contributing factor in many of these fatalities. These findings demonstrate that psychoactive substances are frequently detected in individuals who die from sharp-force injuries and suggest that intoxication, particularly involving multiple substances, may contribute to impaired judgment, aggression, or increased vulnerability to violence. The results underscore the importance of comprehensive post-mortem toxicological investigations and the integration of toxicological patterns into violence-prevention initiatives and public health strategies.
International journal of molecular sciencesAleksandra Zorychta, Marcin Tomsia, Rafał Skowronek, Elżbieta Chełmecka
In forensic toxicology, the analysis of blood and urine is regarded as the so-called "gold standard." However, when forensic experts are confronted with advanced postmortem changes, it becomes necessary to secure and examine alternative matrices. Suitable specimens in cases of advanced putrefactive decomposition may include: skin appendages (hair and nails), bones, bone marrow, cartilage tissue, teeth, antemortem fingerprints, cerebrospinal fluid, vitreous humor, breast milk, meconium, placenta and umbilical cord tissue, oral fluid, sweat, and other evidentiary materials. These tissues, owing to their structure and anatomical location, are more resistant to putrefactive decomposition than body fluids and soft tissues. However, several limitations complicate the reliable analysis of these matrices. These include incomplete drug incorporation, depending on physicochemical properties, the inability to correlate analyte concentrations with pharmacological effects, low xenobiotic levels, and the need for highly sensitive analytical methods. Collectively, these factors contribute to interpretative challenges during forensic expert evaluation and reporting. To address key forensic questions, epigenetic variability analyses (forensic epigenetics) may also be employed, particularly when standard DNA profiling is uninformative. DNA methylation patterns in specific tissues and in individual subjects can be used, among other purposes, to identify the tissue of origin of a human biological trace, to differentiate monozygotic twins, and to predict the age of an unidentified trace donor. Over the past few years, this approach has gained increasing importance; in the context of forensic trace analysis, it offers both advantages and limitations. The aim of this study is to present current scientific evidence regarding the use of alternative biological matrices as potential sources of information of forensic relevance. The paper discusses the characteristics of individual biological materials, with particular emphasis on their analytical properties, potential applications, limitations, and possible advantages in forensic toxicological investigations. Furthermore, the key aspects associated with the application of epigenetic methods in forensic science are presented, with particular focus on their role in individual identification and the reconstruction of circumstances surrounding forensic events. Alternative biological matrices and modern analytical approaches may constitute valuable complementary tools to conventional evidentiary materials used in forensic medicine and criminalistics. Due to their physicochemical properties, stability, and ability to preserve specific biological information, these matrices may provide significant data that enable substance identification, assessment of exposure to psychoactive compounds, reconstruction of event circumstances, and support for identification procedures. However, their application requires consideration of analytical limitations, the specific characteristics of the examined matrix, and the necessity for standardization and validation of diagnostic procedures.
Environmental science & technologyNicoletta D'Alessandro, Luca Mannino, Giusy Del Giudice, Laura Ylä-Outinen, Noora Perho, Laura A Saarimäki, Emanuele Di Lieto, Zeyad Al-Abdulraheem, Marcella T…
Toxicology faces the need to shift from generalized hazard evaluation toward precision approaches that account for the impact of the exposed biological systems. This need is particularly evident for per- and polyfluoroalkyl substances (PFAS), a highly persistent and diverse chemical class whose multiorgan apical toxicities are well documented, yet whose mechanistic understanding remains fragmented. To address this gap, a comprehensive toxicogenomic collection covering multiple PFAS and biological systems is curated, harmonized, and standardized. Through systematic integration of these data with the Adverse Outcome Pathway framework, biological context-aware key event networks that capture the progression from molecular initiating events to apical outcomes are reconstructed. Additionally, new transcriptomic profiles from macrophages exposed to seven PFAS are generated to address the critical but under-investigated immune-related context. Analysis reveals that PFAS toxicity arises from shared early molecular perturbations that diverge across biological systems to produce organ-specific outcomes where immune-related processes consistently emerge as central contributors across multiple contexts. In the liver emerges a conserved mechanistic core underlying PFAS-induced steatosis activated through system-specific pathways shaped by PFAS physicochemical properties and biological context. Overall, this work provides a framework for advancing precision toxicology, enabling rapid, mechanistically grounded, and context-aware PFAS hazard characterization, and supporting the prioritization of uncharacterized PFAS based on shared and context-dependent mechanistic patterns.
Plasticizers are widely used polymer additives whose release into aquatic environments poses ecological risks, while their structural diversity challenges conventional toxicological assessment. To address this data gap, an integrated modeling framework was established for hazard assessment and prioritization of plasticizers. Species-specific QSAR models were developed for six species across three trophic levels, enabling accurate toxicity prediction for 600 plasticizers. Eighty-five plasticizers were identified as high-priority due to potent toxicity (LC50 < 10 μM) across trophic levels, 54.1% of which are alternative plasticizers. Approximately half of these high-priority plasticizers are widely used in polyvinyl chloride plastics, and 21.2% occur in bioplastics, indicating their potential contribution to the chemical risk of conventional plastics and "green" polymers. Across multiple taxa, predicted toxicity differences among plasticizer subclasses were linked to structural features captured by 2D descriptors. By integrating QSAR predictions with interspecies correlation estimation, the hazardous concentrations for 5% of species (HC5: 0.04-274 μg/L) were derived for high-priority plasticizers using species sensitivity distributions, with crustaceans identified as the most sensitive taxa. Linking hazard prediction and prioritization with polymer occurrence enables the systematic identification of high-priority plasticizers and highlights polymers that contain plasticizers of potential concern.
Environmental science & technologyChristina Galafton, Laura J Zantis, Nils Thonemann, Martina G Vijver
Soils act as major plastic pollution sinks, with potential implications for terrestrial ecosystem services. In life cycle assessment (LCA), such impacts are quantified using characterization factors that describe the fate, exposure, and effects of emitted substances. In this study, we develop terrestrial ecotoxicity exposure-and-effect factors (XEFs) for plastics in soil for use in LCA. We compiled 680 effect concentrations (ECs) from published ecotoxicity studies involving plants and invertebrates and converted them to EC10eq values. Following the USEtox framework, species-specific average EC10eq values were used to construct log-normal fitted species sensitivity distribution curves, from which XEFs were derived. The resulting XEF for plastics in soil equals 0.128 [0.07; 0.30] PAF·m3·kg-1. Specific XEFs for nanoplastics ≤1 μm and plastics >1 μm were calculated at 0.921 [0.11; 13.10] and 0.126 [0.07; 0.29] PAF·m3·kg-1, respectively. Specific XEFs for natural and agricultural soil were estimated at 0.159 [0.08; 0.40] and 0.236 [0.09; 0.77] PAF·m3·kg-1, respectively. The derived XEFs are lower than aquatic and sediment XEFs by 4 and 2 orders of magnitude, respectively. Consequently, direct read-across between compartments is not justified. Overall, the proposed XEFs are compatible with the USEtox methodology and enable improved inclusion of plastic-related terrestrial ecotoxicity impacts in LCA.
Environmental science and pollution research internationalBeáta Baranová, Kristína Bednárová, Daniela Gruľová, Qaiser Javed, Mária Sabalová, Barbara Kudláčková, Lenka Svojanovská, Giuseppe Amato, Lucia Caputo, Vincenz…
Allelopathy is associated with biodiversity loss and assemblage changes in Solidago invaded plots. Although plant-allelopathic effects are well documented, little is known about the toxicity of goldenrod allelochemicals to invertebrates, despite its high biomass production and release of bioactive compounds via volatilization, root exudation or during decomposition, whereas the lack of information is especially evident for the new hybrid S. × niederederi. We conducted a laboratory experiment to assess the effect of water extracts and essential oils derived from the invasive Solidago canadensis and hybrid Solidago × niederederi against larvae of the blood-feeding, invasive Asian bush mosquito (Aedes japonicus), glass worms (larvae of non-hematophagous Chaoborus mosquitoes), sludge worms (Tubifex tubifex), and earthworms (Eisenia fetida), which represent aquatic, semi-aquatic and soil-dwelling invertebrates. Impact was determined on the model organism's mortality and lethal concentration values (LC50/LC90). Except for the content of phenolic acids and saponins in the water extracts, and the representation of dominant compounds in the essential oils plus their acetylcholinesterase and butyrylcholinesterase inhibition activity were determined too. Subsequently, possible link to the toxicity was evaluated as well. Used invertebrates exhibited various sensitivities, depending on the extract type: essential oils from both goldenrod species caused high mortality of A. japonicus larvae. We also observed pronounced toxicity of S. canadensis essential oil toward glass worms. In contrast, the essential oils from invader and hybrid had no harmful effect on the earthworms or sludge worms, while both goldenrod's water extracts caused certain mortality of T. tubifex and total mortality of E. fetida. Our analyses suggest that the toxicity of goldenrod's allelochemicals against invertebrates may be linked to the presence of specific phenolic acids and saponins in water extracts, and the inhibitory activity of essential oils against acetylcholinesterase and butyrylcholinesterase consequently. The study thus indicates possible ecological consequences of goldenrod invasion on the impacted faunal communities. Possible usage of Solidago representative's biomass for the bio-based pesticide development, as part of the practically focused plant invasion management, can be deduced as well.
The precise detection and profiling of biotoxins are of paramount importance in analytical and forensic toxicology. Investigating these toxicants within highly chaotic background matrices-ranging from postmortem biological fluids to suspected poisoning vehicles such as complex dietary and environmental samples-requires robust recognition molecules capable of overcoming severe interference. This comprehensive review summarizes recent analytical developments in biotoxin detection, categorizing molecular recognition platforms into three primary types: immunological recognition (antibodies and recombinant derivatives), aptamer-based recognition, and entirely synthetic recognition (molecularly imprinted polymer, MIP). To meet the rigorous ultra-trace demands of medicolegal analysis, we further discuss the strategic integration of these recognition elements with powerful catalytic amplification cascades, highlighting the transition from natural biological enzymes to highly durable nanozymes and DNAzymes. By detailing recent structural optimizations and preparation strategies, this review critically evaluates the respective advantages, matrix tolerances, and limitations of each recognition mode when applied to diverse and challenging analytical samples. Finally, we provide a forward-looking perspective on the translational potential of these specific binding agents, emphasizing how computational rational design and portable integration will overcome practical bottlenecks in modern toxicological investigations.
Recently, abuse and fatal overdoses of over-the-counter (OTC) medications have become serious social and health issues worldwide. However, many drug components in OTC medications that are abused have different chemical structures from those assessed by commercially available rapid drug test kits. In this research, we focused on using dried blood spot (DBS) samples to develop a simple and accurate method for forensic toxicological analysis of drugs of interest in deaths by poisoning. We combined the DBS method with liquid chromatography coupled with electrospray ionization-tandem mass spectrometry to quantify eight OTC drug components (apronal, caffeine, chlorpheniramine, dextromethorphan, diphenhydramine, diprophylline, noscapine, and promethazine). The linearities of the calibration curves were good in the concentration range of 0.1-2.0 μg/mL. The method achieved repeatable and accurate quantification of the eight target drugs with intra- and inter-assay coefficients of variation of below 13.0% and 12.6%, respectively. Comparison with our general practice method showed good positive correlations at the quantifiable concentrations of all drugs. Furthermore, when applied to an actual case, it was possible to diagnose death by drug poisoning, confirming the usefulness of this method.
Molecules (Basel, Switzerland)Flavio Polito, Paola Malaspina, Annarita La Neve, Antonella Smeriglio, Laura Cornara, Vincenzo De Feo, Domenico Trombetta
The valorization of medicinal and aromatic plant by-products represents a sustainable strategy to recover bioactive compounds and reduce agricultural waste. In this study, pruning residues of Laurus nobilis L. and Myrtus communis L. were investigated as sources of essential oils (EOs) for bio-based weed management. Leaves and young stems were subjected to anatomical, micromorphological, and histochemical analyses to ascertain that active secretory structures were maintained. EOs were then obtained by steam distillation and characterized by GC-FID and GC-MS. Phytotoxicity was evaluated on two crop species, Hordeum vulgare L. and Raphanus sativus L., and two weed species, Lolium multiflorum Lam. and Sinapis alba L.; ecotoxicity was assessed using Artemia salina and Daphnia magna. The EOs were mainly composed of oxygenated monoterpenes, with 1,8-cineole as the dominant constituent in both oils. Phytotoxicity was species- and concentration-dependent, with M. communis EO showing the strongest inhibition, particularly against S. alba. Ecotoxicological assays revealed limited toxicity toward A. salina, but concentration-dependent immobilization of D. magna. Overall, these findings support the circular valorization of laurel and myrtle pruning residues as sources of bioactive EOs while highlighting the need for further formulation, selectivity, and environmental safety studies before agricultural application.
International journal of molecular sciencesSaad Lodhi, Marcel Van Herwijnen, Colette Kelly, Harvey Fowler-Williams, Carrie A Duckworth, D Mark Pritchard, Florian Caiment, Theo M C M de Kok, Marcha C T V…
Animal models are standard for safety evaluation, yet physiological differences limit human translation. Doxorubicin, a chemotherapeutic agent, can cause off-target gastrointestinal toxicity and treatment discontinuation. This study evaluated human colonoids as a controlled human-derived epithelial model for doxorubicin-induced gastrointestinal toxicity by comparing transcriptomic responses across human colonoids, mouse colonoids, and male C57BL/6J mouse colon tissue. Within each dataset, doxorubicin-treated conditions were pooled across model-specific exposure levels and time points to estimate broad doxorubicin-associated transcriptional signatures. Using a parallelogram approach, differential expression, co-expression, pathway mapping, and Comparative Toxicogenomics Database benchmarking were applied to compare model concordance and identify doxorubicin-responsive mechanisms. Shared responses converged on cell-cycle regulation, DNA damage response, DNA repair, and apoptosis. Concordance in differentially expressed genes was highest between mouse colonoids and mouse colon, while pathway mapping showed similarities between colonoid systems. A core set of p53-associated genes, including BAX, INKA2, and ZMAT3, was shared across datasets, with additional apoptotic and DNA damage response features observed in colonoids. Comparative Toxicogenomics Database benchmarking supported doxorubicin biology and highlighted underrepresented gastrointestinal toxicity-relevant signals, indicating gaps in intestinal toxicogenomic annotations. Colonoid-based transcriptomics supports human colonoids as controlled epithelial models for mechanistic gastrointestinal toxicity assessment, although missing vascular, immune, and systemic context means clinical validation remains necessary.
World journal of microbiology & biotechnologyFirat Yavuz Öztürk, Fadime Özdemir
Microplastics (MPs) have emerged as pervasive environmental contaminants in aquatic, terrestrial, and atmospheric ecosystems. Once released into the environment, MPs are rapidly colonized by microorganisms, leading to the formation of complex biofilm communities collectively termed the "plastisphere." These biofilms significantly alter the physicochemical properties, transport behavior, ecological interactions, and toxicity of microplastics. This review synthesizes recent findings on the mechanisms of biofilm formation on microplastics, including the roles of polymer type, surface aging, eco-corona formation, and environmental factors such as salinity, temperature, nutrient availability, and hydrodynamics. The composition and ecological functions of plastisphere communities, including bacteria, archaea, fungi, algae, and protists, are discussed with emphasis on extracellular polymeric substances (EPS), quorum sensing, metabolic interactions, and horizontal gene transfer. The review further evaluates the role of biofilm-coated microplastics as vectors for pollutants, antibiotic resistance genes, and pathogenic microorganisms across marine, freshwater, wastewater, soil, and agricultural systems. In addition, the interactions between microplastics and co-contaminants such as heavy metals, pharmaceuticals, PFAS, and organic pollutants are examined in the context of ecotoxicological risks. Current methodological approaches, environmental implications, and regulatory challenges are also addressed. Overall, this review emphasizes the importance of adopting a biofilm-centered perspective for understanding the environmental fate and ecological impacts of microplastics, delves deeper into microplastic-associated biofilms across diverse ecosystems, including marine and freshwater environments, wastewater and urban water systems, and soils and agricultural lands, while only briefly considering less-studied compartments such as the atmosphere, integrating pathogen-specific food safety evidence, nanoplastic-EPS interactions, AMR/HGT mechanisms, and co-contaminant-derived risks, identifying the impacts of microplastics/biofilms themselves or the pathogens they carry on human health, and identifies critical knowledge gaps that require future investigation.