Microcystin-LR (MC-LR) is a potent hepatotoxin that has been shown to cause liver damage even at doses lower than the established Low Observable Adverse Effect Level (LOAEL) of 200 μg/kg in animal models. We have previously observed that low-dose exposure to MC-LR in animals with diet-induced Metabolic Dysfunction-Associated Steatotic Liver Disease (MASLD) and subsequent treatment with antioxidants like N-acetylcysteine (NAC) and the Na+/K+ ATPase-Src kinase inhibitor pNaKtide significantly alleviated hepatic infiltration of immune cells, downregulated markers of inflammation and hepatotoxicity, increased the breakdown of the toxin molecule, and restored phase I and II drug metabolism pathways, including the glutathione pathway. Because the liver is composed of heterogeneous cell types, this study aimed to determine the specific role of hepatocytes in the uptake and metabolism of MC-LR, especially in the setting of MASLD. To address this, we used two well-established hepatocyte cell lines-AML-12 murine hepatocytes and human Hep3B hepatocytes. Preliminary dose comparison studies with AML-12 cells showed that MC-LR at 10 μM concentration showed a significant upregulation in the genetic expression of the markers of hepatotoxicity-OSMR (p ≤ 0.01) and SerpinE (p ≤ 0.0001)-in comparison to Vehicle. Treatment with pNaKtide (1 µM) and/or NAC (10 mM) in the presence of MC-LR significantly reduced the expression of both OSMR (p ≤ 0.0001) and SerpinE (p ≤ 0.01 and p ≤ 0.0001, respectively). To model steatotic hepatocytes characteristic of the MASLD phenotype, Hep3B hepatocytes were first treated with 500 µM of oleic acid (OA) before exposing them to the toxin in the presence and absence of antioxidants. MC-LR exposure, induced markers of inflammation and hepatotoxicity to be elevated significantly in the presence of OA as compared to MC-LR exposure alone. This elevation of the genetic markers of inflammation and hepatotoxicity was significantly attenuated on treatment with pNaKtide (1 µM) and NAC (10 mM). Quantification of human SERPINE1 (PAI1) and 8-OHdG, a stable marker of oxidative stress, in the spent media of Hep3B cells corroborated the trends observed in the genetic markers of hepatotoxicity. These observations support the central role that hepatocytes play in the uptake and metabolism of MC-LR, which is complicated by the presence of MASLD-like conditions and can help in the development of future therapeutic strategies.
Although chronic contamination by silver ions (Ag+) can persist in aquatic systems over long periods of time and can therefore have an impact on population developments, regulatory testing commonly relies on single-generation endpoints. Here, we used Daphnia magna to quantify long-term effects of pg/L to ng/L concentrations of Ag+ across generations and to test whether recovery depends on exposure history. Using 21 d life-cycle assays over up to seven consecutive generations, we quantified survival, key life-history traits, and population fitness (intrinsic rate of natural increase, r). In our study, low environmental concentrations of Ag+ caused minimal mortality, but sublethal effects persisted or multiplied over generations. Notably, continuous exposure led to significant reductions in body length and r at 50 pg/L (nominal LOEC) by the fourth generation exposed, representing population-relevant effects of Ag+ at very low concentrations which should be given consideration in the assessment of both water quality and the chemical itself. Recovery was concentration-dependent: low-concentration-exposed lineages recovered within a few generations, whereas 15 ng/L exposure resulted in persistent deficits even through the recovery period of three generations. Exposure-history patterns indicated that long-term outcomes were dominated by the cumulative number of exposed generations. These findings highlight the limitations of acute and single-generation assays and emphasize the importance of considering information on the effects of chemicals, including Ag+, across multiple generations in risk assessments. They also highlight the need to include expectations regarding recovery after the removal of pollutants in these assessments.
Aim: The aim of this study was to develop and validate a reliable HS-GC-FID/FID method for the determination of ethanol and other low-molecular-weight volatile compounds in biological fluids for forensic applications. Method: The method is based on headspace gas chromatography with dual-column and dual flame ionization detection (HS-GC-FID/FID), using Zebron ZB-BAC1 and ZB-BAC2 columns. The procedure was validated in terms of linearity, limits of detection and quantification, precision, and accuracy, as well as carryover. Results: The method demonstrated linearity over the concentration range of 0.05-5.0‱, with R2 values of 0.997-0.999. The limit of quantification (LOQ) was 0.05‱, and the limit of detection (LOD) was 0.025‱. Precision and accuracy were both below 5%. Retention times (min) on the two columns were as follows: methanol (1.70/1.81), ethanol (2.06/2.29), acetone (2.60/2.59), isopropanol (2.45/2.73), n-propanol (3.24/3.98), and n-butanol (6.35/8.45). The developed method was successfully evaluated through international proficiency testing and is routinely applied in our laboratory for forensic casework. Conclusions: The developed HS-GC-FID/FID method provides accurate and reliable determination of ethanol and other relevant volatile compounds in biological fluids (i.a., blood) and meets the requirements for forensic toxicology. Additionally, the literature review conducted in this study highlights that globally unified principles for forensic alcohol analysis are still lacking, and that certain inappropriate methodological approaches remain in use. The present paper also provides recommendations defining essential methodological requirements to ensure the evidential reliability of ethanol analysis in forensic toxicology.
Glyphosate and its primary metabolite aminomethylphosphonic acid (AMPA) are widely detected in aquatic environments, yet their combined effects on fish remain insufficiently understood. This study used label-free blood plasma proteomic profiling to explore molecular patterns associated with 14-day exposure of juvenile common carp (Cyprinus carpio) to environmentally relevant concentrations of glyphosate (100 µg/L), AMPA (100 µg/L), and their mixture (50 + 50 µg/L). Across the three exposure groups, 41 proteins of interest showed pronounced abundance differences relative to the control based on fold-change selection criteria. These proteins were mainly associated with immune recognition, innate immune and complement-associated functions, coagulation and extracellular protease regulation, lipid/sterol transport, and extracellular matrix organization. In the mixture group, proteins of interest spanned several functional categories, suggesting that combined exposure deserves further attention in future studies of plasma-level responses to glyphosate and AMPA. Overall, these findings provide preliminary insights into blood plasma protein patterns associated with systemic responses of fish to glyphosate, AMPA, and their mixture at environmentally relevant concentrations and highlight the importance of considering parent compounds, metabolites, and their co-occurrence when assessing the potential biological effects of herbicide contamination in aquatic ecosystems.
Invasive alien species (IAS) such as Silurus glanis and Procambarus clarkii represent major ecological threats but may also serve as effective bioindicators of environmental contamination; therefore, this study aimed to evaluate their potential for multi-contaminant monitoring and assess their suitability as alternative feed and food resources within a circular economy framework. Multi-contaminant profiles were investigated in S. glanis and P. clarkii from Avigliana Lakes (NW Italy), analyzing trace elements, rare earth elements (REEs), and organic contaminants in fish muscle, and microplastics (MPs) in intestinal tracts. In S. glanis, total trace element concentrations and ΣREEs were markedly higher in Small Lake than in Great Lake, with ΣREEs reaching 0.445 and 0.056 mg/kg w.w., respectively. Mean concentrations of the regulated elements in Great Lake were 0.017 mg/kg w.w. (As), 0.003 mg/kg w.w. (Cd), and 0.16 mg/kg w.w. (Pb), increasing in Small Lake to 0.19, 0.03, and 1.86 mg/kg w.w., respectively. In P. clarkii, contamination levels were lower, with ΣREEs averaging 0.074 mg/kg w.w. and mean concentrations of As, Cd, and Pb of 0.25, 0.006, and 0.21 mg/kg w.w., respectively. Organic contaminants, including polycyclic aromatic hydrocarbons (PAHs), non-dioxin-like polychlorinated biphenyls (NDL-PCBs), and pesticides, were generally below limits of quantification. MPs were detected in 100% of specimens, with mean concentrations of 4.2 ± 2.15 and 4.4 ± 2.70 MPs per intestinal tract in S. glanis (Great and Small Lake, respectively) and 2.7 ± 2.39 MPs/intestinal tract in P. clarkii. Permutational multivariate analysis of variance (PERMANOVA) indicated significant site-related differences in S. glanis and species-related differences between S. glanis and P. clarkii within Great Lake. Most regulated contaminants were below applicable EU thresholds; however, Pb in S. glanis from Small Lake exceeded the maximum level established for fish muscle intended for human consumption.
Toluene, as a common organic solvent in academic laboratories in university campuses, poses potential exposure concerns to students and staff in university campuses. Hence, by using a computational fluid dynamics simulation, we investigated the dispersion characteristics of toluene at a campus in Guangzhou under meteorological conditions and the impact of newly constructed buildings on toluene concentrations. The numerical simulation results reveal that toluene is readily accumulated in the free movement area under the prevailing east wind, in the administrative area under the prevailing north-northeast wind, and in the teaching area under the prevailing south wind. Therein, the teaching buildings (TB3-TB6) possess the highest average concentration of toluene compared with other functional areas. In the presence of newly constructed buildings, the toluene concentrations are decreased under the south-southeast wind but are aggravated under the southeast wind. As the height increases, under south-southeast winds, the merging of vortex structures continuously reduces toluene concentrations at TB3 and TB4 and the expansion of the wake region rebounds the toluene pollution at TB5 and TB6; under southeast winds, the expanding vertical vortex structures aggravate toluene pollution at TB3 and TB5 but attenuate toluene pollution at TB4 and TB6. Our results reveal that the teaching areas of the target campus represent a critical zone for potential student exposure during summer and require particular attention. This study provides new insights into the coupled effects of prevailing wind conditions and campus morphology on VOC dispersion characteristics and improves the understanding of airflow pollutant interactions in complex campus environments.
Atrial septal defect (ASD) has become increasingly common in the USA and now affects 1 in 11.3 children in some places, but space-time analysis has not been applied to this emerging trend. ASD rate (ASDR) data were obtained from the National Birth Defects Prevention Network 2003-2020. Substance (cigarettes, alcohol, cannabis, analgesics, cocaine) use data were obtained from the National Survey of Drug Use and Health. Income data were obtained from the US Census. Analysis was limited to the Non-Hispanic White population by technical factors. Time-sequential univariate and bivariate maps were prepared for both covariates and outcomes and their combinations. Spatial regression of the ASDR was performed using the R package splm. A total of 7.6% of data was interpolated by linear regression. A total of 110,107 ASD cases were identified amongst 17,751,437 live births in 27 US states across 10 reporting periods. Time series maps showed that ASDR showed concordant patterns with indices of cannabis use rather than other substances. This was confirmed by multivariate spatial regression where cannabis and cannabinoids alone were found to significantly relate to ASDR, with p = 0.00002 for cannabidiol. Cannabis legal status similarly tracked with ASDR. Compared to states where cannabis was not legal, ASDR was more prevalent in cannabis-legal states (OR = 2.73 (2.66, 2.80); E-Value 4.90 (lower C.I. 4.76)). Twenty-seven of 34 (79.4%) E-values were >9 (high range) and 34/34 were > 1.25 (causal threshold). Data show that cannabis, including cannabis legalization, is driving the US ASD epidemic. While most high-ASDR states have high rates of cannabis use, Midwestern states where cannabis is farmed, such as Kentucky, Tennessee and Missouri, do not, suggesting other routes of exposure, potentially implicating environmental contamination. ASD is a bellwether marker for cannabinoid teratogenicity, indicating that communities should carefully control cannabinoid exposure and limit transgenerational cannabinoid genotoxicity more generally.
Despite the undisputable ecosystem importance of honeybees, human activities have a substantial impact on their health. Since foraging is directly linked to a wide range of crops and bee-attracting flowers, plant protection products are at the forefront of chemical scrutiny, along with contamination of pollen, nectar, beehive components and water by other xenobiotics. In this study, a non-targeted Liquid Chromatography-High-Resolution Mass Spectrometry (LC-HRMS) screening was applied to 25 honeybee samples collected after reported death incidents in Greece. This approach led to the tentative annotation of over 50 compounds across various chemical classes, including pesticides, PFAS candidates not included in the EFSA "PFAS-4", pharmaceuticals, antibiotics, industrial chemicals, and natural product constituents. In parallel, targeted pesticide residue analysis using liquid and gas chromatography coupled to tandem mass spectrometry (LC-MS/MS and GC-MS/MS) was performed, covering more than 250 active substances and providing direct quantitative results, revealing 11 active substances in concentrations ranging from <limit of quantification (LOQ) to 0.95 mg/kg, overlapping substantially with the HRMS detection. Overall, this study does not allow concrete causal attribution of mortality to specific chemicals; however, it documents complex co-occurrence patterns (pesticides together with other xenobiotics and plant bioactives), not excluding sublethal and mixture-toxicity effects. Quantified pesticide concentrations were below acute LD50-based thresholds, yet selected samples combined neonicotinoid/pyrethroid/fungicide signatures and other contaminants, supporting the need for mixture-toxicity frameworks and effect-based follow-ups.
Larixyl acetate, a primary component of Larch turpentine, is a naturally occurring compound with a broad spectrum of medicinal properties, including anti-inflammatory effects. It is a potent and selective inhibitor of TRPC6, a widely expressed Ca2+ channel that is involved in many respiratory diseases. Despite its demonstrated efficacy, it lacks a well-defined preclinical and phenotypic safety profile, which limits its therapeutic potential and implementation. In this study, female BALB/c mice were used to assess the toxicity of intranasally administered Larixyl acetate through a subacute model based on OECD Test Guideline 412, followed by a detailed analysis of physical, blood, biochemical, and tissue changes at the administration sites and beyond. Within the study's 30-day timeframe, our results show no statistically significant differences (p > 0.05) in any of the examined toxicity parameters between the controls or three treatment groups (0.5, 1, and 2 mg/kg). While no pharmacokinetic data were obtained to confirm local or systemic exposure of Larixyl acetate, these findings are crucial for establishing a solid foundation for future therapeutic endeavors, especially in the context of TRPC6-driven respiratory diseases.
Styrene, a constituent of polystyrene food-contact materials, can migrate into hot beverages, but data on short-term consumer exposure and associated biological responses remain limited. In this single-arm longitudinal human biomonitoring pilot study, 40 healthy adults consumed tea or coffee daily in Styrofoam cups for approximately two weeks. Biomarkers were measured at baseline, day 6, and day 11, including urinary mandelic acid (MA) and phenylglyoxylic acid (PGA), salivary malondialdehyde (MDA), comet assay parameters in peripheral blood lymphocytes, and micronucleus (MN) frequency in buccal cells. Measured styrene migration into beverages ranged from 3.3 to 7.1 μg/L, below the World Health Organization guideline value. Urinary metabolites and salivary MDA showed substantial interindividual variability and no consistent temporal pattern. In contrast, generalized estimating equation models showed progressive increases in comet assay indicators over the exposure period. Tail intensity and tail moment increased over time, with stronger changes among participants consuming two cups daily. MN frequency did not change significantly. These findings suggest that repeated short-term consumption of hot beverages in polystyrene cups was associated with modest changes in selected early biomarkers of biological effect under consumer-use conditions. The results should be interpreted cautiously in light of the modest sample size, short follow-up, and absence of more specific mechanistic endpoints, but they support further study of repeated low-level exposure to food-contact materials.
The widespread use of modern insecticide formulations underscores the need for mechanistic evaluation of their potential renal toxicity. This study investigated the nephrotoxic effects of Ampligo® 150 ZC, a binary formulation of lambda-cyhalothrin and chlorantraniliprole, in female rabbits under subacute exposure conditions, with particular emphasis on apoptosis-related and epithelial integrity biomarkers, and evaluated the protective effects of thyme essential oil (TEO) and vitamin C. Rabbits were allocated into four groups: control, AP, AP + TEO, and AP + TEO + vitamin C. Ampligo (AP) exposure resulted in significant renal dysfunction, as evidenced by elevated biochemical biomarkers and marked histopathological lesions. At the molecular level, AP induced p53 upregulation alongside Bcl-2 and Cyclin D1 downregulation, suggesting apoptosis induction and cell cycle dysregulation. Moreover, reduced E-cadherin and β-catenin expressions indicated disruption of epithelial junction integrity and impaired renal structural homeostasis. Notably, co-administration of TEO and vitamin C markedly attenuated these alterations, improving biochemical, histopathological, and immunohistochemical parameters. Overall, these findings suggest that AP-driven nephrotoxicity may involve apoptotic and epithelial pathways under subacute exposure conditions, whereas antioxidant co-treatment may mitigate kidney injury, supporting the potential of natural antioxidants as adjuncts against pesticide-induced renal injury.
The sorption behavior of antibiotics in wastewater treatment systems plays a critical role in determining their environmental fate and removal efficiency. In this study, the sorption of 15 antibiotics representing multiple classes was investigated using two sewage sludge samples with different physicochemical characteristics. Batch equilibrium experiments were conducted to evaluate time-dependent sorption behavior and to determine solid-water distribution coefficients (Kd). The results showed that sorption occurred rapidly, with most compounds approaching a stable concentration within 24 h. The Kd values varied widely depending on the compound, ranging from 74 to 737 L/kg. For 13 of the 15 investigated antibiotics, higher Kd values were observed in sludge B than in sludge A, with the largest difference observed for tiamulin (402 ± 53 and 737 ± 76 L/kg for sludge A and sludge B, respectively). Sludge B generally exhibited higher sorption capacity for most compounds than sludge A, despite having a lower specific surface area, indicating that sorption was governed primarily by chemical composition and pore structure rather than surface area alone. Elemental and morphological analyses suggested that differences in metal-associated components and pore structure may contribute to the higher sorption capacity observed in sludge B. However, the specific sorption mechanisms could not be directly confirmed by the present analyses. Comparison with previous studies confirmed that the measured Kd values fall within reported ranges but are generally higher for sulfonamides, suggesting enhanced sorption capacity of the investigated sludge matrices. Application of an equilibrium-based model demonstrated that sorption alone can account for approximately 20-70% of antibiotic removal under typical activated sludge conditions, depending on compound affinity. These findings highlight the importance of sludge-specific properties in controlling antibiotic partitioning and demonstrate that incorporating such characteristics into predictive models can improve the accuracy of environmental fate assessments in wastewater treatment systems.
Toxicological testing for drugs of abuse (DOAs) is an essential tool for healthcare practitioners and law enforcement agencies. Oral fluid (OF) is an alternative biological fluid for detecting recent DOA intake and is widely employed in forensic investigations. In the current study, a relatively novel and "green" fabric phase sorptive extraction (FPSE) procedure for sample preparation was coupled to liquid chromatography-tandem mass spectrometry (LC-MS/MS) to provide simplicity, cost-effectiveness, rapidity, low solvent consumption, and high analytical performance for the quantitative determination of ten commonly encountered DOAs and metabolites: amphetamine, benzoylecgonine, cocaine, codeine, ecgonine methyl ester, methadone, methamphetamine, 3,4-methylenedioxyamphetamine, 6-monoacetylmorphine, and morphine. The FPSE procedure was optimized by testing different filters, pH, extraction time, and solvents. The validated method demonstrated excellent linearity for all analytes, selectivity, acceptable precision, and high sensitivity (ranges for limits of detection (LODs) and quantification (LOQs) were 0.01-2 ng/mL and 0.03-6 ng/mL, respectively). Autosampler and short-term freeze stability exceeded 95% and 90% for all analytes, respectively. Overall, the combination of FPSE with LC-MS/MS provided a sensitive, selective, and environmentally friendly innovative analytical approach for the determination of DOA in OF and is suitable for both screening and confirmatory forensic and clinical applications.
African soils face increasing levels of metal pollution due to industrialization, artisanal mining activities, improper waste management, and enhanced agricultural productivity. However, unlike many organic pollutants, heavy metals do not degrade naturally and therefore persist in environmental systems for prolonged periods. Heavy metals accumulate over many decades in the soil and bioaccumulate through the food chain causing severe health complications such as cancer, kidney problems, and neurological impairment. This paper reviews the current literature on the origin, prevalence, and behavior of the main pollutants Pb, Cd, Cr, As, Hg, and Cu. The major phytoremediation methods including phytoextraction, rhizofiltration, phytostabilization, and phytovolatilization are highlighted alongside in planta screening methods for hyperaccumulating plants including Berkheya coddii (Ni) and Haumaniastrum robertii (Co). The paper evaluates various enhancement techniques such as the use of chelators, Rhizobium inoculations, and genetic modifications. The significance of these approaches in tropical and subtropical climates is discussed. The paper suggests a holistic framework involving empirical kinetic modeling, geospatial machine learning (random forest, kriging), and molecular omics in prediction modeling. Major hurdles in such predictions include lack of field-based verification of the models, biotechnology safety of genetically modified (GM) organisms, and inadequate regulations. Future perspectives emphasize community-driven phytomining, biomass recycling, and resilient phytoremediation solutions.
M8OI is a cytotoxic methylimidazolium ionic liquid solvent through its binding to the ubiquinone binding site on complex I of the mitochondrial electron transport chain. Given the overlap in terms of toxic mechanism of action with the pesticide rotenone, the potential neurotoxic effects of M8OI were examined. In vitro, cytotoxicity and mitochondrial function were assessed in SH-SY5Y cells by measuring MTT reduction and oxygen consumption/extracellular acidification using a Seahorse analyser. SH-SY5Y cells were sensitised to M8OI toxicity by replacing medium glucose with galactose. Glucose protected the cells from M8OI toxicity, whereas galactose showed no clear dose-response protection. M8OI induced a dose-dependent reduction in oxygen consumption rate with a compensatory increase in extracellular acidification rate, consistent with inhibition of mitochondrial oxidative phosphorylation and a shift toward glycolysis. In vivo, rats were orally exposed via drinking water for 20 weeks and assessed using behavioural tests. In addition, the concentrations of M8OI and its metabolites were quantified by LC-MS in rat brain and other tissues. In rats, M8OI concentrations were ~30-fold higher in kidney than brain, and brain levels were at least 100-fold lower than the concentrations that affected SH-SY5Y cell viability in vitro. However, based on open field tests, M8OI exposure suppressed motor activity without any anxious behaviours. The cytotoxicity of M8OI in SH-SY5Y neuroblastoma cells was associated with metabolic mitochondrial dysfunction. However, the neurobehavioural changes observed in orally exposed rats occurred at significantly lower brain concentrations than would be predicted to lead to neural cell death. Nevertheless, direct comparisons between acute in vitro exposures and chronic in vivo outcomes should be interpreted cautiously.
The intensive use of agricultural inputs and the increasing incorporation of nano-materials into crop management practices raise concerns about their ecotoxicological interactions in plant systems. This study evaluated phytotoxicity, cytotoxicity, and genotoxicity in Allium cepa L. under experimental nano-agrochemical exposure scenarios combining two conventional nitrogen fertilizers-ammonium sulfate (AS) and urea-with silver nanoparticles (AgNPs). Biological responses were assessed across fertilizer concentrations (0.03-0.5 g/L), applied individually, simultaneously, and sequentially, to identify modulatory effects of AgNPs on plant proliferative activity and genomic stability. Results showed the relative stability of morphophysiological indicators associated with root growth, whereas cytogenetic biomarkers exhibited selective alterations under specific conditions. Significant increases in genetic damage markers were detected at intermediate ammonium sulfate concentrations, suggesting sublethal phytotoxicity windows not reflected by macroscopic growth parameters. In addition, modulation of the mitotic index and absence of generalized genotoxic effects in most combined or sequential treatments indicate that AgNPs primarily acted as modulators of proliferative responses rather than direct cytotoxic agents. Overall, these findings highlight the dynamic and non-linear nature of nano-agrochemical interactions in plant systems and underscore the importance of multibiomarker approaches for the early detection of genomic instability. The results provide experimental evidence relevant to the environmental risk assessment of nano-enabled fertilization strategies under realistic mixed-exposure scenarios. This study contributes to advancing the ecotoxicological understanding of emerging agricultural technologies and supports the need for further mechanistic research and field-based evaluations to guide the safe and sustainable use of nanomaterials in crop production.
This study investigated the individual and combined effects of mercury (as HgCl2) and Bisphenol S (BPS) on the blue crab Callinectes sapidus, focusing on biomarkers measured in hemolymph, gills, and hepatopancreas and ultrastructure (transmission electron microscopy, TEM) observations. Adult males were exposed for seven days to environmentally relevant concentrations of each contaminant, alone and in mixture. Results revealed clear tissue-specific responses. In hemolymph, both contaminants reduced total hemocyte count and affected immune-related parameters, including hemocyte proliferation and enzymatic activities. In gills, mercury significantly decreased total antioxidant capacity, while both contaminants increased lipid peroxidation, indicating oxidative stress. BPS and the mixture stimulated catalase activity, whereas electron transport system activity increased only under combined exposure. In contrast, the hepatopancreas showed limited biochemical alterations, suggesting a higher resilience to medium-term exposure. TEM observations revealed a general decrease in granularity of hemocytes from crabs exposed to BPS and mixture, suggesting cell degranulation. In addition, infiltration of hemocytes into gills was observed in crabs exposed to experimental conditions, suggesting migration of hemocytes from hemolymph to peripheral tissues, while alterations of the microvilli arrangement were detected in digestive cells of BPS-treated crabs. Overall, the findings suggested that hemolymph and gills were more sensitive to contaminant exposure, while the hepatopancreas appears more resistant, at least under the experimental conditions tested. The study also suggests potential non-additive interactions between mercury and BPS.
Acetamiprid (ACMP) exposure mediates a variety of pathological complications, including testicular toxicity. Berberine (BBR) is a plant-derived alkaloid with potential pharmacological properties. This study sought to evaluate the ameliorative effects of BBR against ACMP-induced testicular toxicity. Male Wistar rats were divided into four groups: control, BBR-treated, ACMP-exposed, and BBR+ACMP co-treated, and were administered with BBR (150 mg/kg b.wt) and ACMP (21.7 mg/kg b.wt) for 21 days. Biochemical and FTIR analyses, RT-PCR, computational analyses, and histopathological examination were conducted to assess alterations in lipid and protein profiles, as well as apoptotic and structural changes. ACMP exposure was associated with oxidative injury, functional alterations (stretching of -OH, -CH2, -NH, C=O, C-N, -COO-, -PO2-), and compositional changes in proteins and lipids. Pre-treatment of BBR (2 h prior) was associated with attenuation of the functional and compositional alterations in proteins and lipids in co-treated rats. RT-PCR and computational analysis showed increased Bax and caspase-3 and decreased Bcl-2 mRNA expression, suggesting a potential modulation of ACMP-induced apoptosis by BBR. Histological examination showed that pre-treatment with BBR prevented ACMP-induced structural alterations, including cellular disorganization and alteration in seminiferous tubules. The study suggested that the BBR may exert ameliorative effects against ACMP-induced testicular toxicity by modulating lipid and protein changes and the anti-apoptotic pathway. Thus, BBR could be used as a potential ameliorative agent against oxidative stress. However, more mechanistic studies are needed for broader biological relevance and validity.
Pesticide formulations may influence toxicological outcomes beyond the intrinsic properties of active ingredients; however, these differences are often overlooked in regulatory risk assessment. Using human liver HepG2 cells, this study compared the cytotoxic and genotoxic effects of two commercial lambda-cyhalothrin formulations that differ in formulation type and in composition: an emulsifiable concentrate (Lambda-Cyhalothrin 5% EC) and a suspension concentrate co-formulated with thiamethoxam (Duer SC). Cytotoxicity was assessed using propidium iodide exclusion, while genotoxicity was evaluated using the cytokinesis-block micronucleus (CBMN) and alkaline comet assays. Lambda-Cyhalothrin 5% EC showed significant cytotoxicity from 500 μM onward, whereas Duer SC induced cytotoxic effects even at lower concentrations beginning at 200 μM, indicating greater cytotoxic potency of the combined formulation. Lambda-Cyhalothrin 5% EC induced a concentration-dependent increase in the number of binucleated cells containing micronuclei, with a significant effect at 50 μM. Duer SC also increased micronucleus frequency but did not differ significantly from the negative control. Proliferation indices remained comparable to controls for both formulations. The comet assay showed that Lambda-Cyhalothrin 5% EC produced significant DNA damage from 10 μM onward, while Duer SC exhibited minimal genotoxic effects. These findings demonstrate that formulation type modifies the toxicological profile of lambda-cyhalothrin.
Domestic cats (Felis catus) are among the most popular pets in the world, with the global domestic cat population generally estimated to exceed 600 million and potentially approach 1 billion when feral populations are included. As hypercarnivores, cats exhibit unique metabolic deficiencies, particularly in phase II conjugation enzymes (e.g., glucuronidation, glycine conjugation), which impair elimination of phenolic xenobiotics including pharmaceuticals, feed additives, and contaminants. Consequently, the European Food Safety Authority (EFSA) Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) recommends an additional default uncertainty factor (UF) of 5 for such compounds. Physiologically based kinetic (PBK) modelling offers a mechanistic approach to refine these default factors using chemical-specific kinetic data and such models for the domestic cat are not currently available to the scientific and risk assessment community. Hence, this manuscript focuses on the development and validation of a generic PBK model for the species Felis catus according to the six-step process from the template of the Organisation for Economic Cooperation and Development (OECD) guidance document on characterisation, validation and reporting of PBK models for regulatory purposes. The model integrates meta-analysed physiological parameters from the peer-reviewed literature and 11 perfusion limited compartments. The model has been validated using chemical-specific inputs for 15 pharmaceuticals using in vitro and in vivo clearances to compare in vivo to in vivo and in vitro to in vivo predictions with the available experimental data for plasma maximum concentration (Cmax) and area-under-the-curve (AUC) values in blood after oral and intravenous exposure. Impact of bioavailability on model performance has also been assessed using conservative default values and reported or estimated values. In addition, global sensitivity analysis using the Sobol method identified the muscle:blood partition coefficient as the dominant parameter influencing model output variance. Overall, the generic PBK cat model performed well and most predictions accounting for bioavailability using in vitro derived clearance yielded 86% of Cmax predictions and 64% of AUC predictions were within 2-3-fold of the experimental data as recommended by the OECD. Future applications and refinements of the model with regard to NGRA of food and feed chemicals are highlighted.