Fruit rarely contains just one pesticide, yet the cumulative risk of cooccurring residues to specific population groups remains poorly understood. Using validated GC‒ECD and UPLC‒MS/MS methods, we quantified 270 pesticides in 305 fruit samples. Residues were detected in 58.7% of the samples, 45.9% of which contained multiple residues; 41 pesticides were found in total, and nine samples (3.0%) exceeded China's maximum residue limits. Single-pesticide assessment identified omethoate as the dominant high-risk compound (RQ of up to 5.37 in children aged 2–12 years). The cumulative assessment classified 1.3% of the samples as high risk, driven mainly by omethoate (54–98% contribution), and 11.1% as moderate risk. Quantum chemical calculations revealed an O–CH3 bond dissociation energy of 77.1 kcal/mol for omethoate, suggesting that intrinsic molecular stability is a plausible mechanism underlying its persistence and risk. Children consistently showed the highest exposure, underscoring the need for age-stratified cumulative risk assessment.

Amphibians are key species in maintaining ecological balance and promoting the development of human activities. However, pesticide use potentially threatens amphibian population survival. The development of agriculture and pesticide exposure are closely linked to the exacerbation of population declines. In recent years, increasing attention has been directed toward the toxic effects of pesticides on amphibians. Nevertheless, the mechanisms underlying pesticide toxicity in amphibians, as well as the challenges faced by research methods, evaluation approaches, and assessment frameworks, remain poorly understood. Herein, we provide an in-depth exploration of the currently known toxic effects of pesticides on amphibians, along with the methods and systems for toxicity research, evaluation, and risk assessment. Furthermore, new strategies are proposed for studying pesticide toxicity, elucidating mechanisms, and improving risk assessment frameworks for amphibians. This work provides valuable insights into the ecological impacts of pesticides on amphibians and offers guidance for the evaluation of such chemicals.

The improper application of pesticides is a critical reason for the population decline of pollination insects. Despite decades of extensive ecotoxicological investigations into pollinators, the majority of studies have primarily focused on the impact of individual pesticides, whereas the combined impacts of multiple pesticide and their underlying mechanisms remain insufficiently understood. This study aimed to investigate the individual and combined toxic effects of imidacloprid (IDP) and epoxiconazole (EXC) on honey bees (Apis mellifera L.) and to further elucidate the associated mechanistic basis by assessing enzymatic biomarkers, gene expression, and gut microbiota. Our acute toxicity results exerted that IDP showed a higher acute toxicity to A. mellifera compared to EXC. There was an acute synergistic effect on the bees when co-exposure to IDP and EXC. The individual and combined exposures significantly affected the activities of CAT, PPO, α-AMS, GST, and CarE, as along with the expressions of genes (caspase-1, CYP4G11, CYP306A1, GSTS3, dorsal-2 and vtg) related to apoptosis, detoxification metabolism, immune response and lifespan. In addition, the 16S rDNA amplicon analysis of honey bees revealed that the abundance and structure of the microbiota were significantly affected, especially after 14 days of exposure. The abundance of the core bacterial genus (Bifidobacterium, Apilactobacillus, and Lactobacillus) have remarkably diminished. Our results would help to elucidate the mechanisms underlying the complex impact of pesticide mixtures on honey bees. Overall, these findings provided worthy reference for effective protection of pollination insects and contributed to the design of guideline for the rational application of pesticide mixtures in agro-ecosystems.

In agricultural ecosystems, pesticides and heavy metals often coexist as understudied threats to ecological health. Although traditional risk assessments focus on single contaminants, the combined effects, particularly on ecologically relevant amphibians such as Pelophylax nigromaculatus, remain poorly understood. Among these pollutants, the neonicotinoid acetamiprid and cadmium are widely detected in aquatic environments, yet their combined hepatotoxicity in adult amphibians has not been adequately investigated. In this study, we aimed to investigate the individual and combined toxic effects of acetamiprid and cadmium in P. nigromaculatus at environmentally relevant concentrations, employing an integrated multi-omics approach in conjunction with biochemical and histopathological evaluations. Individual exposures induced significant metabolic disruption and tissue injury. In contrast, co-exposure triggered a unique adaptive compensation network that maintained redox balance and redirected amino acid and nucleotide metabolism, thereby mitigating inflammatory activation and liver damage. Molecular docking simulations further revealed differential binding interactions of acetamiprid and an acetamiprid-cadmium complex with key compensatory proteins, providing a mechanistic basis for the attenuated toxicity under combined exposures. These results highlight the importance of incorporating mixture toxicity and life-stage-specific responses into ecological risk assessments, reveal unexpected adaptive pathways in amphibians exposed to complex pollutant scenarios, and emphasize the need for more comprehensive evaluations of interactions among environmental contaminants.

The water-level fluctuation zone (WLFZ) serves as a critical buffer that intercepts terrestrial pesticides from entering reservoirs and protects aquatic environmental security. In this study, 78 soil samples were collected via systematic sampling from multiple elevations and typical sites across the Three Gorges WLFZ. Pesticides were extracted by modified QuEChERS and quantified with LC‑MS/MS combining non-targeted screening with targeted quantification to investigate residue profiles, spatial patterns and ecological risks. Six pesticides (prosulfocarb, tolfenpyrad, mandipropamid, pyraclostrobin, acetamiprid, and pyroquilon) were universally detected. Among these, acetamiprid was the most prevailent with a mean concentration of 0.23 ± 0.44 mg kg⁻1. The FLA site exibited the highest average residue level (0.73 mg kg⁻1), substantially exceeding those of the other 11 locations; The maximum concentration of 8.12 mg kg⁻1 was recorded at an elevation of 160 m, compared to the regional average of 0.68 mg kg⁻1. Around 92

In recent years, green manure has attracted increasing attention for its potential in remediating contaminated soils. However, studies investigating how green manure modulates soil microbial communities and alleviates plant metabolic stress under pesticide-contaminated conditions remain limited. In this study, a nicosulfuroncontaminated soil-potato cropping system was established using Vicia villosa as green manure to assess its effects on soil microbial dynamics and plant metabolism. The results showed that green manure significantly enhanced soil enzymatic activities, including urease and sucrase. Compared with the nicosulfuron-only treatment (NI), the green manure-amended treatment (NIGM) promoted the enrichment of functional microbial phyla, such as Proteobacteria and Actinobacteriota, and markedly shortened the pesticide's half-life in soil. Metabolomic analysis revealed that pesticide stress in the NI group inhibited photosynthesis and caused downregulation of the flavonol compound kaempferol. In contrast, the NIGM treatment upregulated metabolites such as N-feruloyloctopamine and cinnamic acid, contributing to the restoration of key pathways including amino acid metabolism and the glutathione cycle. These findings provide new insights into green manure application for enhancing plant stress resilience and promoting sustainable soil remediation.

Thifluzamide (TF) is a widely used fungicide for controlling sheath blight in rice and is frequently detected in aquatic environments. Lead (Pb) may show toxicity to aquatic organisms due to its persistence and bioaccumulation. However, currently studies on the impacts of these two pollutants on aquatic organisms remain limited. To explore their toxic effects and potential risks to amphibians, the present study investigates the single and combined toxicities of TF and Pb at environmentally relevant concentrations on Xenopus laevis tadpoles, based on a multi-omics approach and molecular docking simulations. The results demonstrated that TF and Pb impaired tadpole growth and development, induced oxidative stress and triggered inflammatory responses, and disrupted normal locomotor behavior. Transcriptomic and metabolomic analyses indicated that the signaling pathways (FoxO, MAPK and Cytokine-cytokine receptor interaction) associated with the toxic effects of TF and Pb, as well as metabolic pathways (Histidine metabolism, Purine metabolism, and Alanine, aspartate, and glutamate metabolism) were significantly affected. Molecular docking simulations indicate a higher binding affinity between inflammation-related proteins and their corresponding receptors. Moreover, these adverse effects on tadpoles were more pronounced under combined exposure. In conclusion, we clarified the toxic effects and potential toxic mechanisms of amphibian larvae exposure to Pb and TF. These results provide a scientific foundation for assessing the ecological risks of complex pollutant mixtures in aquatic ecosystems.

Pesticides are recognized as prominent toxicants in aqueous ecosystems, which frequently impact aquatic life. The pyrethroid insecticide lambda-cyhalothrin (LDC) and the triazole fungicide difenoconazole (DFC) are frequently co-detected in many water sources. However, the joint harmful effects of these chemicals on fish are still poorly understood. In this study, a 30-day co-exposure experiment was conducted using LDC and DFC on the hook snout carp (Opsariichthys bidens) to assess their toxic impacts on hepatic tissues. Biochemical assays demonstrated a significant increase in catalase (CAT) and caspase-3 (CASP-3) activities of hepatic cells following exposure to either LDC or DFC alone, as well as their combination. Notably, the combined exposure group exhibited a more pronounced elevation in these enzymatic activities compared to individual exposures, suggesting enhanced oxidative stress and mitochondrial dysfunction. At the molecular level, exposure to both pesticides, either individually or in combination, caused erβ1 and socs3a to be downregulated and cxcl-c1c to be upregulated. Consistent with the biochemical findings, the combined exposure had a more substantial impact on gene expression than the individual exposures, indicating heightened immunotoxic and endocrine-disrupting effects. The synergistic interaction between LDC and DFC suggested that their co-presence exacerbates toxicity in fish liver, revealing underlying mechanisms of oxidative damage, mitochondrial impairment, and immune dysregulation. These findings provide insightful knowledge that could inform chemical regulatory guidelines for pesticide application and management in agricultural settings, aiming to mitigate the environmental impact of these commonly used agrochemicals.

NIs have become prevalent contaminants. Given the limited understanding of residual profiles and toxicological impacts of NIs in different food, assessing their health risks are crucial. This investigation analyze 813 samples of different types from Guizhou Province. Key factors of spatiotemporal distribution patterns, dietary risk of eight NIs were quantified. Acetamiprid, imidacloprid, thiacloprid, thiamethoxam and clothianidin were mainly detected in plant-derived commodities, whereas not have NIs in meat. IMIRPF is correlated with the annual average temperature, the number of tourists, and the amount of pesticide usage. The risk assessment results indicate that although the acute risk associated with NIs residues in these foods is significantly higher than the chronic risk, these risks are acceptable. The results of risk ranking indicated the samples of 91.51 % is low-risk. This study will provide valuable references for the rational use of pesticides, enhancing regulatory strategies, developing comprehensive risk assessment and monitoring frameworks of NIs.

Surface-enhanced Raman spectroscopy (SERS) for simultaneous detection of multiple pesticides in food samples remains a challenge due to the competitive absorption of multiple analytes on SERS substrate. Herein, a combined method of DFT calculation and SERS experiment was utilized to explore the competitive adsorption of thiabendazole and pymetrozine on silver surface. Subsequently, a SERS method was developed for simultaneous detection of thiabendazole and pymetrozine residues in apples. The simulated parameters of pesticide molecules adsorbed on silver cluster showed that competitive adsorption occurred between thiabendazole and pymetrozine on silver surface, which was further validated by the SERS intensities of single and mixed pesticide solutions. The SERS method was successfully established, featuring a satisfactory linearity range of 1.0-30 and 0.5-15 mg/L with the LOD of 0.9 and 0.4 mg/L for thiabendazole and pymetrozine in apple sample, respectively, and the average recovery was 80.02-120.73 %. The findings not only provide a valuable reference for the investigation of competitive adsorption but also offer a reliable method for the simultaneous detection of multiple pesticide residues in food samples.

Current calibration models based on surface-enhanced Raman spectroscopy (SERS) spectra data for pesticide residue detection in foods demonstrate excellent species-specific performance, yet their cross-species applicability remains challenging. This study aimed to build a universal model suitable for in situ detection of pymetrozine residues on two different vegetable species. SERS spectra of cabbage and Chinese cabbage leaves contaminated with pymetrozine were collected, and species-specific and universal models were constructed based on the spectral data using the partial least-squares regression algorithm. The results suggested that species-specific models demonstrated strong predictive performance for estimating the pymetrozine residue concentrations on a specific vegetable but limited accuracy when applied to another vegetable species. However, the universal model exhibited excellent performance for both vegetable species, with an correlation coefficient of prediction set (Rp2) and root-mean-square error of prediction (RMSEP) values of 0.9637 and 0.6225 mg/L for cabbage and 0.9621 and 0.6362 mg/L for Chinese cabbage, respectively. The findings of the study indicate that the universal model is suitable for in situ detection of pymetrozine residue concentrations on two different vegetable species.

Atrazine residues are widely present in agricultural soils and pose a significant threat to sensitive crops such as potato. This study investigated the effects of atrazine and the application of the green manure Vicia villosa on the growth, physiological and biochemical characteristics of potato seedlings, as well as soil environmental factors and the underlying mechanisms. The results indicated that atrazine exposure induced oxidative stress in potato seedling leaves, which increased respiratory activity and enhanced carbohydrate metabolism. Concurrently, oxidative stress disrupted chlorophyll structure. Moreover, atrazine inhibited NADPH synthesis in the nicotinic acid and nicotinamide metabolic pathways, impairing chlorophyll biosynthesis and thereby reducing photosynthetic activity, ultimately compromising carbohydrate biosynthesis. These metabolic disruptions suggest that atrazine exposure severely interferes with energy production, metabolism, and biosynthetic processes in potato seedlings, significantly inhibiting growth and potentially resulting in plant death. In addition, atrazine adversely affected soil environmental conditions by altering soil physicochemical properties and reducing enzyme activities, microbial diversity, and microbial community abundance. The application of Vicia villosa partially alleviated these negative effects; however, the findings also highlight the irreversible damage caused by atrazine to potato seedlings. This study elucidates the phytotoxic effects of atrazine and its potential mechanisms, providing a theoretical basis for addressing the toxic impacts of persistent herbicides in crop production.

With the continuous impact of human activities on the ecological environment, buprofezin and cadmium are frequently detected in soil, sediment, and aquatic environments, posing ecological risks to non-target aquatic organisms. However, limited research exists on the toxic effects and mechanisms of action of these pollutants on aquatic organisms. This study used Xenopus laevis tadpoles as model organisms to experiment with buprofezin and cadmium. Through biochemical parameters and multi omics analysis methods, the single and combined toxicity mechanisms were explored. The experiment used environmentally relevant exposure levels to monitor the growth indicators, movement parameters, oxidative stress biomarkers of tadpoles, and conducted metabolomics and transcriptomics analysis. The results indicate that cadmium inhibits the growth of tadpoles, leading to a decrease in weight, and mixed exposure has a similar effect. Under dark conditions, buprofezin and cadmium significantly alter the swimming behavior of tadpoles, decreasing distance and average speed. Moreover, tadpoles exposed to buprofezin and cadmium experienced oxidative stress, which was reflected in increased levels of malondialdehyde and decreased activities of superoxide dismutase and glutathione S-transferase. Metabolomics and transcriptomics results showed that the combined exposure group produced more differentially accumulated metabolites and differentially expressed genes than the single exposure group. These genes and substances mainly affect the energy metabolism and signal transduction processes of tadpoles. In summary, buprofezin and cadmium interfere with gene expression and alter metabolite levels in tadpoles. This study reveals the combined toxicity of buprofezin and cadmium at environmentally relevant exposure levels. The research results provide toxicological evidence for the risk assessment of environmental pollutants and offer new insights into the effects of complex mixtures.

Agrochemical organosilicone spray adjuvants, widely applied in agroecosystems, are suspected to pose environmental risks based on recent studies. In response to these concerns, we have purified the active components of these adjuvants, specifically a series of oligomers of hydroxy(polyethyleneoxy) propyl-heptamethyl trisiloxane (TSS-H), including ethylene oxide (EO) units ranging from 4 to 12. For analysis of TSS-H in zebrafish and water, a method utilizing high-performance liquid chromatography coupled with high resolution mass spectrometry (HPLCHRMS) was developed. The findings revealed that at total TSS-H concentrations of 0.04, 0.4, and 4 mg/kg, the average recovery rate of TSS-H containing 4 to 12 EO units in zebrafish ranged from 81.2 % to 107.2 %, with relative standard deviations (RSDs) between 0.1 % and 8.4 %. The 96-hour LC50 of TSS-H in zebrafish was 6.71±0.04 mg/L. At exposure concentrations of 0.0671 mg/L and 0.671 mg/L, TSS-H oligomers reached a steady state in zebrafish within 14-21 days, with concentrations ranging from 0.008 to 0.05 mg/kg and 0.018 to 0.348 mg/kg, respectively. Bioconcentration factor (BCF) values for TSS-H oligomers (4-12 EO units) were 1.87-11.59 at the low dose and 0.79-9.19 at the high dose after 14 days. The elimination half-lives of the oligomers were <2.37 days during the elimination phase. Non-targeted metabolomic analysis revealed both TSS-H doses significantly impacted lipid and amino acid metabolic pathways, with disruptions largely associated with abnormal energy metabolism, oxidative stress, and alterations in membrane composition. These findings are critical for assessing the food safety and environmental risks of agrochemical organosilicone spray adjuvants in fish.

Coix seed (C. lacryma-jobi L.) is used as a medicinal and edible grain in China. In this study, nontargeted screening of pesticide residues in 126 coix seed samples was conducted in Guizhou Province, southwestern China. The 39 pesticides screened were detected and quantified using a liquid chromatography-tandem mass spectrometry method after modified QuEChERS pretreatment. The samples were extracted with an acetonitrile-formic acid (98:2;v/v) mixture. Method linearity was confirmed between 0.001-1 mg kg-1, with significant correlation coefficients (R2 >0.980). Mean recoveries were 72.2%-115% and the limit of quantification was 0.01 mg kg-1. Intraday repeatability and inter-day reproducibility ranged from 0.2% to 17.8% and 0.3% to 18.0%, respectively. Risk assessment showed that Hazard Quotient (HQ) and Hazard Index (HI) were well below 1. This method can help to regulate pesticide use in cultivation areas and improve the quality and safety of coix seed products.

Given dimethachlon's known nephrotoxic effects, it is critical to conduct a thorough investigation to identify its metabolites in crops, residue levels, processing factors, toxicity, and bioactivity of both the parent compound and its metabolites. In this study, high-performance liquid chromatography coupled with high-resolution mass spectrometry (HPLC-HRMS) was used to unequivocally identify 4-(3,5-dichloroanilino)-4-oxobutanoic acid (DCBAA) and 3,5-dichloroaniline (3,5-DCA) as metabolites produced by the application of dimethachlon to rice and tomato. During the degradation period, the maximum residue levels of DCBAA and 3,5-DCA in crops were 14.64 % and 19.03 % of the parent compound dimethachlon, respectively. Polishing and cooking paddy rice, as well as washing tomatoes, effectively removed over 87 % of dimethachlon. Processing tomatoes into tomato sauce increased the concentrations of dimethachlon and its two metabolites, while cooking rice selectively increased the concentration of DCBAA. A dietary risk assessment, based on total residues and adjusted for processing factors, indicated that human exposure to dimethachlon was within acceptable limits. Ecological Structure Activity Relationships (ECOSAR) analysis showed the acute toxicity ranking for fish, daphnia and earthworms was: 3,5-DCA > dimethachlon > DCBAA.

Given the widespread presence of imidacloprid in aquatic environments and the limited research on its impact on amphibian renal health, in this study, we investigated the effects of this commonly used neonicotinoid insecticide on kidney function and molecular mechanisms in Xenopus laevis. Employing a 28-day exposure model, histopathological changes and enzymatic responses induced by two concentrations of imidacloprid were examined, along with gene expression alterations and metabolic disruptions at environmentally relevant levels. The results highlighted significant renal histopathological damage and changes in key enzymes involved in oxidative stress and neurotoxicity, such as superoxide dismutase, glutathione S-transferase, and acetylcholinesterase. Transcriptomic and metabolomic analyses elucidated profound alterations in gene expression and metabolic profiles, particularly affecting carbohydrate, amino acid, and purine metabolism pathways. This study demonstrates the dual role of metabolic adaptations-serving both protective functions and potentially leading to long-term detrimental effects under continuous exposure. These findings underscore the need for cautious management of neonicotinoid usage to mitigate environmental impacts on aquatic wildlife, particularly amphibians, and inform conservation strategies.

Dinotefuran has been detected worldwide due to its widespread use, high water solubility and persistence, poses a threat to aquatic organisms. This study investigates the potential hazards of dinotefuran and its enantiomers to Xenopus laevis (X. laevis) tadpoles through bioaccumulation and elimination experiments at environmental concentrations. S-dinotefuran exhibited lower acute toxicity in tadpoles, while R- and Rac-dinotefuran demonstrated moderate toxicity. Despite the low levels of dinotefuran and its enantiomer (0.106-1.20 mg/kg) in the tadpoles, which are eliminated after 14 days, dinetofuran caused growth disruption, oxidative stress, inflammation, and neurotoxicity, indicating persistent toxicity. Key regulatory pathways associated with dinotefuran-induced stereoselective toxicity were identified: R-dinotefuran influenced amino acid metabolism, GnRH pathway, and cytokine-cytokine receptor interaction; Rac-dinotefuran affected the Wnt, TGF-β and NOD-like signaling pathways; and S-dinotefuran significantly affected MAPK, FOXO and C-type lectin receptor signaling pathways. Molecular docking revealed strong binding affinity of R-dinotefuran to acetylcholine, thyrotropin, glutamate, and serotonin (5-HT) receptors. Risk assessments showed that dinotefuran poses acute (1.3 < RQ < 213) and chronic (1 < RQ < 50) risks to aquatic organisms. These findings provide valuable references for the safety assessment of dinotefuran in aquatic amphibians and offer new insights for the ecological assessment of chiral pesticides.

As an alternative to traditional pesticides, sulfoxaflor (SFX) is a sulfoximine insecticide with the same mechanism of action as neonicotinoid insecticides (NNIs). However, increasing evidence suggests that SFX poses a threat to aquatic organisms. To investigate the toxic effects and potential risks in amphibians, bioaccumulation and elimination experiments were conducted at environmentally relevant concentrations. The results indicate that although SFX exhibits low acute toxicity and accumulation, it demonstrates neurotoxicity and endocrine-disruptive properties. SFX alters regulatory patterns of growth-related genes and interferes with the regulation of thyroid hormones and its genes, promoting the tadpoles' growth. Additionally, SFX induces oxidative stress, leading to inflammation and immune regulation in the tadpoles. It also affects neurotransmitter transmission as well as the genes associated with neural synapses, receptor, and signal transmission and interferes with tadpole behavior. These toxic effects persisted until the elimination stage. Compared with other NNIs, SFX has the most binding sites with AChR and a weak interaction, and binding to β-agonists is similar in molecular docking. Risk assessment suggests that SFX has a potential risk and impact on aquatic amphibians, which may be underestimated. The result provides valuable reference and new perspective for the ecological safety assessment and supervision of SFX, NNIs, and insecticides of low acute toxicity.