N-(phosphonomethyl)glycine (glyphosate) is the most sprayed herbicidal chemical in the world. Although glyphosate is considered safe in humans and higher animals due to its targeting of the shikimate pathway found only in plants and microorganisms, recent in vivo and in vitro studies show evidence of toxicity across a range of systems, including the vertebrate brain. Given that developing brains with immature blood-brain barriers can be especially sensitive to environmental contaminants, here we test the effects of embryonic exposure to environmental levels of glyphosate or its commercial formulation, Roundup®. Embryonic zebrafish exposed to an environmental concentration of Roundup (10 µg/L acid equivalent) from 10 to 48 hours postfertilization (hpf) showed defects in morphology, respiratory capacity, and hatching at 48 hpf as well as changes in locomotion, light-startle response, and thigmotaxis behaviors at 5 days postfertilization (dpf), with some effects lasting in the juvenile. To understand neurodevelopmental changes underlying these behavioral abnormalities, we tested for changes to the timing of neurogenesis and conducted bulk RNA sequencing. We found increased neurogenesis and uncovered dysregulation of axonogenesis pathways, which was confirmed using immunohistochemistry. Finally, to test if axonal deficits might manifest as dysregulation of neuronal circuits, we observed changes in phospho-extracellular signal-regulated kinases levels and Ca2+ dynamics as indicators of activity disruptions at 5 dpf following embryonic exposure to Roundup. These results suggest that Roundup at current environmental levels may not be safe for organismal exposure, and glyphosate toxicity warrants closer attention.
To address the challenge of cooperative roundup of maneuvering targets under limited perception, this paper proposes TransMARL, a transformer-based multi-agent reinforcement learning framework for observation-constrained coordination. The roundup task is formulated as a decentralized partially observable Markov decision process (Dec-POMDP), together with a local observation model and a dynamically updated interaction graph. The proposed framework combines a graph feature encoding module with a policy execution module to support decentralized decision-making under partial observability. A task-informed reward function is designed to encourage angular coverage, target approach, formation uniformity, and collision avoidance. In addition, the transformer depth is adaptively adjusted according to the team size as an empirically motivated design choice to balance representational capacity and computational cost. Experimental results in a 2D obstacle-free simulation environment show that, under the evaluated settings, TransMARL achieves competitive and often improved performance relative to the selected baselines, especially under constrained sensing radii. These results suggest that the proposed framework is a practical and scalable approach for cooperative control in observation-constrained multi-agent roundup scenarios, while its broader generalization and formal theoretical characterization remain to be further studied.
The widespread use of glyphosate-based herbicides (GBHs), including formulations such as Roundup, has raised toxicological concern due to their capacity to induce oxidative stress, inflammatory signaling, and mitochondrial dysfunction leading to hepatocellular injury. The present study investigated whether coenzyme Q10 (CoQ10) confers hepatoprotection against GBH exposure through modulation of oxidative stress-driven P2X7/NLRP3 inflammasome signaling and downstream inflammatory and apoptotic pathways. Fifty male ICR mice were randomly assigned to five groups (control, sham, CoQ10, GBH, and GBH + CoQ10) and treated by oral gavage for 49 days with Roundup (500 mg/kg/day), CoQ10 (200 mg/kg/day), or their combination. Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP) levels were assessed along with hepatic oxidative stress markers malondialdehyde (MDA), glutathione (GSH), total antioxidant status (TAS), total oxidant status (TOS), and oxidative stress index (OSI). Liver injury was evaluated histologically, while proliferative and inflammatory responses were analyzed by immunohistochemistry. Hepatic expression of inflammasome- and apoptosis-related genes (P2X7, NLRP3, caspase-1, IL-1β, NF-κB, TNF-α, caspase-3, and Bcl-2) was quantified by RT-qPCR. GBH exposure markedly increased serum liver enzymes, disrupted oxidative balance, and induced necroinflammatory liver injury with Kupffer cell activation and hepatocellular nuclear alterations. These changes were associated with increased PCNA and COX-2 immunoreactivity, activation of the P2X7-NLRP3 inflammasome axis, and a proapoptotic transcriptional profile. CoQ10 coadministration attenuated biochemical, histopathological, inflammatory, and apoptotic alterations, although several parameters remained incompletely normalized. In conclusion, CoQ10 provides partial but meaningful hepatoprotection against GBH-induced liver injury, highlighting P2X7-NLRP3-linked oxidative inflammation as a key mechanism in GBH hepatotoxicity.
Glyphosate is the most widely used pesticide globally, raising concerns about its environmental persistence and biological impacts. Therefore, monitoring pesticide use is essential for assessing agricultural practices and the risks to human health associated with chemical use. This research examined glyphosate contamination in water (40 samples) and soil (28 samples) from northeastern Romania, an important agricultural region. Glyphosate concentrations in environmental water and soil samples were quantified using a spectrophotometric method based on ninhydrin derivatization, with good linearity over the concentration range 1-30 µg/mL (R2 = 0.9981). Glyphosate was detected at concentrations above the LOQ in one water sample. Also, the study proposes a UHPLC-MS/MS method for the confirmation of glyphosate presence in the analyzed sample. Additionally, this study contributes to the characterization of the toxicity profiles of glyphosate and a commercial glyphosate-based formulation (Roundup®) in primary human gingival fibroblast (hGF) cell lines. The commercial product Roundup, containing glyphosate, exhibited cytotoxicity similar to that of the active compound at low and intermediate doses; a significant cytotoxic effect was observed at a maximum concentration of 1 mM, with prolonged exposure. These findings demonstrate minimal cytotoxicity under the examined conditions and underscore the need for dose- and time-dependent assessments to evaluate the biological impact of herbicide formulations.
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Since glyphosate has been widely used in agriculture, it has frequently been detected in water bodies and has posed risks to environmental quality and human health. This study investigated glyphosate adsorption by commercial adsorbents (zeolite and activated carbon) and industrial residues (furnace slag, burning ashes, and foundry sand). Initial studies assessed the influence of pH (4, 7, and 10) and surface treatments with aqueous solutions of CuSO4, SDS, AgNO3, Fe-(NO3)3, CTAB, and ZnO on glyphosate removal. Among all materials and treatments, untreated burning ashes showed the highest removal efficiency and were selected for dosage, kinetic, and isotherm investigations. Glyphosate adsorption onto burning ashes was pH-insensitive and achieved 100% removal with the detection limit of 0.025 mg·L-1. In the dosage study (at an initial glyphosate concentration of 5 mg·L-1), 100% removal was reached when the ash dose was 25 g·L-1 and 87.91% when the dose was 1.25 g·L-1. The dose of 1.25 g·L-1 was defined as optimal not only because it met drinking water regulatory limits but also because it minimized material consumption. Regarding kinetics and equilibrium, some studies indicated that glyphosate adsorption equilibrium in ash was reached after 8 h with a maximum adsorption capacity of 5.39 mg·g-1. The Avrami kinetic model and the Temkin isotherm model exhibited the best fit to the experimental data. Glyphosate and AMPA were quantified by liquid chromatography-mass spectrometry (LC-MS) after derivatization. Results showed that burning ashes are capable of removing >95% of the initial concentration of up to 5 mg·L-1 glyphosate. Conclusions of this study indicate that the use of waste material is a promising and sustainable alternative for the removal of glyphosate from aqueous solutions.
Glyphosate (GLY) and Atrazine (ATZ) rank among the most widely used herbicides globally and frequently co-occur in coastal waters. These substances threaten the physiological integrity of filter-feeding bivalves like Mytilus galloprovincialis, which bioaccumulate contaminants, with consequent implications for the overall health of coastal ecosystems. Mussels were exposed for 14 days under controlled laboratory conditions to individual herbicides and their mixture. Biomarkers of metabolic function (electron transport system, ETS), antioxidant defense (SOD, GPx, TAC), detoxification (GST, CbE), oxidative damage (lipid peroxidation, LPO), and neurotoxicity (AChE) were analyzed in the digestive gland and gills. Furthermore, the transcriptional expression of stress-response genes (MT10, MT20, HSP70) and the DNA-damage marker p53 were analyzed via RT-qPCR to elucidate the molecular mechanisms of toxicity. The combined exposure to GLY and ATZ significantly elevated ETS activity and LPO levels while disrupting antioxidant and detoxification enzymes in both tissues, with responses most evident under the mixture treatment. At the molecular level, the combined treatment induced a significant upregulation of all tested genes, with p53 and HSP70 showing the most pronounced increases, indicating significant genotoxic and proteotoxic impairment. AChE inhibition in these organs indicated sublethal neurotoxicity. These findings reveal the complex toxicological effects of combined herbicide residues at environmentally relevant levels, providing data for ecological risk assessment in contaminated coastal ecosystems.
Here, we present a protocol to describe the bacteriome of the intestinal content of toxin-exposed fallow deer. We describe steps for measuring fecal mycotoxin (deoxynivalenol, zearalenone, fumonisin B1, and aflatoxin B1) levels using liquid chromatography-mass spectrometry, as well as serum glyphosate. We then detail a short-read shotgun DNA sequencing-based bioinformatic pipeline for the toxin level-associated analysis of the bacteriome and resistome and the construction of metagenome-assembled bacterial genomes. This protocol has potential applications in further toxin level-associated metagenome studies. For complete details on the use and execution of this protocol, please refer to Tóth et al.1.
A biomimetic dual-function nanozyme based on a cerium-based metal-organic framework (UiO-66(Ce)-3) with engineered oxygen vacancies was developed for efficient hydrolysis and sensitive detection of glyphosate. Acetic acid-triggered "missing-linker" defects partially disrupted the Ce-O coordination environment, generating oxygen vacancies and exposing Ce3+/Ce4+ redox sites. The Ce4+ centers polarize the phosphate bond, while the adjacent Ce3+ sites facilitate the nucleophilic cleavage of the C-P bond. Under alkaline conditions, UiO-66(Ce)-3 showed excellent organophosphorus hydrolase-like activity with a kcat of 57.77 s-1, an SA of 0.70 U·mg-1, and a kcat/Km of 152.04 × 103 M-1 s-1. The nanozyme effectively mitigated the glyphosate-induced toxicity in plants with a survival rate of 95%. Furthermore, the oxygen vacancies endowed pH-switch enzyme-like activity. With the pH at 4.0, UiO-66(Ce)-3 showed oxidase-like activity, which could realize high-sensitivity detection of glyphosate via point-of-care testing (LOD of 11.66 ng·mL-1) with good stability and reusability. And it showed a good recovery of 93.51%-110.19% in complex matrices, validation against the HPLC. The prepared pH-switchable nanozyme provides a novel approach to the detection and degradation of organophosphorus compounds.
Weeds are a major constraint to peanut (Arachis hypogaea L.) production, causing substantial yield and quality losses. This study was conducted during the 2020 and 2021 growing seasons to evaluate the effects of different pre- and post-emergence herbicides on weeds and peanut yield under field conditions. The experiment was arranged in a randomized complete block design with four replications. Treatments included one non-selective burndown herbicide (glyphosate), two pre-emergence herbicides (pendimethalin and dimethanamid p), four post-emergence herbicides (bentazon, quizalofop-p-ethyl, imazamox, clethodim), applied alone or in selected sequences. Weed control varied significantly with herbicides type, weed species, and evaluation time. Control levels increased markedly at 21 and 35 days after treatments (DAT) but declined by 105 DAT, indicating reduced residual activityand late weed emergence. The post-emergence combinations of bentazon + quizalofop-P-ethyl and bentazon + imazamox provided the highest weed control (88.2% and 88.9%, respectively) and significantly reduced weed dry biomass. Weed interference reduced peanut yield by approximately 60-70% copared with the weed-free control. All effective herbicide treatments significantly reduced weed biomass and prevent yield loss, leading to substiantially higher than the weedy control. Overall, the findings indicate that appropriate herbicide selection and optimized application timing play a critical role in achieving effective weed control and minimizing yield losses in peanut production.
A major concern about growing transgenic glyphosate-tolerant (GT) soybean (Glycine max) is the potential for gene flow to the wild relative Glycine soja, through hybridization and introgression. This could create new and serious weed-control problems and pollute wild soybean germplasm resources. This risk deserves particular attention in northern China, which is the origin center for soybean, a key reservoir of wild genetic diversity, and a major region of soybean cultivation. Our previous research on F1-F3 hybrids of three wild soybeans (IMBT, JLBC-1, HJLHRB-1) from northern China and GT soybean showed hybrid fitness increasing over successive generations, with two distinct segregation patterns observed. Consistent with these two segregation patterns, two types of F4 hybrids were examined: trait-segregating hybrids (IMBT F4) and segregation-distorted hybrids (JLBC-1 F4, HJLHRB-1 F4). Both displayed increased relative composite fitness compared with their wild progenitors, driven by enhanced vegetative growth (vine growth habit, higher biomass) and reproductive capacity (seed number and weight). Specifically, trait-segregating IMBT F4 hybrids exhibited various traits among and within sister groups. Segregation-distorted JLBC-1 F4 and HJLHRB-1 F4 displayed uniform, weedy-like phenotypes. If initial hybridization occurs, GT transgenes could introgress into wild soybean populations to produce fitness-enhanced trait-segregating and segregation-distorted F4. Trait-segregating may have a major role in hybrids adapting to a wider range of ecological niches, whereas segregation distortion may promote establishment in wild soybean habitats. Thus, effective measures are needed to prevent gene flow from GT soybean released in regions sympatric with wild soybean. © 2026 Society of Chemical Industry.
Environmental quality associated with agricultural intensification alters the availability and nutritional quality of floral resources, with consequences for pollinator health. Here, we evaluated honey bee (Apis mellifera) colonies as environmental sentinels in two contrasting agroecosystems of the Argentine Pampas: a crop-dominated landscape and a livestock-dominated landscape. Within each setting, colonies received either protein supplementation or no supplementation. An integrative approach combining biological, pathological, toxicological, and environmental indicators was applied to assess colony responses across seasons. Colony condition was characterized using hemolymph protein concentration, prevalence of Varroa destructor and Nosema spp., brood area development, survival probability, and herbicide residues (glyphosate, glufosinate, and 2,4-D) in hive products. After accounting for year, supplementation, and Varroa treatment, colonies located in the livestock-dominated landscape exhibited higher hemolymph protein levels, greater survival probability, and lower herbicide residues. In contrast, colonies from the crop-intensive landscape showed reduced protein concentrations, increased Nosema prevalence, and detectable herbicide residues in honey and pollen. Principal component analysis identified glufosinate concentration in honey and reduced hemolymph protein levels as the main variables discriminating between environments. Together, these results indicate patterns consistent with an association between landscape composition, herbicide exposure, and honey bee physiology and survival within the studied agroecosystems, highlighting the potential role of environmental quality and floral diversity in shaping pollinator health.
For wastewater remediation, efficiently avoiding harmful co-product formation while achieving carbon source recovery remains a challenge worthy of attention. Herein, we deliberately bridge a polar pyridinic moiety into carbon nitride (PDCN) to construct a lattice distortion structure with interfacial charge asymmetric polarization, achieving complete photodegradation of glyphosate (Gly) wastewater into sarcosine (>99% selectivity) and CO (1166.2 µmol g-1 h-1) within a short timeframe. The introduction of polar pyridine not only causes the C─N─C bond angle of PDCN to distort, generating a local polarization field, but also induces a dipole field, which achieves effective separation and directional transfer of photogenerated carriers, respectively. This directed electron localization enhances O2 adsorption by PDCN's electron-rich pyridine regions and its conversion into reactive •OH for cleaving C─P bonds. Furthermore, PDCN's electron-deficient heptazine region activates Gly's α-C─H bond via electrostatic interactions with phosphate groups, promoting the selective cleavage of C─P bonds in Gly and avoiding the formation of toxic aminomethylphosphonic acid. Glycine and other co-products show no significant impact on the growth of organisms like fathead minnows and mung beans. This work establishes a viable perspective for efficient wastewater purification and carbon recovery enabled by the synergistic interaction of localized polarization and dipole fields.
Genetically modified crops have the advantages of high yield, strong resistance and low cost, and are currently cultivated in many countries around the world. During our research, we discovered a type of semiwild soybean that emerged due to genetic drift at a transgenic test site. However, the potential risks of genetically modified crops in the process of environmental release, such as gene drift, weeding, impact on soil ecological environment, have raised concerns whether genetically modified glyphosate-resistant soybeans can be widely promoted. This study investigated the biological characteristics of genetically modified glyphosate-resistant semiwild soybean from various perspectives. Based on this, the study further evaluated the survival competitiveness of genetically modified glyphosate-resistant semiwild soybean against weeds. These results indicate that this transgenic material retains some characteristics of wild type soybean while exhibiting unique advantages in physiological functions and adaptability.
Zebrafish (Danio rerio) have become the pre-eminent vertebrate model in aquatic ecotoxicology, yet the evidence base on pesticide bioavailability, bioaccumulation, and bioconcentration in this species is characterised by profound methodological fragmentation that limits the derivation of reliable risk estimates. This review critically appraises the primary literature across five major pesticide classes, namely organophosphates, pyrethroids, organochlorines, triazole and succinate dehydrogenase inhibitor (SDHI) fungicides, and herbicides, and argues that the field's central limitation is not a deficit of primary studies but rather a systematic failure to implement methodological standards capable of generating internally consistent, cross-comparable toxicokinetic data. Bioconcentration factors (BCFs) in zebrafish span four orders of magnitude, from less than 2 for glyphosate to over 1300 for chlorfenapyr, with log Kow as the primary predictor; yet BCF values derived from embryo stages cannot be extrapolated to adults given the profound life-stage dependency of CYP450 metabolic capacity. A recurring pattern of nominal-concentration reporting, pure active-ingredient testing, single life-stage design, and single-compound exposure has generated a body of evidence that diverges systematically from real-world pesticide exposure conditions. Quantitative analysis of methodological bias reveals that biotransformation alone can reduce internal concentrations by 5- to 90-fold; sediment partitioning can confine pyrethroid bioavailability in the water column to as little as 0.07% of total added mass; and formulation adjuvants can redirect pesticide distribution entirely between water and sediment compartments. A cross-study quality assessment of the ten primary studies forming the evidence core of this review shows that the majority score 7 or below on a 10-point methodological adequacy scale, with no study simultaneously verifying concentrations, capturing tissue-specific BCFs, tracking metabolites, and incorporating realistic mixture or formulation conditions. This review provides the first integrated, methodologically-focused, quantitative synthesis of these issues in zebrafish, identifies five priority knowledge gaps, and proposes a minimum standards framework for future zebrafish pesticide bioaccumulation studies.
Given the global prevalence of glyphosate as a broad-spectrum herbicide, the monitoring of its residues is essential for protecting environmental and food safety. In this study, Hemin-UiO-66-NH2, with peroxidase (POD)-like activity, was successfully developed by immobilizing hemin, which is the active center of natural horseradish peroxidase, onto UiO-66(Hf)-NH2. Using UiO-66-NH2 as the support matrix, hemin was effectively loaded and stabilized, leading to a remarkable enhancement in its POD-like activity. Hemin-UiO-66-NH2 was found to efficiently catalyze the colorimetric reaction of 3,3',5,5'-tetramethylbenzidine (TMB) in the presence of H2O2. A colorimetric detection method for glyphosate was established based on the specific binding between glyphosate and Hemin-UiO-66-NH2, so that a decrease in the POD-like activity of Hemin-UiO-66-NH2 can be observed. The response curve exhibits a dual-segment linear range from 0.4 to 17.5 μM (R2 = 0.992) and 17.5 to 1000 μM (R2 = 0.993), with detection limits of 0.13 μM (σ/k = 3, n = 11). This work not only provides valuable insights for high-performance POD-like nanozymes but also offers a novel and reliable strategy for the convenient and sensitive detection of glyphosate, demonstrating great potential for applications in environmental monitoring and food safety inspection.
Glyphosate is the most extensively applied systemic herbicide worldwide, yet its safety remains under scrutiny, with ongoing investigations into potential carcinogenicity. Epidemiological studies associate chronic glyphosate exposure with elevated risks of non-Hodgkin lymphoma and possible endocrine disruption, emphasizing the need for sensitive detection methods. Here, we report a handheld enzymatic biosensor, GlyphoSense Chip, for direct, rapid detection of underivatized glyphosate in drinking water. The device integrates a photodiode-based CMOS chip with an engineered glyphosate N-acetyltransferase and a colorimetric reaction, achieving a sensitivity of 38 µV·mL µg-1·s-1 and quantification within one minute. Biosensor response was linear over 0.016-12.5 µg mL-1 (R2 = 0.993) with a detection limit of 0.028 µg mL-1. Recovery analysis in fortified tap water yielded relative standard errors of 1.2-5.8%, and results were statistically indistinguishable from quantitative mass spectrometry (p > 0.05). This work establishes a robust, field-deployable platform for glyphosate monitoring in water resource safety applications.
Glyphosate, an anionic organophosphorus herbicide, is frequently detected in surface waters alongside other emerging contaminants (ECs). Effective removal strategies are essential to reduce their environmental impact. This study synthesized a metal organic framework (MOF) called Zeolitic Imidazolate Framework-8 (ZIF-8), a material recently recognized for its exceptional versatility, and employed it for the removal of glyphosate from aqueous systems via adsorption. ZIF-8 particles were synthesized using zinc nitrate hexahydrate as the metal source, 2-methylimidazole as the ligand, and methanol as the solvent. ZIF-8 was characterized by XRD, BET, zeta potential, and FTIR. Adsorption data were well described by both Langmuir and Freundlich models (R 2 = 0.99), indicating contributions from monolayer and multilayer adsorption mechanisms. The maximum adsorption capacity was 67.67 mg/g at 308 K according to the Langmuir model. The adsorption mechanism involved a combination of physisorption and chemisorption and was influenced by coexisting chloride anions. Thermodynamic analysis indicated that glyphosate adsorption is endothermic and involves chemical interactions between glyphosate and ZIF-8. FTIR spectra showed new bands corresponding to the phosphoryl group after adsorption, confirming glyphosate binding. Overall, this study demonstrates that ZIF-8 is a promising adsorbent for the removal of ECs, offering an effective strategy for environmental protection.
Pesticides and climate change are significant drivers of biodiversity loss, with their interaction likely increasing their detrimental consequences. Ectotherms, such as lizards, are among the most vulnerable non-target organisms given their occurrence in agricultural habitats. Combined exposure to pesticides, and increased temperature can alter the energetic cost for maintenance as most physiological processes, including detoxification, are temperature-depended. However, their combined effects on the rate of metabolic processes of lizards are currently unknown. We conducted a biologically relevant ecotoxicological experiment to assess the combined effects of glyphosate, the most widely used herbicide worldwide, and temperature on the level of expression of standard metabolic rates (SMR) of male wall lizards Podarcis bocagei. Exposure to glyphosate occurred every second day, for 21 days, through contaminated mealworms previously injected with 5 μl of glyphosate sub-lethal dosage (0.5 mg/kg bw). We measured post-exposure SMRs through open-flow respirometry at three different temperatures relevant for the species (18, 25, 29 °C). Across the tested temperature range, SMR increased with temperature, glyphosate-exposed animals displayed elevated SMRs and heavier animals showed greater increase of energy expenditure. The combined mass and exposure effects may pose a threat for populations occurring in agricultural environments, as the largest animals have the highest reproductive output. This evidence is valuable for future risk assessment and conservation prioritization of species which bioenergetic processes may be particularly sensitive to glyphosate exposure.