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Adverse Effects of Glyphosate and Microcystin-LR on Fish Health: Evidence from Structural and Functional Impairments in Zebrafish Gills

Yidan Zhang,Han Hu,Linmei Song,Zhihui Liu,Junguo Ma,Xiaoyu Li

Simple SummaryThe coexistence of two environmental pollutants, glyphosate (GLY, a widely used herbicide) and microcystin-LR (MC-LR, a toxic byproduct from harmful algal blooms) has become an issue that cannot be ignored. The adverse effects of both individual and combined exposures to GLY and MC-LR on aquatic organisms warrant increasing attention. This study demonstrates that exposure to GLY and MC-LR, either individually or in combination, can induce toxic effects on fish gill structure, potentially impairing their physiological function and ultimately threatening fish survival. Overall, the findings of this study not only enhance our understanding of the detrimental impacts of these pollutants on aquatic animals but also provide valuable insights for ecological and health risk assessments.Glyphosate (GLY) and microcystin-LR (MC-LR) frequently co-occur in natural water bodies. In this study, a subacute exposure test was conducted on zebrafish treated with 3.5 mg/L GLY and 35 μg/L MC-LR, individually and in combination, for 21 d to determine their effects on the gills of zebrafish and their potential mechanisms. The hematoxylin and eosin staining and scanning electron microscopy examination results showed that GLY and MC-LR exposure caused structural damage to gills. Biochemical analysis revealed Na+-K+-ATPase activity decreased, and the levels of reactive oxygen species, 8-hydroxy-2′-deoxyguanosine, and malondialdehyde increased, inducing oxidative damage to DNA and lipids of gills. Meanwhile, the inflammatory and immune function of the gill was significantly influenced, as evidenced by the alteration of the expression of tumor necrosis factor-α, interleukin-1β, complement 3, and immunoglobulin M. RNA-seq results revealed that GLY and/or MC-LR treatment induced transcriptional changes in the fish gills, which may affect various biological functions, and the lipid metabolism disruption potentially involved in the aforementioned process. Integrating histopathological, biochemical, and transcriptomic analyses, this study revealed that both individual and combined exposures to GLY and MC-LR had adverse effects on zebrafish gills, with combined exposure appears to result in more pronounced adverse effects, potentially compromising fish health.

Preparation of a β-CD@Fe3O4/KHA/CMC/AM/MAH hydrogel and its slow-release herbicidal properties against glyphosate

Jie Song,Yingnan Zhou,Lijun Chen,Zhiwen Yuan,Yuhua Niu,Xi Li,Liangxian Huang

This study employed β-cyclodextrin (β-CD) for the hydrophilic modification of iron tetraoxide (Fe3O4) magnetic materials. Subsequently, potassium humate (KHA) and carboxymethyl cellulose (CMC) were used as raw materials. A β-CD@Fe3O4/KHA/CMC/AM/MAH hydrogel with magnetic properties which can be easily recycled was synthesized for the slow release of glyphosate (GLY). FTIR, XRD, TG and VSM analyses demonstrated that the hydrogel possesses good thermal stability and magnetic properties with a saturation magnetization strength of 29.77 emu g−1. The equilibrium solubility of the hydrogel in pH = 7.4 and pH = 8.5 buffer was 24.58 g g−1 and 25.83 g g−1, respectively. The maximum release of GLY was recorded at 63.31% and 67.78%, with the drug release behavior under different loading gels following the Fickian diffusion mechanism. A pot planting experiment indicated that the water content of plants treated with the β-CD@Fe3O4/KHA/CMC/AM/MAH drug-loaded gel decreased by 51.69%. Additionally, soluble protein levels decreased to 2.69 mg g−1, chlorophyll content decreased to 0.486 mg g−1, SOD levels decreased to 43.81 U g−1, and MDA increased to 4.02 μmol g−1, effectively inhibiting the growth of grass and providing a theoretical basis for achieving slow drug release.

Co-exposure to polystyrene nanoplastics and glyphosate promotes intestinal apoptosis in mice via intestinal barrier impairment and immunoinflammatory dysregulation

Beining Wu,Han Liu,Ruoyu Dong,Haoyu Xu,Jingyi Qi,Huimei Liang,Chen Guo,Huan Zeng,Jinghua Zhao,Houhui Song,Yongchun Yang,Wei Wang

Environmental contaminants such as polystyrene nanoplastics (PSNPs, 1–1000 nm) and glyphosate pose significant environmental and public health risks. This study aimed to investigate the intestinal toxicity and molecular mechanisms induced by PSNPs and/or glyphosate. Mice were exposed to PSNPs (<100 nm), glyphosate, or a combination of both for 35 days via intragastric administration (PSNPs: 0.5 mg/d; glyphosate: 50 mg/kg-bw/day; PSNPs + glyphosate: 0.5 mg/d +50 mg/kg-bw/day). The control group received same volume of distilled water. Our findings revealed that exposure to PSNPs and/or glyphosate aggravated pathological alterations, including inflammatory cell infiltration, severe mitochondrial cristae fracture, and an approximately 50 % reduction in goblet cells in the intestine. Moreover, exposure to PSNPs and/or glyphosate caused a critical 75 % inhibition of FOXP3 and dissociation of tight junctions in the intestine (reflected by a 50 % decrease in Occludin, and a 20 %–50 % decrease in ZO-1). These changes were accompanied by significant alterations in beneficial gut microbiota, metabolic profiles, bile acid metabolism disorders, and a pronounced elevation in 3-β-deoxycholic acid, a metabolite tied to bile acid receptor signaling and barrier dysfunction. Although exposure to glyphosate led to the most significant upregulation of the pro-inflammatory factors TNF-α and the pro-apoptosis proteins Cleave-caspase-3, co-exposure did not exacerbate cell apoptosis in animal tissue experiments, which is contrasts with the cell-based findings. MODE-K (mouse intestinal epithelial) cells were treated with PSNPs (0.75 mg/mL) or glyphosate (0.5 mg/mL). In vitro experiments showed that PSNPs aggravated the disrupted Treg/Th17 immune-inflammatory balance, impaired intestinal barrier function (with a 50 % reduction in ZO-1 and Occludin), and increased cell apoptosis, caused by glyphosate. This study advances our understanding of the health risks posed by endocrine-disrupting chemical mixtures and provides critical insights into the molecular mechanisms of PSNP-glyphosate-induced intestinal toxicity. These findings lay the groundwork for future research aimed at mitigating the pathophysiological impacts of environmental pollutants.

A turn-on fluorescence sensor based on Cu2+ modulated DNA-templated silver nanoclusters for glyphosate detection and mechanism investigation

Yixia Yang,Behafarid Ghalandari,Liyun Lin,Xiao Sang,Wenqiong Su,Adeleh Divsalar,Xianting Ding

The abuse application of glyphosate can result in a potential hazard for environment and human, however its ultrasensitive detection remains challenging. Herein, a Cu 2+ modulated DNA-templated silver nanoclusters (DNA-AgNCs) sensor was constructed to sensitively determine glyphosate based on the turn-on fluorescence strategy. The fluorescence quenching of DNA-AgNCs occurred with the existence of Cu 2+ . Upon the presence of glyphosate, the functional groups on the surface of glyphosate could chelate with Cu 2+ , following the fluorescence recovery of DNA-AgNCs. Through the stoichiometric methods, we unveil that Cu 2+ -trigged fluorescence quenching mode is a combination of static and dynamic quenching with the static mode being predominant. In DNA-AgNCs/Cu 2+ system, the carboxylate, amine, and phosphonate groups of glyphosate interact with Cu 2+ through chelation, in which the carboxylate oxygen, the phosphonate oxygen atoms, and the monoprotonated secondary amine nitrogen atom and Cu 2+ form chelate rings. This fluorescence sensor showed a desired linearity of glyphosate analysis under the optimum conditions, ranging from 15 to 100 μg/L with a low detection down to 5 μg/L. Moreover, the proposed sensor was successfully utilized to measure glyphosate in real samples, indicating a promising application in pesticide residues detection.

Efficient degradation of organic phosphorus in glyphosate wastewater by catalytic wet oxidation using modified activated carbon as a catalyst

Bo Xing,Honglin Chen,Xiaoming Zhang

A ratio fluorescence sensor based on rhodamine B embedded metal-organic framework for glyphosate detection in agri-food products

Chao-Qun Wan,Yue-Hong Pang,Yong-Wei Feng,Xiao-Fang Shen

Glyphosate, a broad-spectrum and high-efficiency herbicide, could accumulate in the human body through the consumption of agri-food products. Herein, a ratio fluorescence sensor based on rhodamine B-embedded amino-functionalized iron-based metal-organic framework (MOF, NH 2 -MIL-88(Fe)@RhB) bonded with Cu 2+ was developed for rapid detection of glyphosate. The synthesized NH 2 -MIL-88(Fe) was a biconical prism and had a cavity for the embedding of RhB as a reference compound. In the presence of Cu 2+ , Lewis interactions with NH 2 -MIL-88(Fe)@RhB cause the fluorescence signal to be turned off. When glyphosate was added, the signal was turned on due to chelation with Cu 2+ and hydrogen bonding interactions with NH 2 -MIL-88(Fe)@RhB. Under optimal conditions, the developed sensor exhibited a linear range of 0.60–45 μmol L −1 with a response time of less than 1 min. The sensor was applied in the analysis of agri-food products (tea, soybean, wheat, cucumber), with recoveries between 97.93% and 109.06%, indicating its promising application in agri-food safety.

A Turn-On Fluorescence Sensor Based on Nitrogen-Doped Carbon Dots and Cu2+ for Sensitively and Selectively Sensing Glyphosate

Ziqiang Li,Shuang Liang,Li Zhou,Fengjian Luo,Zhengyun Lou,Zongmao Chen,Xinzhong Zhang,Mei Yang

Glyphosate has excellent herbicidal activity, and its extensive use may induce residue in the environment and enter into humans living through the food chain, causing negative impact. Here, water-soluble 1.55 nm size nitrogen-doped carbon quantum dots (NCDs) with strong blue fluorescence were synthesized using sodium citrate and adenine. The maximum excitation and emission wavelengths of NCDs were 380 nm and 440 nm, respectively. The above synthesized NCDs were first used for the construction of a fluorescence sensor for glyphosate detection. It was found that Cu2+could quench the fluorescence of NCDs effectively through the photoinduced electron transfer (PET) process, which was confirmed using fluorescence lifetime measurements. Additionally, the fluorescence was restored with the addition of glyphosate. Hence, a sensitive turn-on fluorescence sensor based on NCDs/Cu2+for glyphosate analysis was developed. The LODs of glyphosate for water and rice samples were recorded as 0.021 μg/mL and 0.049 μg/mL, respectively. The sensor was applied successfully for ultrasensitive and selective detection of glyphosate in environmental water and rice samples with satisfied recoveries from 82.1% to 113.0% using a simple sample pretreatment technique. The proposed strategy can provide a significant potential for monitoring glyphosate residue in water and agricultural product samples.Keywords:carbon quantum dots;fluorescence;glyphosate detection;water;rice

Sensitive and Label-Free Colorimetric Detection of Glyphosate Based on the Suppression Peroxidase-Mimicking Activity of Cu(II) Ions

Qing Li,Yumeng Guo,Xiangyi He,Guangli Li

The sensitive and accurate determination of glyphosate (Glyp) is urgently demanded because it is closely correlated with human health and environmental safety. In this work, we proposed a sensitive and convenient colorimetric assay by employing copper ion peroxidases for the detection of Glyp in the environment. Free Cu(II) ions displayed high peroxidase activity and can catalytically oxidize the colorless 3,3′,5,5′-tetramethylbenzidine (TMB) into blue oxTMB, resulting in an obviously visible discoloration reaction. Once the Glyp is added, the ability of copper ions to mimic peroxidase can be largely suppressed because of the generation of Glyp–Cu2+chelate. The favorable selectivity and sensitivity were demonstrated in the colorimetric analysis of Glyp. Furthermore, this rapid and sensitive method was successfully applied in the accurate and reliable determination of glyphosate in the real sample, holding promising applications in pesticide determination in the environment.Keywords:label-free;colorimetric analysis;copper ions;peroxidase-mimicking activity;glyphosate

An advanced ratiometric molecularly imprinted sensor based on metal ion reoxidation for indirect and ultrasensitive glyphosate detection in fruit

Xiaoyu Su,Kaiyi Zheng,Xiaoyu Tian,Xuan Zhou,Xiaobo Zou,Xuechao Xu,Zongbao Sun,Wen Zhang

An indirect and ultrasensitive ratiometric molecularly imprinted (MIP) sensor, based on metal ion reoxidation, is introduced for glyphosate (GLY) determination in fruit. As high-performance signal amplification substrates, carbon nanotubes (MWCNTs) and gold nanoparticles (AuNPs) are conveniently modified on GCE. The artificial antibody-MIP membrane, presents typical three-dimensional structure to GLY template. Built-in reference methylene blue (MB) is directly electropolymerized on MWCNTs-Au/GCE. Particularly, Cu 2+ and GLY interestingly form chelate complex, and the Cu 2+ ( I Cu ) in Cu(Ⅱ)-GLY-complex can be reoxidized, and indirectly quantizes GLY. The reference signal ( I MB ) presents noteworthy stability with different GLY levels, and the ratiometric readout ( I Cu / I MB ) is recognized as a more trustworthy indicator to quantize GLY. Proposed sensor presents broad range as 1.73 ∼ 400 ng/mL, and limit of detection is well found as 0.24 ng/mL (S/N = 3). Finally, as-fabricated method is verified with standard HPLC in real-fruit-sample, and the errors and recovery rates are calculated as 3.4% ∼ 6.7% and 94.4% ∼ 104.6%, respectively.

Construction of a N-CDs/AuNCs@ZIF-8-assisted ratiometric fluorescent nanosensor for glyphosate detection in edible and medicinal malt

Doudou Lei,Lingling Li,Pengyue Song,QingBin Xu,Lihua Huang,Xiao Ma,Lidong Zhou,Weijun Kong

Glyphosate (Gly) is a widely-used herbicide in food production, while, the residue of which due to the long-term and excessive overspray poses serious threats to public health. The development of reliable methods for its sensitive detection is of great urgency. In this study, a novel ratiometric fluorescent nanosensor by encapsulating N-doped carbon dots (N-CDs) and gold nanoclusters (AuNCs) in zeolitic imidazole framework-8 (ZIF-8) as the dual-emissive fluorescence probes at 410 and 650 nm, respectively, was constructed for Gly detection. Due to the adsorption property of ZIF-8, the N-CDs/AuNCs@ZIF-8 nanoprobes accumulated Cu 2+ to quench the red fluorescence of AuNCs, and the blue fluorescence of N-CDs was stable. While thiocholine, a product of acetylthiocholine, hydrolyzed by acetylcholinesterase could coordinate with Cu 2+ , resulting in significant fluorescence recovery of AuNCs. This phenomenon was utilized for the quantitation of Gly, due to its inhibitory effect on acetylcholinesterase activity. By calculating the fluorescence intensity ratio ( I 650 /I 410 ), Gly in real sample could be accurately determined in a concentration range of 2–100 ng/mL with a limit of detection of 1.92 ng/mL because of the anti-interference and the self-correction ability of the two fluorescence signals. The N-CDs/AuNCs@ZIF-8 nanoprobes-assisted ratiometric fluorescent nanosensor exhibited unique merits of rapid construction, simple operation, high specificity, and good accuracy for Gly in edible and medicinal malt samples with recoveries of 93.7–108.2 %. This study presents a multiple tool for the versatile sensing of trace pesticides in more food matrices, which can be extended to a full range of environmental and food safety applications.

Ratiometric fluorescence probe based on boric acid-modified carbon dots and alizarin red for sensitive and rapid detection of glyphosate

Yang Jingqi,Wang Xinru,Yu Ying,Cao Yujuan,Guo Manli,Hu Xiaogang,Wang Li,Lin Bixia

By combining boric acid-modified carbon dots ( p -CDs) and alizarin red (ARS), a double emission probe p -CDs@ARS with fluorescence at 410 nm and 600 nm is designed for the detection of glyphosate. When Cu 2+ is added, it binds with ARS to cause ARS release from p -CDs@ARS, which decreases the fluorescence at 600 nm. However, in the presence of glyphosate, glyphosate competes to the binding of Cu 2+ , releasing ARS to bind with p -CDs again. Therefore, the fluorescence of 600 nm recovers. Based on this, the fluorescence of 410 nm and 600 nm act as the reference and response signal, respectively, achieving the ratiometric fluorescence detection of glyphosate. The linear range of glyphosate detection is 0.5–50 µM with a limit of detection at 0.37 µM which is well below the maximum residue limit for glyphosate in food. When the probe is used to detect the glyphosate residue in Pearl River water and cucumber, the detection results are well consistent with those detected by HPLC. The established method based on p -CDs@ARS has the advantages that the assembly of ratiometric fluorescence probe is simple, and the detection speed is fast. Additionally, a typical INHIBIT logical system has been successfully constructed based on glyphosate, Cu 2+ , and the fluorescence signal of p -CDs@ARS. Graphical

A Rapid Detection Method for Atrazine, Diquat, and Glyphosate in Water Using Radar Fingerprint Spectra of Algal Photosynthetic Fluorescence Parameters

Ziqi Ye,Tingting Gan,Gaofang Yin,Nanjing Zhao,Xiaoxuan Tan,Letong Li,Xingchi Li,Yunfei Luo,Wenqing Liu

As typical pesticide pollutants in water environments, herbicides pose a serious threat to the safety of aquatic ecosystems. However, effective methods for rapid screening are still lacking at present. This study developed a stepwise prediction method for atrazine, diquat, and glyphosate using radar fingerprint spectra derived from algal photosynthetic fluorescence responses. Microcystis aeruginosa was selected as the indicator alga from four freshwater algae. Using a strategy of “clustering dimension reduction and comprehensive scoring”, 11 parameters were selected from 26 JIP test parameters, and the optimal exposure time was determined to be 60   min. Radar fingerprint spectra were constructed from the selected parameters, and radial, geometric, and template features were extracted. Four classification and four regression algorithms were evaluated across seven feature sets using nested cross validation. The selected combinations were integrated into a stepwise prediction method following an “identification first, then quantification” strategy. Nested cross validation yielded an identification accuracy of 91.11% and R² values of 0.979, 0.790, and 0.982 for atrazine, diquat, and glyphosate, respectively. The final method achieved an independent validation accuracy of 96.67%, and the corresponding LOQs were 1   μg∙L -1 , 1   mg∙L -1 , and 6   mg∙L -1 , respectively. In filtered actual water samples individually spiked with one target herbicide, mean recoveries ranged from 94.20% to 105.65%. This study provides a novel method for translating algal physiological stress responses into structured spectral features, thereby providing methodological support for rapid detection of herbicides and for emergency monitoring and early warning of water pollution.