Commercial Glyphosate-based Herbicides Research Articles

Impact of glyphosate and glyphosate-based herbicides on phyllospheric Methylobacterium

Symbiotic Methylobacterium comprise a significant portion of the phyllospheric microbiome, and are known to benefit host plant growth, development, and confer tolerance to stress factors. The near ubiquitous use of the broad-spectrum herbicide, glyphosate, in farming operations globally has necessitated a more expansive evaluation of the impacts of the agent itself and formulations containing glyphosate on important components of the plant phyllosphere, including Methylobacterium.This study provides an investigation of the sensitivity of 18 strains of Methylobacterium to glyphosate and two commercially available glyphosate-based herbicides (GBH). Nearly all strains of Methylobacterium showed signs of sensitivity to the popular GBH formulations WeatherMax® and Transorb® in a modified Kirby Bauer experiment. However, exposure to pure forms of glyphosate did not show a significant effect on growth for any strain in both the Kirby Bauer test and in liquid broth, until polysorbate-20 (Tween20) was added as a surfactant. Artificially increasing membrane permeability through the introduction of polysorbate-20 caused a 78–84% reduction in bacterial cell biomass relative to controls containing glyphosate or high levels of surfactant only (0–9% and 6–37% reduction respectively). Concentrations of glyphosate as low as 0.05% w/v (500 µg/L) from both commercial formulations tested, inhibited the culturability of Methylobacterium on fresh nutrient-rich medium.To better understand the compatibility of important phyllospheric bacteria with commercial glyphosate-based herbicides, this study endeavours to characterize sensitivity in multiple strains of Methylobacterium, and explore possible mechanisms by which toxicity may be induced.

Glyphosate-Based Herbicide Formulations and Their Relevant Active Ingredients Affect Soil Springtails Even Five Months after Application

Glyphosate is the most widely used active ingredient (AI) in glyphosate-based herbicides (GBHs) worldwide and is also known to affect a variety of soil organisms. However, we know little about how the effects of glyphosate AIs differ from those of GBHs that also contain so-called inert co-formulants. We conducted a greenhouse experiment using the model cover crop white mustard (Sinapis alba) to investigate the effects of three GBHs (Roundup PowerFlex, Roundup LB Plus, and Touchdown Quattro) and their respective glyphosate AIs (glyphosate potassium, isopropylamine, and diammonium salt) on epedaphic springtails (Sminthurinus niger; Collembola) activity in soils with low (3.0%) or high (4.1%) organic matter content (SOM). Springtail activity was assessed using pitfall traps. Most GBHs and AIs reduced springtail activity compared to mechanical removal of mustard in the short-term and even up to 5 months after application. GBHs and AIs differed considerably in their effects on springtail activity, and effects were modified by SOM content. Our results highlight the need to (i) distinguish between the effects of glyphosate AIs and commercial GBH formulations, (ii) disclose all ingredients of GBHs, as co-formulants also affect non-target organisms, and (iii) include soil properties in ecotoxicological risk assessments for soil organisms to better characterize the situation in the field.

Mutagenic effects of a commercial glyphosate-based herbicide formulation on the soil filamentous fungus Aspergillus nidulans depending on the mode of exposure.

Glyphosate-based herbicides (GBH) are the most used pesticides worldwide. This widespread dissemination raises the question of non-target effects on a wide range of organisms, including soil micro-organisms. Despite a large body of scientific studies reporting the harmful effects of GBHs, the health and environmental safety of glyphosate and its commercial formulations remains controversial. In particular, contradictory results have been obtained on the possible genotoxicity of these herbicides depending on the organisms or biological systems tested, the modes and durations of exposure and the sensitivity of the detection technique used. We previously showed that the well-characterized soil filamentous fungus Aspergillus nidulans was highly affected by a commercial GBH formulation containing 450g/L of glyphosate (R450), even when used at doses far below the agricultural application rate. In the present study, we analysed the possible mutagenicity of R450 in A. nidulans by screening for specific mutants after different modes of exposure to the herbicide. R450 was found to exert a mutagenic effect only after repeated exposure during growth on agar-medium, and depending on the metabolic status of the tested strain. The nature of some mutants and their ability to tolerate the herbicide better than did the wild-type strain suggested that their emergence may reflect an adaptive response of the fungus to offset the herbicide effects. The use of a non-selective molecular approach, the quantitative random amplified polymorphic DNA (RAPD-qPCR), showed that R450 could also exert a mutagenic effect after a one-shot overnight exposure during growth in liquid culture. However, this effect was subtle and no longer detectable when the fungus had previously been repeatedly exposed to the herbicide on a solid medium. This indicated an elevation of the sensitivity threshold of A. nidulans to the R450 mutagenicity, and thus confirmed the adaptive capacity of the fungus to the herbicide.

The effect of commercial herbicide and bioherbicide of Ketapang (Terminalia catappa L.) on germination and chlorophyll content of cayenne pepper (Capsicum frutescens L.)

Herbicides that have less impact on non-target crops are better suited for weed control. The use of a commercial glyphosate-based herbicide (Roundup®) is a farmer's choice for controlling these weeds. The negative effects of glyphosate residues on cultivated crops, environment and human health have led farmers to switch to use bioherbicides. Ketapang (Terminalia catappa L.) leaves have been studied having the potential as bioherbicide but their effects on non-target plant have been less studied. This study aimed to examine the effects of Roundup® and Ketapang leaf extract on germination and chlorophyll content of cayenne pepper as non-target plant. This experimental study was conducted using completely randomized design (CRD) with 3 replications. Cayenne pepper was sown for 14 days. Parameters observed were germination, morphology, weight, and chlorophyll content of cayenne pepper sprouts. The data were analyzed using SPSS ver. 16 and tested with One-Way ANOVA or Kruskal Wallis tests at p-value of 0.05 depended on parametric assumption tests. There were significant differences between the Roundup® solution treatment with the control on all parameters (germination parameters, weight, and chlorophyll content of cayenne pepper sprouts) (p<0.05). The effect was getting worse seen in cayenne pepper seeds given high doses of Roundup® solution. On the contrary, there were no significant differences between the treatments of the aqueous extract of Ketapang leaves with the control on all parameters (p>0.05), except the vigor index. There were morphological changes in the radicle and root of cayenne pepper sprouts exposed to them. Rotted radicles were highly visible in Roundup®-exposed sprouts.

Assessment of Genetic Damage Induced via Glyphosate and Three Commercial Formulations with Adjuvants in Human Blood Cells.

There is considerable controversy regarding the genotoxicity of glyphosate (N-(phosphonomethyl) glycine). It has been suggested that the genotoxicity of this herbicide is increased by the adjuvants added to commercial formulations based on glyphosate. The effect of various concentrations of glyphosate and three commercial glyphosate-based herbicides (GBH) on human lymphocytes was evaluated. Human blood cells were exposed to glyphosates of 0.1, 1, 10 and 50 mM as well as to equivalent concentrations of glyphosate on commercial formulations. Genetic damage (p < 0.05) was observed in all concentrations with glyphosate and with FAENA and TACKLE formulations. These two commercial formulations showed genotoxicity that was concentration-dependent but in a higher proportion compared to pure glyphosate only. Higher glyphosate concentrations increased the frequency and range of tail lengths of some migration groups, and the same was observed for FAENA and TACKLE, while in CENTELLA the migration range decreased but the frequency of migration groups increased. We show that pure glyphosate and commercial GBH (FAENA, TACKLE and CENTELLA) gave signals of genotoxicity in human blood samples in the comet assay. The genotoxicity increased in the formulations, indicating genotoxic activity also in the added adjuvants present in these products. The use of the MG parameter allowed us to detect a certain type of genetic damage associated with different formulations.

Degradation of commercial glyphosate-based herbicide via advanced oxidative processes in aqueous media and phytotoxicity evaluation using maize seeds

Glyphosate is a herbicide that acts as a broad-spectrum, non-selective, post-emergence systemic pest controller. Its continuing, increasing, and excessive use in many countries in recent years poses a significant threat to the environment and human health due to the prevalence of this herbicide in water bodies and its impact on non-target organisms. In this context, it is essential to develop processes aimed at the non-selective degradation of glyphosate and its by-products. In this study, various advanced oxidative processes were applied: Fenton, electro-Fenton, photoelectro-oxidation, and photoelectro-Fenton, with the objective of oxidizing glyphosate in the commercial product Roundup®. The resultant oxidation products and the phytotoxicological effect on maize seed germination were also analyzed. Following each treatment, chemical oxygen demand (COD), total organic carbon (TOC), glyphosate degradation, and oxidation by-product formation were analyzed. The treated solutions were used to germinate maize seeds for 7 days in a germination chamber applying a photoperiod of 12 h at 24 °C. The % of germination, protein and hydrogen peroxide (H2O2) content, lipid peroxidation extent (MDA), and superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) activities were determined. The photoelectro-Fenton treatment was the most effective in degrading glyphosate, operating synergistically to break glyphosate bonds, thereby generating non-toxic short-chain molecules. Maize seed germination was satisfactory (> 50 %), but the persistent formation of reactive oxygen species (ROS) led to increased antioxidant activities of SOD, CAT, and POD enzymes acting in a compensatory manner against ROS, thus sustaining the photosynthetic apparatus. Hormesis, a stimulatory effect of glyphosate, was also observed in the presence of low concentrations of glyphosate.

Glyphosate-based herbicide induces long-lasting impairment in neuronal and glial differentiation.

Glyphosate‐based herbicides (GBH) are among the most sold pesticides in the world. There are several formulations based on the active ingredient glyphosate (GLY) used along with other chemicals to improve the absorption and penetration in plants. The final composition of commercial GBH may modify GLY toxicological profile, potentially enhancing its neurotoxic properties. The developing nervous system is particularly susceptible to insults occurring during the early phases of development, and exposure to chemicals in this period may lead to persistent impairments on neurogenesis and differentiation. The aim of this study was to evaluate the long‐lasting effects of a sub‐cytotoxic concentration, 2.5 parts per million of GBH and GLY, on the differentiation of human neuroepithelial stem cells (NES) derived from induced pluripotent stem cells (iPSC). We treated NES cells with each compound and evaluated the effects on key cellular processes, such as proliferation and differentiation in daughter cells never directly exposed to the toxicants. We found that GBH induced a more immature neuronal profile associated to increased PAX6, NESTIN and DCX expression, and a shift in the differentiation process toward glial cell fate at the expense of mature neurons, as shown by an increase in the glial markers GFAP, GLT1, GLAST and a decrease in MAP2. Such alterations were associated to dysregulation of key genes critically involved in neurogenesis, including PAX6, HES1, HES5, and DDK1. Altogether, the data indicate that subtoxic concentrations of GBH, but not of GLY, induce long‐lasting impairments on the differentiation potential of NES cells.

Differential effects of two prevalent environmental pollutants on host-pathogen dynamics

Chemical pollutants are a major factor implicated in freshwater habitat degradation and species loss. Microplastics and glyphosate-based herbicides are prevalent pollutants with known detrimental effects on animal welfare but our understanding of their impacts on infection dynamics are limited. Within freshwater vertebrates, glyphosate formulations reduce fish tolerance to infections, but the effects of microplastic consumption on disease tolerance have thus far not been assessed. Here, we investigated how microplastic (polypropylene) and the commercial glyphosate-based herbicide, Roundup®, impact fish tolerance to infectious disease and mortality utilising a model fish host-pathogen system. For uninfected fish, microplastic and Roundup had contrasting impacts on mortality as individual stressors, with microplastic increasing and Roundup decreasing mortality compared with control fish not exposed to pollutants. Concerningly, microplastic and Roundup combined had a strong interactive reversal effect by significantly increasing host mortality for uninfected fish (73% mortality). For infected fish, the individual stressors also had contrasting effects on mortality, with microplastic consumption not significantly affecting mortality and Roundup increasing mortality to 55%. When combined, these two pollutants had a moderate interactive synergistic effect on mortality levels of infected fish (53% mortality). Both microplastic and Roundup individually had significant and contrasting impacts on pathogen metrics with microplastic consumption resulting in fish maintaining infections for significantly longer and Roundup significantly reducing pathogen burdens. When combined, the two pollutants had a largely additive effect in reducing pathogen burdens. This study is the first to reveal that microplastic and Roundup individually and interactively impact host-pathogen dynamics and can prove fatal to fish.

Influence of a glyphosate-based herbicide on growth parameters and aflatoxin B1 production by Aspergillus section Flavi on maize grains

Glyphosate-based herbicides (GBH) are the main pesticides applied worldwide on maize production. Glyphosate-resistant weeds led to the repeated application of high doses of the pesticide. In addition to environmental conditions, the presence of GBH affects the development of Aspergillus species and aflatoxin B1 (AFB1) production under in vitro conditions. The aim of this work was to evaluate the influence of a commercial GBH on growth and AFB1 production by Aspergillus flavus and Aspergillus parasiticus strains under different water activity (aW) conditions. The following concentrations of active ingredient glyphosate were evaluated: 20, 50, 200 and 500mM. The lag phase prior to growth and growth rate did not change at 20 and 50mM (that is, at field recommended doses) at 0.98 and 0.95 aW; however, at increasing GBH concentrations, between 200 and 500mM, the growth rate decreased at all aW conditions. In general, as the GBH concentration increased, AFB1 production decreased. However, a significant increase in toxin accumulation was found only at one of the aW conditions (0.95) at 21 days with 50mM of GBH in A. flavus and 20 and 50mM of GBH in A. parasiticus. These results show that, even though Aspergillus section Flavi growth did not increase, AFB1 production increased on maize grains at GBH concentrations similar to those of field recommended doses under favorable water availability and temperature conditions.

Can glyphosate and commercial glyphosate-based herbicide formulations alter human microbiomes?

Glyphosate exerts its herbicidal action by inhibiting 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) of the shikimate pathway, thus blocking aromatic amino acid biosynthesis. Glyphosate may also affect shikimate pathway positive microorganisms and is patented as an antibiotic. We have recently shown that glyphosate and a commercial formulation of glyphosate-based herbicide can indeed inhibit EPSPS and the shikimate pathway in the gut microbiome of rats. However, it is uncertain whether glyphosate exposure could lead to perturbations of human microbiota including those of the gut. We have thus conducted bioinformatics analyses of publicly available datasets for effects of glyphosate on human microbiomes, especially the gut. Data from the Human Microbiome Project for 6 major body sites shows that the shikimate pathway is found in almost all individuals and all locations. Comparison of the abundance of the shikimate pathway in paired metagenomes and metatranscriptomes indicated that in the human gut microbiome most bacteria do not possess a complete shikimate pathway, which is mostly transcriptionally inactive. This suggests that gut bacteria are mostly aromatic amino acid auxotrophs and thus relatively resistant to potential growth inhibition by glyphosate. We also classified E. coli EPSPS homologues as Class I (sensitive to glyphosate) and Class II (resistant to glyphosate). Among 44 subspecies reference genomes, accounting for 72% of the total assigned microbial abundance in 2,144 human faecal metagenomes, 9 subspecies have Class II EPSPS. The study of gut metagenomes also indicated that glyphosate might be degraded by Proteobacteria in the human gut microbiome using the carbon-phosphorus lyase pathway. Overall, there is limited experimental evidence available for the effects of glyphosate on the human microbiomes including the gut. Further investigations using more advanced molecular profiling techniques including transcriptomics and metabolomics are needed to ascertain whether glyphosate and glyphosate-based herbicides can alter the function of microbiomes with consequent health implications.

Interactive effect of glyphosate-based herbicides and organic soil layer thickness on growth and reproduction of the tropical earthworm Pontoscolex corethrurus (Müller, 1857)

Glyphosate-based herbicides (GBHs) have negative effects on the fitness parameters of earthworms; however, these effects may vary from one commercial product to another and between different recommended doses. Additionally, frequent applications of GBHs may decrease the thickness of the topsoil layer due to increased soil erosion through vegetation reduction. To investigate the effects of frequent applications of GBHs, and their interactive effect on the performance of P. corethrurus, we performed a two-factor experiment exposing in vitro the earthworm to two commercial GBHs (factor I, FAENA FUERTE (FF, Monsanto Co.) and GLYFOS (GLY, Cheminova Co., with three levels)) and soil thickness (factor II, with two levels); having six treatments in total, each with four replicates. Both herbicides were applied at their recommended doses; a total of 11 times at intervals of 14 days in soil layers 2.5 and 5 cm deep. Earthworm biomass and cocoon production were measured every 12 days for 168 days. Application of GLY resulted in significant biomass decreases of between 51 ± 11.54% (here and further S.E. is provided) and 100 ± 0.00% compared to a control. For FF, the final biomass decreased significantly by between 19 ± 7.33% and 24 ± 8.44% at both doses, independently of soil thickness. Additionally, a significant biomass reduction of 41 ± 7.93% was detected when soil thickness was low at highest dose. At a soil depth of 5 cm, GLY significantly decreased cocoon production by between 69 ± 7.86% and 83 ± 10.53% at each dose compared to the control. In contrast, the different doses of FF did not affect the number of cocoons produced, although there was a significant decrease of 72 ± 7.18% when the soil layer was reduced. The growth rate increased significantly under exposure to both GBH at all tested doses. Our results suggest that cumulative doses of GLY are more detrimental for P. corethrurus than those of FF and that soil thickness may help buffer some of the negative effects of GBHs. In order to reduce negative impacts on earthworms in the field, we therefore suggest maintaining the topsoil layer thickness and adjusting the doses of GBHs to the lowest end of the recommended range.

Commercial glyphosate-based herbicides effects on springtails (Collembola) differ from those of their respective active ingredients and vary with soil organic matter content

Glyphosate-based herbicides (GBH) are currently the most widely used agrochemicals for weed control. Environmental risk assessments (ERA) on nontarget organisms mostly consider the active ingredients (AIs) of these herbicides, while much less is known on effects of commercial GBH formulations that are actually applied in the field. Moreover, it is largely unknown to what extent different soil characteristics alter potential side effects of herbicides. We conducted a greenhouse experiment growing a model weed population of Amaranthus retroflexus in arable field soil with either 3.0 or 4.1% soil organic matter (SOM) content and treated these weeds either with GBHs (Roundup LB Plus, Touchdown Quattro, Roundup PowerFlex) or their respective AIs (isopropylammonium, diammonium or potassium salts of glyphosate) at recommended dosages. Control pots were mechanically weeded. Nontarget effects were assessed on the surface activity of the springtail species Sminthurinus niger (pitfall trapping) and litter decomposition in the soil (teabag approach). Both GBHs and AIs increased the surface activity of springtails compared to control pots; springtail activity was higher under GBHs than under corresponding AIs. Stimulation of springtail activity was much higher in soil with higher SOM content than with low SOM content (significant treatment x SOM interaction). Litter decomposition was unaffected by GBHs, AIs or SOM levels. We suggest that ERAs for pesticides should be performed with actually applied herbicides rather than only on AIs and should also consider influences of different soil properties.

Transcriptome profiling of the fungus Aspergillus nidulans exposed to a commercial glyphosate-based herbicide under conditions of apparent herbicide tolerance

Glyphosate-based herbicides, such as Roundup®, are the most widely used non-selective, broad-spectrum herbicides. The release of these compounds in large amounts into the environment is susceptible to affect soil quality and health, especially because of the non-target effects on a large range of organisms including soil microorganisms. The soil filamentous fungus Aspergillus nidulans, a well-characterized experimental model organism that can be used as a bio-indicator for agricultural soil health, has been previously shown to be highly affected by Roundup GT Plus (R450: 450 g/L of glyphosate) at concentrations far below recommended agricultural application rate, including at a dose that does not cause any macroscopic effect. In this study, we determined alterations in the transcriptome of A. nidulans when exposed to R450 at a dose corresponding to the no-observed-adverse-effect level (NOAEL) for macroscopic parameters. A total of 1816 distinct genes had their expression altered. The most affected biological functions were protein synthesis, amino acids and secondary metabolisms, stress response, as well as detoxification pathways through cytochromes P450, glutathione-S-transferases, and ABC transporters. These results partly explain the molecular mechanisms underlying alterations in growth parameters detected at higher concentrations for this ascomycete fungus. In conclusion, our results highlight molecular disturbances in a soil fungus under conditions of apparent tolerance to the herbicide, and thus confirm the need to question the principle of “substantial equivalence” when applied to plants made tolerant to herbicides.

Metabolic switch in energy metabolism mediates the sublethal effects induced by glyphosate-based herbicide on tadpoles of a farmland frog Microhyla fissipes

Glyphosate-based herbicides (GBHs) are widely-used agricultural chemicals, bringing potential detriments to aquatic organisms. Currently, our understanding of sublethal effects and underlying toxicologic mechanisms of GBHs are still limited, especially in amphibians. Here, the sublethal effects of a commercial GBH (KISSUN®) on tadpoles of a farmland dwelling frog, Microhyla fissipes, were investigated. The 10-d LC50 of “KISSUN®” GBH was 77.5 mg/L. Tadpoles exposed to 60–120 mg/L showed increased preference to higher temperature. After 10 days exposure, obvious growth suppression was observed in survived GBH-stressed tadpoles, characterized by dosage depended decrement in body mass, body width, total length, etc. GBH-stressed tadpoles also showed decreased tail length/snout-vent length ratio and smaller tail muscle fiber diameter. Comparative transcriptomics (control, 60 mg/L and 90 mg/L groups) was conducted to analyze the underlying molecular processes. GBH-stressed tadpoles showed downregulated transcription of ribosomal proteins and cytoskeleton proteins, which could explain their suppressed whole body and tail muscle growth. Moreover, GBH-stressed tadpoles showed transcriptional downregulation of carbohydrate and lipid catabolism, but upregulation of amino acid catabolism. It suggested a metabolic switch from carbohydrate and lipid to amino acid in these tadpoles. Accordingly, there was a trade-off between protein synthesis and energy production in respect to amino acid allocation, and it provided a metabolic explanation for why protein synthesis was downregulated and growth was suppressed in GBH-stressed tadpoles. In combination with existing literatures, we speculated that GBH might directly target the enzymes in carbohydrate and lipid catabolism, and this metabolic effect of GBH might be common to fish and amphibians. In conclusion, our study provided a systematic insight into the sublethal symptoms of GBH-stressed tadpoles, and a metabolic switch from carbohydrate and lipid to amino acid likely underlay some common toxic symptoms of GBHs on both fish and tadpoles.

Roundup-Induced AMPK/mTOR-Mediated Autophagy in Human A549 Cells.

The extensive use of pesticide caused an amount of pressure on the environment and increased the potential human health risk. Glyphosate-based herbicide (GBH) is one of the most widely used pesticides based on a 5-enolpyruvylshikimate-3-phosphate synthase target, which does not exist in vertebrates. Here, we study autophagic effects of the most famous commercial GBH Roundup (RDP) on human A549 cells in vitro. Intracellular biochemical assay indicated opening of mitochondrial permeability transition pore, LC3-II conversion, up-regulation of beclin-1, down-regulation of p62, and the changes in the phosphorylation of AMPK and mTOR induced by RDP in A549 cells. Further experimental results indicated that all the effects induced by RDP were related to its adjuvant polyethoxylated tallow amine, not its herbicidal active ingredient glyphosate isopropylamine salt. All these results showed that RDP has the ability to induce AMPK/mTOR-mediated cell autophagy in human A549 cells. This study would provide a theoretical basis for understanding RDP's autophagic effects on human A549 cells and attract attention on the potential human health risks induced by the adjuvant.

The response of the algae Fucus virsoides (Fucales, Ochrophyta) to Roundup® solution exposure: A metabolomics approach

Glyphosate, as a broad-spectrum herbicide, is frequently detected in water and several studies have investigated its effects on several freshwater aquatic organisms. Yet, only few investigations have been performed on marine macroalgae. Here, we studied both the metabolomics responses and the effect on primary production in the endemic brown algae Fucus virsoides exposed to different concentration (0, 0.5, 1.5 and 2.5 mg L−1) of a commercial glyphosate-based herbicide, namely Roundup®. Our results show that Roundup® significantly reduced quantum yield of photosynthesis (Fv/Fm) and caused alteration in the metabolomic profiles of exposed thalli compared to controls. Together with the decrease in the aromatic amino acids (phenylalanine and tyrosine), an increase in shikimate content was detected. The branched-amino acids differently varied according to levels of herbicide exposure, as well as observed for the content of choline, formate, glucose, malonate and fumarate. Our results suggest that marine primary producers could be largely affected by the agricultural land use, this asking for further studies addressing the ecosystem-level effects of glyphosate-based herbicides in coastal waters.

Evaluation of the cytotoxic effects of glyphosate herbicides in human liver, lung, and nerve

Glyphosate-based herbicides are broad-spectrum pesticides widely used in the world, which is considered a highly safe pesticide due to their target specificity, but recently, there has been an ongoing controversy regarding their carcinogenicity and possible side effects of glyphosate on human health. Commercial glyphosate-based herbicides (GBHs) consist of declared active ingredient (glyphosate salts) and a number of formulants such as ethoxylated formulants (4130®, 3780®, and A-178®). The aim of our study is to investigate whether the toxicity of GBHs is related to formulants. The effects of GBHs on human health were studied at the cellular level based on their toxicity to liver, lungs and nerve tissue. The inhibitory toxicity to cell viability by GBHs was examined with cell-based systems using three human cell lines: HepG2, A549, and SH-SY5Y. Data obtained showed that all tested ethoxylated formulants and their mixtures with declared active ingredient glyphosate isopropylamine salt (GP) have significant inhibitory effect on cell proliferation, while the declared active ingredient has no significant toxicity. Our study demonstrates that the toxic effect of GBH is primarily due to the use of formulants. This result suggests that GP is relatively safe and a new approach for the assessment of toxicity should be made.

Evaluation of estrogen receptor alpha activation by glyphosate-based herbicide constituents

The safety, including the endocrine disruptive capability, of glyphosate-based herbicides (GBHs) is a matter of intense debate. We evaluated the estrogenic potential of glyphosate, commercial GBHs and polyethoxylated tallowamine adjuvants present as co-formulants in GBHs. Glyphosate (≥10,000 μg/L or 59 μM) promoted proliferation of estrogen-dependent MCF-7 human breast cancer cells. Glyphosate also increased the expression of an estrogen response element-luciferase reporter gene (ERE-luc) in T47D-KBluc cells, which was blocked by the estrogen antagonist ICI 182,780. Commercial GBH formulations or their adjuvants alone did not exhibit estrogenic effects in either assay. Transcriptomics analysis of MCF-7 cells treated with glyphosate revealed changes in gene expression reflective of hormone-induced cell proliferation but did not overlap with an ERα gene expression biomarker. Calculation of glyphosate binding energy to ERα predicts a weak and unstable interaction (−4.10 kcal mol−1) compared to estradiol (−25.79 kcal mol−1), which suggests that activation of this receptor by glyphosate is via a ligand-independent mechanism. Induction of ERE-luc expression by the PKA signalling activator IBMX shows that ERE-luc is responsive to ligand-independent activation, suggesting a possible mechanism of glyphosate-mediated activation. Our study reveals that glyphosate, but not other components present in GBHs, can activate ERα in vitro, albeit at relatively high concentrations.

Spectroscopic Detection of Glyphosate in Water Assisted by Laser-Ablated Silver Nanoparticles.

Glyphosate is one of the most widely used herbicides in the world. Its safety for both human health and aquatic biomes is a subject of wide debate. There are limits to glyphosate’s presence in bodies of water, and it is usually detected through complex analytical procedures. In this work, the presence of glyphosate is detected directly through optical interrogation of aqueous solution. For this purpose, silver nanoparticles were produced by pulsed laser ablation in liquids. Limits of detection of 0.9 mg/L and 3.2 mg/L were obtained with UV-Vis extinction and Surface Enhanced Raman spectroscopies, respectively. The sensing mechanism was evaluated in the presence of potential interferents as well as with commercial glyphosate-based herbicides.

Impact of litter contaminated with glyphosate-based herbicide on the performance of Pontoscolex corethrurus, soil phosphatase activities and soil pH

The bioavailability of glyphosate from plant residues may be high in the soil matrix. In order to determine the impact of litter contaminated with glyphosate (GLY) on the growth and cocoon production of the earthworm Pontoscolex corethrurus as well as on acid and basic phosphatase activities and soil pH, a commercial glyphosate-based herbicide was applied to three types of litter mixed with soil. Earthworms from a GLY-free coffee plantation were grown for 132days in microcosms in a two-factorial design. The first factor was glyphosate (GLY; 21.6g of glyphosate per kg of litter, with two levels (−GLY, +GLY), and the second factor was type of litter (Coffea arabica, Musa cavendishii, Inga vera). All studied factors (except days*litter*glyphosate interaction) had significant effects on earthworm biomass (P<0.05). Treatments with Coffea and Musa litter -GLY had significantly higher biomass (P<0.05), whereas treatments with Inga litter -GLY and +GLY had the lowest biomass. A decline in earthworm biomass was detected in all treatments with glyphosate (+GLY) after 72days. No significant effect on cocoon production was detected for the main factors or their interaction. Litter type showed a marginal effect (P=0.06); the average (±SE) number of cocoons decreased from Musa (5.8±2.9) to Coffea (1.3±0.56) and Inga (0.17±0.17). Soil pH significantly (P=0.006) decreased at the end of the experiment from 4.4±0.17 in -GLY to 3.9±0.06 in +GLY. Acid phosphatase activity increased in treatments with Musa and Inga +GLY (P<0.05), while alkaline phosphatase activity in the same substrates was higher in treatments with -GLY (P<0.05). We conclude that repeated application of litter contaminated with glyphosate negatively affects earthworm vitality and increases soil acidity and acid phosphatase activity.