Pure Glyphosate Research Articles

Glyphosate-based herbicides alter soil carbon and phosphorus dynamics and microbial activity

Glyphosate is among the most used herbicides worldwide. However, both the active ingredient and additives of the commercial formulations may adversely affect chemical and biological processes in soil. Therefore, the aim of this work was to evaluate the effect of glyphosate and the commercial formulation (CH) on soil C, N and P dynamics, and microbial enzyme activity in two soil management conditions: a nopal plot (NP) with an history of 5 years of glyphosate application, and in an abandoned plot (AP) with a history of previous agricultural management without glyphosate applications. We performed three experiments: a) glyphosate and CH effects on phosphorus sorption by performing a non-biological P fixation experiment; b) glyphosate and CH effects on microbial activity, C, N and P dynamic, and the biodiversity of soil microorganisms in terms of biomarker fatty acids after a 27 day incubation experiment; and c) glyphosate and CH degradation was tested isolating bacteria from the incubated soil samples and growing in a defined media with potassium phosphate, glyphosate or CH as sole source of P. Our results suggested that other compounds from CH may be affecting P sorption, because when the herbicide was applied, P sorption was reduced in the NP, but in AP soil no significant effects were observed. In contrast, in the AP soil both glyphosate and the CH reduced C mineralization. Besides, CH application reduced Pmic concentration and laccase activity, but increased the specific activity of phosphonatase and phosphodiesterase in the AP soil. Additionally, the microbial community in NP soil revealed higher diversity index than the AP soil, but major changes in microbial groups were shown only when CH was applied. In vitro assay with native bacterial isolates confirmed their susceptibility to the commercial herbicide in both NP and AP soils. In conclusion, our results demonstrated changes on some processes related to soil C and P dynamics when pure glyphosate and a commercial glyphosate were applied, along with changes in microbial activity and community structure, being stronger the effects with the commercial formulation used. Further research should be done for test the effects of glyphosate on microbial stoichiometry and enzyme stoichiometry.

Autism-like Behaviors in Male Juvenile Offspring after Maternal Glyphosate Exposure.

ObjectiveExposure to the herbicide glyphosate during pregnancy and lactation may increase the risk for autism spectrum disorder (ASD) in offspring. Recently, we reported that maternal exposure of formulated glyphosate caused ASD-like behaviors in juvenile offspring. Here, we investigated whether maternal exposure of pure glyphosate could cause ASD-like behaviors in juvenile offspring.MethodsWater or 0.098% glyphosate was administered as drinking water from E5 to P21 (weaning). Behavioral tests such as grooming test and three-chamber social interaction test in male offspring were performed from P28 to P35.ResultsMale offspring showed ASD-like behavioral abnormalities (i.e., increasing grooming behavior and social interaction deficit) after maternal exposure of glyphosate.ConclusionThe findings suggest that the exposure of glyphosate during pregnancy and lactation may cause ASD-like behavioral abnormalities in male juvenile offspring. It is likely that glyphosate itself, but not the other ingredients, may contribute to ASD-like behavioral abnormalities in juvenile offspring.

Performance and mineral status of weaning pigs fed diets with different levels of glyphosate and tryptophan

Glyphosate-based herbicides (GBH), like Roundup, are used worldwide for weed control in genetically modified feed crops and pre-harvest desiccation of conventional crops, leaving glyphosate residues in feed made from these sources. Glyphosate possesses herbicidal and antimicrobial properties due to inhibition of the shikimate pathway, involved in aromatic amino acid synthesis in plants and microorganisms, including animal gut bacteria. Furthermore, glyphosate possesses mineral chelating properties. The objective of the present study was to investigate potential effects of dietary glyphosate residues (here glyphosate-amended feed) on livestock, focusing on performance and mineral status of weaning pigs. Two experiments were conducted with pigs from 20 litters (4 females and 4 males per litter), weaned on day 28 of age. In experiment I, 104 pigs from 13 litters were assigned to one of four treatments on the weaning day and fed a standard weaning diet amended with glyphosate as follows: no glyphosate (CON), 20 mg glyphosate/kg as Glyphomax HL (GM20) (Albaugh UK,Ltd., London, UK), 20 mg glyphosate/kg as IPA salt (IPA20) (Monsanto Company, St. Louis, MO, USA) and 200 mg glyphosate/kg as IPA salt (IPA200). The EU maximum residue level (MRL) of glyphosate is 20 mg/kg for a range of common feed ingredients. In experiment II, 56 pigs from seven litters were assigned to one of two treatments on the weaning day and fed a weaning diet similar to that used in experiment I, but without supplementation of crystalline tryptophan. The glyphosate amendments were: no glyphosate (CON-II) and 200 mg glyphosate/kg as IPA salt (IPA200-II). In both experiments, half of the pigs were euthanised on day 9/10 (referred to as day 9) and the other half on day 35 after commencement of the treatments. Individual weight, feed intake and faeces score were registered during the experiment, and small intestine digesta, blood and urine were sampled immediately after euthanising. Analysed levels of glyphosate in the diets were close to the intended levels; only, the glyphosate level in the control diets was not zero, indicating that glyphosate-free diets are difficult to obtain. The concentration of glyphosate in small intestine digesta, serum and urine reflected the treatment levels, e.g. at day 9, serum concentrations were 0.06, 10.9, 11.7 and 68.2 µg/L for CON, GM20, IPA20 and IPA200, respectively. Neither feed intake, growth rate, serum mineral concentrations nor faeces score were significantly affected by level or source (pure glyphosate salt or GBH) of the dietary glyphosate.

Recovery trend to co-exposure of iron oxide nanoparticles (γ-Fe2O3) and glyphosate in liver tissue of the fish Poecilia reticulata

Citrate-coated iron oxide nanoparticles (IONPs) have potential use in environmental remediation, with possibilities in decontaminating aquatic environments exposed to toxic substances. This study analyzed IONPs associated to Roundup Original, a glyphosate-based herbicide (GBH), and pure glyphosate (GLY), through ultrastructural and histopathological biomarkers in liver tissue, from females of Poecilia reticulata exposed to: iron ions (0.3 mg/L) (IFe) and IONPs (0.3 mgFe/L) associated with GLY (0.65 mg/L) and GBH (0.65 mgGLY/L (IONP + GBH1) and 1.30 mgGLY/L (IONP + GBH2)) for a period of 7, 14 and 21 days, followed by an equal post-exposure period only in reconstituted water. For the assays, the synthetized IONPs had crystalline and rounded shape with an average diameter of 2,90 nm, hydrodynamic diameter 66,6 mV, zeta potential −55,4 and diffraction profile of maghemite (γ-Fe2O3). The data obtained by biomarkers indicated a high inflammatory response in all treatments. These same parameters, considered during the post-exposure period indicated recovery in reaction patterns of circulatory disturbances and regressive changes, resulting in average reductions of 37,53 points in IFe, 21 points in IONP + GBH1, 15 points in IONP + GBH2 and 11 points in IONP + GLY in total histopathological index of liver after 21 days post-exposure. However, although the cellular and tissue responses were significant, there was no change in the condition factor and hepatosomatic index, denoting resilience of the experimental model.

Repeated annual application of glyphosate reduces the abundance and alters the community structure of soil culturable pseudomonads in a temperate grassland

Grasslands include some of the most endangered and less protected ecosystems on the planet, such as the Pampa of South America. In the Flooding Pampa where native grasslands were not replaced by crops, glyphosate is frequently sprayed in late-summer every year to increase winter forage productivity for cattle without knowing the impacts on both aboveground and belowground grassland genetic resources. Here, we evaluated the effect of this practice on the abundance and community structure of members of the genus Pseudomonas, one of the most diverse bacterial taxa of the plant rhizosphere broadly recognized by their benefits on plant growth and protection, and on soil remediation. In a field experiment, we analyzed the impact of a single application or of a repeated annual application during 4 consecutive years of the recommended dose of commercial glyphosate (Glacoxan®, 3 l ha-1 and 1440 g active ingredient ha-1). In a greenhouse experiment, we tested whether exposure of bare soil to either commercially formulated or pure glyphosate has a direct impact on the resident pseudomonas community in the absence of plants. In the field, we found that the repeated annual herbicide application resulted in a reduction in the abundance and a shift in the community structure of culturable pseudomonads, with evidence of an enrichment of glyphosate-metabolizing lineages. On the other hand, a single application of glyphosate to bare soil under greenhouse conditions did not affect the abundance nor the community structure of pseudomonads. Considering that these plant-beneficial bacteria are natural inhabitants of the rhizosphere and that their fitness is promoted by plant root exudates, the observed alterations are most likely associated with the drop in the vegetation cover directly provoked by the effect of glyphosate. For the first time, we report on the consequences that a reiterated glyphosate application has on a diverse soil reservoir of bacteria that contributes to crucial ecosystem functions in a temperate natural grassland under animal production. Our findings strengthen the notion that this agronomical practice has an unavoidable imprint on non-target microorganisms that are relevant components of the plant-soil feedbacks and that may be instrumental for ecosystem restoration. We conclude that it is necessary to develop sustainable management practices to avoid affecting key functional microorganisms and exploit their contributions to society.

Effects of glyphosate-based herbicides and their active ingredients on earthworms, water infiltration and glyphosate leaching are influenced by soil properties

BackgroundGlyphosate-based herbicides (GBHs) are among the most often used pesticides. The hundreds of GBHs used worldwide consist of the active ingredient (AI) glyphosate in form of different salts, possibly other AIs, and various mostly undisclosed co-formulants. Pesticide risk assessments are commonly performed using single AIs or GBHs at standard soil conditions without vegetation. In a greenhouse experiment, we established a weed population with common amaranth (Amaranthus retroflexus) to examine the effects of three GBHs (Roundup LB Plus, Roundup PowerFlex, Touchdown Quattro) and their corresponding AIs (salts of glyphosate isopropylammonium, potassium, diammonium) on the activity and physiological biomarkers (glutathione S-transferase, GST; acetylcholine esterase, AChE) of an ecologically relevant earthworm species (Lumbricus terrestris). GBHs and AIs were applied at recommended doses; hand weeding served as control. Experiments were established with two soil types differing in organic matter content (SOM; 3.0% vs. 4.1%) and other properties.ResultsEarthworm activity (casting and movement activity) decreased after application of glyphosate formulations or active ingredients compared to hand weeding. We found no consistent pattern that formulations had either higher or lower effects on earthworm activity than their active ingredients; rather, differences were substance-specific. Earthworm activity was little affected by soil organic matter levels. Biomarkers remained unaffected by weed control types; GST but not AChE was decreased under high SOM. Water infiltration after a simulated heavy rainfall was interactively affected by weed control types and SOM. Leachate amount was higher after application of formulations than active ingredients and was higher under low SOM. Glyphosate concentrations in soil and leachate were strongly affected by application of formulations or active ingredients and varied with SOM (significant weed control type x SOM interaction).ConclusionsWe found that both commercial formulations and pure active ingredients can influence earthworms with consequences on important soil functions. Glyphosate products showed increased, reduced or similar effects than pure glyphosate on particular soil functions; soil properties can substantially alter this. Especially at lower SOM, heavy rainfalls could lead to more glyphosate leaching into water bodies. A full disclosure of co-formulants would be necessary to further decipher their specific contributions to these inconsistent effects.

Involvement of mitochondrial fission in renal tubular pyroptosis in mice exposed to high and environmental levels of glyphosate combined with hard water

Chronic interstitial nephritis in agricultural communities (CINAC) has reached epidemic proportions. The combination of glyphosate and hard water has been postulated to play a potent aetiological role in CINAC. Therefore, dynamin-related protein 1 (Drp1)-mediated aberrant mitochondrial fission and subsequent activation of the nucleotide-binding oligomerization domain (NOD)-like receptor protein 3 (Nlrp3)/caspase1 pathway may be involved in the pathogenesis of nephropathy. In the present study, mice were sub-chronically exposed to high doses and environmental levels of glyphosate (100 mg/kg body weight (mg/kg·bw) glyphosate in Roundup and 0.7 mg/L pure glyphosate, respectively) and hard water (2500 mg/L CaCO3 and 250 mg/L Ca2+, respectively) in drinking water. Moreover, Mdivi-1 (Md-1, 10 mg/kg·bw) was intraperitoneally injected to inhibit Drp1 on the basis of the high-dose experiment. Histopathological examination, biochemical analysis, ELISA, western blotting and fluorescent staining were used to analyse renal structure, renal tubular pyroptosis and mitochondrial fission/fusion alterations. The results showed dramatic proximal tubular injury, particularly in the combined groups. Moreover, significant increases in the protein expression levels of calmodulin (CaM), calmodulin-dependent protein kinase II (CaMKII), Drp1/p-Drp1-Ser616 and the Txnip/Nlrp3/caspase1 signalling pathway, and alterations in oxidative stress were observed in the combined groups, and these effects were attenuated by the Drp1 inhibitor Md-1. Intriguingly, there may be a synergistic effect of glyphosate and hard water on renal injury. Taken together, these results suggest that the combination of glyphosate and hard water, even at environmental exposure levels, enhances pyroptosis and ongoing tubulointerstitial inflammation through excessive Drp1-mediated mitochondrial fission.

Glyphosate-based herbicides affect behavioural patterns of the livebearer Jenynsia multidentata.

Roundup® is one of the most widely marketed glyphosate-based herbicides in the world. There are many different formulations of this brand that differ from each other in glyphosate concentration, salts and adjuvants, including surfactants, which are labelled as "inert" compounds. Several studies have shown that these formulations are highly toxic to fish, even compared with pure glyphosate. However, mechanisms underlying this toxicity are not fully understood. In this context, this study evaluated the effects of exposure to Roundup Original® (RO), Roundup Transorb® (RT), and Roundup WG® (RWG) on the behavioural patterns of the livebearer Jenynsia multidentata. This fish naturally inhabits agricultural areas in southern Brazil and Argentina where glyphosate is used extensively. In the experiment, animals were exposed to the herbicides for 96 h, at the environmentally relevant concentration of 0.5 mg/L of glyphosate. Swimming performance, anxiety, aggressiveness, long-term memory and male sexual activity were recorded. The formulation RWG negatively affected swimming performance, thigmotaxia and long-term memory consolidation. Conversely, RT reduced the sexual performance of males. These results confirm that Roundup® formulations are extremely harmful and also that they have different targets of toxicity, affecting behaviours that are essential for fish survival.

Oral or Topical Exposure to Glyphosate in Herbicide Formulation Impacts the Gut Microbiota and Survival Rates of Honey Bees.

Honey bees are important agricultural pollinators that rely on a specific gut microbiota for the regulation of their immune system and defense against pathogens. Environmental stressors that affect the bee gut microbial community, such as antibiotics and glyphosate, can indirectly compromise bee health. Most of the experiments demonstrating these effects have been done under laboratory conditions with pure chemicals. Here, we investigated the oral and topical effects of various concentrations of glyphosate in a herbicide formulation on the honey bee gut microbiota and health under laboratory and field conditions. Under all of these conditions, the formulation, dissolved in sucrose syrup or water, affected the abundance of beneficial bacteria in the bee gut in a dose-dependent way. Mark-recapture experiments also demonstrated that bees exposed to the formulation were more likely to disappear from the colony, once reintroduced after exposure. Although no visible effects were observed for hives exposed to the formulation in field experiments, challenge trials with the pathogen Serratia marcescens, performed under laboratory conditions, revealed that bees from hives exposed to the formulation exhibited increased mortality compared with bees from control hives. In the field experiments, glyphosate was detected in honey collected from exposed hives, showing that worker bees transfer xenobiotics to the hive, thereby extending exposure and increasing the chances of exposure to recently emerged bees. These findings show that different routes of exposure to glyphosate-based herbicide can affect honey bees and their gut microbiota.IMPORTANCE The honey bee gut microbial community plays a vital role in immune response and defense against opportunistic pathogens. Environmental stressors, such as the herbicide glyphosate, may affect the gut microbiota, with negative consequences for bee health. Glyphosate is usually sprayed in the field mixed with adjuvants, which enhance herbicidal activity. These adjuvants may also enhance undesired effects in nontargeted organisms. This seems to be the case for glyphosate-based herbicide on honey bees. As we show in this study, oral exposure to either pure glyphosate or glyphosate in a commercial herbicide formulation perturbs the gut microbiota of honey bees, and topical exposure to the formulation also has a direct effect on honey bee health, increasing mortality in a dose-dependent way and leaving surviving bees with a perturbed microbiota. Understanding the effects of herbicide formulations on honey bees may help to protect these important agricultural pollinators.

Exposure to glyphosate during pregnancy induces neurobehavioral alterations and downregulation of Wnt5a-CaMKII pathway

Glyphosate-based formulations are the most popular herbicide used around the world. These herbicides are widely applied in agriculture to control weeds on genetically modified crops. Although there is much evidence showing that glyphosate-based herbicides induce toxic effect on reproductive and hepatic systems, and also cause oxidative damage on cells, studies from recent years revealed that the nervous system may represent a key target for their toxicity. In the present work, we evaluated the effect of glyphosate (without adjuvants) in neonate rats after gestational exposure. Particularly, we examined whether glyphosate during gestation affected the nervous system function at early development. Pregnant Wistar rats were treated with 24 or 35 mg/kg of pure glyphosate every 48 h and neurobehavioral studies were performed. Our results indicated that gestational exposure to glyphosate induced changes in reflexes development, motor activity and cognitive function, in a dose-dependent manner. To go further, we evaluated whether prenatal exposure to glyphosate affected the Ca+2-mediated Wnt non-canonical signaling pathway. Results indicated that embryos exposed to glyphosate showed an inhibition of Wnt5a-CaMKII signaling pathway, an essential cascade controlling the formation and integration of neural circuits. Taken together, these findings suggest that gestational exposure to glyphosate leads to a downregulation of Wnt/Ca+2 pathway that could induce a developmental neurotoxicity evidenced by deficits at behavioral and cognitive levels in rat pups.

Glyphosate and its formulation Roundup impair pig oocyte maturation

Glyphosate, formulated as glyphosate-based herbicides (GBHs) including the best-known formulation Roundup, is the world's most widely used herbicide. During the last years, the growing and widespread use of GBHs has raised a great concern about the impact of environmental contamination on animal and human health including potential effect on reproductive systems. Using an in vitro model of pig oocyte maturation, we examined the biological impact of both glyphosate and Roundup on female gamete evaluating nuclear maturation, cytoplasmic maturation and developmental competence of oocytes, steroidogenic activity of cumulus cells as well as intracellular levels of glutathione (GSH) and ROS of oocytes. Our results indicate that although exposure to glyphosate and Roundup during in vitro maturation does not affect nuclear maturation and embryo cleavage, it does impair oocyte developmental competence in terms of blastocyst rate and cellularity. Moreover, Roundup at the same glyphosate-equivalent concentrations was shown to be more toxic than pure glyphosate, altering steroidogenesis and increasing oocyte ROS levels, thus confirming that Roundup adjuvants enhance glyphosate toxic effects and/or are biologically active in their side-effect and therefore should be considered and tested as active ingredients.

Effects of Roundup and its main component, glyphosate, upon mammalian sperm function and survival

The wide use of glyphosate-based herbicides (GBHs) has become a matter of concern due to its potential harmful effects on human health, including men fertility.This study sought to investigate, using the pig as a model, the impact of pure glyphosate and its most known commercial formulation, Roundup, on sperm function and survival. With this purpose, fresh commercial semen doses were incubated with different concentrations (0–360 µg/mL) of glyphosate (GLY; exp. 1) or Roundup, at the equivalent GLY concentration (exp. 2), at 38 °C for 3 h. Glyphosate at 360 µg/mL significantly (P < 0.05) decreased sperm motility, viability, mitochondrial activity and acrosome integrity but had no detrimental effect at lower doses. On the other hand, Roundup did significantly (P < 0.05) reduce sperm motility at ≥ 5 µg/mL GLY-equivalent concentration; mitochondrial activity at ≥ 25 µg/mL GLY-equivalent concentration; and sperm viability and acrosome integrity at ≥ 100 µg/mL GLY-equivalent concentration as early as 1 h of incubation. In a similar fashion, GLY and Roundup did not inflict any detrimental effect on sperm DNA integrity. Taken together, these data indicate that, while both glyphosate and Roundup exert a negative impact on male gametes, Roundup is more toxic than its main component, glyphosate.

Impact of field-realistic doses of glyphosate and nutritional stress on mosquito life history traits and susceptibility to malaria parasite infection.

Glyphosate is the world's most widely used herbicide. The commercial success of this molecule is due to its nonselectivity and its action, which would supposedly target specific biosynthetic pathways found mainly in plants. Multiple studies have however provided evidence for high sensitivity of many nontarget species to glyphosate and/or to formulations (glyphosate mixed with surfactants). This herbicide, found at significant levels in aquatic systems through surface runoffs, impacts life history traits and immune parameters of several aquatic invertebrates' species, including disease‐vector mosquitoes. Mosquitoes, from hatching to emergence, are exposed to aquatic chemical contaminants. In this study, we first compared the toxicity of pure glyphosate to the toxicity of glyphosate‐based formulations for the main vector of avian malaria in Europe, Culex pipiens mosquito. Then we evaluated, for the first time, how field‐realistic dose of glyphosate interacts with larval nutritional stress to alter mosquito life history traits and susceptibility to avian malaria parasite infection. Our results show that exposure of larvae to field‐realistic doses of glyphosate, pure or in formulation, did not affect larval survival rate, adult size, and female fecundity. One of our two experimental blocks showed, however, that exposure to glyphosate decreased development time and reduced mosquito infection probability by malaria parasite. Interestingly, the effect on malaria infection was lost when the larvae were also subjected to a nutritional stress, probably due to a lower ingestion of glyphosate.

Mitochondrial metabolism is central for response and resistance of Saccharomyces cerevisiae to exposure to a glyphosate-based herbicide

Glyphosate-based herbicides, the most extensively used herbicides in the world, are available in an enormous number of commercial formulations with varying additives and adjuvants. Here, we study the effects of one such formulation, Credit41, in two genetically diverse yeast strains. A quantitative trait loci (QTL) analysis between a sensitive laboratory strain and a resistant strain linked mitochondrial function to Credit41 resistance. Two genes encoding mitochondrial proteins identified through the QTL analysis were HFA1, a gene that encodes a mitochondrial acetyl CoA carboxylase, and AAC3, which encodes a mitochondrial inner membrane ATP/ADP translocator. Further analysis of previously studied whole-genome sequencing data showed that, although each strain uses varying routes to attain glyphosate resistance, most strains have duplications of mitochondrial genes. One of the most well-studied functions of the mitochondria is the assembly of Fe–S clusters. In the current study, the expression of iron transporters in the transcriptome increased in cells resistant to Credit41. The levels of iron within the cell also increased in cells exposed to Credit41 but not pure glyphosate. Hence, the additives in glyphosate-based herbicides have a significant contribution to the negative effects of these commercial formulations on biological systems.

Disruption of cytochrome P450 enzymes in the liver and small intestine in chicken embryos in ovo exposed to glyphosate.

Glyphosate is the active component of several commercial formulations as in Roundup®. The present study was investigated the toxic effects of pure glyphosate or Roundup® on the liver and small intestine of chick embryos. On day 6, a total of 180 fertile eggs injected with deionized water (control group), 10mg pure glyphosate, or 10mg of the active ingredient glyphosate in Roundup®/kg egg mass. The results showed an increase in relative weights of the liver in embryos that treated with Roundup®. Furthermore, oxidative stress was observed in the embryos treated with glyphosate or Roundup®, increased total superoxide dismutase, and content of malondialdehyde in the liver and intestine; moreover, decrease of glutathione peroxidase in the liver with increased in the intestine compared with the control. Besides, glutamic-pyruvic transaminase was increased in Roundup® group compared with other groups. Moreover, histopathological alterations in the liver and intestine tissues were observed in treated groups. Suppression of hepatic CYP1A2, CYP1A4, CYP1B1, and MDR1 mRNA expression after exposed to Roundup®. Furthermore, inhibition of CYP1A4 in the duodenum, CYP1A4, and MRP2 in the jejunum in embryos exposed to glyphosate or Roundup®. In addition, glyphosate treatment caused an increase of CYP3A5, CYP1C1, and IFNY mRNA expression in the jejunum and CYP1A2 expression in the ileum, while IFN-Y gene increase in embryos treated with Roundup®. In conclusion, in ovo exposure to glyphosate caused histopathological alterations and induced oxidative stress in the liver and small intestines. Moreover, the expression of cytochrome P450, MDR1, and MRP2 transporters was also modulated in the liver and small intestines for chick embryos.

Ecotoxicological effects of new C-substituted derivatives of N-phosphonomethylglycine (glyphosate) and their preliminary evaluation towards herbicidal application in agriculture.

In this paper, comparison of ecotoxicological and herbicidal effect of newly synthesized N‑[(phosphono)(aryl)methyl]glycines 1a-g (C-substituted glyphosate derivatives) with pure glyphosate (N-phosphonomethylglycine) (2) was demonstrated. All of tested glyphosate derivatives (1a-g) in contrast to glyphosate, were found to be completely safe for oat (Avena sativa) and classified as not harmful for marine bacteria Aliivibrio fischeri. Compounds 1a-g were also found rather harmless to radish (Raphanus sativus) as compared to N-phosphonomethylglycine, but they were moderately toxic against freshwater crustaceans Heterocypris incongruens. One of synthesized compounds, namely N-[(phosphono)(4-hydroxyphenyl)methyl]glycine (1f) was found to possess stronger herbicidal properties against gallant soldier (Galinsoga parviflora) and common sorrel (Rumex acetosa) when compared to pure glyphosate and demonstrated total death of these weeds being ranked 1 in the European Weed Research Council (EWRC) scale. Considering lower phytotoxicity of compound 1f against cultivated plants and tested microorganisms when compared to pure glyphosate, this aminophosphonate may be good candidate for further, more comprehensive study toward its agrochemical application, especially that this active agent demonstrated much stronger herbicidal properties than N-phosphonomethylglycine.

Zebrafish (Danio rerio) ability to activate ABCC transporters after exposure to glyphosate and its formulation Roundup Transorb®

The emergent demand for food production has increased the widespread use of pesticides, especially glyphosate-based herbicides as they can protect different types of crops, especially transgenic ones. Molecules of glyphosate have been found in water bodies around the world, and its presence can cause negative effects on non-target organisms, such as fish. Glyphosate toxicity appears to be systemic in fish but does not affect their organs equally. Also, its formulations can be more toxic than pure glyphosate. In this sense, we investigated if these variations in toxicity could be related to ATP binding cassette subfamily C (ABCC) transporters and the cellular detoxification capacity, following exposure to herbicides. Thus, adults of Danio rerio were exposed (24 and 96 h) to glyphosate and Roundup Transorb® (RT) at an environmental concentration of 0.1 mg/L, and the activity of ABCC proteins and gene expression of five isoforms of ABCC were analyzed. Glyphosate and RT exposure increased ABCC protein activity and gene expression up to 3-fold when compared to controls, indicating the activation of detoxification mechanisms. Only in the brain of D. rerio, the exposure to RT did not stimulate the activity of ABCC proteins, neither the expression of genes abcc1 and abcc4 that responded to the exposure to pure glyphosate. These results may suggest that the brain is more sensitive to RT than the other target-tissues since the mechanism of detoxification via ABCC transporters were not activated in this tissue as it was in the other.

Developmental and lethal effects of glyphosate and a glyphosate-based product on Xenopus laevis embryos and tadpoles.

Effects of pure glyphosate and a glyphosate-based product were evaluated comparatively using two embryonic development stages of Xenopus laevis as model system. When pure glyphosate was applied in pH adjusted media, lethal or developmental effects were not observed at concentrations up to 500mgL-1. The 96h LC50 values for the commercial herbicide, in contrast, were 32.1 and 35.1mg active ingredientL-1 for embryos and tadpoles, respectively. Since pure glyphosate has no effect on the selected biomarkers, it is thought that developmental toxic effects caused by glyphosate-based products are increased mainly due to formulation additives.

Glyphosate residues in soil affect crop plant germination and growth

Glyphosate-based herbicides (GBH) are the most widely used pesticides globally. Their persistence in soils and effects on non-target organisms have become a concern in agricultural and natural ecosystems. We experimentally studied, whether residues of GBH (Roundup Gold) or pure glyphosate in soils affect the germination or sprouting and growth of crop plants after the safety period. The seed germination of faba bean, oat and turnip rape, and sprouting of potato tubers was delayed in the greenhouse experiments in soils treated with GBH or with pure glyphosate. The total shoot biomass of faba bean was 28%, oat 29% and turnip rape 58% higher in control compared to GBH soils four weeks after sowing. In the beginning of the growing season, the plant growth in the field experiment supported the observations in the greenhouse experiment. However, at the end of the field experiment, potato shoot biomass was 25% and tuber biomass 14% greater in GBH soil compared to control soil. Potato tubers tended to gather low amounts of glyphosate (0.02 mg/kg) and its metabolite AMPA (0.07 mg/kg). Grazing by barnacle geese was three times higher in oats growing in the GBH soils compared to control oats in the field. Our results draw attention to complex indirect effects of GBH on crop plant seedling establishment and resistance to herbivores.

The surfactant polyethoxylated tallowamine (POEA) reduces lifespan and inhibits fecundity in Drosophila melanogaster- In vivo and in vitro study

In order to develop an understanding of the role of adjuvants in a popular glyphosate-based herbicide – Roundup® Concentrate Plus (RCP), on non-target organisms, the effects of pure glyphosate [N-(phosphonomethyl)-glycine], RCP and a non-ionic surfactant – polyethoxylated tallowamine (POEA) were studied in the fruit fly Drosophila melanogaster. Acute exposure to sub-lethal concentrations of RCP (15 μg/mL) and POEA (45 μg/mL) reduced (p < 0.001) lifespan of female flies compared to untreated controls or glyphosate (100 μg/mL). Negative geotaxis responses in female flies were reduced (p < 0.05) following acute exposure to sub-lethal concentrations of RCP and POEA whereas glyphosate did not significantly affect this response compared to untreated flies. Acute exposure to sub-lethal concentrations of RCP and POEA elevated (p < 0.05) protein carbonyl levels while markedly (p < 0.01) inhibiting carbonyl reductase activity whereas glyphosate treatment did not significantly affect protein carbonyl levels or carbonyl reductase activity. Fecundity was reduced (p < 0.05) following exposure to sub-lethal concentrations of RCP and POEA whereas glyphosate did not affect fecundity. In vitro treatment of ovarian stem sheath (OSS) cells with sub-lethal concentrations of RCP and POEA revealed decreased cell viability and enhanced caspase activity indicative of pro-apoptotic processes after 48 h compared to untreated controls. Glyphosate however was non-toxic at the concentration used. The results suggest that RCP and the surfactant POEA are more toxic than pure glyphosate and inhibit fecundity in Drosophila by impairing cell viability through enhanced apoptosis.