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PubMed2005

Oral bioavailability of glyphosate: studies using two intestinal cell lines.

Glyphosate is a commonly used nonselective herbicide that inhibits plant growth through interference with the production of essential aromatic amino acids. In vivo studies in mammals with radiolabeled glyphosate have shown that 34% of radioactivity was associated with intestinal tissue 2 h after oral administration. The aim of our research was to investigate the transport, binding, and toxicity of glyphosate to the cultured human intestinal epithelial cell line, Caco-2, and the rat small intestinal crypt-derived cell line, ileum epithelial cells-18 (IEC-18). An in vitro analysis of the transport kinetics of [14C]-glyphosate showed that 4 h after exposure, approximately 8% of radiolabeled glyphosate moved through the Caco-2 monolayer in a dose-dependent manner. Binding of glyphosate to cells was saturable and approximately 4 x 10(11) binding sites/cell were estimated from bound [14C]. Exposure of Caco-2 cells to > or =10 mg/ml glyphosate reduced transmembrane electrical resistance (TEER) by 82 to 96% and increased permeability to [3H]-mannitol, indicating that paracellular permeability increased in glyphosate-treated cells. At 10-mg/ml glyphosate, both IEC-18 and Caco-2 cells showed disruption in the actin cytoskeleton. In Caco-2 cells, significant lactate dehydrogenase leakage was observed when cells were exposed to 15 mg/ml of glyphosate. These data indicate that at doses >10 mg/ml, glyphosate significantly disrupts the barrier properties of cultured intestinal cells.

Actins↗

PubMed2004

The correlation of the spraying volume with herbicide adherence and herbicide penetration in glyphosate treatments.

The relationship between the spraying volume and glyphosate effectiveness has been widely studied over the years. Likewise, the effect of different adjuvants on glyphosate leaf adherence/penetration is currently well understood. However, not much is known about the way spraying volumes may influence these two herbicide properties or, in other words, which factor (adherence or penetration) is the limiting factor when spraying at low or high volumes of treatment. Using grass-weed Lolium rigidum and broad-leaved weed Portulaca oleracea as experimental systems, the effect of low (200 L ha(-1)) and high (800 L ha(-1)) spraying volumes and six commercial adjuvants (poly-l-p-menthene, a mixture of methyl oleate and palmitate, alkylglycol ester, dodecylbenzene ammonium sulphonate, and two paraffinic oils) plus ammonium sulfate on the effectiveness of five commercial glyphosate formulations has been studied under laboratory controlled conditions. Dose-response assays showed significant differences between low and high spraying volume treatments as expected, the former being much more effective than the latter. However, and in all commercial formulations and weed species tested, the best results at low spraying volumes were always obtained when formulating glyphosate with oils or their derivatives, with a minimal improvement observed in glyphosate/surfactant mixtures. On the contrary, both the use of ionic or non-ionic surfactants as glyphosate adjuvants gave the best results at high spraying volumes, with only a marginal success observed on paraffinic oils and fatty acidesters. Data point out glyphosate absorption as being a two-step process made up of adherence first and then penetration, both events being essential for glyphosate effectiveness. Under these terms, high performance glyphosate-rich low spraying volumes succeed by themselves in soaking leaf surfaces, penetration therefore being the main and only limiting factor, while low performance glyphosate-poor high spraying volumes fail to either soak or penetrate leaf cuticles. The use of penetrative agents such as oils is therefore advised as a single method to increase glyphosate effectiveness at low spraying volumes, whereas glyphosate treatments at high spraying volumes are better improved by using surfactant agents or surfactant + oil mixtures.

Biological Transport↗

PubMed1992

NTP technical report on the toxicity studies of Glyphosate (CAS No. 1071-83-6) Administered In Dosed Feed To F344/N Rats And B6C3F1 Mice.

Glyphosate is a systemic, broad-spectrum, post-emergence herbicide used for non-selective weed control. It was selected for study because of its widespread use, potential for human exposure, and the lack of published reports concerning comprehensive toxicity or carcinogenicity evaluations. Chemical disposition, 13-week toxicity, and mutagenicity studies of glyphosate were conducted. In disposition studies, male F344/N rats were administered an oral dose (5.6 or 56 mg/kg) of 14C-glyphosate. Blood, urine, fecal, and tissue samples were collected and analyzed for radioactivity. Within 72 hours after glyphosate dosing, 20-30% of the administered radioactivity was eliminated via urine, 70-80% via feces, and about 1% of the radioactivity remained in the tissues. Studies following oral, intravenous, and intraperitoneal administration of glyphosate indicated that the urinary radioactivity represented the amount of glyphosate absorbed and that the fecal radioactivity represented the amount unabsorbed from the gastrointestinal tract. In the 13-week toxicity studies, groups of 10 male and female F344/N rats and B6C3F1 mice were administered glyphosate in feed at 0, 3125, 6250, 12500, 25000, or 50000 ppm. Glyphosate administration induced increases in serum bile acids, alkaline phosphatase, and alanine aminotransferase activities in rats, suggesting mild toxicity to the hepatobiliary system. Clinical pathology measurements were not performed with mice. No histopathologic lesions were observed in the livers of rats or mice. There was no evidence of adverse effects on the reproductive system of rats or mice. Cytoplasmic alteration was observed in the parotid and submandibular salivary glands of rats and parotid salivary glands in mice. The salivary gland effects of glyphosate were demonstrated to be mediated through an adrenergic mechanism which could be blocked by the adrenergic antagonist, propanolol. Glyphosate was not mutagenic in Salmonella, and did not induce micronuclei in mice. The no-observed-adverse-effect level (NOAEL) for the salivary gland lesions was 3125 ppm in the diet for mice. A NOAEL could not be determined from the rat study. Synonyms: Glyphosate, technical grade; Glycine, N-(phosphonomethyl); N-phosphono-methyl glycine; N-(phosphonomethyl)glycine; MON 0573; MON 2139.

Journal Article↗

PubMed2004

Sorption, mineralization and mobility of N-(phosphonomethyl)glycine (glyphosate) in five different types of gravel.

Sorption, mineralization and mobility of glyphosate were studied in six substrates: the five types of gravel most frequently used as surfacing in Denmark and a sandy agricultural soil from Simmelkaer that served as a reference soil. Cumulative mineralization of [methyl-14C]glyphosate in batch studies was highest in coarse gravel, amounting to 14% after 4 days at 30 degrees C and 32% after 31 days. Mineralization was slowest in the sandy reference soil, amounting to only 2% after 31 days. The adsorption coefficient (Kd) of glyphosate in gravel ranged from 62 to 164 litre kg(-1), while that in the sandy reference soil was 410 litre kg(-1). The results indicate that the relatively low Kd in gravel allows a relatively high rate of glyphosate mineralization by the biomass. When Kd is high, in contrast, mineralization is slow. Lowering the temperature to 10 degrees C decreased mineralization by 50% in one of two gravels. The leaching of glyphosate was screened in simple columns of gravel or soil in which precipitation events (20 mm over a 2-h period) were simulated on three occasions, starting either immediately after or 2 days after application of glyphosate. [14C]Glyphosate was applied as a tracer mixed with the commercial product Roundup Garden at the recommended rate of 2.4 kg glyphosate ha(-1), equivalent to 1 microg g(-1) soil. The highest concentration of [14C] compounds (expressed in terms of glyphosate concentration) in leachate from the columns exceeded 1300 microg litre(-1), and was detected in rounded gravel after the first rain event. No glyphosate was detected in leachate from the sandy reference soil.

Adsorption↗

PubMed2001

Influence of glyphosate on amino acid composition of Egyptian broomrape.

The parasitic plant broomrape is entirely dependent on its host for reduced carbon and nitrogen and is also susceptible to inhibition by glyphosate that is translocated to the parasite through a host. Studies were conducted to examine the effect of broomrape parasitism on amino acid concentrations of two hosts: common vetch that is tolerant of low levels of glyphosate and oilseed rape that has been genetically engineered for glyphosate resistance. The influence of glyphosate on the amino acid content of broomrape and the two hosts was also examined. Amino acid concentrations in leaves and roots of parasitized common vetch plants were generally similar to those of the corresponding tissues of nonparasitized plants. Amino acid concentrations in broomrape were lower than those of the parasitized common vetch root. For common vetch, glyphosate applied at rates that selectively inhibited broomrape growth did not alter individual amino acid concentrations in the leaves, but generally increased amino acid levels at 0.18 kg ha-1. Glyphosate application also increased the amino acid concentrations, with the exception of arginine, of broomrape growing on common vetch and did not generally influence concentrations in leaves or roots of common vetch. In oilseed rape, parasitization by broomrape generally led to higher amino acid concentrations in leaves but lower concentrations in roots of parasitized plants. Broomrape had higher amino acid concentrations than roots of the parasitized oilseed rape. Glyphosate applied at 0.25 and 0.5 kg ha-1 generally increased the amino acid concentrations in oilseed rape leaves, but the 0.75 kg ha-1 application caused the amino acid concentrations to decrease compared to those of untreated plants. In oilseed rape root the general trend was an increase in the concentration of amino acids at the two highest rates of glyphosate. Individual amino acid concentrations in broomrape attachments growing on oilseed rape were generally increased following glyphosate application of 0.25 kg ha-1. These results indicate that low rates of glyphosate alter amino acid profiles in both host and broomrape and raise questions about the regulation of amino acid metabolism in the parasite.

Amino Acids↗

PubMed1999

Influence of incubation temperature on the behavior of triethylamine-extractable glyphosate (N-phosphonomethylglycine) in four soils.

The behavior of glyphosate, extracted from four soils using aqueous triethylamine, was investigated at two temperatures. For each soil, and at both temperatures, there was a marked loss in the amount of extractable glyphosate immediately after addition of the herbicide to soil. This rapid loss of glyphosate was ascribed to adsorption of the herbicide into a nonextractable form. For three of the four soils used when incubated at 25 degrees C, the rates of loss of extractable glyphosate were similar to previously measured rates of degradation of this herbicide in these soils. However, loss of extractable glyphosate from the Culgoa clay loam was due not only to substrate degradation but also to slow sorption of glyphosate into the nonextractable form in this soil over the experimental period. For the Rutherglen and Walpeup soils, when incubated at 10 degrees C, the rates of loss of extractable glyphosate were half of the previously measured rate of degradation of this herbicide in these soils. However, there was no measured loss of extractable glyphosate from the Wimmera clay. As previous work has shown glyphosate to decompose readily in these soils at this temperature, these findings suggest that desorption of glyphosate may occur at a rate greater than degradation at this temperature and, hence, that temperature may play a pivotal role in sorption processes. Investigations with these soils when sterilized by gamma-irradiation showed that for the Walpeup, Wimmera, and Rutherglen soils, sorption was complete soon after the addition of the herbicide; however, for the Culgoa soil, further adsorption occurred over the entire experimental period.

Ethylamines↗

PubMed2004

Toxicity of glyphosate-based pesticides to four North American frog species.

Glyphosate-based herbicides are among the most widely used pesticides in the world. We compared the acute toxicity of the glyphosate end-use formulation Roundup Original to four North American amphibian species (Rana clamitans, R. pipiens, R. sylvatica, and Bufo americanus) and the toxicity of glyphosate technical, the polyethoxylated tallowamine surfactant (POEA) commonly used in glyphosate-based herbicides, and five newer glyphosate formulations to R. clamitans. For R. clamitans, acute toxicity values in order of decreasing toxicity were POEA > Roundup Original > Roundup Transorb > Glyfos AU; no significant acute toxicity was observed with glyphosate technical material or the glyphosate formulations Roundup Biactive, Touchdown, or Glyfos BIO. Comparisons between the four amphibian species showed that the toxicity of Roundup Original varied with species and developmental stage. Rana pipiens tadpoles chronically exposed to environmentally relevant concentrations of POEA or glyphosate formulations containing POEA showed decreased snout-vent length at metamorphosis and increased time to metamorphosis, tail damage, and gonadal abnormalities. These effects may be caused, in some part, by disruption of hormone signaling, because thyroid hormone receptor beta mRNA transcript levels were elevated by exposure to formulations containing glyphosate and POEA. Taken together, the data suggest that surfactant composition must be considered in the evaluation of toxicity of glyphosate-based herbicides.

Animals↗

PubMed1988

Import of a precursor protein into chloroplasts is inhibited by the herbicide glyphosate.

Import of the precursor to 5-enolpyruvylshikimate-3-phosphate synthase (pEPSPS) into chloroplasts is inhibited by the herbicide glyphosate. Inhibition of import is maximal at glyphosate concentrations of >/=10 mum and occurs only when pEPSPS is present as a ternary complex of enzyme-shikimate-3-phosphate-glyphosate. Glyphosate alone had no effect on the import of pEPSPS since it is not known to interact with the enzyme in the absence of shikimate-3-phosphate. Experiments with wild-type and glyphosate-resistant mutant forms of pEPSPS show that inhibition of import is directly proportional to the binding constants for glyphosate. Inhibition of import is thus a direct consequence of glyphosate binding to the enzyme-shikimate-3-phosphate complex. The potential for non-specific effects of glyphosate on the chloroplast transport mechanism has been discounted by showing that import of another chloroplast-designated protein was unaffected by high concentrations of glyphosate and shikimate-3-phosphate. The mechanism of import inhibition by glyphosate is consistent with a precursor unfolding/refolding model.

Journal Article↗

PubMed1984

Glyphosate Inhibition of 5-Enolpyruvylshikimate 3-Phosphate Synthase from Suspension-Cultured Cells of Nicotiana silvestris.

Treatment of isogenic suspension-cultured cells of Nicotiana silvestris Speg. et Comes with glyphosate (N-[phosphonomethyl]glycine) led to elevated levels of intracellular shikimate (364-fold increase by 1.0 millimolar glyphosate). In the presence of glyphosate, it is likely that most molecules of shikimate originate from the action of 3-deoxy-d-arabino-heptulosonate 7-phosphate (DAHP) synthase-Mn since this isozyme, in contrast to the DAHP synthase-Co isozyme, is insensitive to inhibition by glyphosate. 5-Enolpyruvylshikimate 3-phosphate (EPSP) synthase (EC 2.5.1.19) from N. silvestris was sensitive to micromolar concentrations of glyphosate and possessed a single inhibitor binding site. Rigorous kinetic studies of EPSP synthase required resolution from the multiple phosphatase activities present in crude extracts, a result achieved by ion-exchange column chromatography. Although EPSP synthase exhibited a broad pH profile (50% of maximal activity between pH 6.2 and 8.5), sensitivity to glyphosate increased dramatically with increasing pH within this range. In accordance with these data and the pK(a) values of glyphosate, it is likely that the ionic form of glyphosate inhibiting EPSP synthase is COO(-)CH(2)NH(2) (+)CH(2)PO(3) (2-), and that a completely ionized phosphono group is essential for inhibition. At pH 7.0, inhibition was competitive with respect to phosphoenolpyruvate (K(i) = 1.25 micromolar) and uncompetitive with respect to shikimate-3-P (K(i)' = 18.3 micromolar). All data were consistent with a mechanism of inhibition in which glyphosate competes with PEP for binding to an [enzyme:shikimate-3-P] complex and ultimately forms the dead-end complex of [enzyme:shikimate-3-P:glyphosate].

Journal Article↗

PubMed2005

Effect of freezing and thawing on microbial activity and glyphosate degradation in two Norwegian soils.

Little research has been done on pesticide dissipation in cold climates and there is a need to focus on the influence of climate on pesticide degradation in soil. Glyphosate, N-(phosphonomethyl)glycine, is a herbicide frequently used for controlling perennial weeds through application after harvest and was used as a model compound for this study. The effect of freeze-thaw activity on the availability of glyphosate in soil, and consequently its mineralization by soil microorganisms, was studied through laboratory incubations of repacked soil cores treated with 14C-labelled glyphosate and subjected to different freeze-thaw treatments. Winter simulation regimes applied were constant thaw (+5 degrees C), constant freezing (-5 degrees C), unstable conditions with short fluctuations (24 h of -5 degrees C followed by 24 h of +5 degrees C), and long duration fluctuations (3 weeks of -5 degrees C followed by 3 weeks of +5 degrees C). Distribution of 14C-glyphosate was followed during the incubation through measurements of the mineralized fraction (14CO2), soil water fraction, KOH extractable fraction, and non-extractable fraction. Microbial parameters used to characterize the soils were estimates of size of microbial biomass, overall microbial activity and microbial diversity. The constant freezing treatment exhibited the lowest amount of glyphosate mineralization. The constant thawed treatment and the treatments with fluctuating temperature exhibited significantly increased mineralization. These results were in accordance with the observed concentration of glyphosate in soil water; the higher the activity, the lower the concentration. The amount of glyphosate extractable with KOH and the resulting non-extractable fraction, however, were not significantly affected by soil type or temperature regime. The glyphosate mineralization pattern was comparable with the overall microbial activity in the soils. Observed different levels of diversity might explain some of the difference in total glyphosate mineralization between soils.

Biodegradation, Environmental↗

PubMed2006

Comparison of efficacy, absorption and translocation of three glyphosate formulations on velvetleaf (Abutilon theophrasti).

Growth analysis, absorption and translocation studies were conducted to compare a 1-aminomethanamide dihydrogen tetraoxosulfate (GLY-A) formulation of glyphosate with two isopropylamine (GLY-IPA-1, GLY-IPA-2) formulations of glyphosate on velvetleaf. The two isopropylamine formulations differed by the presence of a surfactant in the formulation, GLY-IPA-1 containing surfactant whereas GLY-IPA-2 did not. Four- to six-leaf velvetleaf was treated with GLY-A and GLY-IPA-1 and GLY-IPA-2 (0, 50, 67, 89, 119, 158, 280, 420, 560 and 840 g AE ha(-1)) with and without ammonium sulfate (AMS; 20 g L(-1)). GLY-A and GLY-IPA-2 included a non-ionic surfactant (2.5 mL L(-1)) in the spray solution at all herbicide concentrations. No additional surfactant was added to GLY-IPA-1. The IC50 value for GLY-A was 88 g AE ha(-1) compared with 346 and 376 g AE ha(-1) for GLY-IPA-1 and GLY-IPA-2 respectively in the absence of AMS. When AMS (20 g L(-1)) was added to the spray solution, the estimated IC50 values were 143, 76 and 60 g AE ha(-1) for GLY-IPA-1, GLY-IPA-2 and GLY-A respectively. Absorption of 14C-glyphosate into the third leaf of five- to six-leaf velvetleaf was three- to sixfold greater 72 h after treatment (HAT) when applied as GLY-A compared with GLY-IPA-1 and GLY-IPA-2 respectively in the absence of AMS. AMS (20 g L(-1)) increased absorption of 14C-glyphosate in all glyphosate formulations two- to threefold, but differences among the formulations remained. Approximately three- and sixfold more 14C-glyphosate applied as GLY-A had translocated out of the treated leaf compared with GLY-IPA-1 and GLY-IPA-2 respectively by 72 HAT. Adding AMS (20 g L(-1)) increased translocation of 14C-glyphosate out of the treated leaf approximately 2.5-fold for all three formulations. The increased efficacy of GLY-A versus GLY-IPA-1 and GLY-IPA-2 on velvetleaf is due to the greater rate of absorption and subsequent translocation of glyphosate out of the treated leaf. AMS increased the efficacy of all three formulations by increasing absorption and translocation of glyphosate in the plant.

Absorption↗

PubMed2006

Cosorption of zinc and glyphosate on two soils with different characteristics.

Agricultural application of large amounts of glyphosate [N-(phosphonomethyl)-glycine] may affect soil metal behaviors to some extend, because glyphosate can react with many kinds of metals to form metal complexes. Cosorption of Zn and glyphosate on a Red soil (RS, Udic Ferrosols) and a Wushan soil (WS, Anthrosol) was studied. In comparison with the WS, the RS has less adsorption capacity for Zn and higher for glyphosate. The presence of glyphosate decreased Zn adsorption on the two soils, which are resulted from the decreased equilibrium solution pH caused by the added glyphosate, and also the formation of water-soluble complexes of glyphosate with solution Zn(2+) that had lower affinity to soil surface in comparison with Zn(2+) itself. Such effect is more significant on the RS than on the WS, mainly because of the less adsorption quantity of Zn on the former one. On the contrary, the presence of Zn increased the adsorption quantities of glyphosate on the RS and WS, which is resulted from the decreasing pH value of the equilibrium solution caused by Zn(2+) exchange with H(+) ions of soil surface. Such results suggest that glyphosate in field may increase the mobility and bioavailability of Zn and correspondingly increase its environmental risk.

Adsorption↗

PubMed2005

Urban contributions of glyphosate and its degradate AMPA to streams in the United States.

Glyphosate is the most widely used herbicide in the world, being routinely applied to control weeds in both agricultural and urban settings. Microbial degradation of glyphosate produces aminomethyl phosphonic acid (AMPA). The high polarity and water-solubility of glyphosate and AMPA has, until recently, made their analysis in water samples problematic. Thus, compared to other herbicides (e.g. atrazine) there are relatively few studies on the environmental occurrence of glyphosate and AMPA. In 2002, treated effluent samples were collected from 10 wastewater treatment plants (WWTPs) to study the occurrence of glyphosate and AMPA. Stream samples were collected upstream and downstream of the 10 WWTPs. Two reference streams were also sampled. The results document the apparent contribution of WWTP effluent to stream concentrations of glyphosate and AMPA, with roughly a two-fold increase in their frequencies of detection between stream samples collected upstream and those collected downstream of the WWTPs. Thus, urban use of glyphosate contributes to glyphosate and AMPA concentrations in streams in the United States. Overall, AMPA was detected much more frequently (67.5%) compared to glyphosate (17.5%).

Atrazine↗

PubMed2003

Effect of glyphosate on the microbial activity of two Brazilian soils.

Glyphosate [N-(phosphonomethyl)-glycine] is a broad-spectrum, non-selective, post-emergence herbicide that is widely used in agricultural. We studied, in vitro, changes in the microbial activity of typical Hapludult and Hapludox Brazilian soils, with and without applied glyphosate. Glyphosate was applied at a rate of 2.16 mg glyphosate kg(-1) of soil and microbial activity was measured by soil respiration (evolution of CO(2)) and fluorescein diacetate (FDA) hydrolysis over a period of 32 days. We found an increase of 10-15% in the CO(2) evolved and a 9-19% increase in FDA hydrolyses in the presence of glyphosate compared with the same type of soil which had never received glyphosate. Soil which had been exposed to glyphosate for several years had the strongest response in microbial activity. Most probable number (MPN) counts showed that after 32 days incubation the number of actinomycetes and fungi had increased while the number of bacteria showed a slight reduction. After the incubation period, high pressure liquid chromatography (HPLC) detected the glyphosate metabolite aminomethyl phosphonic acid (AMPA), indicating glyphosate degradation by soil microorganisms.

Bacteria↗

PubMed2006

Effects of surface sorption on microbial degradation of glyphosate.

Sorption may affect the bioavailability and biodegradation of pesticides in soils. The aim of this study was to test the effect of surface sorption on microbial utilization of the herbicide glyphosate as a source of phosphorus, nitrogen, or carbon. We added goethite to a humus soil to manipulate the soil's glyphosate sorption capacity. The addition of glyphosate generally either decreased microbial CO2 production or produced no effect. Additions of glyphosate, in combination with glucose and N, did not change the respiration rate in comparison with the same treatment but without glyphosate. In contrast, glyphosate additions combined with glucose and P decreased microbial growth, whereas the combination with goethite counteracted the negative effect. The different treatments were examined using attenuated total reflectance Fourier transform (ATR-FTIR) spectroscopy; the results suggest that glyphosate was de-carboxylated in the sorbed state. Stimulating microbial growth by the addition of glucose and nitrogen resulted in further oxidation of glyphosate and only phosphate was detectable on the goethite surface after 13 days incubation. Our results show that sorbed glyphosate is microbially degradable, and it retards microbial activity. This study emphasizes the importance of combining quantitative measurements with a molecular-level examination, to better understand biogeochemical processes.

Biodegradation, Environmental↗

PubMedDate not supplied

Ecological risk assessment for aquatic organisms from over-water uses of glyphosate.

Although the herbicide glyphosate is most widely used in agriculture, some is used for the control of emergent aquatic weeds in ditches, wetlands, and margins of water bodies, largely as the formulation Rodeo. This article presents an ecological risk assessment (ERA) of glyphosate and some of the recommended surfactants as used in or near aquatic systems. Glyphosate does not bioaccumulate, biomagnify, or persist in a biologically available form in the environment. Its mechanism of action is specific to plants and it is relatively nontoxic to animals. As a commercial product, glyphosate may be formulated with surfactants that increased efficacy but, in some cases, are more toxic to aquatic organisms than the parent material. For this risk assessment, three model exposure scenarios--static or low-flow systems such as ponds, flowing waters such as streams, and systems subjected to tidal flows such as estuaries--were chosen and application rates from 1 to 8 kg glyphosate/ha were modeled. Additional measured exposure data from several field studies were also used. As acute exposures are most likely to occur, acute toxicity data were used as effect measures for the purposes of risk assessment. Toxicity data were obtained from the literature and characterized using probabilistic techniques. Risk assessments based on estimated and measured concentrations of glyphosate that would result from its use for the control of undesirable plants in wetlands and over-water situations showed that the risk to aquatic organisms is negligible or small at application rates less than 4 kg/ha and only slightly greater at application rates of 8 kg/ha. Less is known about the environmental fate and toxicology of the surfactants commonly used in combination with the Rodeo formulation of glyphosate. The surfactants used for this purpose were judged not to be persistent nor bioaccumulative in the environment. Distributional analysis of measured deposition concentrations of LI 700, suggest that this surfactant presents an insignificant acute risk to aquatic organisms. Assuming similar applications rates, significant ecological effects would not be expected from the use of some other surfactants such as Induce or X-77. Risks from the use of glyphosate +MON 0818 (Roundup) were slightly greater than those from glyphosate and surfactants such as LI 700; however, in over-water uses, risks were still considered small. Similar small risks were observed for measured concentrations of glyphosate in surface waters resulting from aerial application of Vision (a formulation equivalent to Roundup) to forestry areas in Canada. Concentrations measured after ground application presented a greater risk, but the data were sparse and the assessment is more uncertain.

Animals↗

PubMed2005

Identification of a glyphosate-resistant mutant of rice 5-enolpyruvylshikimate 3-phosphate synthase using a directed evolution strategy.

5-enolpyruvylshikimate 3-phosphate synthase (EPSPS) is a key enzyme in the shikimate pathway and is targeted by the wide-spectrum herbicide glyphosate. Here, we describe the use of a selection system based on directed evolution to select glyphosate-resistant mutants of EPSPS. Using this system, the rice (Oryza sativa) EPSPS gene, mutagenized by Error-Prone polymerase chain reaction, was introduced into an EPSPS-deficient Escherichia coli strain, AB2829, and transformants were selected on minimal medium by functional complementation. Three mutants with high glyphosate resistance were identified in three independent glyphosate selection experiments. Each mutant contained a C(317)-->T transition within the EPSPS coding sequence, causing a change of proline-106 to leucine (P106L) in the protein sequence. Glyphosate resistance assays indicated a 3-fold increase in glyphosate resistance of E. coli expressing the P106L mutant. Affinity of the P106L mutant for glyphosate and phosphoenolpyruvate was decreased about 70-fold and 4.6-fold, respectively, compared to wild-type EPSPS. Analysis based on a kinetic model demonstrates that the P106L mutant has a high glyphosate resistance while retaining relatively high catalytic efficiency at low phosphoenolpyruvate concentrations. A mathematical model derived from the Michaelis-Menten equation was used to characterize the effect of expression level and selection conditions on kinetic (Ki and Km) variation of the mutants. This prediction suggests that the expression level is an important aspect of the selection system. Furthermore, glyphosate resistance of the P106L mutant was confirmed in transgenic tobacco (Nicotiana tabacum), demonstrating the potential for using the P106L mutant in transgenic crops.

3-Phosphoshikimate 1-Carboxyvinyltransferase↗

PubMed2005

Cancer incidence among glyphosate-exposed pesticide applicators in the Agricultural Health Study.

Glyphosate is a broad-spectrum herbicide that is one of the most frequently applied pesticides in the world. Although there has been little consistent evidence of genotoxicity or carcinogenicity from in vitro and animal studies, a few epidemiologic reports have indicated potential health effects of glyphosate. We evaluated associations between glyphosate exposure and cancer incidence in the Agricultural Health Study (AHS), a prospective cohort study of 57,311 licensed pesticide applicators in Iowa and North Carolina. Detailed information on pesticide use and other factors was obtained from a self-administered questionnaire completed at time of enrollment (1993-1997). Among private and commercial applicators, 75.5% reported having ever used glyphosate, of which > 97% were men. In this analysis, glyphosate exposure was defined as a) ever personally mixed or applied products containing glyphosate; b) cumulative lifetime days of use, or "cumulative exposure days" (years of use times days/year); and c) intensity-weighted cumulative exposure days (years of use times days/year times estimated intensity level). Poisson regression was used to estimate exposure-response relations between glyphosate and incidence of all cancers combined and 12 relatively common cancer subtypes. Glyphosate exposure was not associated with cancer incidence overall or with most of the cancer subtypes we studied. There was a suggested association with multiple myeloma incidence that should be followed up as more cases occur in the AHS. Given the widespread use of glyphosate, future analyses of the AHS will allow further examination of long-term health effects, including less common cancers.

Adult↗