Lowdown on Roundup Part IV

Aristilde et. al, 2017

Ludmilla Aristilde, Michael L. Reed, Rebecca A. Wilkes, Tracy Youngster, Matthew A. Kukurugya, Valerie Katz, and Clayton R. S. Sasaki, “Glyphosate-Induced Specific and Widespread Perturbations in the Metabolome of Soil Pseudomonas Species,” Frontiers in Environmental Science, 2017, 5:34, DOI: 10.3389/fenvs.2017.00034.


Previous studies have reported adverse effects of glyphosate on crop-beneficial soil bacterial species, including several soil Pseudomonas species. Of particular interest is the elucidation of the metabolic consequences of glyphosate toxicity in these species. Here we investigated the growth and metabolic responses of soil Pseudomonas species grown on succinate, a common root exudate, and glyphosate at different concentrations. We conducted our experiments with one agricultural soil isolate, P. fluorescens RA12, and three model species, P. putida KT2440, P. putida S12, and P. protegens Pf-5. Our results demonstrated both species- and strain-dependent growth responses to glyphosate. Following exposure to a range of glyphosate concentrations (up to 5 mM), the growth rate of both P. protegens Pf-5 and P. fluorescens RA12 remained unchanged whereas the two P. putida strains exhibited from 0 to 100% growth inhibition. We employed a 13C-assisted metabolomics approach using liquid chromatography-mass spectrometry to monitor disruptions in metabolic homeostasis and fluxes. Profiling of the whole-cell metabolome captured deviations in metabolite levels involved in the tricarboxylic acid cycle, ribonucleotide biosynthesis, and protein biosynthesis. Altered metabolite levels specifically in the biosynthetic pathway of aromatic amino acids (AAs), the target of toxicity for glyphosate in plants, implied the same toxicity target in the soil bacterium. Kinetic flux experiments with 13C-labeled succinate revealed that biosynthetic fluxes of the aromatic AAs were not inhibited in P. fluorescens Pf-5 in the presence of low and high glyphosate doses but these fluxes were inhibited by up to 60% in P. putida KT2440, even at sub-lethal glyphosate exposure. Notably, the greatest inhibition was found for the aromatic AA tryptophan, an important precursor to secondary metabolites. When the growth medium was supplemented with aromatic AAs, P. putida S12 exposed to a
lethal dose of glyphosate completely recovered in terms of both growth rate and selected metabolite levels. Collectively, our findings led us to conclude that the  glyphosateinduced specific disruption of de novo biosynthesis of aromatic AAs accompanied by widespread metabolic disruptions was responsible for dose-dependent adverse effects of glyphosate on sensitive soil Pseudomonas species.  FULL TEXT

Balbuena et. al, 2015

María Sol Balbuena, Léa Tison, Marie-Luise Hahn, Uwe Greggers, Randolf Menzel, and Walter M. Farina, “Effects of sub-lethal doses of glyphosate on honeybee navigation,” Journal of Experimental Biology,


Glyphosate (GLY) is a herbicide that is widely used in agriculture for weed control. Although reports about the impact of GLY in snails, crustaceans and amphibians exist, few studies have investigated its sublethal effects in non-target organisms such as the honeybee Apis mellifera, the main pollen vector in commercial crops. Here, we tested whether exposure to three sublethal concentrations of GLY (2.5, 5 and 10 mg l−1: corresponding to 0.125, 0.250 and 0.500 μg per animal) affects the homeward flight path of honeybees in an open field. We performed an experiment in which forager honeybees were trained to an artificial feeder, and then captured, fed with sugar solution containing traces of GLY and released from a novel site either once or twice. Their homeward trajectories were tracked using harmonic radar technology. We found that honeybees that had been fed with solution containing 10 mg l−1 GLY spent more time performing homeward flights than control bees or bees treated with lower concentrations. They also performed more indirect homing flights. Moreover, the proportion of direct homeward flights performed after a second release from the same site increased in control bees but not in treated bees. These results suggest that, in honeybees, exposure to levels of GLY commonly found in agricultural settings impairs the cognitive capacities needed to retrieve and integrate spatial information for a successful return to the hive. Therefore, honeybee navigation is affected by ingesting traces of the most widely used herbicide worldwide, with potential long-term negative consequences for colony foraging success.  FULL TEXT

Bento et. al, 2016

Bento CP, Yang X, Gort G, Xue S, van Dam R, Zomer P, Mol HG, Ritsema CJ, Geissen V, “Persistence of glyphosate and aminomethylphosphonic acid in loess soil under different combinations of temperature, soil moisture and light/darkness,” Science of the Total Environment, 2016, DOI: 10.1016/j.scitotenv.2016.07.215.


The dissipation kinetics of glyphosate and its metabolite aminomethylphosphonic acid (AMPA) were studied in loess soil, under biotic and abiotic conditions, as affected by temperature, soil moisture (SM) and light/darkness. Nonsterile and sterile soil samples were spiked with 16mgkg-1 of glyphosate, subjected to three SM contents (20% WHC, 60% WHC, saturation), and incubated for 30days at 5°C and 30°C, under dark and light regimes. Glyphosate and AMPA dissipation kinetics were fit to single-first-order (SFO) or first-order-multicompartment (FOMC) models, per treatment combination. AMPA kinetic model included both the formation and decline phases. Glyphosate dissipation kinetics followed SFO at 5°C, but FOMC at 30°C. AMPA followed SFO dissipation kinetics for all treatments. Glyphosate and AMPA dissipation occurred mostly by microbial activity. Abiotic processes played a negligible role for both compounds. Under biotic conditions, glyphosate dissipation and AMPA formation/dissipation were primarily affected by temperature, but also by SM. Light regimes didn’t play a significant role. Glyphosate DT50 varied between 1.5 and 53.5days, while its DT90 varied between 8.0 and 280days, depending on the treatment. AMPA persisted longer in soil than glyphosate, with its DT50 at 30°C ranging between 26.4 and 44.5days, and its DT90 between 87.8 and 148days. The shortest DT50/DT90 values for both compounds occurred at 30°C and under optimal/saturated moisture conditions, while the largest occurred at 5°C and reaching drought stress conditions. Based on these results, we conclude that glyphosate and AMPA dissipate rapidly under warm and rainy climate conditions. However, repeated glyphosate applications in fallows or winter crops in countries where cold and dry winters normally occur could lead to on-site soil pollution, with consequent potential risks to the environment and human health. To our knowledge, this study is the first evaluating the combined effect of temperature, soil moisture and light/dark conditions on AMPA formation/dissipation kinetics and behaviour.

Bonini et al., 2009

E.A. Bonini, M.L.L. Ferrarese, R. Marchiosi, P.C. Zonetti, O. Ferrarese-Filho, “A simple chromatographic assay to discriminate between glyphosate-resistant and susceptible soybean (Glycine max) cultivars,” European Journal of Agronomy, 2009, 31:3, 1730176, DOI: 10.1016/e.eja.2009.03.006.


In glyphosate-susceptible soybean (Glycine max L. Merrill), the herbicide glyphosate [(N-phosphonomethyl)glycine] inhibits the enzyme 5-enolpyruvylshikimate-3-phosphate (EPSP) synthase, causing a massive accumulation of the metabolite shikimate. This phenomenon does not occur in glyphosate-resistant soybean due the presence of a gene encoding glyphosate-insensitive EPSP synthase. This study proposes a simple and reliable assay as an alternative tool for differentiating glyphosate-resistant from susceptible soybean cultivars. The assay is based on a single extraction of leaf or root tissue. Shikimate is quantified by reversed-phase high–performance liquid chromatography at 220 nm and isocratic elution with phosphoric acid. After glyphosate treatment, tissues of glyphosate-susceptible plants were found to massively accumulate shikimate, whereas the tissues of glyphosate-resistant plants did not accumulate the metabolite.  FULL TEXT

Brodeur et. al, 2014

Julie Céline Brodeur, María Belén Poliserpi, María Florencia D’Andrea, Marisol Sánchez, “Synergy between glyphosate- and cypermethrin-based pesticides during acute exposures in tadpoles of the common South American Toad Rhinella arenarum,” Chemosphere, 2014, 112:70-76, DOI: 10.1016/j.chemosphere.2014.02.065.


The herbicide glyphosate and the insecticide cypermethrin are key pesticides of modern management in soy and corn cultures. Although these pesticides are likely to co-occur in ephemeral ponds or aquatic systems supporting amphibian wildlife, the toxicological interactions prevailing in mixtures of these two pesticides have been little studied. The current study evaluated the toxicity of equitoxic and nonequitoxic binary mixtures of glyphosate- and cypermethrin-based pesticides to tadpoles of the common South American toad, Rhinella arenarum. Two different combinations of commercial products were tested: glyphosate Glifosato Atanor + cypermethrin Xiper and glyphosate Glifoglex + cypermethrin Glextrin. When tested individually, the formulations presented the following 96 h-LC50s: Glifosato Atanor 19.4 mg ae L1 and Glifoglex 72.8 mg ae L1 , Xiper 6.8 mg L1 and Glextrin 30.2 mg L1. Equitoxic and non-equitoxic mixtures were significantly synergic in both combinations of commercial products tested. The magnitude of the synergy (factor by which toxicity differed from concentration addition) was constant at around twofold for all tested proportions of the glyphosate Glifoglex + cypermethrin Glextrin mixture; whereas the magnitude of the synergy varied between 4 and 9 times in the glyphosate Glifosato Atanor + cypermethrin Xiper mixture. These results call for more research to be promptly undertaken in order to understand the mechanisms behind the synergy observed and to identify and quantify the extent of its environmental impacts.

Brodeur et. al, 2016

Julie Celine Brodeur, Solene Malpel, Ana Belen Anglesio, Diego Cristos, María Florencia D’Andrea, María Belen Poliserpi, “Toxicities of glyphosate- and cypermethrin-based pesticides are antagonic in the tenspotted livebearer fish (Cnesterodon decemmaculatus),” Chemosphere, 2016, 155:429-435, DOI:  10.1016/j.chemosphere.2016.04.075.


Although pesticide contamination of surface waters normally occurs in the form of mixtures, the toxicity and interactions displayed by such mixtures have been little characterized until now. The present study examined the interactions prevailing in equitoxic and non-equitoxic binary mixtures of formulations of glyphosate (Glifoglex®) and cypermethrin (Glextrin®) to the tenspotted livebearer (Cnesterodon decemmaculatus), a widely distributed South American fish. The following 96 h-LC50s were obtained when pesticide formulations were tested individually: Glifoglex® 41.4 and 53 mg ae glyphosate/L; Glextrin® 1.89 and 2.60 mg cypermethrin/L. Equitoxic and non-equitoxic mixtures were significantly antagonic in all combinations tested. The magnitude of the antagonism (factor by which toxicity differed from concentration addition) varied between 1.37 and 3.09 times in the different non-equitoxic mixtures tested. Antagonism was due to a strong inhibition of cypermethrin toxicity by the glyphosate formulation, the toxicity of the cypermethrin-based pesticide being almost completely overridden by the glyphosate formulation. Results obtained in the current study with fish are radically opposite to those previously observed in tadpoles where synergy was observed when Glifoglex® and Glextrin® were present in mixtures.  FULL TEXT

Cakmak et. al, 2009

Ismail Cakmak, Atilla Yazici, Yusuf Tutus, Levent Ozturk, “Glyphosate reduced seed and leaf concentrations of calcium, manganese, magnesium, and iron in non-glyphosate resistant soybean,” European Journal of Agronomy, 2009, 31:3, 114-119, DOI: 10.1016/e.eja.2009.07.001.


Greenhouse experiments were conducted to study the effects of glyphosate drift on plant growth and concentrations of mineral nutrients in leaves and seeds of non-glyphosate resistant soybean plants (Glycine max, L.). Glyphosate was sprayed on plant shoots at increasing rates between 0.06 and 1.2% of the recommended application rate for weed control. In an experiment with 3-week-old plants, increasing application of glyphosate on shoots significantly reduced chlorophyll concentration of the young leaves and shoots dry weight, particularly the young parts of plants. Concentration of shikimate due to increasing glyphosate rates was nearly 2-fold for older leaves and 16-fold for younger leaves compared to the control plants without glyphosate spray. Among the mineral nutrients analyzed, the leaf concentrations of potassium (K), phosphorus (P), copper (Cu) and zinc (Zn) were not affected, or even increased significantly in case of P and Cu in young leaves by glyphosate, while the concentrations of calcium (Ca), manganese (Mn) and magnesium (Mg) were reduced, particularly in young leaves. In the case of Fe, leaf concentrations showed a tendency to be reduced by glyphosate. In the second experiment harvested at the grain maturation, glyphosate application did not reduce the seed concentrations of nitrogen (N), K, P, Zn and Cu. Even, at the highest application rate of glyphosate, seed concentrations of N, K, Zn and Cu were increased by glyphosate. By contrast, the seed concentrations of Ca, Mg, Fe and Mn were significantly reduced by glyphosate. These results suggested that glyphosate may interfere with uptake and retranslocation of Ca, Mg, Fe and Mn, most probably by binding and thus immobilizing them. The decreases in seed concentration of Fe, Mn, Ca and Mg by glyphosate are very specific, and may affect seed quality.  FULL TEXT

Casabe et al., 2007

Norma Casabé, Lucas Piola, Julio Fuchs, María Luisa Oneto, Laura Pamparato, Silvana Basack, Rosana Giménez, Rubén Massaro, Juan C. Papa and Eva Kesten, “Ecotoxicological Assessment of the Effects of Glyphosate and Chlorpyrifos in an Argentine Soya Field,” Journal of Soils and Sediment, 2007, DOI: 10.1065/JSS2007.04.224.


BACKGROUND, AIM, AND SCOPE: Continuous application of pesticides may pollute soils and affect non-target organisms. Soil is a complex ecosystem; its components can modulate the effects of pesticides. Therefore, it is recommended to evaluate the potential environmental risk of these compounds in local conditions. We performed an integrated field-laboratory study on an Argentine soya field sprayed with glyphosate and chlorpyrifos under controlled conditions. Our aim was to compare the sensitivity of a series of endpoints for the assessment of adverse effects of the extensive use of these agrochemicals.

MATERIALS AND METHODS: A RR soya field in a traditional farming area of Argentina was sprayed with glyphosate (GLY) or chlorpyrifos (CPF) formulations at the commercially recommended rates, according to a randomized complete block design with 3 replicates. In laboratory assays, Eisenia fetida andrei were exposed to soil samples (0–10 cm depth) collected between the rows of soya. Endpoints linked to behavior and biological activity (reproduction, avoidance behavior and bait-lamina tests) and cellular/subcellular assays (Neutral Red Retention Time –NRRT; DNA damage – Comet assay) were tested. Field assays included litterbag and bait-lamina tests. Physico/chemical analyses were performed on soil samples.

RESULTS: GLY reduced cocoon viability, decreasing the number of juveniles. Moreover, earthworms avoided soils treated with GLY. No effects on either reproduction or on avoidance were observed at the very low CPF concentration measured in the soils sampled 10 days after treatment. Both pesticides caused a reduction in the feeding activity under laboratory and field conditions. NRRT was responsive to formulations of CPF and GLY. Comet assay showed significantly increased DNA damage in earthworms exposed to CPF treated soils. No significant differences in DNA migration were observed with GLY treated soils. Litterbag field assay showed no differences between treated and control plots.     DISCUSSION: The ecotoxicological effects of pesticides can be assessed by monitoring the status of communities in real ecosystems or through the use of laboratory toxicity tests. Litterbag field test showed no influence of the treatments on the organic matter breakdown, suggesting a scarce contribution of soil macrofauna. The bait-lamina test, however, seemed to be useful for detecting the effects of GLY and CPF treatments on the activity of the soil fauna. CPF failed to give significant differences with the controls in the reproduction test and the results were not conclusive in the avoidance test. Although the field population density of earthworms could be affected by multiple factors, the effects observed on the reproduction and avoidance tests caused by GLY could contribute to its decrease, with the subsequent loss of their beneficial functions. Biomarkers measuring effects on suborganism level could be useful to predict adverse effects on soil organisms and populations. Among them, NRRT, a lysosomal destabilization biomarker, resulted in demonstrating more sensitivity than the reproduction and avoidance tests. The Comet assay was responsive only to CPF. Since DNA damage can have severe consequences on populations, it could be regarded as an important indicator to be used in the assessment of soil health.

CONCLUSIONS: Reproduction and avoidance tests were sensitive indicators of GLY exposure, with the former being more labor intensive. Bait-lamina test was sensitive to both CPF and GLY. NRRT and Comet assays revealed alterations at a subcellular level, and could be considered complementary to the biological activity tests. Because of their simplicity, some of these bioassays seemed to be appropriate pre-screening tests, prior to more extensive and invasive testing.

RECOMMENDATIONS AND PERSPECTIVES: This study showed deleterious effects of GLY and CPF formulations when applied at the nominal concentrations recommended for soya crops. Further validation is needed before these endpoints could be used as field monitoring tools in Argentine soya soils (ecotoxicological risk assessment – ERA tools).  FULL TEXT

Coupe et. al, 2011

Richard H Coupe, Stephen J Kalkhoff, Paul D Capelc, and Caroline Gregoired, “Fate and transport of glyphosate and aminomethylphosphonic acid in surface waters of agricultural basins,” Pest Management Science, 2011, 68:1, 16-30, DOI: 10.1002/ps.2212.


BACKGROUND: Glyphosate [N-(phosphonomethyl)glycine] is a herbicide used widely throughout the world in the production of many crops and is heavily used on soybeans, corn and cotton. Glyphosate is used in almost all agricultural areas of the United States, and the agricultural use of glyphosate has increased from less than 10 000 Mg in 1992 to more than 80 000 Mg in 2007. The greatest intensity of glyphosate use is in the midwestern United States, where applications are predominantly to genetically modified corn and soybeans. In spite of the increase in usage across the United States, the characterization of the transport of glyphosate and its degradate aminomethylphosphonic acid (AMPA) on a watershed scale is lacking.

RESULTS: Glyphosate and AMPA were frequently detected in the surface waters of four agricultural basins. The frequency and magnitude of detections varied across basins, and the load, as a percentage of use, ranged from 0.009 to 0.86% and could be related to three general characteristics: source strength, rainfall runoff and flow route.

CONCLUSIONS: Glyphosate use in a watershed results in some occurrence in surface water; however, the watersheds most at risk for the offsite transport of glyphosate are those with high application rates, rainfall that results in overland runoff and a flow route that does not include transport through the soil. FULL TEXT

Cuhra et. al, 2014

M. Cuhra, T. Traavik, and T. Bohn, “Life cycle fitness differences in Daphnia magna fed Roundup-Ready soybean or convetional soybean or organic soybean,” Aquaculture Nutrition, 2014, DOI: 10.1111/ani.12199.


A lifelong feeding study with soybean from different production systems was carried out in the crustacean Daphnia magna (water flea), an acknowledged model organism for ecotoxicological studies. Experimental diets were prepared with soybean meal from different agriculture production systems: (i) genetically modified Roundup-Ready soy (Glyphosate-Tolerant), (ii) conventional soy and (iii) soy from organic agriculture (agriculture with neither synthetic pesticides nor synthetic fertilizers). Overall, feed produced from organic soybeans resulted in the highest fitness (higher survival, better growth and fecundity) in the model organism. Animals fed Roundup-Ready soybean consistently performed less well compared to animals fed either conventional or organic soybeans. We conclude that accumulation of herbicide residues in Roundup-Ready soy and related nutritional differences between the soy types may have caused the observed fitness differences. The results accentuate the need for further research clarifying qualitative aspects, including potential large-scale consequences for food and feed quality, of this dominant crop.  FULL TEXT