Effects of glyphosate herbicide ingestion on kidney function in rats on a balanced diet / Efeitos da ingestão do herbicida glifosato na função renal de ratos em uma dieta balanceada

Abstract Introduction: Glyphosate is the most widely used herbicide worldwide and in Brazil. There is currently increasing concern about the effects of glyphosate on human health. The Brazilian Institute for Consumer Protection showed data on the presence of glyphosate in some of Brazil's most consumed ultra-processed products. Currently, regulations on the upper limit for these residues in ultra-processed foods have yet to be established by the National Health Surveillance, and ultra-processed food consumption is independently associated with an increased risk of incident chronic kidney disease. Methods: Since an unbalanced diet can interfere with kidney function, this study aims to investigate the effect of daily intake of 5 mg/kg bw glyphosate in conjunction with a balanced diet and the possible impact on renal function in rats. Kidney function, kidney weight, markers of renal injury, and oxidative stress were evaluated. Results: There was a decrease in kidney weight. The main histopathological alterations in renal tissues were vacuolation in the initial stage and upregulation of the kidney injury marker KIM-1. Renal injury is associated with increased production of reactive oxygen species in mitochondria. Conclusion: This study showed changes in the kidney of rats exposed to a balanced diet with glyphosate, suggesting a potential risk to human kidney. Presumably, ultra-processed food that contain glyphosate can potentiate this risk. The relevance of these results lies in drawing attention to the need to regulate glyphosate concentration in ultra-processed foods in the future.

RESUMO Introdução: O glifosato é o herbicida mais utilizado no mundo e no Brasil. Atualmente, há uma preocupação crescente com os efeitos do glifosato na saúde humana. O Instituto Brasileiro de Defesa do Consumidor apresentou dados sobre a presença de glifosato em alguns dos produtos ultraprocessados mais consumidos no Brasil. Atualmente, as regulamentações sobre o limite máximo desses resíduos em alimentos ultraprocessados ainda não foram estabelecidas pela Vigilância Sanitária Nacional, e o consumo de alimentos ultraprocessados está indepen­dentemente associado a um risco maior de doença renal crônica incidente. Métodos: Como uma dieta desbalanceada pode interferir na função renal, este estudo tem como objetivo investigar o efeito da ingestão diária de 5 mg/kg pc de glifosato em conjunto com uma dieta equilibrada e o possível impacto na função renal em ratos. Foram avaliados função renal, peso dos rins, marcadores de lesão renal e estresse oxidativo. Resultados: Houve redução no peso dos rins. As principais alterações histopatológicas nos tecidos renais foram vacuolização no estágio inicial e regulação positiva do marcador de lesão renal KIM-1. A lesão renal está associada à produção aumentada de espécies reativas de oxigênio nas mitocôndrias. Conclusão: Esse estudo mostrou alterações nos rins de ratos expostos a uma dieta balanceada com glifosato, sugerindo um risco potencial ao rim humano. Presumivelmente, alimentos ultraprocessados que contenham glifosato podem potencializar esse risco. A relevância desses resultados está no fato de chamar a atenção para a necessidade de regulamentar a concentração de glifosato em alimentos ultraprocessados no futuro.

Residuos de glifosato en cereales de desayuno de origen internacional y nacional comercializados en Saltillo, Coahuila / Glyphosate residues in breakfast cereals of national and international origin marketed in Saltillo, Coahuila

Resumen Objetivo: Detectar y cuantificar residuos de glifosato en cereales de desayuno comercializados en la ciudad de Saltillo, Coahuila. Materiales y Métodos: Se analizaron tres muestras de 12 marcas de cereal distintas. Para la extracción de plaguicidas se utilizó un equipo Soxhlet y la cuantificación se realizó por medio de Cromatografía Líquida de Alta Eficiencia (HPLC), utilizando un equipo marca Agillent modelo 1100 Series acoplado a un detector UV-Vis. Resultados: Se detectó glifosato en el 100% de las muestras analizadas, se obtuvieron concentraciones de entre 170 a 2 400 mg/kg, las muestras que presentaron mayores concentraciones fueron las de marcas internacionales. Conclusiones: Se detectaron residuos de glifosato en las 24 muestras analizadas de cereales de desayuno, las cuales sobrepasaron los LMR establecidos en el CODEX ALIMENTARIUS, por lo que es importante realizar más estudios con el fin de monitorear y asegurar la inocuidad de los productos procesados que son consumidos.

Abstract Objective: To detect and quantify glyphosate residues in breakfast cereals marketed in the city of Saltillo, Coahuila. Materials and Methods: Two samples of 12 different cereal brands were analyzed. A Soxhlet apparatus was used for pesticide extraction and quantification was performed by High Performance Liquid Chromatography (HPLC) using an Agillent model 1100 Series equipment coupled to a UV- Vis detector. Results: Glyphosate was detected in 100% of the samples analyzed, with concentrations ranging from 170 to 2 400 mg/kg; the samples with the highest concentrations were those of international brands. Conclusions: All the concentrations detected in the commercial cereal samples exceeded the MRLs established in CODEX ALIMENTARIUS, so it is important to carry out more studies in order to monitor and ensure the safety of the processed products that are consumed.

Amazonian soil fungi are efficient degraders of glyphosate herbicide; novel isolates of Penicillium, Aspergillus, and Trichoderma / Os fungos do solo amazônico são degradadores eficientes do herbicida glifosato; novos isolados de Penicillium, Aspergillus e Trichoderma

Abstract Pesticide residues that contaminate the environment circulate within the hydrological cycle can accumulate within the food chain and cause problems to both environmental and human health. Microbes, however, are well known for their metabolic versatility and the ability to degrade chemically stable substances, including recalcitrant xenobiotics. The current study focused on bio-prospecting within Amazonian rainforest soils to find novel strains fungi capable of efficiently degrading the agriculturally and environmentally ubiquitous herbicide, glyphosate. Of 50 fungal strains isolated (using culture media supplemented with glyphosate as the sole carbon-substrate), the majority were Penicillium strains (60%) and the others were Aspergillus and Trichoderma strains (26 and 8%, respectively). All 50 fungal isolates could use glyphosate as a phosphorous source. Eight of these isolates grew better on glyphosate-supplemented media than on regular Czapek Dox medium. LC-MS revealed that glyphosate degradation by Penicillium 4A21 resulted in sarcosine and aminomethylphosphonic acid.

Resumo Resíduos de agrotóxicos que contaminam o meio ambiente circulam no ciclo hidrológico, podendo se acumular na cadeia alimentar e causar problemas tanto à saúde ambiental quanto humana. Por sua vez, microrganismos são bem conhecidos por sua versatilidade metabólica e capacidade de degradar substâncias quimicamente estáveis, incluindo xenobióticos recalcitrantes. O estudo atual se concentrou na bioprospecção nos solos da floresta amazônica para encontrar novas linhagens de fungos capazes de degradar com eficiência o herbicida onipresente na agricultura e no meio ambiente, o glifosato. Entre os 50 fungos isolados (usando meio de cultura suplementado com glifosato como única fonte de carbono), a maioria eram isolados do gênero Penicillium (60%) e os outros eram isolados de Aspergillus e Trichoderma (26 e 8%, respectivamente). Todos os 50 isolados de fungos foram capazes de usar glifosato como fonte de fósforo. Oito desses isolados cresceram melhor em meio suplementado com glifosato do que em meio Czapek Dox regular. LC-MS revelou que a degradação do glifosato por Penicillium 4A21 resultou nos metabólitos sarcosina e ácido aminometilfosfônico.

Investigation on health status of workers exposed to glyphosate / 中华劳动卫生职业病杂志

Objective: To analyze the concentrations of glyphosate and its metabolites in occupational exposed workers and their possible effects on human health, so as to provide a reference for improving the safe use of glyphosate and toxicity research. Methods: From April to December 2020, 247 workers directly exposed to glyphosate in 5 enterprises were selected as the contact group, and 237 workers who were not exposed to glyphosate and other pesticides in the same enterprise were selected as the control group. Questionnaire survey and occupational health examination were conducted on objects, and the concentrations of glyphosate and its metabolites in the air of workplaces and biological samples were detected. The correlation between the concentrations and the difference of health examination between the two groups were analyzed. Results: The urine glyphosate concentration (0.022-47.668 mg/L), the rate of exceeding the standard (60.32%, 149/247) and the urine aminomethyl phosphonic acid concentration (<0.010-1.624 mg/L) in the contact group were higher than those in the control group [urine glyphosate concentration (<0.020-4.482 mg/L), the rate of exceeding the standard (2.53%, 6/237) and the urine aminomethyl phosphonic acid concentration (<0.010-0.524 mg/L) ], respectively (P<0.001). The exceeding standard rate of glyphosate concentration in the workplace was 33.67% (33/98). The concentration of glyphosate in the workplace was positively correlated with the concentrations of glyphosate and aminomethylphosphonic acid in urine (r(s)=0.804, 0.238, P<0.001), and the concentration of glyphosate in urine was positively correlated with the concentration of aminomethylphosphonic acid in urine (r(s)=0.549, P<0.001). The alanine aminotransferase (ALT), white cell ratio, creatinine, uric acid, the abnormal rates of ALT and total protein (TP) in the contact group were higher than those in the control group, and TP was lower than that in the control group, the differences were statistically different (P<0.05). The abnormal rates of overall liver function, overall renal function, blood routine test, urine routine test, electrocardiogram, liver B ultrasound and blood lipid in the contact group were significantly higher than those in the control group (P<0.05) . Conclusion: The concentration of glyphosate in the workplace is related to the concentrations of glyphosate and aminomethyl phosphonic acid in the urine of workers, and exposure to glyphosate may have some harmful effects on human health.

Amazonian soil fungi are efficient degraders of glyphosate herbicide; novel isolates of Penicillium, Aspergillus, and Trichoderma / Os fungos do solo amazônico são degradadores eficientes do herbicida glifosato; novos isolados de Penicillium, Aspergillus e Trichoderma

Pesticide residues that contaminate the environment circulate within the hydrological cycle can accumulate within the food chain and cause problems to both environmental and human health. Microbes, however, are well known for their metabolic versatility and the ability to degrade chemically stable substances, including recalcitrant xenobiotics. The current study focused on bio-prospecting within Amazonian rainforest soils to find novel strains fungi capable of efficiently degrading the agriculturally and environmentally ubiquitous herbicide, glyphosate. Of 50 fungal strains isolated (using culture media supplemented with glyphosate as the sole carbon-substrate), the majority were Penicillium strains (60%) and the others were Aspergillus and Trichoderma strains (26 and 8%, respectively). All 50 fungal isolates could use glyphosate as a phosphorous source. Eight of these isolates grew better on glyphosate-supplemented media than on regular Czapek Dox medium. LC-MS revealed that glyphosate degradation by Penicillium 4A21 resulted in sarcosine and aminomethylphosphonic acid.

Resíduos de agrotóxicos que contaminam o meio ambiente circulam no ciclo hidrológico, podendo se acumular na cadeia alimentar e causar problemas tanto à saúde ambiental quanto humana. Por sua vez, microrganismos são bem conhecidos por sua versatilidade metabólica e capacidade de degradar substâncias quimicamente estáveis, incluindo xenobióticos recalcitrantes. O estudo atual se concentrou na bioprospecção nos solos da floresta amazônica para encontrar novas linhagens de fungos capazes de degradar com eficiência o herbicida onipresente na agricultura e no meio ambiente, o glifosato. Entre os 50 fungos isolados (usando meio de cultura suplementado com glifosato como única fonte de carbono), a maioria eram isolados do gênero Penicillium (60%) e os outros eram isolados de Aspergillus e Trichoderma (26 e 8%, respectivamente). Todos os 50 isolados de fungos foram capazes de usar glifosato como fonte de fósforo. Oito desses isolados cresceram melhor em meio suplementado com glifosato do que em meio Czapek Dox regular. LC-MS revelou que a degradação do glifosato por Penicillium 4A21 resultou nos metabólitos sarcosina e ácido aminometilfosfônico.

Amazonian soil fungi are efficient degraders of glyphosate herbicide; novel isolates of Penicillium, Aspergillus, and Trichoderma

Abstract Pesticide residues that contaminate the environment circulate within the hydrological cycle can accumulate within the food chain and cause problems to both environmental and human health. Microbes, however, are well known for their metabolic versatility and the ability to degrade chemically stable substances, including recalcitrant xenobiotics. The current study focused on bio-prospecting within Amazonian rainforest soils to find novel strains fungi capable of efficiently degrading the agriculturally and environmentally ubiquitous herbicide, glyphosate. Of 50 fungal strains isolated (using culture media supplemented with glyphosate as the sole carbon-substrate), the majority were Penicillium strains (60%) and the others were Aspergillus and Trichoderma strains (26 and 8%, respectively). All 50 fungal isolates could use glyphosate as a phosphorous source. Eight of these isolates grew better on glyphosate-supplemented media than on regular Czapek Dox medium. LC-MS revealed that glyphosate degradation by Penicillium 4A21 resulted in sarcosine and aminomethylphosphonic acid.

Resumo Resíduos de agrotóxicos que contaminam o meio ambiente circulam no ciclo hidrológico, podendo se acumular na cadeia alimentar e causar problemas tanto à saúde ambiental quanto humana. Por sua vez, microrganismos são bem conhecidos por sua versatilidade metabólica e capacidade de degradar substâncias quimicamente estáveis, incluindo xenobióticos recalcitrantes. O estudo atual se concentrou na bioprospecção nos solos da floresta amazônica para encontrar novas linhagens de fungos capazes de degradar com eficiência o herbicida onipresente na agricultura e no meio ambiente, o glifosato. Entre os 50 fungos isolados (usando meio de cultura suplementado com glifosato como única fonte de carbono), a maioria eram isolados do gênero Penicillium (60%) e os outros eram isolados de Aspergillus e Trichoderma (26 e 8%, respectivamente). Todos os 50 isolados de fungos foram capazes de usar glifosato como fonte de fósforo. Oito desses isolados cresceram melhor em meio suplementado com glifosato do que em meio Czapek Dox regular. LC-MS revelou que a degradação do glifosato por Penicillium 4A21 resultou nos metabólitos sarcosina e ácido aminometilfosfônico.

Exposure of Artemia salinato glyphosate and bioremediation by isotherapy

Artemia salina, an ecotoxicity bioindicator, isa microcrustacean belonging to the order Anostraca. Glyphosate is an herbicide widely used to control weeds. However, its intoxication can cause serious damage to human health and the balance of the environment, given its effects as an endocrine disruptor.Objective:verify the possible protection of the artemia exposed to glyphosate by the addition of its isotherapic into water, by means of the evaluation of the behavioral and morphological features of nauplii and of the physical properties of remedies andtreated water, to elucidate the involved mechanisms. Methods:Artemia salinacysts were kept in culture bottles containing artificial seawatercontaining glyphosate at LC 10 (lethal concentration 10%), to promote hatching within 48 hours. The isotherapic preparations were inserted in each bottle in a 10% of the total water volume. Part of the nauplii was distributed in transparent tubes, being 10 nauplii per tube and 6 tubes per group, for behavioral analysis, and part were collected for a detailed morphological analysis, under an optical microscope. About 80 to 270 nauplii were analyzed per group. The reserved water was divided into aliquots for physicochemical analysis, that is, evaluation of the water dipole behavior by Cartwright ́s method.Results:Gly 6cH presented selective effects on nauplii hatching (p=0.02) and on defected/healthy ratio (p=0.001), representing some protective action. This result was dependent of the salinity of water and presented correspondence with the effects on solvatochromic dyes, indicating that charges and ions can be critical factors involved in the mechanism of action. We concluded that the use of isotherapics could be a plausible tool to reduce the environmental impact of the indiscriminate use of glyphosate, since these results can be reproduced in further studies.

Herbicide efficacy in the fall management of Richardia brasiliensis, Commelina benghalensis, Conyza sumatrensis AND Digitaria insularis

Other herbicides, alone or in combination with glyphosate, may be effective in controlling Richardia brasiliensis, Commelina benghalensis, Conyza sumatrensis e Digitaria insularis. Therefore, the aim of this study was to evaluate the efficacy of herbicides, alone or in combination, in the control of these weeds in the off-season. Three composite experiments were conducted by applying herbicides, alone or in combinations. Experiments 1 and 2 were conducted in Palotina, State of Paraná (PR), Brazil. Experiment 3 in Iporã, PR, Brazil. The three during fall 2017 in fallow areas after soybean harvest. The experiment was a randomized complete block design with 4 replications and weed control at 7, 14, 21, 28, 35, and 42 days after application (DAA). The control of R. brasiliensis and C. benghalensis was evaluated in the three experiments, the control of C. sumatrensis, in experiments 1 and 2, while the control of D. insularis was evaluated only in experiment 3. Data were subjected to analysis of variance and F-test (p < 0.05). The treatment means values of experiments 1 and 2 were compared by Tukey's test (p < 0.05), and the treatment mean values of experiment 3 were grouped by the Scott and Knott test (p < 0.05). Some herbicide combinations were effective in controlling R. brasiliensis, C. benghalensis, in a single or sequential application. Herbicide combinations were effective in controlling C. sumatrensis with sequential application. Herbicide combinations in a single application did not provide satisfactory final control (≥ 80%) for D. insularis.

Exposure to glyphosate-based herbicide during early stages of development increases insulin sensitivity and causes liver inflammation in adult mice offspring

ABSTRACT Objective: To investigate the effect of pre and postnatal exposure to a glyphosate-based herbicide on glucose metabolism and liver histology in adult F1 mice offspring. Methods: Female mice (C57Bl/6) received 0.5% of glyphosate (Roundup Original DI®) in drinking water or purified water (Glyphosate Group and Control Group respectively) during pregnancy and lactation. Offspring (F1) were submitted to glucose and insulin tolerance tests and euthanized on postnatal day 150. Body and plasma parameters, and liver histology were analyzed. Results: Exposure to glyphosate reduced maternal body weight gain during pregnancy and lactation, with no impacts on litter size. Pre and postnatal exposure to glyphosate did not affect body parameters but increased glucose tolerance on postnatal day 60. In spite of glucose tolerance normalization by postnatal day 143, this effect was associated with higher insulin sensitivity relative to mice in the Control-F1 Group. Mice in the Glyphosate-F1 Group had mild and moderate lobular inflammation in the liver. Conclusion: Maternal exposure to glyphosate affected insulin sensitivity and caused hepatic inflammation in adult F1 mice offspring.

Determination of glyphosate in breast milk of lactating women in a rural area from Paraná state, Brazil

The aim of this study was to verify the presence of glyphosate in breast milk and to characterize maternal environmental exposure. Sixty-seven milk samples were collected from lactating women in the city of Francisco Beltrão, Paraná, living in urban (n=26) and rural (n=41) areas, at the peak of glyphosate application in corn and soy crops in the region (April and May 2018). To characterize the study population, socio-epidemiological data of the women were collected. To determine glyphosate levels, a commercial enzyme immunosorbent assay kit was used. Glyphosate was detected in all breast milk samples analyzed with a mean value of 1.45 µg/L. Despite some descriptive differences, there were no statistically significant differences (P<0.05) between the categories of the variables tested. Also, glyphosate was detected in drinking water samples from the urban area and in artesian well water from the rural area of the region where the studied population lived. The estimation of the total amount of glyphosate ingested by breastfeeding babies in a period of 6 months was significant. These results suggest that the studied lactating population was contaminated with glyphosate, possibly through continued environmental exposure.

Clinical characteristics of 83 patients with acute glyphosate herbicide poisoning / 中华急诊医学杂志

Objective:To analyze the clinical characteristics of patients with acute glyphosate herbicide poisoning and the differences in the severity of poisoning.Methods:A retrospective analysis was performed on patients with acute glyphosate herbicide poisoning admitted to the First Affiliated Hospital of Zhengzhou University from January 2014 to December 2020. The general information, exposure time, poisoning dose, poisoning cause, poisoning route, clinical manifestations, laboratory examination results during hospitalization, treatment measures, hospital stays and prognosis of the patients were collected. The patients were graded according to the poisoning severity scoring standard of Chinese Expert Consensus on Diagnosis and Treatment of Acute Poisoning in 2016. The highest severity score during hospitalization was used as the final grade. According to the final grade, asymptomatic and mild patients were included in the mild group, and moderate, severe and death patients were included in the severe group. The independent sample T test or Mann-Whitney U test was used for measurement data, and χ2 test or Fisher's exact test was used for counting data. The differences of general data and clinical data between the two groups were compared. Results:According to the inclusion and exclusion criteria, 83 patients with acute glyphosate herbicide poisoning were selected as the study subjects. All patients survived, mainly mild poisoning (56.6%), with a male to female ratio of 33∶50, and an average age of 39 years. The number of poisoning cases increased yearly (the highest in 2019), and most cases occurred in spring and summer. The main cause of poisoning was suicide (71.1%), direct oral administration (83.1%) was the primary route of poisoning, and the dominating clinical manifestations were digestive symptoms (71.1%). Laboratory tests showed increased white blood cell count (WBC), neutrophil percentage (NEUT %) and D-dimer, and decreased hemoglobin and potassium. Compared with the mild group, patients in the severe group were older [(51±17) years vs. (35±19) years], had a higher proportion of suicide and direct oral administration, a longer hospital stay [8.0 (4.8, 12.0) d vs. 3.0 (2.0, 5.5) d], a higher dose of poisoning [200.0 (50.0, 200.0) mL vs. 30.0 (11.3, 57.5) mL], and higher NEUT % within 24 h of admission [(83.4±10.4) vs. (73.2±12.8)]. The increase of WBC, NEUT %, aspartate aminotransferase, prothrombin time, D-dimer and the decrease of serum potassium were more common in the severe group than the mild group, with statistical significance (all P<0.05). Conclusions:The number of patients with acute glyphosate herbicide poisoning is increasing yearly. Generally, the condition is mild and the prognosis is satisfying. The severity is more serious in the middle-aged and elderly patients andthose with direct oral administration, high toxic dose, and high NEUT % within 24 h of admission. Severe poisoning is more likely to cause changes in laboratory indicators.

Ultrafiltração na remoção de glifosato e ácido aminometilfosfônico em águas de abastecimento / Ultrafiltration for removal of glyphosate and aminomethylphosphonic acid in drinking water

RESUMO O glifosato é um herbicida não seletivo utilizado no controle de ervas daninhas em ambientes agrícolas e urbanos. É encontrado em mananciais associado ao seu principal metabólito, o ácido aminometilfosfônico, sendo necessária a remoção destes em estações de tratamento de água. O objetivo do trabalho foi avaliar a ultrafiltração direta na rejeição do glifosato e do ácido aminometilfosfônico em água de manancial superficial fortificada nas concentrações de 630, 800 e 1.250 μg L-1 e variação de pH entre 4 e 10. Para o glifosato, os aumentos do pH e da concentração e a presença de matéria orgânica natural contribuíram para a taxa de rejeição. A massa de glifosato adsorvida foi de 1,2 μg cm-2, sendo a adsorção na superfície e nos poros da membrana o principal mecanismo de rejeição. A ultrafiltração direta não foi efetiva para rejeição do ácido aminometilfosfônico. Para concentração do glifosato de 630 μg L-1 e pH entre 6,1 e 7,2, a ultrafiltração direta produziu permeado com concentração inferior a 500 μg L-1, atendendo ao padrão de potabilidade brasileiro, Portaria Gabinete do Ministro/Ministério da Saúde nº 888, publicada em 7 de maio de 2021.

ABSTRACT Glyphosate is a non-selective herbicide used to control weeds in agricultural and urban environments. It is found in water sources associated with its main metabolite, the aminomethylphosphonic acid, and it must be removed in the public water treatment. The aim of this work was to evaluate the direct ultrafiltration in the removal of glyphosate and aminomethylphosphonic acid in fortified surface water at concentrations of 630, 800, and 1,250 μg L-1 and pH variation between 4 and 10. For glyphosate, the increase in pH, concentration, and the presence of natural organic matter contributed to the rejection rate. The herbicide mass adsorbed was 1.2 μg cm-2, being adsorption onto the membrane surface/pores the main mechanism of herbicides retention. The UF process was not effective for removal of aminomethylphosphonic acid. For glyphosate concentration of 630 μg L-1, pH of 6.1-7.2, the UF was effective to produce the permeated in concentration of less than 500 μg L-1, as recommended by the Brazilian Drinking Water Legislation of the Ministry of Health, Portaria Gabinete do Ministro/Ministério da Saúde nº 888, publicada em 7 de maio de 2021.

The Effect of Glyphosate on Human Sperm: In Vitro Approximation / Efecto del glifosato sobre los espermatozoides humanos: una aproximación in vitro

Introduction Glyphosate is an herbicide used to eradicate illicit crops; however, its use is controversial due to different health problems associated with it. The present study aims to evaluate the effects of glyphosate on human sperm in vitro. Methods Twenty-two semen samples from healthy normozoospermic men were included; 11 semen samples were incubated with Panzer (INVESA S.A., Antiquia, Colombia) and 11 with Roundup (Monsanto Company, MO, USA). The changes in motility and viability were observed. Functional seminal parameters were evaluated as well. Results The samples exposed to glyphosate showed less motility and viability; a decrease in the potential of the mitochondrial membrane was observed, and an increase in the lipoperoxidation of the membrane was evidenced. Conclusion Based on the present results, we concluded that glyphosate has cytotoxic potential for exposed people and may affect their fertility.

Introducción El glifosato es un herbicida utilizado ampliamente para la erradicación de cultivos ilícitos; sin embargo, su uso es polémico debido a diferentes problemas de salud asociados con él. El objetivo del presente estudio fue evaluar los efectos del glifosato sobre los espermatozoides humanos in vitro. Métodos Se incluyeron 22 muestras de semen de hombres sanos normozoospérmicos, de las cuales 11 se incubaron con Panzer y 11 con Roundup, y se evaluaron los cambios en la movilidad y la viabilidad espermática, además de valorar los parámetros seminales funcionales. Resultados Las muestras expuestas al glifosato presentaron una menor movilidad y viabilidad, una disminución en el potencial de la membrana mitocondrial, y un aumento en la lipoperoxidación de la membrana. Conclusiones El glifosato es potencialmente citotóxico para las personas que estén expuestas, y puede afectar su fertilidad.

Destino ambiental y efectos ecológicos de los tres herbicidas más utilizados en Colombia / Environmental fate and ecological effects of the three most commonly used herbicides in Colombia / Destino ambiental e efeitos ecológicos dos três herbicidas mais comumente usados na Colômbia

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Abstract Knowledge of the environmental and transport fate of pesticides is integral to characterizing environmental exposures. Agencies such as the Environmental Protection Agency (EPA) 27 integrate the characterization of an exposure with ecological effects to determine the risks of pesticide use. The three most widely used herbicides in Colombia are glyphosate, propanil, and glufosinate ammonium. Although glyphosate had, since the early 1970's, been considered an environmentally friendly herbicide, the weight of the evidence of studies beginning in the 1990s show that inadvertent applications in wetlands can be fatal to the larvae of amphibians. Although glyphosate formulations are typically considered moderately toxic (LC50 = 1-10 ppm) for most aquatic organisms, for many amphibian species they are highly toxic (LC50 ˂ 1 ppm). This concern is compounded by the fact that Colombia has an incredibly high diversity of amphibians (746 species). In addition, chronic exposures of some amphibian species to sublethal concentrations, which are more likely found in the environment, have been shown to cause decreased survival, intersex gonads, tail damage, prolongation of time to metamorphosis, and liver lipidosis. In Colombia, aerial applications of Glyphos-Cosmo-Flux have been used since 1997 to fumigate illicit coca crops. As of 2019, the International Union for Conservation of Nature (IUCN) Red List had 285 species of amphibians in Colombia considered threatened. We found that 132 of the endangered (EN) and critically endangered (CR) amphibian species were present in 11 of the 32 Colombian departments with illegal coca plantations. Therefore, studies are needed to evaluate the impact of the coca eradication program on the amphibian population of the country. Propanil is the main herbicide used in rice fields. Despite its rapid environmental degradation ((1 day in aerobic conditions), studies on the impact of propanil have found that at projected application rates (4.5 kg/ha), within 48 hours after application, it reduces dissolved oxygen to levels critically important for survival of fish. At very low concentrations of 50 ppb or less of propanil, this effect correlates with lower phytoplankton production rather than higher community respiration. Despite being mild to moderately toxic at acute exposures, chronic exposure data also show that concentrations greater than 9 ppb affect the growth, reproduction, and development of fish and invertebrates. With respect to glufosinate ammonium, although it is highly likely to contaminate surface (runoff) and groundwater due to its physicochemical properties, data suggest that it is practically non-toxic to aquatic and terrestrial species. The acute hazard ratios were well below the level of concern, and therefore the EPA determined that the acute risk to non-target terrestrial and aquatic organisms is very low.

Resumo O conhecimento do destino ambiental e de transporte dos pesticidas é parte integrante da caracterização das exposições ambientais. Agências como a Agência de Proteção Ambiental (EPA) 27 integram a caracterização de uma exposição com efeitos ecológicos para determinar os riscos do uso de pesticidas. Os três herbicidas mais usados na Colômbia são glifosato, propanil e glufosinato de amônio. Embora o glifosato tenha sido considerado um herbicida ambientalmente correto desde o início da década de 1970, foi evidenciado por estudos iniciados na década de 1990 que aplicações inadvertidas em pântanos podem ser fatais para larvas de anfíbios. Embora as formulações de glifosato sejam tipicamente consideradas moderadamente tóxicas (CL50 = 1-10 ppm) para a maioria dos organismos aquáticos, para muitas espécies de anfíbios elas são altamente tóxicas (CL50 ˂ 1 ppm). Essa preocupação é agravada pelo fato de que a Colômbia tem uma diversidade incrivelmente alta de anfíbios (791 espécies). Além disso, foi demonstrado que exposições crônicas de algumas espécies de anfíbios a concentrações subletais, que são mais prováveis no ambiente, causam diminuição da sobrevivência, gônadas intersex, dano na cauda, prolongamento do tempo até a metamorfose e lipidose hepática. Na Colômbia, aplicações aéreas de Glyphos-Cosmo-Flux têm sido usadas desde 1997 para fumigar plantações ilícitas de coca. Em 2019, a Lista Vermelha da União Internacional para a Conservação da Natureza (IUCN) tinha 285 espécies de anfíbios na Colômbia consideradas ameaçadas. Descobrimos que 132 das espécies de anfíbios em perigo (EN) e criticamente em perigo (CR) estavam presentes em 11 dos 32 departamentos colombianos onde há mais plantações ilegais de coca. Portanto, são necessários estudos para avaliar o impacto do programa de erradicação da coca na população de anfíbios do país. O propanil é o principal herbicida usado nos campos de arroz. Apesar de sua rápida degradação ambiental ((1 dia em condições aeróbicas), estudos sobre o impacto do propanil descobriram que nas taxas de aplicação projetadas (4,5 kg / ha), ele reduz o oxigênio dissolvido a níveis criticamente importantes para a sobrevivência dos peixes em 48 horas. após a aplicação. Este efeito se correlaciona com a produção de fitoplâncton mais baixa, em vez de maior respiração da comunidade em concentrações muito baixas de 50 ppb ou menos de propanil. Apesar de ser leve a moderadamente tóxico em exposições agudas, os dados de exposição crônica também mostram que concentrações maiores que 9 ppb afetam o crescimento, a reprodução e o desenvolvimento de peixes e invertebrados. Com relação ao glufosinato de amônio, embora tenha grande probabilidade de contaminar a superfície (escoamento) e as águas subterrâneas devido às suas propriedades físico-químicas, os dados sugerem que é praticamente atóxico para as espécies aquáticas e terrestres. As razões de risco agudo estavam bem abaixo do nível de preocupação e, portanto, a EPA determinou que o risco agudo para organismos terrestres e aquáticos não visados é muito baixo.

Importance of sequential herbicide application in the control of Elephantopus mollis / A importância da aplicação sequencial no controle de Elephantopus mollis

RESUMO: Popularmente conhecida como pata de elefante ou sussuaiá, Elephantopus mollis, é uma espécie que recentemente surgiu como uma planta daninha em lavouras de cultivos de grãos. Objetivou-se com este estudo avaliar a sensibilidade diferencial de biótipos de E. mollis oriundos de áreas agrícolas e não agrícolas e determinar as melhores combinações de herbicidas aplicados em diferentes estádios de desenvolvimento para o controle desta espécie. Foram realizados três experimentos. No experimento de sensibilidade diferencial foram utilizados dez herbicidas em dois biótipos em duas fases de desenvolvimento. O experimento de dose resposta foi conduzido com oito doses de sete herbicidas. O experimento a campo foi realizado em uma área que apresenta problemas com a planta daninha para validar os resultados dos experimentos anteriores. O biótipo oriundo de área agrícola apresentou menor sensibilidade aos herbicidas quando comparado ao biótipo oriundo de área não agrícola. Os herbicidas de contato inicialmente apresentam controle superior, mas logo a planta apresenta rebrote em função de gemas presentes no colo da planta. A aplicação isolada de 2,4-D com doses entre 1005 e 1675 g e.a ha-1 possui controle de 25%. Quando o 2,4-D é seguido da aplicação de paraquat (400 g i.a ha-1) o controle passa a ser entre 51 e 68%. O melhor tratamento para o controle de plantas desenvolvidas é a mistura de 2,4-D + glyphosate (1340 + 1080 g e.a ha-1) com aplicação sequencial de paraquat.

ABSTRACT: Elephantopus mollis, popularly known as elephant paw or sussuaiá, is a species that has recently emerged as a weed affecting grain crops. This study aimed to evaluate the differential sensitivity of biotypes of E. mollis obtained from agricultural and nonagricultural areas and to determine the best combinations of herbicides applied at different stages of development for controlling this species. Three experiments were conducted. In the differential sensitivity experiment, 10 herbicides were used in 2 biotypes in 2 stages of development. The dose-response experiment was conducted using 8 doses of 7 herbicides. The field experiment was conducted in an area with weed issues to validate the results of previous experiments. The biotype from the agricultural area showed lower sensitivity to herbicides than the biotype from the nonagricultural area. The contact herbicides initially presented better control; however, the weed shortly showed re-growth due to the presence of buds in the plant crown. Isolated application of 2.4-D amine at doses between 1005 and 1675 g a.e. ha−1 demonstrated 25% control. When 2,4-D amine was followed by paraquat application (400 g a.i. ha−1), the control observed was between 51% and 68%. The best treatment for the control of completely developed weeds is 2,4-D amine + glyphosate mixture (1340 + 1080 g a.e. ha−1), followed by sequential paraquat application.

Integrated management of tough lovegrass (Eragrostis plana Nees): associating chemical control tools and plant physiology / Manejo integrado de capim-annoni (Eragrostis plana Nees): associando ferramentas do controle químico e fisiologia da planta

ABSTRACT: The objective of the study was to evaluate the efficiency of herbicides use in the control of tough lovegrass according to the availability of solar radiation and the presence or absence of flooded. Two experiments were conducted in a completely randomized design, in a 2x2x4 and 2x2x5 factorial scheme, where factor A was equivalent to the environment (natural or reduced radiation); factor B to water condition (with a 2 cm flooded or without); and, factor C the herbicides: cyhalofop butyl (315 g ha-1), glyphosate (1080 g ha-1), cyhalofop butyl + glyphosate (315 g ha-1 + 1080 g ha-1) and control (without application) in the first experiment. In the second experiment, the herbicides used were cyhalofop butyl (315 g ha-1), glyphosate (1080 g ha-1), sethoxydim (184g ha-1), imazethapyr (106 g ha-1) and control (without application). Glyphosate efficiently controls plants of tough lovegrass with four tillers (>90%), being superior to cyhalofop, imazethapyr and sethoxydim; independently, of resource conditions. Reduction in the availability of solar radiation generates less shoot dry mass production from the weed, and improves the control only by imazethapyr and cyhalofop. In general, a flooded condition does not affect tough lovegrass control by herbicides.

RESUMO: O objetivo do trabalho foi avaliar a eficiência de uso de herbicidas no controle de capim-annoni em função da disponibilidade de radiação solar e da presença ou ausência de lâmina de água. Para isso, foram conduzidos dois experimentos em delineamento inteiramente casualizado, em esquema fatorial 2x2x4 e 2x2x5, em que o fator A equivaleu ao ambiente (radiação natural ou reduzida); fator B à condição hídrica (com lâmina d'água de 2 cm ou sem); e, o fator C aos herbicidas: cyhalofop butyl (315 g ha-1), glyphosate (1080 g ha-1), cyhalofop butyl + glyphosate (315 g ha-1+ 1080 g ha-1) e testemunha (sem aplicação) no primeiro experimento. No segundo experimento, os herbicidas utilizados foram: cyhalofop butyl (315 g ha-1), glyphosate (1080 g ha-1), sethoxydim (184 g ha-1), imazethapyr (106 g ha-1) e testemunha (sem aplicação). Glyphosate controla eficientemente plantas de capim-annoni com quatro afilhos (>90%), sendo superior a cyhalofop butyl, imazethapyr e sethoxydim, independente das condições. A redução da disponibilidade de radiação solar gera menor produção de matéria seca da parte aérea pelo capim-annoni, podendo inclusive, contribuir para maior eficiência de controle somente por imazethapyr e cyhalofop. De maneira geral, a condição de lâmina d'água não afeta o controle de capim-annoni pelos herbicidas.

Study of muscle fibers of the extensor digitorium longus and soleus muscles of C57BL/6 females exposed to glyphosate during pregnancy and lactation / Estudo das fibras musculares dos músculos extensor longo dos dedos e sóleo de fêmeas C57BL/6 expostas ao glifosato durante a prenhez e a lactação

ABSTRACT Objective To evaluate the morphology and morphometry of the muscles extensor digitorium longus and soleus of C57BL/6 females, who were exposed to glyphosate during pregnancy and lactation. Methods Twelve female mice from the C57BL/6 lineage were used. After detection of pregnancy, they were divided into a Control Group, which received only water, and a Glyphosate Group, which received water with 0.5% glyphosate during pregnancy and lactation. Both groups received ad libitum standard diet. After weaning, the females were euthanized and weighed; naso-anal length was measured, and fats were collected and weighed. The muscles extensor digitorium longus and soleus were collected, and their length and weight were measured. Then, the muscles were fixed in Methacarn to perform the histological study of muscle fibers. Results Glyphosate Group presented lower weight gain during pregnancy and also lower final body weight and naso-anal length; however, the other body parameters evaluated did not present a significant difference in relation to the Control Group. Significant differences were also not observed in the analysis of muscle fibers and connective tissue. Conclusion Exposure to 0.5% glyphosate during pregnancy and lactation resulted in lower weight gain during pregnancy, final weight, and naso-anal length. Despite not directly altering the morphology of muscle tissue, these results may indicate enough exposure to interfere with animal metabolism.

RESUMO Objetivo Avaliar a morfologia e a morfometria dos músculos extensor longo dos dedos e sóleo de fêmeas C57BL/6 expostas ao glifosato durante a prenhez e lactação. Métodos Foram utilizados 12 camundongos fêmeas da linhagem C57BL/6. Após detecção da prenhez, foram separadas em Grupo Controle, que recebeu somente água, e Grupo Glifosato, que recebeu água com 0,5% de glifosato durante a prenhez e lactação. Ambos os grupos receberam dieta padrão ad libitum. Após o desmame, as fêmeas foram eutanasiadas e pesadas; o comprimento nasoanal foi mensurado, e as gorduras foram coletadas e pesadas. Os músculos extensor longo dos dedos e sóleo foram coletados, e seu comprimento e peso foram mensurados. Em seguida, os músculos foram fixados em Methacarn para a realização do estudo histológico das fibras musculares. Resultados O Grupo Glifosato apresentou menor ganho de peso durante a prenhez e também menor peso corporal final e comprimento nasoanal, entretanto os demais parâmetros corporais avaliados não apresentaram diferença significativa em relação ao Grupo Controle. Na análise das fibras musculares e do tecido conjuntivo, também não foram observadas diferenças significativas. Conclusão A exposição a 0,5% de glifosato durante a prenhez e lactação resultou em menor ganho de peso na gestação, peso final e comprimento nasoanal, o que pode indicar que, apesar de não alterar a morfologia do tecido muscular diretamente, a exposição foi suficiente para interferir no metabolismo dos animais.

Cloning and Characterization of 5-enopy ruvylshikimate-3-phosphate Synthase from Fragaria vesca

Abstract To discover and isolate a glyphosate-resistant gene from Fragaria vesca through gene mining. An open reading frame (ORF) of 1563 bp encoding EPSPSwas amplified from Fragaria vesca (FvEPSPS). FvEPSPS (Genebank: XP004306932.1) encodes a polypeptide of 520 amino acids and it has hightly homologous with EPSPS from other plants. qRT-PCR analysis showed that the FvEPSPS was expressed extensively in all tissues including leaves, roots and stems, with higher expression in leaves. Furthermore, transgenic Arabidopsis Thaliana exhibited 10 mM glyphosate to resistance. Therefore, this research offers a new glyphosate-resistant gene for development of transgenic crops.

Isolation of culturable mycota from Argentinean soils exposed or not-exposed to pesticides and determination of glyphosate tolerance of fungal species in media supplied with the herbicide / Aislamiento de la microbiota cultivable de suelos argentinos expuestos y no expuestos a plaguicidas y determinación de la tolerancia a glifosato de especies fúngicas en medios suplementados con el herbicida

Abstract The current agricultural system has led to the development of glyphosate (GP)-resistant weeds, causing an increase in GP doses and applications. Native mycota of pesticide-contaminated sites are the major source of pesticide-degrading microorganisms. The aims of the present study were to isolate the GP-tolerant culturable mycota in two soils with different pesticide exposure from Córdoba, Argentina, and to evaluate the growth parameters in native fungal isolates in the presence of GP and the effective dose that caused 50% growth reduction. The results showed that the genera Fusarium, Aspergillus, Mucor, Penicil-lium and Sterilia were the prevalent fungi isolated from soils both exposed and not-exposed to pesticides. The highest value (>100 mM) of effective concentration of herbicide that caused 50% growth inhibition (EC50), was found for Trichoderma isolates. Sterilia spp. had EC50 val-ues of 100mM, while Aspergillus spp. and Mucor had EC50 values between 50 and 100mM. The growth rate evaluation varied according to the isolates and GP concentrations. The data showed that all Aspergillus spp., Trichoderma spp., Mucor and three Sterilia spp. had the best growth performance in media supplied with GP after a variable acclimation period. This study provides valuable data for further studies that would allow to know the metabolic capacity of these fungal species that can be potential candidates for GP removal from contaminated environments.

Resumen El sistema agrícola actual condujo ai desarrollo de malezas resistentes a glifos-ato (GP), lo que deriva en el aumento de las dosis y el número de aplicaciones requeridas. La microbiota nativa de sitios contaminados con plaguicidas constituye la fuente principal de microorganismos capaces de degradar este herbicida. Los objetivos del presente estudio fueron aislar la microbiota cultivable tolerante a GP a partir de dos suelos agrícolas con diferente grado de exposición a plaguicidas, ambos de la provincia de Córdoba, Argentina, y evaluar en los hongos nativos aislados los parámetros de crecimiento en presencia de GP y la dosis efectiva que produce un 50% de reducción del crecimiento (EC50). Los géneros Fusarium, Aspergillus, Mucor, Penicillium y Sterilia fueron los de mayor prevalencia, tanto en suelos expuestos como no expuestos a plaguicidas. La mayor EC50 hallada (> 100 mM) correspondió a dos aislamientos de Trichoderma. En dos aislamientos de Sterilia, los valores de EC50 fueron de 100 mM, mientras que en los dos de Aspergillus y en uno de Mucor fueron de entre 50 y 100 mM. La velocidad de crecimiento varió según la concentración de GP empleada y el aislamiento considerado. Todos los aislamientos de Aspergillus, Trichoderma y Mucor, y tres aislamientos de Sterilia mostraron el mejor rendimiento en cuanto al crecimiento en los medios con GP después de un período de aclimatación variable. Este trabajo proporciona datos preliminares valiosos referidos a la capacidad metabólica de estos aislados y su potencial para remover GP en ambientes contaminados.

Effect of Roundup 41% (glyphosate) on blood serum biochemical parameters of freshwater fish, Rasbora daniconius

Aim: The present study was carried out to investigate the impact of Roundup 41% (glyphosate) on serum enzymes, like aminotransferases activity (ALT and AST), alkaline phosphates (ALP), total protein and glucose of fresh water fish, Rasbora daniconius.Methodology: LC50 of glyphosate for R. daniconius was calculated by static bioassays and recorded as 5.6 ppm at 96 hr. The fish were exposed to sub-lethal concentration (1/10th 96 hr LC50) of glyphosate for a period of 7, 14, 21 and 28 days. The serum ALT, AST, ALP, glucose and total protein were determined by standard methods. Results: Statistically important differences were observed in alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), total protein and glucose levles. ALT, AST and ALP values were significantly higher (p<0.05) in treated group, compared with the control group. There was also a significant increase (p<0.05) in the level of serum glucose while the reduced amount of protein content as compared to control fish. Interpretation: Increased metabolic enzymes, protein metabolism and hyperglycemia in the serum may be a possible indicator of liver damage caused by Roundup 41% (glyphosate) exposure in fish, Rasbora daniconius.