Ecotoxicological effects of commercial herbicides on the reproductive system of aquatic arthropod Limnocoris submontandoni (Hemiptera: Naucoridae) / Efeitos ecotoxicológicos de herbicidas comerciais no sistema reprodutivo do artrópode aquático Limnocoris submontandoni (Hemiptera: Naucoridae)

Abstract Worldwide, conventional agriculture makes extensive use of pesticides. Although the effects of herbicides are relatively well known in terms of environmental impacts on non-target organisms, there is very little scientific evidence regarding the impacts of herbicide residues on aquatic arthropods from tropical conservation areas. This study evaluates for the first time the toxicity of the herbicides ametryn, atrazine, and clomazone on the aquatic insect Limnocoris submontandoni (Hemiptera: Naucoridae). The lethal concentration (LC50) of herbicides was evaluated for these insects, as well as the effect of the herbicides on the insects' tissues and testicles. The estimated LC50 was 1012.41, 192.42, and 46.09 mg/L for clomazone, atrazine, and ametryn, respectively. Spermatocyte and spermatid changes were observed under the effect of atrazine, and effects on spermatogenesis were observed for some concentrations of clomazone, with apparent recovery after a short time. Our results provide useful information on the effects of herbicide residues in aquatic systems. This information can help minimize the risk of long-term reproductive effects in non-target species that have been previously overlooked in ecotoxicology studies.

Resumo Em todo o mundo, a agricultura convencional faz uso extensivo de pesticidas. Embora os efeitos dos herbicidas sejam relativamente bem conhecidos em termos de impactos ambientais em organismos não-alvo, há pouca evidência científica sobre os impactos de resíduos de herbicidas em artrópodes aquáticos de áreas de conservação tropicais. Este estudo avalia pela primeira vez a toxicidade dos herbicidas ametryn, atrazine e clomazone sobre o inseto aquático Limnocoris submontandoni (Hemiptera: Naucoridae). A concentração letal (LC50) de herbicidas foi avaliada para esses insetos, bem como o efeito dos herbicidas nos tecidos e testículos dos insetos. A LC50 estimada foi de 1012,41, 192,42 e 46,09 mg/L para clomazone, atrazine e ametryn, respectivamente. Alterações nos espermatócitos e espermátides foram observadas sob o efeito de atrazine, e efeitos na espermatogênese foram observados para algumas concentrações de clomazone, com aparente recuperação após um curto período de tempo. Nossos resultados fornecem informações úteis sobre os efeitos de resíduos de herbicidas em sistemas aquáticos. Essas informações podem ajudar a minimizar o risco de efeitos reprodutivos de longo prazo em espécies não-alvo que foram negligenciadas anteriormente em estudos de ecotoxicologia.

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.

Intoxicação cutânea grave por Paraquat / Serious skin poisoning by Paraquat

O Paraquat é um herbicida não seletivo altamente tóxico, sendo responsável por altas taxas de letalidade acidentais ou provocadas, devido, principalmente, à insuficiência respiratória. Apesar de a intoxicação por via oral ser a principal e mais grave, o contato prolongado com a substância em uma grande área corporal pode gerar uma toxicidade similar e levar ao óbito ­ fato pouco elucidado na literatura. Este é o relato de caso de um homem de 22 anos, que foi admitido em um hospital devido a queixas de mal-estar, náuseas, febre, cefaleia, dor abdominal, diarreia, queimaduras e dispneia. A suspeita diagnóstica inicial foi de hantavirose, leptospirose, dengue e tromboembolismo pulmonar e, posteriormente, foi comprovado intoxicação exógena com agrotóxico por via inalatória e cutânea. Exames de imagem revelaram fibrose pulmonar difusa e ele também apresentou alterações renais, hepáticas e coagulatórias. Como não há antídoto específico, foi empregado tratamento sintomático e suportivo, com uso de carvão ativado, antibióticos, corticoides, antioxidantes e hemodiálise. No entanto, o paciente teve uma piora progressiva do quadro, vindo a óbito devido à síndrome da angústia respiratória aguda e fibrose pulmonar. O Paraquat, embora proibido no Brasil em 2020, continua sendo utilizado de forma ilegal. Além disso, seu substituto, o Diquat, possui toxicidade semelhante. Assim, é fundamental que os profissionais da saúde reconheçam o diagnóstico da intoxicação por tais substâncias e suas diferentes vias de exposição. Também são necessárias novas medidas de fiscalização das substâncias e maior investimento em educação em saúde para evitar exposições acidentais, assim como relatado (AU).

Paraquat is a highly toxic non-selective herbicide and is responsible for high accidental or provoked lethality rates, mainly due to respiratory failure. Although oral poisoning is the primary and most severe, prolonged contact with the substance in a large body area can lead to similar toxicity and death ­ a fact that is little elucidated in the literature. This is the case report of a 22-year-old man admitted to a hospital due to complaints of malaise, nausea, fever, headache, abdominal pain, diarrhea, burns and dyspnea. The initial diagnostic suspicion was hantaviruses, leptospirosis, dengue and pulmonary thromboembolism, and there was subsequently proven exogenous intoxication with pesticides by inhalation and cutaneous route. Imaging tests revealed diffuse pulmonary fibrosis and he also had renal, hepatic and coagulation alterations. As there is no specific antidote, symptomatic and supportive treatment was performed using activated charcoal, antibiotics, corticosteroids, antioxidants and hemodialysis. However, the patient had a progressive worsening of the condition and died due to acute respiratory distress syndrome and pulmonary fibrosis. Paraquat, although banned in Brazil in 2020, continues to be used illegally. In addition, its substitute, the Diquat, has similar toxicity. Thus, it is essential that health professionals recognize the diagnosis of intoxication by such substances and their different exposure routes. New control measures for these substances and greater investment in health education are also needed to prevent accidental exposure, as reported (AU).

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.

Consultorio de Salud Ambiental en un Hospital Privado de Comunidad de la Ciudad de Buenos Aires / Environmental Health consulting office in a Private Community Hospital of the Buenos Aires City

En los últimos años surge el concepto de Una Sola Salud que reconoce la interdependencia sistémica al observar que los cambios en la salud humana se expresan de manera sincrónica e indivisible de la salud del ambiente. Nuevas enfermedades y daños crónicos inespecíficos ocurren a la par de la pérdida de biodiversidad y vitalidad. En las últimas décadas venimos observando el surgimiento de "adaptaciones sistémicas" que requieren un abordaje desde la clínica y la toxicología a nivel individual y desde la epidemiología de la complejidad a nivel poblacional. Luego de un largo recorrido de investigaciones, el Hospital Italiano formalizó el consultorio de Salud Ambiental con la intención de brindar respuesta a la demanda de pacientes que atribuyen síntomas y signos a la polución ambiental. (AU)

In recent years, the concept of One Health has emerged, recognizing the systemic interdependence and the changes in human health that are expressed synchronously and indivisible from the environment. New diseases and nonspecific chronic damage are occuring in parallel with the loss of biodiversity and vitality.In recent decades we have observed the appearance of "systemic adaptations" that require a clinical and toxicological approach at the individual level, and address the population level from an epidemiological and complexity science paradigm. After many years of research, the Hospital Italiano de Buenos Aires formalized the Environmental Health consulting office and the measurement of glyphosate levels, giving answer to the demand of patients who associate their signs and symptoms to environmental pollution. (AU)

Does doses and time of 2, 4-D application interfere in the physiology and wheat grains yield components?

The identification of the application stage and correct dose of 2,4-dichlorophenoxyacetic acid (2,4-D) herbicide is important so that wheat is not harmed. In view of this, the objective of this study was to evaluate the effect of 2,4-D doses applied at different development stages of wheat crop. The experiment was conducted in a randomized block design, arranged in a 4 × 5 factorial scheme, with four replications. In factor A, the application stages (before tillering, tillering, first node and booting) were allocated and the doses of 2,4-D (0, 349, 698, 1047 and 1396 g.ha­1) were allocated in factor B. The variables evaluated were phytotoxicity at 7, 14, 21, 28 and 35 days after application of the treatments (DAT), photosynthetic activity, CO2 internal concentration, stomatal conductance, efficient water use and carboxylation efficiency. The number of spikes·m­2, spike length and number of full and sterile grains were determined in the preharvest. Thousand grain mass, grain yield and hectoliter weight were determined after harvest. The results demonstrate that the herbicide caused phytotoxicity to wheat, being greater in increasing doses and mainly before tillering, causing grain sterility and decreased productivity. The other yield components did not present difference when increasing the dose and application in different stages as well as the physiological variables. The increase of the 2,4-D doses applied before tillering and in the booting stage caused linear decrease in wheat grain yield.

Mortalidade de abelhas sem ferrão Scaptotrigona bipunctata sob os efeitos dos herbicidas paraquat e diquat / Mortality of Scaptotrigona bipunctata stingless bees under the effects of paraquat and diquat herbicides / Mortalidad de abejas sin aguijón Scaptotrigona bipunctata bajo los efectos de los herbicidas paraquat y diquat

As abelhas exercem essencial atividade polinizadora, além de fundamentais na manutenção ecossistêmica, também polinizam diversos sistemas agrícolas. São crescentes os relatos sobre o desaparecimento de abelhas, possivelmente em decorrência do intenso uso de agrotóxicos, os quais não possuem seus efeitos sobre a biodiversidade, de fato, conhecidos. A maioria dos estudos nessa área é voltada para a compreensão dos efeitos de inseticidas, porém, os herbicidas também podem influenciar na mortandade dos insetos polinizadores. Referente aos herbicidas paraquat e diquat, amplamente utilizados na agricultura, pouco se conhece sobre os efeitos que atingem organismos não-alvo, como as abelhas. Este trabalho avaliou a mortalidade das abelhas Scaptotrigona bipunctata submetidas aos herbicidas paraquat e diquat, via contato e ingestão. Dois experimentos foram conduzidos no delineamento de blocos casualizados em esquema fatorial 2 x 2 com tratamento adicional, em três repetições. Os tratamentos foram: dois herbicidas (paraquat e diquat), duas doses (50 e 100%) e um tratamento adicional sem exposição ao produto (controle). Cada parcela consistiu em dez indivíduos, expostos aos tratamentos e incubados à 28oC durante 72 horas em ambiente controlado. Para o experimento de ingestão, ambas as doses dos herbicidas resultaram em maior mortalidade de abelhas em relação à condição controle. Ademais, a dose 100% resultou em maior toxicidade comparada à dose 50%. Para a contaminação via contato, não houve aumento na mortalidade das abelhas perante a exposição. Conclui-se que a contaminação por ingestão com os herbicidas paraquat e diquat pode afetar a sobrevivência das abelhas sem ferrão, podendo em longo prazo alterar a viabilidade e a dinâmica das populações das abelhas Scaptotrigona bipunctata.(AU)

Bees have essential pollinating activities and are fundamental to the maintenance of the ecosystem since they also pollinate various agricultural systems. There are growing reports about the disappearance of bees, possibly due to the intense use of pesticides, which do not have their effects on biodiversity fully known. Most studies in this area are aimed at understanding the effects of insecticides; however, herbicides can also influence the mortality of pollinating insects. Regarding the paraquat and diquat herbicides, widely used in agriculture, little is known about the effects on non-target organisms, such as bees. This work evaluated the mortality of Scaptotrigona bipunctata bees submitted to the paraquat and diquat herbicides via contact and ingestion. Two experiments were conducted in a randomized block design in a 2 x 2 factorial scheme with additional treatment, in three replications. The treatments were: two herbicides (paraquat and diquat), two doses (50 and 100%), and an additional treatment without contamination (control). Each plot consisted of ten individuals exposed to treatments and incubated at 28 ºC for 72 hours in a controlled environment. For the ingestion experiment, both doses of the herbicides resulted in higher bee mortality when compared to the control condition. In addition, the 100% dose resulted in greater toxicity when compared to the 50% dose. For contact contamination, there was no increase in bee mortality on exposure. It can be concluded that contamination by ingestion of paraquat and diquat can affect the survival of stingless bees, which in the long term may alter the viability and population dynamics of Scaptotrigona bipunctata bees.(AU)

Las abejas ejercen una actividad polinizadora esencial, además de ser fundamentales en el mantenimiento de los ecosistemas, también polinizan varios sistemas agrícolas. Cada vez hay más informes sobre la desaparición de abejas, posiblemente como consecuencia del uso intenso de pesticidas, que no tienen sus efectos sobre la biodiversidad, de hecho, conocidos. La mayoría de los estudios en esta área tienen como objetivo comprender los efectos de los insecticidas, sin embargo, los herbicidas también pueden influir en la mortalidad de los insectos polinizadores. Con respecto a los herbicidas paraquat e diquat, ampliamente utilizados en la agricultura, se sabe poco sobre los efectos que afectan a organismos no objetivo, como las abejas. Esta investigación evaluó la mortalidad de abejas Scaptotrigona bipunctata sometidas a herbicidas paraquat y diquat, por contacto e ingestión. Dos experimentos fueron conducidos en el delineamiento de bloques casualizados en esquema factorial 2 x 2 con tratamiento adicional, en tres repeticiones. Los tratamientos fueron: dos herbicidas (paraquat y diquat) dos dosis (50 y 100%) y un tratamiento adicional sin exposición al producto (control). Cada parcela estuvo compuesta por diez individuos, expuestos a tratamientos e incubados a 28ºC durante 72 horas en ambiente controlado. Para el experimento de ingestión, ambas dosis de herbicidas resultaron en mayor mortalidad de abejas en comparación con la condición de control. Además, la dosis del 100% resultó en mayor toxicidad en comparación con la dosis del 50%. Para la contaminación por contacto, no hubo un aumento en la mortalidad de las abejas debido a la exposición. Se concluye que la contaminación por ingestión con herbicidas paraquat y diquat puede afectar la supervivencia de las abejas sin aguijón, lo que a largo plazo puede alterar la viabilidad y la dinámica poblacional de las abejas Scaptotrigona bipunctata.(AU)

Biomarker responses (serum biochemistry) in pregnant female wistar rats and histopathology of their neonates exposed prenatally to pesticides

Experiments were conducted to investigate the effects on health of pregnant female rats exposed to pesticides glyphosate and carbendazim. Glyphosate is used as herbicide and carbendazim as a fungicide; all are commercially available readily for various agricultural and domestic purposes. The hypothesis tested in this investigation is that pesticide exposure during pregnancy causes changes in biomarker responses like serum glucose level, total protein, total cholesterol, triglycerides, SGOT, SGPT, and billirubin level. Significant changes were observed in all above biomarker responses, when compared with the reference. Histopathology of skin and kidney of rat neonates showed marked damage. Degenerative changes and vacuolization with eroded capsule were observed in kidney sample and thinning of epidermis in skin sample was seen in pesticides exposed neonates of rats. The serum biochemistry and histopathological findings are valuable markers for observing the changes caused by pesticide exposure.

Determinação de paraquate em batatas / Determination of paraquat in potatoes

O Paraquate (1,1'-dimetil-4,4'-bipiridina- dicloreto) é herbicida amplamente utilizado em vários países para diferentes culturas. O objetivo é determinar a concentração de Paraquate em batatas comercializadas em diferentes estabelecimentos da zona leste de São Paulo. Foram coletadas 12 (doze) amostras de batatas adquiridas no comércio varejista (sacolões, ou seja, do de frutas, verduras e legumes; supermercados e feiras livres) da zona leste do município de São Paulo. A quantificação do Paraquate foi baseada na reação de complexação com o ditionito de sódio, gerando composto de cor azulada, cuja absorvância foi lida em espectrofotômetro em comprimento de onda de 600nm. Foi construída a curva padrão e a determinada a equação da reta (y = 1,6448x e R2= 0,9945). O limite de tolerância do herbicida em alimentos é de 0,2 partes por milhão ou 0,2 mg/kg, enquanto que a ingestão diária aceitável (IDA) é de 0,004 mg/kg de peso corpóreo. Assim, pode-se observar que os valores encontrados em três amostras estão acima do limite máximo permitido, enquanto quatro apresentaram concentrações muito próximas ao limite. Os resultados permitem inferir que existe a necessidade de intensificação na fiscalização nos locais de comercialização de alimentos produzidos com a utilização de agrotóxicos.(AU)

Paraquat (1,1'-dimethyl-4,4'- bipyridine-dichloride) is herbicide widely used in several countries in different plantations. The objective is to determine the concentration of Paraquat in potatoes, marketed in different establishments in the eastern zone of São Paulo. Twelve (12) samples of potatoes purchased from the retail trade ("sacolões", ie fruit, vegetable and vegetable markets, supermarkets and free markets) were collected from the eastern part of the city of São Paulo. The quantification of Paraquat was based on the reaction of complexation with the sodium dithionite, generating compound of blue color, whose absorbance was read in a spectrophotometer at wavelength of 600 nm. The standard curve was constructed and the equation of the line was determined (y = 1,6448x e R2 = 0,9945). The tolerance limit of the herbicide in foods is 0.2 parts per million or 0.2 mg/kg, while the acceptable daily intake (ADI) is 0.004 mg/kg body weight. Thus, it can be observed that the values found in three samples are above the maximum allowed limit, while four of them presented concentrations very close to the limit. The results allow inferring that there is a need for intensification in the inspection in the commercial places of food produced with the use of pesticides.

Effects of monosulfuron-ester on metabolic processes of nitrogen-fixing cyanobacteria Anabaena flos-aquae and Anabaena azotica

Abstract Presence of the relatively new sulfonylurea herbicide monosulfuron-ester at 0.03-300 nmol/L affected the growth of two non-target nitrogen-fixing cyanobacteria (Anabaena flos-aquae and Anabaena azotica) and substantially inhibited in vitro Acetolactate synthase activity, with IC50 of 3.3 and 101.3 nmol/L for A. flos-aquae and A. azotica, respectively. Presenting in 30-300 nmol/L, it inhibited protein synthesis of the cyanobacteria with less amino acids produced as its concentration increased. Our findings support the view that monosulfuron-ester toxicity in both nitrogen-fixing cyanobacteria is due to its interference with protein metabolism via inhibition of branch-chain amino acid biosynthesis, and particularly Acetolactate synthase activity.

Efeito protetor da melatonina sobre intoxicações por herbicidas / Protective effect of melatonin on poisoning by herbicides

O uso inadequado de herbicidas pode resultar em intoxicações agudas e, às vezes, crônicas por exposição em longo prazo a baixos níveis desses agentes tóxicos, podendo o herbicida atuar também como agentes teratogênicos, mutagênicos, cancerígenos e desreguladores endócrinos, com o aparecimento de doenças neurodegenerativas e distúrbios reprodutivos. Estudos têm revelado que a melatonina tem propriedades antioxidantes, anti-inflamatórias e imunomoduladoras e atua na reprodução. Essa indolamina está entre os agentes que têm se mostrado benéfico em intoxicações por herbicidas, porém não há relatos do uso de melatonina contra intoxicações por Glifosato-Roundup®, muito menos em associação com o Paraquat. Dessa forma, o maior interesse no tratamento das intoxicações por herbicidas, tem-se concentrado em medidas que impeçam ou minimizem as lesões celulares provocadas nos diversos sistemas biológicos. Assim, a melatonina, como antioxidante conhecido, pode ser mais uma alternativa contra as intoxicações por herbicidas associados e/ou individuais.

The inadequate use of herbicides may cause serious and sometimes chronic poisoning due to long exposure to low levels of toxic agents. Herbicides may also be teratogenic, mutagenic, cancerigenous agents and endocrine disruptors, with the occurrence of neurodegenerative diseases and reproduction disorders. Several studies have shown that melatonin has antioxidant, anti-inflammatory and immune-modulating qualities, besides affecting the reproduction system. It is among the agents which are beneficent in poisoning by herbicides even though no reports are extant on the use of melatonin against poisoning by Glyphosate-Roundup® alone or associated with Paraquat. Solutions that prevent or minimize cell lesions caused by several biological systems have been focused upon in the treatment for poisoning with herbicides. Thus, melatonin, a known antioxidant, may be an alternative against the poisoning by single or associated herbicides.

Acute toxicity and sublethal effects of the mixture glyphosate (Roundup® Active) and Cosmo-Flux®411F to anuran embryos and tadpoles of four Colombian species / Toxicidad aguda y efectos subletales de la mezcla glifosato (Roundup® Activo) y Cosmo-Flux®411F en embriones y renacuajos de cuatro especies de anuros colombianos

Glyphosate is the most widely used herbicide in the world with application in agriculture, forestry, industrial weed control, garden and aquatic environments. However, its use is highly controversial for the possible impact on not-target organisms, such as amphibians, which are vanishing at an alarming and rapid rate. Due to the high solubility in water and ionic nature, the glyphosate requires of surfactants to increase activity. In addition, for the control of coca (Erythroxylum coca) and agricultural weeds in Colombia, formulated glyphosate is mixed and sprayed with the adjuvant Cosmo-Flux®411F to increase the penetration and activity of the herbicide. This study evaluates the acute toxic and sublethal effects (embryonic development, tadpole body size, tadpole swimming performance) of the mixture of the formulated glyphosate Roundup® Active and Cosmo-Flux®411F to anuran embryos and tadpoles of four Colombian species under 96h laboratory standard tests and microcosms, which are more similar to field conditions as they include soil, sand and macrophytes. In the laboratory, embryos and tadpoles of Engystomops pustulosus were the most tolerant (LC50=3 904µg a.e./L; LC50=2 799µg a.e./L, respectively), while embryos and tadpoles of Hypsiboas crepitans (LC50=2 203µg a.e./L; LC50=1 424µg a.e./L, respectively) were the most sensitive. R. humboldti and R. marina presented an intermediate toxicity. Embryos were significantly more tolerant to the mixture than tadpoles, which could be likely attributed to the exclusion of chemicals by the embryonic membranes and the lack of organs, such as gills, which are sensitive to surfactants. Sublethal effects were observed for the tadpole body size, but not for the embryonic development and tadpole swimming performance. In microcosms, no toxicity (LC50 could not be estimated), or sublethal responses were observed at concentrations up to fourfold (14.76kg glyphosate a.e./ha) the highest field application rate of 3.69kg glyphosate a.e./ha. Thus, toxicity was less in the microcosms than in laboratory tests, which may be attributed to the presence of sediments and organic matter which rapidly adsorb glyphosate and surfactants such as POEA. It is concluded that the mixture of glyphosate (Roundup® Active) and Cosmo-Flux®411F, as used in the field, has a negligible toxic effect to embryos and tadpoles of the species tested in this study.

El glifosato es el herbicida más usado en el mundo con aplicaciones para la agricultura, control de malezas forestales, industriales, en jardines y ambientes acuáticos. Sin embargo, su uso es altamente controversial por el posible impacto sobre organismos no blanco, como los anfibios, los cuales están desapareciendo a una tasa alarmantemente rápida. Debido a su alta solubilidad en agua y naturaleza iónica, el glifosato requiere de surfactantes para incrementar su actividad. Además, para el control de Erythroxylum coca y de malezas en la agricultura en Colombia, el glifosato formulado es mezclado y rociado con el coadyuvante Cosmo-Flux®411F para incrementar la penetración y actividad del herbicida. Este estudio evalúa los efectos tóxicos agudos y subletales (desarrollo embrionario, tamaño corporal y desempeño natatorio de los renacuajos) de la mezcla del glifosato formulado Roundup® Activo con el Cosmo-Flux®411F en embriones y renacuajos de cuatro especies de anuros colombianos, bajo pruebas de 96h en condiciones estándar de laboratorio y microcosmos, que son más similares a las condiciones de campo al incluir tierra, arena y macrófitas. En laboratorio, los embriones y renacuajos de Engystomops pustulosus fueron los más tolerantes (CL50=3 904µg a.e./L; CL50=2 799µg a.e./L, respectivamente), mientras que los embriones y renacuajos de Hypsiboas crepitans fueron los más sensibles (CL50=2 203µg a.e./L; CL50=1 424µg a.e./L, respectivamente). R. humboldti y R. marina presentaron una toxicidad intermedia. Los embriones fueron más tolerantes a la mezcla que los renacuajos, lo cual podría ser atribuido a la exclusión de los químicos por las membranas embrionarias y a la falta de órganos, como las branquias, que son más sensibles a los surfactantes. Se observaron efectos subletales en el tamaño corporal de los renacuajos, pero no en el desarrollo embrionario ni el desempeño natatorio de los renacuajos. En microcosmos no se observaron efectos tóxicos ni respuestas subletales a concentraciones hasta cuatro veces (14.67kg glifosato a.e./ha) la tasa de aplicación más alta de 3.69kg glifosato a.e./ha. Por lo tanto, la toxicidad fue menor en los microcosmos que en las pruebas de laboratorio, lo que puede ser atribuido a la presencia de sedimentos y materia orgánica que absorbe rápidamente el glifosato y surfactantes como el POEA. Se concluye que la mezcla del glifosato (Roundup® Activo) y Cosmo-Flux®411F, como se aplica en campo, tiene un efecto tóxico bajo en los embriones y renacuajos de las especies estudiadas.

Longterm melatonin administration alleviates paraquat mediated oxidative stress in Drosophila melanogaster / La administración de melatonina a largo plazo mitiga el estrés oxidativo mediado por paraquat en Drosophila melanogaster

We investigated the effect of melatonin (MEL) in the activities of cytosolic superoxide dismutase (SOD) and catalase as well as in the levels of H2O2 and mitochondrial malondialdehyde (MDA) in paraquat-intoxicated Drosophila melanogaster. Paraquat (40 mM) was administrated for 36 h. Three groups of flies intoxicated with paraquat were used: PQ (exposed during 36h to paraquat), PQ-MEL (exposed during 36h to paraquat and then treated with MEL [0.43 mM] for 12 days) and PQ-Control (maintained in standard corn meal for 12 days). Two additional groups without pre-intoxication with PQ were added: Control (maintained in standard corn meal) and MEL (treated with MEL for 12 days). Immediately after PQ intoxication the concentration of MDA (17.240 ± 0.554 nmoles MDA/mg protein) and H2O2 (3.313 ± 0.086 nmol hydrogen peroxide/mg protein) and the activities of SOD and catalase (419.667 ± 0.731 and 0.216 ± 0.009 Units/mg of protein, respectively) in the PQ group were significantly increased with respect to Control. After 12 days of intoxication with PQ, the PQ-Control flies showed increases in H2O2 (4.336 ± 0.108) and MDA levels (8.620 ± 0.156), and in the activities of SOD and catalase (692.570 ± 0.433 and 0.327 ± 0.003, respectively) as compared to PQ-MEL (p<0.001). Treatment with MEL extended the life span of the groups PQ-MEL and MEL when compared to their corresponding controls. Motor activity decreased significantly in PQ-Control and PQ-MEL flies, suggesting that the damage caused by PQ affected the nervous system of flies. Our findings showed that oxidative damage caused by paraquat was observed even after 12 days and that melatonin mitigates this damage.

Investigamos el efecto de la melatonina (MEL) en la actividad de la superóxido dismutasa citosólica (SOD) y la catalasa, así como en las concentraciones del H2O2 y del malondialdehido mitocondrial (MDA) en la toxicidad inducida por paraquat (PQ) en Drosophila melanogaster. El paraquat (40 mM) fue administrado durante 36h. Tres grupos de moscas se utilizaron después de la intoxicación con paraquat: PQ (expuestas a paraquat durante 36 h), PQ-MEL (expuestas durante 36 horas a PQ y luego tratadas con MEL [0,43 mM] por 12 días) y PQ-Control (mantenidas en medio estándar por 12 días). Se incluyeron dos grupos adicionales sin pre-intoxicación con PQ: Control (mantenido en medio estándar) y MEL (tratado con MEL por 12 días). Inmediatamente después de la intoxicación con PQ, las concentraciones de MDA (17,240 ± 0,554 nmol de MDA/mg de proteína), H2O2 (3,313 ± 0,086 nmol de H2O2/mg de proteína) y las actividades de la SOD y catalasa (419,667 ± 0,731 y 0,216 ± 0,009 unidades/mg de proteína, respectivamente) se incrementaron significativamente con respecto al Control. Doce días después de la intoxicación con PQ, las moscas PQ-Control mostraron un aumento en la concentración de H2O2 (4,336 ± 0,108), de los niveles de MDA (8,620 ± 0,156) y en las actividades de la SOD y la catalasa (692,570 ± 0,433 y 0,327 ± 0,003, respectivamente) en comparación con el grupo PQ-MEL (p<0,001). El tratamiento con MEL extendió el tiempo de vida de los grupos PQ-MEL y MEL en comparación con sus correspondientes controles. La actividad motora disminuyó significativamente en las moscas de los grupos PQ-Control y PQ-MEL, lo que sugiere que el PQ afectó el sistema nervioso de las moscas. Nuestros hallazgos demostraron que el daño oxidativo causado por paraquat en las moscas fue observado aún después de 12 días de intoxicadas y que la melatonina logró mitigar este daño.

Cytogenetic study of the effect of bentazon and glyphosate herbicide on mice

Genotoxic effect of acute and chronic doses of Bentazon and Glyphosate herbicides were estimated using micronucleus [MN], nuclear budding, mitotic index [MI] and chromosomal aberration assays. Five groups of Swiss male albino mice were used in acute exposure of 1/5 and 1/10 LD50 of both herbicides after 24, 96 hours of exposure compared to the control. For chronic exposure 1/20 of LD50 of each herbicide was for two months. Genotoxicity of acute doses at different duration of Bentazon and Glyphosate were confirmed through significant increased number of aberrant cells, different types of chromosomal aberration, micronucleus and nuclear budding. Chronic dose of Bentazone induced more genotoxic effects than Glyphosate

Avaliação da toxicidade de herbicidas usados emcana-de-açúcar para o Paulistinha (Danio rerio) / Toxicity evaluation of herbicides used in sugarcane crops toZebrafish (Danio rerio)

Na natureza, os organismos são constantemente expostos a mais de um agente tóxico e, apesar do fenômeno de interaçõesquímicas ser conhecido há tempos, são poucos os estudos realizados que privilegiam a observação dos efeitosdecorrentes da exposição a duas ou mais substâncias. O objetivo deste trabalho foi avaliar o efeito combinado damistura dos herbicidas Gesapax 500® (ametrina 500 g/L) e Velpar K® (diuron 468 g/kg + hexazinone 132 g/kg) sobreo paulistinha (Danio rerio). O ensaio foi baseado no teste FET da OECD, com duração de 96 horas. As concentraçõestestadas foram 0, 21,22, 29,52, 41,08, 57,17 e 79,56 mg/L de Gesapax 500 vs. 0, 15,21, 21,17, 29,46, 40,99 e 57,04mg/L de Velpar K. Os testes foram conduzidos em triplicata e avaliados diariamente. As CL50-96h determinadas foram41,705 ± 8,373 mg/L para o Gesapax 500 e 55,460 ± 20,826 mg/L para o Velpar K. O modelo da mistura que melhordescreve a relação entre os dois componentes é a ação independente, sendo a toxicidade dose-dependente, ou seja,em baixas doses ocorre antagonismo e, em altas doses, sinergismo. Os endpoints edemas, atraso no desenvolvimentoembrionário (delay) e na absorção do saco vitelínico e diminuição na frequência de cardíaca foram observados apartir das concentrações mais baixas da mistura. Pelos dados obtidos, concluiu-se que a mistura de Gesapax 500 eVelpar K é medianamente tóxica para o paulistinha e que os endpoints avaliados foram úteis na determinação de suatoxicidade.

In nature, organisms are constantly exposed to more than one toxic agent, and although the phenomenon of chemical interactionsare known for some time, there are few previous studies that emphasize observation of the effects resulting fromexposure to two or more substances. The aim of this study was to evaluate the combined effect of the mixture of herbicideGesapax 500® (ametrina 500 g/L) and Velpar K® (diuron 468 g/kg + hexazinone 132 g/kg) on zebrafish (Danio rerio). The testwas based on the Fish Embryo Toxicity (FET) from OECD, lasting 96 hours. The concentrations tested were: 0, 21.22, 29.52,41.08, 57.17 and 79.56 mg/L Gesapax 500 vs. 0, 15.21, 21.17, 29.46, 40.99 and 57.04 mg/L Velpar K. Tests were conductedin triplicate and evaluated daily. LC50-96h was determined to 41.705 ± 8.373 mg/L to Gesapax 500 and 55.460 ± 20.826 mg/Lto Velpar K. The mixture model that best describes the relationship between the two components is independent action andtoxicity of the mixture is dose-dependent, occurring antagonism at low doses and synergism at high doses. The endpointsedema, delay in general development and in yolk sac absorption and decrease in frequency of heart-beat rate were observedfrom the lower concentrations of the mixture. From the data obtained, it is concluded that the mixture of Velpar K and Gesapax500 is moderately toxic to zebrafish and that the endpoints evaluated were useful in determining its toxicity.

Alterations in the duodenum myenteric neurons of Wistar ratsafter ingesting of 2, 4 dichlorophenoxyacetic acid

The 2,4 dichlorophenoxyacetic acid (2,4-D) is a systemic herbicide whose effects in animal organic systemshave been examined in previous studies, being the neurotoxicity considered the predominant effect. However,the studies that detect the 2,4-D neurotoxicity have merely focused in the central nervous system, andtherefore, little is known about the effect of this herbicide in the enteric nervous system. This study aimedto verifying the 2,4-D effects on the myenteric neurons in duodenum of Wistar rats. Ten 60-day-old maleWistar rats (Rattus norvegicus) were divided in two groups: control group (C) that did not receive 2,4-D andexperimental group (E) that received 5.0 mg of 2,4-D/kg for 15 days. At the end of experimental period, theanimal were euthanized, the duodenum was collected and processed for NADPH-diaphorase histochemicalanalysis in order to expose the nitrergic myenteric neurons (NADPH-dp). In the light microscopy analysis, thewhole-mount preparation obtained from duodenum of each animal were image-captured in 120 and 40 fields,for quantitative and morphometric analyses of myenteric neurons, respectively. The neuronal density was notaffected when comparing the two groups, but an increase (p > 0.05) of 8.5% was observed in the cell bodyarea of neurons in the E group. In conclusion, the ingestion of 2,4-D at a dosage of 5.0 mg/kg body weightfor 15 days does not change the neuronal density, but promotes the hypertrophy of NADPH-dp myentericneurons in duodenum of the rats of this study.

Toxicidad en peces de herbicidas formulados con glifosato / Toxicity in fishes of herbicides formulated with glyphosate

En nuestro país existe una gran extensión de hectáreas cultivadas con soja transgénica, la misma ha sido modificada genéticamente para soportar la acción de un herbicida denominado glifosato. Debido a la gran cantidad de formulaciones comerciales que incluyen glifosato es de importancia analizar el impacto ambiental producido por éstas. La evaluacion de la toxicidad aguda de dos herbicidas comerciales formulados con glifosato y de una solución del mismo; frente a peces de la especie Poecilia reticulata "lebistes" acusa que una de las soluciones produce mortalidad del 100 % de los especimenes a 100 μl/l (equivalente a 48 mg/l de principio activo); la otra a 50 μl/l (equivalente a 24 mg/l de principio activo) y la solución formulada con glifosato puro no produce mortalidad aún a concentraciones de 400 mg/l. Utilizando dosis sub letales en función de los datos obtenidos en el ensayo de toxicidad aguda se determinó que a largo plazo especimenes de Cyprinus carpio haematopterus "carpa koi", manifestaron severas alteraciones hematológicas principalmente frente a una de las formulaciones evaluadas.

Nowadays, transgenic soya, modified in order to withstand the impact of the herbicide glyphosate, in one of the main crops grown in Argentina. Due to the large number of commercial formulations that include this drug, is important to analyze both, the acute and chronic environmental impact that they cause. Here the acute toxicity of two commercial herbicides glyphosatebased toward the fish Poecilia reticulate "guppy" was evaluated and compared with pure glyphosate solutions. Interestingly, while commercial herbicides formulations induce a 100% of mortality at concentration ranged between 50 and 100 μl/l, the pure glyphosathe does not present mortality even at doses higher than 400 mg/l. When some long term effects toward Cyprinus carpio haematopterus "koi" were determined by using the sub-lethal doses already calculated it was demonstrated that one of the commercial herbicides induces severe haematological alterations.

Intoxicaciones por paraquat reportadas en el Centro Toxicológico de la Región Centro Occidental "Antonio M. Pineda de Venezuela: período 1997-2000 / Paraquat intoxications reported in the Toxicological Center Antonio M. Pineda: from Venezuela 1997-2000

En Venezuela, es frecuente la utilización de plaguicidas como el paraguat en la actividad agrícola, por ello las intoxicaciones y fallecimientos que se producen anualmente por su mal uso, y manejo es importante. Se analizaron retrospectivamente los casos de intoxicaciones por paraquat registrados en el Centro Toxicológico de la Región Centro Occidental de Venezuela, durante los años 1997-2000, para evaluar frecuencia, causas, vías de ingreso y evaluación de los pacientes intoxicados. La investigación fue descriptiva, de registros médicos. Se registraron 79 pacientes; 44 pacientes del género masculino y 29 del género femenino y 6 no identificados en los registros. El promedio de edad fue de 24,22 ± 12,47 años. La principal vía de ingreso del tóxico fue oral 83.5%, y las causas de intoxicación identificadas fueron: 75,9% ocupacional y 22,8% accidental. El 49,3% de los pacientes fue dado de alta., mientras que el 8,8% falleció

In Venezuela, it is frequent the use of pesticides such as paraquat in agriculture, so poisonings and deaths that occur annually for their improper use and handling its important. We retrospectively analyzed the cases of paraquat poisoning recorded in the Poison Center of the Midwestern Region of Venezuela during the years 1997-2000, to assess prevalence, causes, access routes and evolution of poisoned patients. The research was descriptive, of medical records. There were 79 patienrs, 44 males and 29 females and 6 are not identified in the records. The average age was 24.22 ± 12.47 years. The main route of the toxic was oral 83.5%, and the causes of poisoning identified were: occupational 75.9% and accidental 22.8%. The 49.3% of patients were discharged, while 8.8% died

Down-regulation of survivin suppresses uro-plasminogen activator through transcription factor JunB

Survivin, a member of the inhibitors of apoptosis protein family, is expressed during development and in various human cancers. However, the clinical relevance of survivin in cancer is still a matter of debate. Genes induced by hepatocyte growth factor (HGF) were screened using cDNA microarray technology in the stomach cancer cell lines, NUGC3 and MKN28. The levels of JunB, survivin, and uro-plasminogen activator (uPA) were up-regulated in cells treated with HGF in a dose-dependent manner. HGF-induced up regulation of JunB, survivin, and uPA was inhibited by pre-treatment with a MEK inhibitor (PD 98059). HGF-induced up-regulation of uPA was repressed by survivin knockdown. HGF enhanced the binding activity of JunB to the survivin promoter in control cells, but not in the JunB-shRNA cells. Transfection with survivin-shRNA resulted in a decrement of cell proliferation, as determined with MTT assays. In an in vitro invasion assay, significantly fewer cells transfected with survivin shRNA than control cells were able to invade across a Matrigel membrane barrier. In conclusion, survivin appeared to play an important role in the up-regulation of uPA induced by HGF via JunB and might contribute to HGF-mediated tumor invasion and metastasis, which may serve as a promising target for gastric cancer therapy.