RESUMO

The organophosphate and carbamate pesticides methyl-parathion and carbaryl have a common action mechanism: they inhibit acetylcholinesterase enzyme by blocking the transmission of nerve impulses. However, they can alter the expression of exocytotic membrane proteins (SNARE), by modifying release of neurotransmitters and other substances. This study evaluated the adverse effects of the pesticides methyl-parathion and carbaryl on expression of SNARE proteins: Syntaxin-1, Syntaxin-4 and SNAP-23 in freshwater rotifer Brachionus calyciflorus. Protein expression of these three proteins was analyzed before and after exposure to these two pesticides by Western Blot. The expression of Syntaxin-1, Syntaxin-4 and SNAP-23 proteins in B. calyciflorussignificantly decreases with increasing concentration of either pesticides. This suggests that organophosphates and carbamates have adverse effects on expression of membrane proteins of exocytosis by altering the recognition, docking and fusion of presynaptic and vesicular membranes involved in exocytosis of neurotransmitters. Our results demonstrate that the neurotoxic effect of anticholinesterase pesticides influences the interaction of syntaxins and SNAP-25 and the proper assembly of the SNARE complex.

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RESUMO

AbstractThe organophosphate and carbamate pesticides methyl-parathion and carbaryl have a common action mechanism: they inhibit acetylcholinesterase enzyme by blocking the transmission of nerve impulses. However, they can alter the expression of exocytotic membrane proteins (SNARE), by modifying release of neurotransmitters and other substances. This study evaluated the adverse effects of the pesticides methyl-parathion and carbaryl on expression of SNARE proteins: Syntaxin-1, Syntaxin-4 and SNAP-23 in freshwater rotifer Brachionus calyciflorus. Protein expression of these three proteins was analyzed before and after exposure to these two pesticides by Western Blot. The expression of Syntaxin-1, Syntaxin-4 and SNAP-23 proteins in B. calyciflorussignificantly decreases with increasing concentration of either pesticides. This suggests that organophosphates and carbamates have adverse effects on expression of membrane proteins of exocytosis by altering the recognition, docking and fusion of presynaptic and vesicular membranes involved in exocytosis of neurotransmitters. Our results demonstrate that the neurotoxic effect of anticholinesterase pesticides influences the interaction of syntaxins and SNAP-25 and the proper assembly of the SNARE complex.

ResumoOs pesticidas organofosforados e carbamatos metil- paration e carbaril tem um mecanismo de ação comum: eles inibem a enzima acetilcolinesterase, bloqueando a transmissão dos impulsos nervosos. No entanto, eles podem alterar a expressão de proteínas de membrana de exocitose (SNARE), através da modificação da libertação de neurotransmissores e outras substâncias. Este estudo avaliou os efeitos adversos dos pesticidas metil- paration e carbaril sobre a expressão de proteínas SNARE: Sintaxina -1, Sintaxina-4 e SNAP-23 em rotíferos de água doce Brachionus calyciflorus. A expressão destas três proteínas foi analisada antes e depois da exposição a estes dois pesticidas por Western Blot. A expressão das proteínas Sintaxina-1, Sintaxina-4 e SNAP-23 em B. calyciflorus diminui significativamente com o aumento da concentração de ambos os pesticidas. Isto sugere que os organofosfatos e carbamatos têm efeitos adversos sobre a expressão de proteínas de membrana de exocitose, alterando o reconhecimento, de encaixe e fusão de membranas pré-sinápticas e vesiculares envolvidas na exocitose de neurotransmissores. Nossos resultados demonstram que o efeito neurotóxico de pesticidas anticolinesterásicos influencia a interação de sintaxinas e SNAP-25 e a montagem correta do complexo SNARE.

Assuntos

RESUMO

The organophosphate and carbamate pesticides methyl-parathion and carbaryl have a common action mechanism: they inhibit acetylcholinesterase enzyme by blocking the transmission of nerve impulses. However, they can alter the expression of exocytotic membrane proteins (SNARE), by modifying release of neurotransmitters and other substances. This study evaluated the adverse effects of the pesticides methyl-parathion and carbaryl on expression of SNARE proteins: Syntaxin-1, Syntaxin-4 and SNAP-23 in freshwater rotifer Brachionus calyciflorus. Protein expression of these three proteins was analyzed before and after exposure to these two pesticides by Western Blot. The expression of Syntaxin-1, Syntaxin-4 and SNAP-23 proteins in B. calyciflorussignificantly decreases with increasing concentration of either pesticides. This suggests that organophosphates and carbamates have adverse effects on expression of membrane proteins of exocytosis by altering the recognition, docking and fusion of presynaptic and vesicular membranes involved in exocytosis of neurotransmitters. Our results demonstrate that the neurotoxic effect of anticholinesterase pesticides influences the interaction of syntaxins and SNAP-25 and the proper assembly of the SNARE complex.(AU)

Os pesticidas organofosforados e carbamatos metil- paration e carbaril tem um mecanismo de ação comum: eles inibem a enzima acetilcolinesterase, bloqueando a transmissão dos impulsos nervosos. No entanto, eles podem alterar a expressão de proteínas de membrana de exocitose (SNARE), através da modificação da libertação de neurotransmissores e outras substâncias. Este estudo avaliou os efeitos adversos dos pesticidas metil- paration e carbaril sobre a expressão de proteínas SNARE: Sintaxina -1, Sintaxina-4 e SNAP-23 em rotíferos de água doce Brachionus calyciflorus. A expressão destas três proteínas foi analisada antes e depois da exposição a estes dois pesticidas por Western Blot. A expressão das proteínas Sintaxina-1, Sintaxina-4 e SNAP-23 em B. calyciflorus diminui significativamente com o aumento da concentração de ambos os pesticidas. Isto sugere que os organofosfatos e carbamatos têm efeitos adversos sobre a expressão de proteínas de membrana de exocitose, alterando o reconhecimento, de encaixe e fusão de membranas pré-sinápticas e vesiculares envolvidas na exocitose de neurotransmissores. Nossos resultados demonstram que o efeito neurotóxico de pesticidas anticolinesterásicos influencia a interação de sintaxinas e SNAP-25 e a montagem correta do complexo SNARE.(AU)

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In southern Brazilian apple (Malus spp.) orchards, predominantly organophosphates are used to control the oriental fruit moth, Cydia molesta (Busck) (Lepidoptera: Tortricidae), but control failures often occur. Therefore the susceptibility of three C. molesta Brazilian populations was investigated to five insecticides of different groups and modes of action, in comparison with a susceptible laboratory strain mass reared in southern France for >10 yr. At the same time, comparative biochemical and genetic analysis were performed, assessing the activities of the detoxification enzymatic systems and sequencing a gene of insecticide molecular target to find out markers associated with resistance. The three Brazilian populations were significantly resistant to chlorpyrifos ethyl compared with the reference strain. One of the field populations that had been frequently exposed to deltamethrin treatments showed significant decreasing susceptibility to this compound, whereas none of the three populations had loss of susceptibility to tebufenozide and thiacloprid compared with the reference strain. All three populations had slight but significant increases of glutathione transferase and carboxylesterases activities and significant decrease of specific acetylcholinesterase activities compared with the reference. Only the most resistant population to chlorpyriphos exhibited a significantly higher mixed function oxidase activity than the reference. The acetylcholinesterase of females was significantly less inhibited by carbaryl in the Brazilian populations than in the reference strain (1.7-2.5-fold), and this difference was not expressed in the male moth. However, no mutation in the MACE locus was detected. These biological and molecular characterizations of adaptive response to insecticides in C. molesta provide tools for early detection of insecticide resistance in field populations of this pest.

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Studies were done to see the effects of five toxic baits based on metaldehyde and/or carbaryl on the taxonomic composition of soil meso and macrofauna, and determined the density of beneficial organisms in soybean under no tillage. Six treatments were tested and soil monoliths were taken before application and at 45 and 75 days after application (DAA). Before application, 1601 individuals were recorded, belonging 1086 to mesofauna and 515 to macrofauna. In soil mesofauna such as Annelida: Enchytraeidae (44%), Nematoda (33%) and Arthropoda (23%) were found. In the macrofauna Arthropoda 93% and Annelida: Oligochaeta, Megadrilli 7% were observed. No differences were observed among the treatments (p>0.05) in the total mesofauna density, at 45 and 75 DAA. The density of enchytraeids, mites, collembolans and symphylids showed no differences at45 and 75 DAA (p > 0.05). The total macrofauna density showed differences (p < 0.05) at 45 and 75 DAA. The highest density was obtained with 4 kg ha(-1) MataBibos Acay at 45 and 75 DAA. Earthworm and diplopod densities at45 and 75 DAA showed no differences (p>0.05). The diversity of meso and macrofauna was not affected at 45 and 75 DAA. Bait application did not affect either the abundance or the diversity of soil beneficial invertebrates.

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O objetivo da presente investigaçäo foi determinar os efeitos de doses tóxicas de agentes anticolinesterasicos carbamatos, fisostigmina e carbaril, na pressäo arterial e freqüência cardíaca de ratos, bem como a açäo protetora do reativador de colinesterase, a pralidoxima, contra estes efeitos. A acetilcolina injetada intravenosamente, em ratos, causou uma imediata queda dose-dependente na Pressäo Arterial Média (PAM), seguida por um aumento na PAM que apresentava duraçäo maior que a queda. Houve também uma queda na Freqüência Cardíaca (F.C). Injeçäo intravenosa de carbaril em rato normal causou uma queda na PAM dentro de 5 segundos, seguida pelo aumento significante na mesma em relaçäo ao valor do controle, 5 minutos após a injeçäo Carbaril näo provocou nenhum efeito na F.C. Quando o carbaril foi precedido pela injeçäo de pralidoxima, as alteraçöes na PAM foram similares àquelas observadas após a injeçäo de carbaril isoladamente, porém a F.C. diminuiu de maneira significante nos grupos que receberam doses maiores. Fisostigma injetada intravenosamente em ratos normais näo apresentou efeito imediato (5 segundos) na PAM, porém ocorreu um aumento significante na PAM 5 minutos após a injeçäo. Nenhuma alteraçäo significante ocorreu na F.C. Em animais previamente tratados com pralicoxima, a fisostigmina causou uma queda imediata na PAM que foi seguida por um aumento significante em relaçäo ao controle, 5 minutos após a injeçäo. Uma queda na F.C. também ocorreu. Administraçäo prévia de pralitoxima potenciou os efeitos cardiovasculares causados pela fisostigmina, porém näo os efeitos do carbaril.

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Motor, sensory and thermoregulatoty function were examined in aging rats (12 months) following two schedules of repated po adminstration of the carbamate insecticide carbaryl and these effects were assessed in terms of blood cholinesterase activity. Administration of carbaryl (50 mg/Kg) by gavage daily for 30 days resulted in a resultad in a reduction of locomotor activity in thre open-field and an inhibition of cholinesterase activity within 30 min after the last treatment. Twenty-for h later, only the locomotor effect was evident. After 90 days of exposure to carbaryl in drinking water, no significant effects were observed. These findings suggest that repeated administration of carbaril to aging rats can induce an impairment of motor function and a reduction of cholinesterase activity, while tolerance develops in some other parameters

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The influence of two carbamine pesticides i.e., manebe and carbaryl upon the hepatic microsomal enzymes induction in the rat was studied. Both substances, when administered by themselves, affect only slightly liver weight, P 450 cytochrome rates and bilirubin glucuronosyltransferase, in the microsome fraction of the hepatic homogenate. It seems, however, that carbaryl is involved in producing a slight induction, whereas manebe acts inversely. Yet, manebe changes largely the induction effects of phenobarbital when associated with the latter. In the animal treated simultaneously with manebe and phenobarbital, the increase in the rate of hepatic microsomal P 450 cytochrome as well as the variations in the distribution of fatty acids in phospholipids, are significantly lower than in the animal solely treated with phenobarbital.

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