Use of molecular and In Silico bioinformatic tools to investigate pesticide binding to insect (Lepidoptera) phenoloxidases (PO): Insights to toxicological aspects

In the present work, the promising bioinformatic tools, based on structure–affinity analysis, allowed to screen several pesticides supposed to bind to the insect immune Phenoloxidases (PO). First, the binding of aminoparathion, a reference compound, to the PO was structurally (3D) validated in accordance with previous reports. Second, using the same docking conditions, a range of pesticides was checked for their binding ability to the crystal 3D structure (PDB 3HSS) of the insect Manduca sexta (Lepidoptera) PO. The obtained data showed that many of the tested pesticides were able to bind, in silico, to M. sexta PO. The combination of in vitro (chemical and biochemical) and in silico (automated docking) approaches was found advantageous to elucidate the behavior of phenolic pesticides as substrate-analogues when binding to the active site of insect POs. Our findings emphasize new ecotoxicological aspects of pesticide residues in the agro-chemical and environmental circles.     

土壤中残留对硫磷和甲基对硫磷的高效液相色谱分析

本文介绍一种简介、快速的前处理和反相高效液相色谱法对土壤样品中残留的对硫磷和甲基对硫磷进行定量分析，样品添加回收率分别为８２％和８６％，最小检测浓度为０．１ｍｇ．ｋｇ＾－１。     

Alterations of protein profile in zebrafish liver cells exposed to methyl parathion: a membrane proteomics approach

Methyl parathion (MP) is an extensively used organophosphorus pesticide, which has been associated with a wide spectrum of toxic effects on environmental organisms. The aim of this study is to investigate the alterations of membrane protein profiles in zebrafish liver (ZFL) cell line exposed to MP for 24 h using proteomic approaches. Two-dimensional gel electrophoresis revealed a total of 13 protein spots, whose expression levels were significantly altered by MP. These differential proteins were subjected to matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry analysis, and nine proteins were identified to be membrane proteins, among which seven were up-regulated, while two were down-regulated. In addition, the mRNA levels corresponding to these differential membrane proteins were further analyzed by quantitative real-time PCR. And the differential expression of arginase-2 was specially validated via Western blotting. Regarding the physiological functions, these proteins are involved in molecular chaperon, cytoskeleton system, cell metabolism, signal transduction, transport and hormone receptor respectively, suggesting the complexity of MP-mediated toxicity to ZFL cell. These data could provide useful insights for better understanding the hepatotoxic mechanisms of MP and develop novel protein biomarkers for effectively monitoring MP contamination level in aquatic environment.     

Comparison of homologous and heterologous formats in nanocolloidal gold-based immunoassays for parathion residue determination

The effectiveness of homologous and heterologous formats in a nanocolloidal gold-based immunoassay for pesticide residue determination was investigated. Parathion, one of the most toxic organophosphorus pesticides, was used as the target analyte. One-step homologous and heterologous test strips based on a nanocolloidal gold-labeled monoclonal antibody were developed for the rapid detection of parathion residues. The results showed that the heterologous format was more effective than the homologous format, being more sensitive, more specific to parathion and more tolerant of matrix interferences. The best competitive hapten was found to have a moderate heterology and the opposite electronic distribution to the immunizing hapten. The detection limits for parathion using the preferred heterologous strip were 1 μg/L in water samples and 5 μg/kg in soil and food samples.     

海水中对硫磷的毛细管气相色谱分析

介绍了用配备火焰热离子检测器(FTD)的毛细管气相色谱法检测海水中对硫磷的分析方法.通过对本方法各实验参数验证及与其他方法的简单比较,表明本方法可以适用于海水中对硫磷的检测.本方法检出限为1.95 μg/L.同时,于2005年7月28日至7月31日在大亚湾生态监控区采集海水样品进行检测,检测结果全部为未检出.     

有机磷农药对大蒜素含量的影响及其原因

在不同浓度乐斯本、对硫磷灌根胁迫条件下,测定大蒜生长过程中大蒜素含量及农药的残留量.结果表明,低浓度胁迫条件下,大蒜素含量的变化趋势为先上升后下降再上升;高浓度时,表现为先下降后上升;大蒜素含量与两种农药使用浓度有明显的负剂量效应关系,相关系数均大于0.85,乐斯本、对硫磷残留量分别在用药后5 d、7 d未检出.通过化学分析得知,有机磷农药与大蒜素或蒜氨酸(大蒜素的前身)间可能发生两种作用:1)有机磷农药水解成碱性,与蒜氨酸或蒜酶发生酸碱中和反应;2)蒜氨酸与有机磷农药分子中都有双键,两者间发生了双键聚合反应.研究结果为大蒜的安全生产及加速有机磷农药降解剂的研究与开发提供了理论依据.     

Photochemical nitro-nitrite rearrangement in methyl parathion decay under tropical conditions

This work presents a study of the abiotic degradation of commercially available methyl parathion in aqueous solution at two different concentrations (88 mg/L and 200 μg/L). The effects of solar irradiation and the presence of humic acids were evaluated and revealed a synergistic response between them. The half-life of methyl parathion ranged from 4.9 to 37 days, and the experimental data also show that photochemical processes were the most relevant in this case. The only byproduct found in samples submitted to shadowed conditions was 4-nitrophenol. On the other hand, 4-nitrophenol, methyl paraoxon and a new degradation product (O,O-dimethyl O-p-hydroxyphenyl phosphorothioate) were detected when the samples were exposed directly to sunlight. This newly identified compound was prepared in the laboratory by thiophosphorylation of hydroquinone, and coelution experiments with authentic samples provided unambiguous confirmation of the presence of O,O-dimethyl O-p-hydroxy phenylphosphorothioate in samples.     

IMPACT OF FOUR PESTICIDES ON THE GROWTH AND METABOLIC ACTIVITIES OF TWO PHOTOSYNTHETIC ALGAE

The acute toxicity was determined for soil algae Chlorella kesslerei and Anabaena inaequalis, exposed to pesticides lindane, pentachlorophenol (PCP), isoproturon (IPU), and methyl parathion (MP). Toxicity markers included growth inhibition, chlorophyll biosynthesis, and total carbohydrate content, as a function of dose and time. Concentration response functions (EC₅₀) were estimated by probit data transformation and weighted linear regression analyses. Lindane's toxicity to Chlorella increased sharply with time (EC₅₀=7490, 10.3, 0.09 mg L^?1; 24, 48, 72 h), but remained nearly constant through 72 h with Anabaena (8.7?6.7 mg L^?1; 24–72 h). PCP at low concentrations stimulated algal growth and chlorophyll a production, an effect reversed at higher doses. Anabaena was less tolerant of PCP and MP than was Chlorella. The 96-h static EC₅₀ values for Chlorella were: 0.003, 34, 0.05, and 291 mg L^?1 for lindane, PCP, isoproturon, and MP, respectively; for Anabaena, these were 4.2, 0.13, 0.21, and 19 mg L^?1. Carbohydrate production responses were similar to those of cell density (growth) and chlorophyll biosynthesis, with MP having the lowest adverse impact. The overall relative toxicity among the four tested pesticides was: for Chlorella, lindane>IPU?PCP?MP; and for Anabaena, PCP>IPU>lindane>MP. The results confirm that toxicants such as these pesticides may affect individual (though related) species to significantly different degrees.     

The rate of soil reduction as affected by levels of methyl parathion and 2, 4‐D

Abstract

A simple technique was demonstrated for determining the potential for synthetic organics to stress microbial populations. Oxidized Crowley and Cecil soil materials were amended with varying concentrations of 2,4‐D and methyl parathion, flooded, and then analyzed for changes in pH, redox potential, and levels of soluble plus exchangeable Fe, Mn, and Zn, all of which may be directly or indirectly influenced by the activity of soil microorganisms. At the concentrations tested (up to 75 ppm), there was little effect of 2,4‐D, but methyl parathion apparently did affect microbial activity contributing to changes in the measured soil properties upon flooding. This approach may be a useful technique for screening various compounds for their potential to stress microbial activity that, for many researchers, would be easier than direct observations of microbial parameters such as population numbers and classifications, and enzyme levels.     

Organophosphorus insecticide residues in farm ditches of the Lower Fraser Valley of British Columbia

Abstract

Farm ditches flowing into three important rivers in the Lower Fraser Valley of British Columbia, Canada, were sampled periodically at seven locations from July to December in 1991, to determine the occurrence and levels of seven organophosphorus (OP) insecticides. Based oh sales records for the year, the uses of OP insecticides in this area were as follows: malathion > diazinon > parathion > dimethoate > azinphos‐methyl > fensulfothion, but no sales of chlorfenvinphos. Residues of parathion, diazinon, fensulfothion, dimethoate and chlorfenvinphos were detected at levels ranging from 1 ‐ 7,785 >μg/kg in cropped soils collected from areas adjacent to the sites for sampling ditch water and sediments. Malathion and azinphos‐methyl were not detected in any of the substrates studied, demonstrating their rapid degradation in the environment. Diazinon and dimethoate were consistently found in ditch water at seven locations, with an average concentration of 0.07 μg/L and 0.27 μg/L, respectively. Fensulfothion and parathion, with an average concentration of 0.08 μg/L and 0.17 μg/L, respectively, were sporadically found in ditch water at two locations. In ditch sediments, diazinon was detected at three locations and fensulfothion at two. The average concentrations of these two insecticides were 16 μg/kg and 9 jug/kg, respectively. The potential impact on aquatic organisms of these OP insecticides in ditches is discussed.