In situ chlorpyrifos (CPF) degradation by Acrobeloides maximus: Insights from chromatographic analysis.

The objective of this study was to evaluate the efficiency of nematodes in zooremediation of chlorpyrifos (CPF), an organophosphate pesticide. The nematode population Acrobeloides maximus (A. maximus) was employed for bioremediation, converting CPF into non-toxic residues. Optimal growth conditions for mass production of A. maximus were achieved by maintaining a temperature of 25 °C, pH 8, and supplementing the culture medium with plant nutrients. The nematodes were then immobilized within sodium alginate beads. The efficacy of the degradation process was assessed using various analytical techniques, including UV-Visible spectroscopy, HPTLC, FTIR, and LC-MS, confirming the successful breakdown of CPF. The bioreactor demonstrated a complete degradation efficiency of CPF exceeding 99%. Additionally, LC-MS analysis was conducted to elucidate the degradation pathway based on the formation of intermediates. These results underscore the potential of A. maximus as a sustainable organism for addressing environmental contamination arising from CPF pesticide.

Insight into the environmental fate, hazard, detection, and sustainable degradation technologies of chlorpyrifos-an organophosphorus pesticide.

Pesticides play a critical role in terms of agricultural output nowadays. On top of that, pesticides provide economic support to our farmers. However, the usage of pesticides has created a public health issue and environmental hazard. Chlorpyrifos (CPY), an organophosphate pesticide, is extensively applied as an insecticide, acaricide, and termiticide against pests in various applications. Environmental pollution has occurred because of the widespread usage of CPY, harming several ecosystems, including soil, sediment, water, air, and biogeochemical cycles. While residual levels in soil, water, vegetables, foodstuffs, and human fluids have been discovered, CPY has also been found in the sediment, soil, and water. The irrefutable pieces of evidence indicate that CPY exposure inhibits the choline esterase enzyme, which impairs the ability of the body to use choline. As a result, neurological, immunological, and psychological consequences are seen in people and the natural environment. Several research studies have been conducted worldwide to identify and develop CPY remediation approaches and its derivatives from the environment. Currently, many detoxification methods are available for pesticides, such as CPY. However, recent research has shown that the breakdown of CPY using bacteria is the most proficient, cost-effective, and sustainable. This current article aims to outline relevant research events, summarize the possible breakdown of CPY into various compounds, and discuss analytical summaries of current research findings on bacterial degradation of CPY and the potential degradation mechanism.

Dual-mode supramolecular fluorescent probe for rapid and on-site detection of chlorpyrifos in the environment.

Chlorpyrifos (CPF) as a classic organophosphorus pesticide has been widely used in agricultural applications to control insects and worms. CPF in the environment can cause deaths of diverse kinds of aquatic organism and bring a high risk to human health. Therefore, the development of effective analytical method for CPF is of great importance. In this work, a novel dual-mode albumin (ALB)-based supramolecular probe FD@ALB was designed and prepared for rapid detection of CPF in the environment. The limit of detection is 0.57 µM (∼ 0.2 ppm) with a wider detection range up to 200 µM, which is satisfactory for application. The sensing mechanism can be ascribed to CPF-induced phosphorylation of ALB, thus leading to a change in the binding microenvironment of FD dye. Moreover, the paper-based test strips were used in conjunction with the FD@ALB, realizing the portable detection of CPF. This method was demonstrated to be suitable for on-site detection of CPF in various kinds of environmental samples, including water, soil, and food samples, with the aid of a smartphone. To the best of our knowledge, this is the first analytical method achieving a combination of the rapid and ratiometric detection of CPF in the environment.

Elucidating molecular characterization of chlorpyrifos and profenofos degrading distinct bacterial strains for enhancing seed germination potential of Gossypium arboreum L.

Chlorpyrifos (CP) and profenofos (PF) are organophosphate pesticides (OPs) widely used in agriculture and are noxious to both fauna and flora. The presented work was designed to attenuate the toxicity of both pesticides in the growth parameters of a cotton crop by applying plant growth-promoting rhizobacteria (PGPR), namely Pseudomonas aeruginosa PM36 and Bacillus sp. PM37. The multifarious biological activities of both strains include plant growth-promoting traits, including phosphate solubilization; indole-3-acetic acid (IAA), siderophore, and HCN production; nitrogen fixation; and enzymatic activity such as cellulase, protease, amylase, and catalase. Furthermore, the molecular profiling of multi-stress-responsive genes, including acdS, ituC, czcD, nifH, and sfp, also confirmed the plant growth regulation and abiotic stress tolerance potential of PM36 and PM37. Both strains (PM36 and PM37) revealed 92% and 89% of CP degradation at 50 ppm and 87% and 81% at 150 ppm within 7 days. Simultaneously 94% and 98% PF degradation was observed at 50 ppm and 90% and 92% at 150 ppm within 7 days at 35 °C and pH 7. Biodegradation was analyzed using HPLC and FTIR. The strains exhibited first-order reaction kinetics, indicating their reliance on CP and PF as energy and carbon sources. The presence of opd, mpd, and opdA genes in both strains also supported the CP and PF degradation potential of both strains. Inoculation of strains under normal and OP stress conditions resulted in a significant increase in seed germination, plant biomass, and chlorophyll contents of the cotton seedling. Our findings indicate that the strains PM36 and PM37 have abilities as biodegraders and plant growth promoters, with potential applications in crop sciences and bioremediation studies. These strains could serve as an environmentally friendly, sustainable, and socially acceptable solution to manage OP-contaminated sites.

Chlorpyrifos Oxon Activates Glutamate and Lysine for Protein Cross-linking.

Chronic low-dose exposure to organophosphorus (OP) toxicants is correlated with an increase in the risk of impaired cognition and neurodegenerative diseases. A mechanism to explain this relationship is needed. We suggest that the formation of organophosphate-induced high-molecular-weight protein aggregates that disrupt cell function may be the missing link. It has been demonstrated that such aggregation can be promoted by OP-labeled lysine. Alternatively, OP-labeled glutamate may be the initiator. To test this hypothesis, we treated MAP-rich tubulin Sus scrofa and human transglutaminase with chlorpyrifos oxon. Trypsin-digested proteins were subjected to liquid chromatography-tandem mass spectrometry followed by Protein Prospector searches to identify diethyl phosphate adducts and cross-linked peptides. We report the presence of diethyl phosphate adducts on the side chains of glutamate, lysine, and tyrosine, as well as cross-links between glutamate and lysine. Glutamate-lysine cross-linking could be initiated either by diethyl phosphate-activated glutamate or by diethyl phosphate-activated lysine to form stable isopeptide bonds between and within proteins. It was concluded that organophosphate-induced high-molecular-weight protein aggregates could promote brain dysfunction.

In situ formed silicon-based nanoparticles enabled highly efficient dual-mode biosensing of chlorpyrifos.

Chlorpyrifos is widely used in agriculture but their residue is a threat to food safety and human health. The study aims to find an accurate and sensitive detection method for chlorpyrifos. Silicon-based nanoparticles (Si BNPs) with fluorescent and colorimetric dual signals are in-situ formed through the alkaline phosphatase (ALP) triggered reaction. The dual-mode sensor for chlorpyrifos can be facilely fabricated because the existence of chlorpyrifos would inhibit the activity of ALP and thus affect the production of Si BNPs. Under optimal condition, the fluorescence intensity and absorption intensity linearly correlate with the logarithm of chlorpyrifos concentration over wide range of 0.5-5000 ng/mL and 1-5000 ng/mL with low detection limit by the two detection modes, respectively. The dual-mode sensor is further applied to the detection of spiked chlorpyrifos in practical samples with satisfactory recovery, rendering it as a promising candidate to current methodologies for the simple and accurate detection of chlorpyrifos.

Organophosphorus Pesticides Promote Protein Cross-Linking.

Exposure to organophosphorus pesticides (OP) can have chronic adverse effects that are independent of inhibition of acetylcholinesterase, the classic target for acute OP toxicity. In pure proteins, the organophosphorus pesticide chlorpyrifos oxon induces a cross-link between lysine and glutamate (or aspartate) with loss of water. Tubulin is particularly sensitive to OP-induced cross-linking. Our goal was to explore OP-induced cross-linking in a complex protein sample, MAP-rich tubulin from Sus scrofa and to test 8 OP for their capacity to promote isopeptide cross-linking. We treated 100 µg of MAP-rich tubulin with 100 µM chlorpyrifos, chlorpyrifos oxon, methamidophos, paraoxon, diazinon, diazoxon, monocrotophos, or dichlorvos. Each sample was separated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and stained with Coomassie blue. Five gel slices (at about 30, 50, 150, and 300 kDa, and the top of the separating gel) were removed from the lanes for each of the eight OP samples and from untreated control lanes. These gel slices were subjected to in-gel trypsin digestion. MSMS fragmentation spectra of the tryptic peptides were examined for isopeptide cross-links. Sixteen spectra yielded convincing evidence for isopeptide cross-linked peptides. Ten were from the chlorpyrifos oxon reaction, 1 from dichlorvos, 1 from paraoxon, 1 from diazinon, and 3 from diazoxon. It was concluded that catalysis of protein cross-linking is a general property of organophosphorus pesticides and pesticide metabolites. Data are available via ProteomeXchange with identifier PXD034529.

Persistence behavior of chlorpyrifos and biological toxicity mechanism to cucumbers under greenhouse conditions.

Chlorpyrifos, a broadly utilized insecticide, inhibits many cellular and physiological processes in plants. Here, the phyto-toxicity of chlorpyrifos on cucumber plants, as well as the dissipation kinetics of chlorpyrifos in leaves, were investigated. Those results showed that chlorpyrifos accumulated primarily in the leaves under normal agrochemical spraying conditions with the half-lives among 2.48-4.59 days. Residues of the primary metabolite, 3,5,6-trichloro-2-pyridinol (TCP), rapidly accumulated in plant tissues and soil with chlorpyrifos degradation. The application amount of chlorpyrifos had a significant effect on the persistence of chlorpyrifos and TCP in both plant and soil environments. Chlorpyrifos generated excessive reactive oxygen species (ROS) and malondialdehyde (MDA), which led to oxidative damage. High chlorpyrifos stress even inhibited antioxidant enzymes. The photosynthetic system and gas exchange were suppressed, which ultimately lead to inefficient light use under chlorpyrifos stress. Morphological results revealed that chlorpyrifos induced membrane damage and harmed organelles such as mitochondria and chloroplast. Noninvasive micro-test technology (NMT) showed that chlorpyrifos promoted intracellular Ca2+ influx and efflux of H+ and K+. The Ca2+ influx was significantly stimulated after both high and low chlorpyrifos treatment with the minimum value of - 336.33 pmol·cm-2·s-1 at 258 s and - 155.68 pmol·cm-2·s-1 at 288 s, respectively. Chlorpyrifos stress reversed the H+ influx to an efflux in cucumber mesophyll with the mean value of 0.45 ± 0.03 pmol·cm-2·s-1 and 0.19 ± 0.03 pmol·cm-2·s-1 in cucumber plants under low and high chlorpyrifos stress. High chlorpyrifos stress dramatically increase K+ efflux in cucumber leaves by 13.68 times higher than the control. We suggest that ion homeostasis destruction, accompanied by ROS, resulted in oxidative damage to the mesophyll cell of cucumber seedlings.

Impact of Micronutrient Supplementation on Pesticide Residual, Acetylcholinesterase Activity, and Oxidative Stress Among Farm Children Exposed to Pesticides.

The present interventional study aimed to assess the impact of micronutrient supplementation on pesticide-residues concentrations, vitamins, minerals, acetylcholinesterase activity and oxidative stress among 129 farm children (9-12 years, n = 66 and 13-15 years, n = 63) involved in farming activities in Ranga Reddy district, Telangana, India. Our data showed the presence of five organophosphorus pesticide residues (chlorpyrifos, diazinon, malathion, monocrotophos, and phosalone) among children before-supplementation (both age-groups); while post-supplementation, only two pesticide residues (chlorpyrifos and diazinon) were detected indicating improved metabolic rate. Vitamin E, copper, magnesium and zinc levels were also improved in both the age-groups and manganese levels were significantly increased only among children of 13-15 years age group. Further, post-supplementation also showed an improvement in acetylcholinesterase activity and a decrease in lipid peroxidation among both the age groups of children. However, further research for ascertaining the ameliorating effect of micronutrients in preventing adverse effects of organophosphorus pesticides must be conducted.

The growing concern of chlorpyrifos exposures on human and environmental health.

Chlorpyrifos (CP) and its highly electrophilic intermediates are principal toxic metabolites. The active form of CP i.e. chlorpyrifos oxon (CP-oxon) is responsible for both the insecticidal activity and is also of greater risk when present in the atmosphere. Thus, the combined effects of both CP, CP-oxan, and other metabolites enhance our understanding of the safety and risk of the insecticide CP. They cause major toxicities such as AChE inhibition, oxidative stress, and endocrine disruption. Further, it can have adverse hematological, musculoskeletal, renal, ocular, and dermal effects. Excessive use of this compound results in poisoning and potentially kills a non-target species upon exposure including humans. Several examples of reactive metabolites toxicities on plants, aquatic life, and soil are presented herein. The review covers the general overview on reactive metabolites of CP, chemistry and their mechanism through toxic effects on humans as well as on the environment. Considerable progress has been made in the replacement or alternative to CP. The different strategies including antidote mechanisms for the prevention and treatment of CP poisoning are discussed in this review. The approach analyses also the active metabolites for the pesticide activity and thus it becomes more important to know the pesticide and toxicity dose of CP as much as possible.

Structure identification and toxicity evaluation of one newly-discovered dechlorinated photoproducts of chlorpyrifos.

Chlorpyrifos (CPF) is an extensively used organophosphorus pesticide. Recently, it has attracted increasing attention due to environmental health problems caused by it. Although numerous studies have discovered the dechlorinated photoproduct of CPF, its structure and toxicity remain largely unknown. In this study, we systematically investigated the structure and toxicity of dechlorinated photoproduct of CPF. The CPF degradation experiment was performed, and its products were identified by ultra high performance liquid chromatography-orbitrap fusion tribid mass spectrometer (UHPLC-Orbitrap Fusion TMS). Additionally, bond dissociation energy (BDE) calculations and photoproduct chemical synthesis were employed to determine the structure of dechlorinated photoproduct of CPF. The toxicity of CPF photoproduct was evaluated through the Ecological Structure Activity Relationships (ECOSAR) Class Program, the Toxicity Estimation Software Tool (T.E.S.T.) software, and acute toxicity testing. The results indicated that the dechlorinated photoproduct of CPF was identified as O,O-Diethyl-O-(3,5-dichloro-2-pyridyl) phosphorothioate (Dechloro-CPF), which was produced in large quantity within the first 30 min of photodegradation experiment. The acute and chronic toxicity values of Dechloro-CPF were obviously higher than those for the other two photoproducts. The median lethal dose (LD50) of Dechloro-CPF was 31.6 mg/kg for female mice and 58.4 mg/kg for male mice. This study reveals the photodegradation mechanism of CPF and confirms that Dechloro-CPF was dechlorinated photoproduct of CPF with potential acute toxicity to aquatic species and mammalian (including human). Our findings will contribute to a more comprehensive risk evaluation of CPF in food and the environment.

Degradation mechanism and toxicity assessment of chlorpyrifos in milk by combined ultrasound and ultraviolet treatment.

The aim of this study was to compare the degradation kinetics of chlorpyrifos by treatment with ultrasound (US), ultraviolet radiation (UV) and a combination of both (US/UV), to evaluate the toxicity of the degradation products and the effect of the treatments on milk quality. US/UV markedly accelerated the degradation of chlorpyrifos. The half-life of chlorpyrifos by US/UV was 6.4 min, which was greatly shortened compared to the treatment with US or UV alone. Five degradation products were identified by GC-MS, and a degradation pathway for chlorpyrifos was proposed, based on density functional theory calculations. According to the luminescent bacteria test and predictions from a structure/activity relationship model, the toxicity of the degradation products was lower than that of chlorpyrifos. In addition, US/UV treatment had little effect on the quality of the treated milk. Therefore, US/UV can be used as a potential non-thermal processing method to degrade pesticide residues in milk.

Characterization of organophosphate pesticide sorption of potato peel biochar as low cost adsorbent for chlorpyrifos removal.

There has been a growing interest in the scientific world in the production of biochar from natural organic wastes as potential sustainable precursors for bioremediation. Potato peel biochar was produced by slow pyrolysis method under oxygen-limited conditions and used as bio adsorbent in bioremediation of commercial pesticide having Chlorpyrifos as an active component. Chlorpyrifos is an organophosphate pesticide, highly neurotoxic, and primarily targets the central nervous system of pests and insects. The excess residues of chlorpyrifos are hazardous to environmental flora and fauna. Chlorpyrifos was treated against biochar at varying physical parameters and further optimized by using response surface methodology through Box-Behnken design (BBD). 72.06% of pesticide removal was observed post 24 h of treatment against a pesticide concentration of 1346.85 µg/ml with a biochar concentration of 1.04 mg/ml under room temperature at pH 5.04. Biochar was characterized by proximate and ultimate analysis, FTIR, and SEM-EDX. Characterization by SEM-EDX showed the surface morphology and minerals on the peel and biochar. Microgram of potato peel shows pores of larger size than biochar having many cavities with different dimensions. In the plant system, growth morphology, nutritional status, polyphenols, total antioxidant content, and free radical scavenging activity were assessed. Enhancement in presence of biochar was recorded in growth morphology and plant biomolecules including photosynthetic pigments. Better translocation of the nutrient is recorded in biochar treated plants, as evidenced by the low amount of carbohydrate and protein in treated leaves. Biocompatibility assessment of chlorpyriphos in fish erythrocytes showed 43.26% hemolysis by pesticide-treated biochar. The practical use of this approach can also be best utilized if applied to those geographical regions where the soil pH is acidic. Biochar is a marketable bio-product, which can have a positive impact in agriculture, industries, and the energy sector creating a bio-based economy with reduced environmental pollution.

Disruption of androgen receptor signaling by chlorpyrifos (CPF) and its environmental degradation products: a structural insight.

Androgen-disruptors are chemicals that interfere with the biosynthesis, metabolism or function of endogenous androgens affecting normal male reproductive development and health. Several epidemiological studies have indicated a link between exposure to androgen disrupting chemicals with reduced sperm counts and increased infertility. The actions of androgens within target cells are transduced by the androgen receptors (ARs). Chlorpyrifos (CPF), a chlorinated organophosphorus pesticide, is known to cause impairment in both male and female reproductive systems. Recent publications have shown molecular interactions of CPF and its environmental degradation products with human progesterone receptor and human estrogen receptor. Exposure to CPF causes a marked reduction in sperm counts with lowering in serum testosterone level, which suggests possible molecular interaction of CPF with AR. The investigation to reveal the possibility and the extent of binding of CPF and some of its degradation products (chlorpyrifos-oxon [CPYO], desethyl chlorpyrifos [DEC], trichloromethoxypyridine [TMP] and trichloropyridinol [TCP]) with AR using molecular docking simulation are reported. The findings of the present docking, binding energy and molecular dynamics studies reveal that CPF and its degradation products may bind to ARs and act as a potent androgen disruptor.Communicated by Ramaswamy H. Sarma.

Effect of N-acetylcysteine on attenuation of chlropyrifos and its methyl analogue toxicity in male rats.

The attenuating effect of 150 mg/kg of N-acetylcysteine (NAC) against the oral administration of 7.88 and 202.07 mg/kg/day for 14 days of either chlropyrifos-ethyl (CPE-E) or chlropyrifos-methyl (CPF-M), respectively, in male rat was investigated using biochemical and genetic markers. Biomarkers such as acetylcholinesterase (AChE), butyrylcholinesterase (BuChE), paraoxonase (PON), adenosine 5'-triphosphatase (ATP-ase), glutathione-S-transferase (GST), catalase (CAT), glutathione reduced (GSH) in serum showed a significant decline in their levels, while calcium (Ca+2), cytochrome C reduction (CYC-R), lipid peroxidation (LPO), nitric oxide (NO) levels showed a significant increase in serum of treated rats. Regarding the genotoxic parameters, when rats are treated either with CPE-E or CPF-M, liver DNA, chromosomal aberration (CA), and micronucleated polychromatic erythrocytes (MnPCE) significantly increased, while the mitotic index (MI) and polychromatic erythrocytes (PCE)/ normochromatic erythrocytes (NCE) ratio were significantly decreased. However, the administration of NAC following the intoxication of CPF-E or CPF-M attenuated the tested biochemical and genotoxic markers. It can be concluded that NAC can be used to ameliorate the toxicity of certain organophosphorus compounds such as CPF-E and CPF-M.

Efficient and selective extraction of azamethiphos and chlorpyrifos residues from mineral water and grape samples using magnetic mesoporous molecularly imprinted polymer.

A magnetic mesoporous molecularly imprinted polymers was synthesized on the surface of magnetic nanoparticles silanized with 3-(trimethoxysilyl) propyl methacrylate to introduce reactive methacrylate groups. Subsequently, methacrylic acid monomers were grafted onto the surface of this adsorbent functionalized via polymerization by precipitation. Magnetic mesoporous molecularly imprinted polymer was properly characterized by different techniques and applied as adsorbent in magnetic solid phase extraction for selective determination of two organophosphorus pesticides, azamethiphos and chlorpyrifos, in mineral water and grape samples. After sample preparation optimization, recoveries of 99.56% and 98.86% were obtained for azamethiphos and chlorpyrifos, respectively. The magnetic solid phase extraction coupled to HPLC-UV presented limit of quantification of 5 ng mL-1, linearity ranged of 5 to 1000 ng mL-1, in addition to adequate accuracy, precision and robustness. The pesticides showed stability in the matrix and were satisfactorily quantified in real mineral water and grape samples.

Effects of Chlorpyrifos on Serine Hydrolase Activities, Lipid Mediators, and Immune Responses in Lungs of Neonatal and Adult Mice.

Chlorpyrifos (CPF) is an organophosphate (OP) pesticide that causes acute toxicity by inhibiting acetylcholinesterase (AChE) in the nervous system. However, endocannabinoid (eCB) metabolizing enzymes in brain of neonatal rats are more sensitive than AChE to inhibition by CPF, leading to increased levels of eCBs. Because eCBs are immunomodulatory molecules, we investigated the association between eCB metabolism, lipid mediators, and immune function in adult and neonatal mice exposed to CPF. We focused on lung effects because epidemiologic studies have linked pesticide exposures to respiratory diseases. CPF was hypothesized to disrupt lung eCB metabolism and alter lung immune responses to lipopolysaccharide (LPS), and these effects would be more pronounced in neonatal mice due to an immature immune system. We first assessed the biochemical effects of CPF in adult mice (≥8 weeks old) and neonatal mice after administering CPF (2.5 mg/kg, oral) or vehicle for 7 days. Tissues were harvested 4 h after the last CPF treatment and lung microsomes from both age groups demonstrated CPF-dependent inhibition of carboxylesterases (Ces), a family of xenobiotic and lipid metabolizing enzymes, whereas AChE activity was inhibited in adult lungs only. Activity-based protein profiling (ABPP)-mass spectrometry of lung microsomes identified 31 and 32 individual serine hydrolases in neonatal lung and adult lung, respectively. Of these, Ces1c/Ces1d/Ces1b isoforms were partially inactivated by CPF in neonatal lung, whereas Ces1c/Ces1b and Ces1c/BChE were partially inactivated in adult female and male lungs, respectively, suggesting age- and sex-related differences in their sensitivity to CPF. Monoacylglycerol lipase (MAGL) and fatty acid amide hydrolase (FAAH) activities in lung were unaffected by CPF. When LPS (1.25 mg/kg, i.p.) was administered following the 7-day CPF dosing period, little to no differences in lung immune responses (cytokines and immunophenotyping) were noted between the CPF and vehicle groups. However, a CPF-dependent increase in the amounts of dendritic cells and certain lipid mediators in female lung following LPS challenge was observed. Experiments in neonatal and adult Ces1d-/- mice yielded similar results as wild type mice (WT) following CPF treatment, except that CPF augmented LPS-induced Tnfa mRNA in adult Ces1d-/- mouse lungs. This effect was associated with decreased expression of Ces1c mRNA in Ces1d-/- mice versus WT mice in the setting of LPS exposure. We conclude that CPF exposure inactivates several Ces isoforms in mouse lung and, during an inflammatory response, increases certain lipid mediators in a female-dependent manner. However, it did not cause widespread altered lung immune effects in response to an LPS challenge.

Computational and Experimental Approaches to Decipher the Binding Mechanism of General Odorant-Binding Protein 2 from Athetis lepigone to Chlorpyrifos and Phoxim.

Insect resistance to insecticides is an increasingly serious problem, and the resistant mechanisms are complicated. The resistance research based on the chemosensory pathway is one of the hot problems at present, but the specific binding mechanism of chemosensory genes and insecticides remains elusive. The binding mechanism of AlepGOBP2 (belong to insect chemosensory gene) with two insecticides was investigated by computational and experimental approaches. Our calculation results indicated that four key residues (Phe12, Ile52, Ile94, and Phe118) could steadily interact with these two insecticides and be assigned as hotspot sites responsible for their binding affinities. The significant alkyl-π and hydrophobic interactions involved by these four hotspot residues were found to be the driving forces for their binding affinities, especially for two residues (Phe12 and Ile94) that significantly contribute to the binding of chlorpyrifos, which were also validated by our binding assay results. Furthermore, we also found that the AlepGOBP2-chlorpyrifos/phoxim complexes can be more efficiently converged in the residue-specific force field-(RSFF2C) and its higher accuracy and repeatability in protein dynamics simulation, per-residue free energy decomposition, and computational alanine scanning calculations have also been achieved in this paper. These findings provided useful insights for efficient and reliable calculation of the binding mechanism of relevant AlepGOBPs with other insecticides, facilitating to develop new and efficient insecticides targeting the key sites of AlepGOBP2.

Interaction of organophosphate pesticide chlorpyrifos with alpha-2-macroglobulin: Biophysical and molecular docking approach.

Organophosphate class of pesticides causes neurotoxicity and carcinogenicity in humans. Once inside the human body, these pesticides often interact with plasma proteins, such as alpha-2-macroglobulin (α2M) which is the key anti-proteinase. Our work focuses on the structural and functional alteration of α2M by chlorpyrifos (CPF), a member of organophosphates. We explored the binding interaction between alpha-2-macroglobulin and CPF by using UV absorption and fluorescence spectroscopy (steady state and synchronous), circular dichroism and molecular docking approach. The functional activity of α2M was analyzed by anti-proteinase trypsin inhibitory assay which showed dose-dependent decrease in alpha-2-macroglobulin antiproteolytic potential. UV absorption studies and fluorescence quenching experiments suggested the formation of a complex between α2M and CPF. The CD spectra suggested a reduction in the beta helical (ß helix) content of α2M. Analysis of thermodynamic parameters suggested the process is spontaneous and endothermic with the ΔG and ΔH values being -5.501 kJ/mol, 11.49 kJ/mol, respectively. CPF binds with Ile-1390, Pro-1391, Leu-1392, Lys-1393, Val-1396, Lys-1397, Arg-1407, Thr-1408, Glu-1409, Val-1410, Asp-282, Glu-281 of α2M as suggested by molecular docking.

Effect of dielectric barrier discharge plasma on the degradation of malathion and chlorpyrifos on lettuce.

BACKGROUND: Pesticides have been widely used to control pests on agricultural products in China, and large amounts of pesticide residues have caused a serious threat to human health. Thus, developing a high-efficiency pesticide degradation method for fresh vegetables represents a great challenge. The present study investigated the effects of dielectric barrier discharge (DBD) plasma on the degradation of malathion and chlorpyrifos in aqueous solutions and on lettuces. RESULTS: DBD treatment significantly degraded malathion and chlorpyrifos in water and on lettuce. After cold plasma treatment at 80 kV for 180 s, the degradation efficiency of malathion (0.5 µg mL-1 ) and chlorpyrifos (1.0 µg mL-1 ) in aqueous solutions reached 64.6% and 62.7%, respectively. The degradation intermediates were explored by HPLC-mass spectrometry and the DBD plasma degradation pathways of malathion and chlorpyrifos were proposed. There was no significant damage to the quality of lettuces, including color and chlorophyll content, after plasma treatment. Ascorbic acid decreased significantly during long-term treatment with DBD plasma. To ensure the quality of lettuces during processing, the treatment time was shortened to 120 s. Under this condition, the degradation efficiency of malathion (0.5 mg kg-1 ) and chlorpyrifos (1.0 mg kg-1 ) on lettuces was found to be 53.1% and 51.4%. More importantly, we noted that cold plasma treatment significantly inactivated the microorganisms on lettuces. CONCLUSION: The results of the present study show that cold plasma is an effective and safe method for the degradation of organic pesticide residues on fresh vegetables at the same time as retaining the original quality. © 2020 Society of Chemical Industry.