Biodegradation of chlorpyrifos pollution from contaminated environment - A review on operating variables and mechanism.

Chlorpyrifos (CPF) is a highly toxic phosphate-rich organic pesticide (OP), identified as an emerging contaminant and used extensively in agricultural production. CPF persistence in the environment and its potential health hazards has become increasingly concerning worldwide in recent years due to exponential rise in food demand. Biodegradation of chlorpyrifos by microbial cultures is a promising approach to reclaiming contaminated soil and aquatic environments. The purpose of this review is to summarize the current understanding of microbiological aspects of xenobiotic chlorpyrifos biodegradation, including microbial diversity, metabolic pathways, and factors that modulate it. In both aerobic and anaerobic environments, CPF is biochemically broken down by a broad spectrum of bacteria and fungi. Hydrolysis, dehalogenation, and oxidation of chlorpyrifos are all enzymatic reactions that lead to its degradation. Biodegradation rate and efficiency are strongly influenced by parametric variables such as co-substrates abundance, pH, temperature, and initial chlorpyrifos concentration. The review provides evidence that microbial biodegradation is a viable method for remediating chlorpyrifos-contaminated sites in a sustainable and safe manner.

Toxic effects of organophosphate pesticide monocrotophos in aquatic organisms: A review of challenges, regulations and future perspectives.

In recent times, usage of pesticide, herbicides and synthetic fertilizers in farming lands has made the environment worse. The pesticide residues and toxic byproducts from agricultural lands were found to contaminate the aquatic ecosystem. The misuse of synthetic pesticide not only affects the environment, but also affects the health status of aquatic organisms. The organophosphate pesticide pollutants are emerging contaminants, which threatens the terrestrial and aquatic ecosystem. Monocrotophos (MCP) is an organophosphate insecticide, utilized on crops including rice, maize, sugarcane, cotton, soybeans, groundnuts and vegetables. MCP is hydrophilic in nature and their solubilizing properties reduce the soil sorption which leads to groundwater contamination. The half-life period of MCP is 17-96 and the half-life period of technical grade MCP is 2500 days if held stable at 38 °C in a container. MCP causes mild to severe confusion, anxiety, hyper-salivation, convulsion and respiratory distress in mammals as well as aquatic animals. The MCP induced toxicity including survival rate, behavioural changes, reproductive toxicity and genotoxicity in different aquatic species have been discussed in this review. Furthermore, the ultimate aim of this review is to highlight the international regulations, future perspectives and challenges involved in using the MCP.

Assessment of the Potential Phytotoxicity of Chlorpyrifos in the Wetland Macrophyte Bidens laevis (L.).

Chlorpyrifos (CPF) has been used worldwide, but its possible negative effects on macrophytes have been scarcely studied. The main goal of the present work was to assess the potential phytotoxic effects of CPF on different stages (seed and seedling) of the wetland macrophyte Bidens laevis. During the germination of seeds, stimulation of radicle growth at low concentrations of CPF (10 µg/L) and inhibition of its elongation at 80 µg/L CPF were observed. In seedlings, concentrations ≤ 160 µg/L CPF did not exhibit adverse effects on growth after 7 days of exposure, despite the decrease of photosynthetic pigments and carotenoids observed at 40 µg/L CPF compared to the control. Environmentally relevant concentrations of CPF altered neither oxidative stress biomarkers nor pigment contents in seedlings exposed for 48 h, suggesting CPF would be non-toxic to B. laevis in natural scenarios.

Acetylcholinesterase Inhibition in Rats and Humans Following Acute Fenitrothion Exposure Predicted by Physiologically Based Kinetic Modeling-Facilitated Quantitative In Vitro to In Vivo Extrapolation.

Worldwide use of organophosphate pesticides as agricultural chemicals aims to maintain a stable food supply, while their toxicity remains a major public health concern. A common mechanism of acute neurotoxicity following organophosphate pesticide exposure is the inhibition of acetylcholinesterase (AChE). To support Next Generation Risk Assessment for public health upon acute neurotoxicity induced by organophosphate pesticides, physiologically based kinetic (PBK) modeling-facilitated quantitative in vitro to in vivo extrapolation (QIVIVE) approach was employed in this study, with fenitrothion (FNT) as an exemplary organophosphate pesticide. Rat and human PBK models were parametrized with data derived from in silico predictions and in vitro incubations. Then, PBK model-based QIVIVE was performed to convert species-specific concentration-dependent AChE inhibition obtained from in vitro blood assays to corresponding in vivo dose-response curves, from which points of departure (PODs) were derived. The obtained values for rats and humans were comparable with reported no-observed-adverse-effect levels (NOAELs). Humans were found to be more susceptible than rats toward erythrocyte AChE inhibition induced by acute FNT exposure due to interspecies differences in toxicokinetics and toxicodynamics. The described approach adequately predicts toxicokinetics and acute toxicity of FNT, providing a proof-of-principle for applying this approach in a 3R-based chemical risk assessment paradigm.

Residue levels of organophosphate pesticides and dialkylphosphates in agricultural products in Japan.

High urinary levels of dialkylphosphates (DAPs), which are common structures of organophosphate pesticides (OPs), have been associated with several adverse health outcomes in human biomonitoring studies. Previous studies have indicated that dietary OP exposure and ingestion of environmentally degraded DAP, which is inactive with acetylcholinesterase, can lead to an increase in urinary DAP levels in the general population. However, the specific food sources contributing to the intake of OPs and DAPs have not been identified. In this study, we analyzed the levels of OPs and preformed DAPs in various food items. DAP levels were markedly high in certain fruits, such as persimmon, apple juice, kiwi, and mandarin. In contrast, only moderate levels of OPs were detected in these foods. Furthermore, the levels of OPs and DAPs were positively associated with vegetables, whereas no such association was observed in fruits. Increased consumption of certain fruits presumably leads to a marked increase in urinary DAP levels in individuals despite limited exposure to OPs, resulting in reduced reliability of urinary DAPs as a marker of OP exposure. Therefore, the possible effects of dietary habits and the resulting intake of preformed DAPs should be considered when interpreting biomonitoring data of urinary DAPs. Additionally, DAP levels in most organic foods were much lower than those in conventional foods, suggesting that the reduction in urinary DAPs by organic diet intervention may be mainly attributed to the reduced intake of preformed DAPs rather than reduced exposure to OPs. Therefore, urinary DAP levels may not be suitable indicators for evaluating ingested OP exposure.

Organophosphate pesticide exposure and biomarkers of liver injury/liver function.

BACKGROUND AND AIMS: There is little epidemiological evidence linking the exposure of organophosphate pesticides (OPs) to liver function or liver injury in the general population. We used data from the National Health and Nutrition Examination Survey 1999-2012 to investigate the relationship of urinary OPs with biomarkers of liver function/liver injury. METHODS: The exposures were the concentrations of urinary OP metabolites (dimethyl phosphate [DMP], dimethyl thiophosphate [DMTP], diethyl phosphate [DEP] and diethyl thiophosphate [DETP]). The health outcomes were biomarkers of liver function/liver injury. The multivariable linear regression model, restricted cubic splines (RCSs) analysis and weighted quantile sum (WQS) regression were used to evaluate the relationship between individual or overall exposure of OPs and outcomes. RESULTS: Regressions of RCSs suggested linear and positive associations of OP metabolites with aspartate aminotransferase/alanine aminotransferase (AST/ALT) ratio (DMP and DEP) and fibrosis-4 (FIB-4) index (DMP, DEP and DMTP) (all p-non-linear values >.05). However, L-shaped relationships were found between OP metabolites (DMTP and DETP) and blood albumin and total protein (TP) concentrations (both p and non-linear values <.05). The positive associations of urinary DMP, DEP and DMTP with AST/ALT ratio, and with FIB-4 score were more pronounced among non-smokers than smokers, among alcohol drinkers than non-drinkers and among those with a body mass index (BMI) of ≥25 than participants with a BMI of <25. However, most of the interaction p values were more than .05, indicating no significant interactions between covariates and OPs on outcomes mainly including AST/ALT, FIB-4, ALB and TP levels. Finally, the WQS indices were positively associated with AST/ALT ratio (p = .014) and FIB-4 score (p = .002). CONCLUSIONS: Our study added novel evidence that exposures to OPs might be adversely associated with the biomarkers of liver function/liver injury. These findings indicated the potential toxic effect of OP exposures on the human liver.

Urinary organophosphate metabolite concentrations and birth sizes among women conceiving through in vitro fertilization in Shanghai, China.

BACKGROUND: Few studies have examined the adverse birth sizes of preconception exposure to organophosphate pesticides (OPs) in women undergoing in vitro fertilization (IVF). OBJECTIVES: We investigated the relationship of preconception OP exposure with birth sizes among Chinese women undergoing IVF. METHODS: This study included 302 couples seeking infertility treatment in the China National Birth Cohort Study, from Shanghai, China, who gave birth to singleton infants between 2018 and 2021. Clinical data were collected from medical records. We measured the concentrations of six nonspecific dialkyl phosphates (DAP) metabolites of OPs [diethylthiophosphate (DETP), diethylphosphate (DEP), diethyldithiophosphate (DEDTP), dimethyldithiophosphate (DMTP), dimethylphosphate (DMP), and dimethyldithiophosphate (DMDTP)] in maternal urine. DMDTP and DEDTP were precluded from further analyses due to the low detection rates. Generalized linear models (GLMs) and weighted quantile sum (WQS) regression analyses were performed to examine the individual and joint effects of OP exposures on gestational age, birth weight, body length, and ponderal index. Odder ratio (OR) of preterm birth were estimated using logistic regression models. RESULTS: Women in the highest as compared with lowest quartile of DEP had shorter gestational age (ß = - 0.68; 95% CI = -1.24, -0.11). The association was modified by sex, with boys showing larger decreases in gestational age (ß = - 0.86; 95% CI = -1.60, -0.13). No associations were found between other DAP metabolites and birth sizes. Results from linear models with individual DAP metabolites were corroborated by the WQS regression where DEP had the largest contribution to the overall mixture effect on gestational age (weight = 0.70). Moreover, DEP concentration was associated with an elevated risk of preterm birth (OR = 1.35, 95% CI = 1.11, 2.25). CONCLUSION: Preconception DEP concentration was associated with shortened gestational age and increased risk of preterm birth, and the association was more pronounced among boys than girls.

Alterations in the non-neuronal cholinergic system induced by in-vitro exposure to diazoxon in spleen mononuclear cells of Nile tilapia (O. niloticus).

Organophosphate pesticides as diazinon disrupt the neuroimmune communication, affecting the innate and adaptive immune response of the exposed organisms. Since the target molecule of diazinon is typically the acetylcholinesterase enzyme (AChE), the existence of a non-neuronal cholinergic system in leukocytes makes them susceptible to alterations by diazinon. Therefore, the aim of this work was to evaluate the activity of AChE, acetylcholine (ACh) concentration, and the expression of nicotinic ACh receptors (nAChR) and muscarinic ACh receptors (mAChR) in spleen mononuclear cells (SMNC) of Nile tilapia (O. niloticus) exposed in vitro to diazoxon, a diazinon metabolite. SMNC were exposed in-vitro to 1 nM, 1 µM, and 10 µM diazoxon for 24 h. The enzyme activity of AChE was then evaluated by spectrophotometry, followed by ACh quantification by ultra-performance liquid chromatography. Finally, mAChR and nAChR expression was evaluated by RT-qPCR. The results indicate that AChE levels are significantly inhibited at 1 and 10 µM diazoxon, while the relative expression of (M3, M4, and M5) mAChR and (ß2) nAChR is reduced significantly as compared against SMNC not exposed to diazoxon. However, ACh levels show no significant difference with respect to the control group. The data indicate that diazoxon directly alters elements in the cholinergic system of SMNC by AChE inhibition or indirectly through the interaction with AChR, which is likely related to the immunotoxic properties of diazinon and its metabolites.

Organophosphate-pesticides induced survival mechanisms and APE1-mediated Nrf2 regulation in non-small-cell lung cancer cells.

Epidemiological and molecular studies have indicated that environmental exposure to organophosphate pesticides (OPPs) is associated with increased cancer risk; however, the underlying molecular mechanisms still need to be explained. Increasing cancer incidence is linked to OPPs-induced oxidative stress (OS). Our study evaluates monocrotophos (MCP) and chlorpyrifos (CP)-induced OS responses and apurinic/apyrimidinic endonuclease 1 (APE1) role in human non-small-cell lung cancer (NSCLC) cells. Our prior study has implicated OPPs-induced base excision repair (BER)-pathway dysregulation and APE1-mediated regulation of transcription factor (TF) c-jun in A549 cells. We further investigated the effects of MCP and CP on apoptosis, proliferation, and APE1's redox-regulation of nuclear factor-like 2 (Nrf2). Data demonstrates that MCP and CP at subtoxic concentrations induced reactive oxygen species generation and oxidative DNA base damage 8-oxo-dG lesions in NCI-H1299 cells. CP moderately upregulated the apoptosis-inducing factor (AIF) in A549 cells, however, it did not trigger other pro-apoptotic factors viz. caspase-9 and caspase-3, suggesting early caspase-independent apoptosis. However, dose-dependent AIF-downregulation was observed for MCP treatment. Furthermore, CP and MCP treatments upregulated proliferating cell nuclear antigen levels. Immunofluorescent confocal imaging showed the colocalization of APE1 with Nrf2 in 10 µM CP- and MCP-treated NCI-H1299 cells. Immunoprecipitation confirmed that APE1 and Nrf2 physically interacted, indicating the role of APE1-mediated Nrf2 activation following OPPs treatment. This study suggests that low concentration MCP and CP exposure generates OS along with DNA damage, and modulates apoptosis, and APE1-mediated Nrf2 activation, which might be considered as the possible mechanism promoting lung cancer cell survival, suggesting that APE1 may have the potential to become a therapeutic target for the treatment of NSCLC.

A treatise on Organophosphate pesticide pollution: Current strategies and advancements in their environmental degradation and elimination.

Pesticides have been used in the field of agriculture ever since their role in protection of crops from pests which include four different categories namely insects, mites, rodents and animals has been identified. Organophosphate pesticides are one of the most extensively applied insecticides in the field of agriculture such that around 40% of all the pesticides that are produced and used commercially belong to this category. The main toxicological effect of these pesticides when exposed to a living being encompasses the irremediable inhibition of the acetylcholinesterase (AChE) enzyme which is involved in the neurotransmission of signals and hence its inhibition causes impairment of the respiratory tract and neuromuscular transmission. Apart from being used as a pesticide, organophosphates have also been applied as herbicides to some extent. The residues of these highly toxic chemicals have found route into the underground water system by seeping into the ground, in rivers where the agricultural run off water is disposed, and in the air when sprayed on the crops hence posing a threat to all the living strata exposed to these chemicals in various ways which are discussed further. Many significant studies have been carried out in order to evaluate the health risks associated with these pesticides which commonly include acute neurological disorders. This review emphasizes on the toxicological effects of organophosphate pesticides and the recent methods of detection that are used to identify trace amounts of organophosphate pesticides along with strategies which are used for their degradation.

Exposure to glyphosate and tetrachlorvinphos induces cytotoxicity and global DNA methylation in human cells.

Two organophosphate pesticides-glyphosate and tetrachlorvinphos-have been announced as carcinogens to humans by various authorities, including the European Chemical Agency and the Environmental Protection Agency. We aimed to investigate molecular mechanisms associated with carcinogenicity and to examine changes in global m5C DNA methylation and cytotoxic potential in A549 lung epithelial cells in response to glyphosate and tetrachlorvinphos, and differential gene expression of m5C DNA methyltransferase genes in Sprague Dawley rats to Roundup (commercial formulation of glyphosate). Global m5C level significantly increased after 1500 µM glyphosate exposure for 24 h. We determined that exposure to tetrachlorvinphos did not significantly increase the m5C level in A549 cells for 24 h. Additionally, we did not observe significant DNA methylation alteration for both pesticides after 12 h exposure. In the animal study, we observed that DNA methyltransferase genes (DNMT3b and DNMT3a) showed significantly higher expression in Roundup-exposed rats than the control group in the liver and kidney. We also observed that a significant cytotoxic effect was determined after the treatment of the cells with higher concentrations of glyphosate and tetrachlorvinphos. Our results revealed that DNA methylation could be modified by exposure to glyphosate and that exposure to Roundup was associated with the differential expression level of m5C DNA methylation methyltransferase. Finally, exposure to both pesticides increased cytotoxicity.

Biodegradation of CP/TCP by a constructed microbial consortium after comparative bacterial community analysis of long-term CP domesticated activated sludge.

The objective of this study was to construct a microbial consortium for effective biodegradation of chlorpyrifos (CP) and its hydrolysis product, 3,5,6-Trichloro-2-pyridinol (TCP). A activated sludge from an organophosphorus pesticide factory was domesticated under long-term (20 weeks) CP stress, and the dynamic change in bacterial communities was analyzed by high-throughput sequencing. Then, a microbial consortium MC-BSPK was constructed of Bacillus sp. MC-B, Serratia sp. MC-S, Pseudomonas sp. MC-P, and Klebsiella sp. MC-K, which were significantly enriched during the domestication process. The biodegradation capacities of the microbial consortium MC-BSPK reached 100% for CP within 9 days and 88.61% for TCP within 15 days under the optimized degradation conditions (pH 8.0 and 31 °C). High-performance liquid chromatography (HPLC) revealed that CP could be degraded by the microbial consortium MC-BSPK into TCP, probably through hydrolysis of the P-O ester bond, and further degraded into other small molecules. A bioassay revealed that the virulence of CP toward Drosophila melanogaster W1118 was clearly reduced by the microbial consortium MC-BSPK biodegradation. Thus, the easily constructed microbial consortium MC-BSPK with high CP/TCP degradation capacities has the potential for application in pesticide-contaminated bioremediation.

Ecotoxicity of malathion pesticide and its genotoxic effects over the biomarker comet assay in Daphnia magna.

Malathion is a highly toxic organophosphate insecticide, being one of the most widely used in the world and is generally used for insect control in food production. Thus, ecotoxicological studies have been used to verify its toxic effects on aquatic organisms, such as Daphnia magna and biomarkers, as the comet assay. The comet assay is a microgel electrophoresis method for the detection and quantification of DNA strand breaks in individual cells. Cells were obtained from Daphnia magna after disaggregation of newborn organisms, exposed at concentrations of 0.23 µg L-1 and 0.47 µg L-1 for 48 h. Malathion has shown to cause damage to DNA of the exposed organisms. It was also observed the need of further studies to standardize the comet assay technique for Daphnia magna, once methodologies used present several differences.

Three-dimensional zinc oxide nanostars anchored on graphene oxide for voltammetric determination of methyl parathion.

The two-step microwave method was used to synthesize zinc oxide nanostars linked to graphene oxide (GO) nanosheets. The material was used to modify a screen printed carbon electrode (SPCE) and then explored as a binder-free electrocatalyst for the electrochemical determination of methyl parathion (MP). The morphology and crystallinity of the material were characterized by various techniques. The modified SPCE shows extraordinary electrochemical performances for sensitive determination of MP. Figures of merit include (a) a wide linear dynamic range (0.03-670 µM), (b) a low detection limit (1.2 nM; at S/N = 3), (c) a comparably low working voltage (-0.69 V vs. Ag/AgCl); and (d) an excellent sensitivity (16.5 µA µM-1 cm-2) that surpasses other modified electrodes. The sensor was successfully applied to the determination of MP, even in the presence of other common electroactive interference, in (spiked) fruits and vegetables. Graphical abstractGraphene oxide anchored three-dimensional zinc oxide nanostars were used to coat electrode for the sensing of methyl parathion (MP) by voltammetry.

Reduced graphene oxide nanosheets modified with nickel disulfide and curcumin nanoparticles for non-enzymatic electrochemical sensing of methyl parathion and 4-nitrophenol.

A method was designed for simultaneous voltammetric determination of methyl parathion pesticide (MP) and 4-nitrophenol (4-NP). Curcumin nanoparticles were deposited on reduced graphene oxide nanosheets that were modified with nickel disulfide. The material was placed on a screen-printed carbon electrode and then displayed high electrocatalytic activities toward MP and 4-NP, with a peak potential near -0.9 and - 0.7 V (vs. pseudo Ag/AgCl), respectively. Figures of merit include (a) good electrochemical sensitivities (7.165 and 6.252 µA·µM-1·cm-2), (b) wide linear ranges (from 0.25 to 80 µM), (c) low limits of detection (8.7 and 6.9 nM at S/N = 3) for MP and 4-NP, respectively, and (d) good selectivity, repeatability, reproducibility, and storage stability. The method was applied in the determination of MP and 4-NP in tomato and apple juices and spiked river water. Graphical abstract A novel electrocatalysis platform based on reduced graphene oxide-nickel disulfide nanosheets decorated with curcumin nanoparticles for simultaneous quantification of methyl parathion and 4-nitrophenol in various vegetarian juices and water samples.

Effects of long-term chlorpyrifos exposure on mortality and reproductive tissues of Banded Gourami (Trichogaster fasciata).

This study assessed the long-term toxicity of chlorpyrifos on survival and reproduction of Banded Gourami by using mortality, gonado-somatic index (GSI) and histopathological observations as endpoints. Adult fish were exposed to five different concentrations of chlorpyrifos (0, 15, 50, 150, 500 µg/L) in 15 PVC tanks for 15, 30, 45, 60 and 75 days. Results showed that all male and female fish died after 15 days of 500 µg/L chlorpyrifos exposure. No consistent significant effect was observed for both male and female GSI. Furthermore, results showed dose- and time-dependent histopathological alterations for both ovary and testes. The 60-d No Observed Effect Concentration (NOEC) for most histopathological alterations of Banded Gourami ovary and testes was 50 µg/L, while 60-d NOEC for mortality of both male and female fish was < 15 µg/L. The results show that the long-term exposure to chlorpyrifos not only affect the reproductive tissues of Banded Gourami at exposure concentrations but also cause their mortality. Future studies should evaluate effects at lower concentrations.

In vitro skin decontamination of paraoxon - wet-type cleansing effect of selected detergents.

The aim of this study was to determine optimal conditions for in vitro skin decontamination using water and detergents as decontamination agents and to test the cleansing efficiency of selected detergents. Experiments were performed using a peristaltic pump for showering of pig skin in modified static diffusion cells. Several conditions were tested including different flow rates (from 5 to 33 ml s-1), quantity of rinsing fluid (from 40 to 400 ml) and concentration of detergents (2; 5; 10%). Further, several types of detergents/commercial decontamination agents were evaluated under the selected conditions to find the most effective means of decontamination. The amount of paraoxon removed from the skin surface following wet-type decontamination was detected in the rinsing fluid spectrophotometrically after hydrolysis of paraoxon - a model contaminant. The efficacy of rinsing by water/Spolapon AES 253 increased with flow rate up to 25 ml s-1 and a rinsing volume of 200 ml. Lutensol AT 25 achieved maximum efficacy at the lowest tested concentration (2%). A flow rate of 16 ml s-1, rinsing volume of 100 ml (values from the middle part of the sigmoid curve) and 5% concentration of decontaminant solution were used for further evaluation of detergents as cleansing agents under the selected conditions. Cetylpyridinium bromide (cationic surfactant), carbethopendecinii bromidum (cationic surfactant) and polyoxyethylene-10-tridecyl ether (non-ionic surfactant), SDS (anionic surfactant), althosan MB (cationic surfactant), sodium dodecylbenzene sulphonate (anionic surfactant), neodekont (mixture), tergitol NPX (non-ionic surfactant), Korynt P (non-ionic surfactant) were found to be the most effective. These decontaminants were able to wash away more than 92% of paraoxon from the contaminated skin.

Binding affinity and adhesion force of organophosphate hydrolase enzyme with soil particles related to the isoelectric point of the enzyme.

The binding affinity of organophosphate hydrolase enzyme (OphB) with soil particles in relation to the isoelectric point (pI) was studied. Immobilization of OphB with soil particles was observed by confocal microscopy, Fourier transform infrared spectroscopy (FT-IR), and Atomic force microscopy (AFM). The calculated pI of OphB enzyme was increased from 8.69 to 8.89, 9.04 and 9.16 by the single, double and triple mutant of OphB enzyme, respectively through the replacement of negatively charged aspartate with positively charged histidine. Practically, the binding affinity was increased to 5.30%, 11.50%, and 16.80% for single, double and triple mutants, respectively. In contrast, enzyme activity of OphB did not change by the mutation of the enzyme. On the other hand, adhesion forces were gradually increased for wild type OphB enzyme (90 pN) to 96, 100 and 104 pN for single, double and triple mutants of OphB enzyme, respectively. There was an increasing trend of binding affinity and adhesion force by the increase of isoelectric point (pI) of OphB enzyme.

Solvent-dependent binding interactions of the organophosphate pesticide, chlorpyrifos (CPF), and its metabolite, 3,5,6-trichloro-2-pyridinol (TCPy), with Bovine Serum Albumin (BSA): A comparative fluorescence quenching analysis.

Analysis of the interaction of pesticides and their metabolites with the cellular proteins has drawn considerable attention in past several years to understand the effect of pesticides on environment and mankind. In this study, we have investigated the binding interaction of Bovine Serum Albumin (BSA) with a widely used organophosphorous insecticide chlorpyrifos (CPF), and its stable metabolite, 3,5,6-trichloro-2-pyridinol (TCPy) to provide a comparative analysis of the two molecules by employing various spectroscopic techniques viz., UV-vis absorption, Circular Dichroism (CD), and Fluorescence spectroscopy. The fluorescence quenching studies of BSA emission in two different solvents viz., water and methanol in presence of CPF and TCPy have led to the revelation of several interesting facts about the pesticide-protein interaction. It has been found that both the molecules cause static quenching of BSA emission as seen from the Stern-Volmer constant (Ksv) irrespective of the solvent used for the analysis. While TCPy is a stronger quencher in water, it exhibits comparable quenching capacity with CPF in methanol. The solvent dependent differential binding interaction of the two molecules finally indicates possibility of diverse bio-distribution of the pesticides within human body. The UV-vis and CD spectra of BSA in presence of the test molecules have unravelled that the molecules formed ground state complex that are highly reversible in nature and have minimal effect on the protein secondary structure. Furthermore it is also understood that structural changes of BSA in presence of CPF is significantly higher compared to that in presence of TCPY.

Methamidophos induces cytotoxicity and oxidative stress in human peripheral blood mononuclear cells.

Previous studies have shown that organophosphate pesticide (OP) exposure is associated with oxidative stress. Methamidophos (MET) is an OP widely used in agriculture, which is regarded as a highly toxic pesticide and it is a potent inhibitor of acetylcholinesterase. The aim of this study was to evaluate whether MET can induce oxidative stress at low concentrations in primary cultures of human peripheral blood mononuclear cells (PBMCs). PBMCs from healthy individuals were exposed to MET (0-80 mg/L) for 0-72 h. We performed the MTT and neutral-red assays to assess the cytotoxicity. As indicators of oxidative stress, the levels of reactive oxygen species (ROS) were assessed using flow cytometry, and the malondialdehyde (MDA) and reduced glutathione (GSH) levels were determined. MET decreased the viability of PBMCs in a dose-dependent manner. At concentrations of 3, 10, or 20 mg/L for 24 h, MET increased the ROS production significantly compared with the vehicle control. Similarly, MET increased the levels of MDA at the same concentrations that increased ROS (10 and 20 mg/L); however, no changes in GSH levels were observed. These results suggest that MET increased the generation of oxidative stress in PBMCs. © 2015 Wiley Periodicals, Inc. Environ Toxicol 32: 147-155, 2017.