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1.
The aim of the work was to investigate the in vitro oxidation of diazinon and malathion, organophosphorous pesticides (OPs) containing phosphorthioate group, catalyzed by enzyme myeloperoxidase (MPO). The oxidation was performed in the presence of hydrogen peroxide. The products were identified as oxon derivatives (phosphates), where the sulfur atom from thioate group was substituted by an oxygen atom. No hydrolysis products were detected after enzyme - induced oxidation. The oxidation efficiency was controlled using acethylcholinesterase (AChE) bioassay for determination of oxon derivatives concentration. The influence of OPs concentration, incubation time of OPs with MPO, as well as MPO concentration on the yield of oxo forms was investigated. Kinetic constants of MPO in oxidation of malathion and diazinon were estimated. The maximum concentration of oxo forms was achieved after 10 min incubation of OPs in 50 mM phosphate buffer (pH 6.0) with 100 nM MPO.  相似文献   

2.
Inhibitory effects of five organophosphorus pesticides (diazinon, malathion, chlorpyrifos, azinphos-methyl and phorate) and their oxo-analogs on human myeloperoxidase (MPO) activity were investigated. While inspecting separately peroxidase and chlorination activity, it was observed that investigated OPs affect peroxidase activity, but not chlorination activity. Among investigated pesticides, malathion and malaoxon have showed the highest power to inhibit MPO peroxidase activity with IC50 values of the order of 3 × 10−7 and 5 × 10−9 M, respectively. It was proposed that inhibition trend is rendered by molecular structure which invokes steric hindrance for OPs interaction with MPO active center responsible for peroxidase activity. In addition, it was concluded that physiological function of MPO is not affected by any of the investigated OPs.  相似文献   

3.
The in vitro inhibition potency of some organophosphates (OPs) and carbamates (CAs) which are widely used to control plant-parasitic nematodes on acetylcholinesterase (AChE) of Meloidogyne javanica, Heterodera avenae and Tylenchulus semipenetrans, the major pathogens responsible for the damage of a wide range of crops in Al-Qassim region, Saudi Arabia was examined. AChE of H. avenae activity was 1.58- and 1.51-fold greater than that of T. semipenetrans or M. javanica, respectively. The order of inhibition potency of the tested compounds against T. semipenetrans AChE was: carbofuran > paraoxon > oxamyl > fenamiphos > phorate-sulfoxide > aldicarb, where the corresponding concentrations that inhibited 50% of the nematode AChE activity (I50) were 5 × 10−8, 7 × 10−7, 7.5 × 10−7, 2 × 10−6, 2 × 10−4 and 2 × 10−3 M, respectively. Paraoxon, fenamiphos and carbofuran exhibited high inhibition potency against M. javanica AChE where the I50 values were below 1 nM. Phorate-sulfoxide and aldicarb were potent inhibitors of M. javanica AChE with I50 values of 3.8 and 8 nM, respectively, while oxamyl exhibited low inhibition potency with I50 of 15 nM. Fenamiphos and paraoxon showed the highest I50 values of <100 μM against H. avenae followed by oxamyl (I50 < 1 mM), whereas paraoxon, carbofuran and aldicarb showed low potency with I50 values >1 mM. All the tested compounds exhibited high inhibition potency to AChE of M. javanica than T. semipenetrans or H. avenae. Except phorate-sulfoxide in M. javanica the inhibition pattern and implied mechanism for all the tested compounds for the three nematodes is suggested to be a linear mixed type (a combination of competitive and non-completive type).  相似文献   

4.
The susceptibility to phosalone and biochemical characteristics of acetylcholinesterase (AChE) were compared between susceptible (SS) strain and four field populations of Colorado potato beetle (CPB) collected from commercial potato fields of Hamedan Province in west of Iran. Bioassays involving topical application of phosalone to fourth instars revealed up to 252 fold resistance in field populations compared with the SS strain. Synergism studies showed that although esterase and/or glutathione S-transferase metabolic pathways were present and active against phosalone, they were not selected for and did not have a major role in resistance. It is likely that piperonyl butoxide (PBO) reduced phosalone toxicity by inhibiting bio-activation of phosalone. The affinity (Km) and hydrolyzing efficiency (Vmax) of AChE to selected substrates, namely, acetylthiocholine iodide (ATC), propionylthiocholine iodide (PTC), and butyrylthiocholine iodide (BTC) were examined. AChE inhibition by higher substrate concentration was evident only in the SS strain. In resistant field populations, Aliabad (Aa), Bahar (B) and Dehpiaz (Dp), substrate inhibition at higher concentrations was not seen. There was no definite optimal concentration found for any of the substrates examined. When ATC, PTC, and BTC were used as substrate, the reaction rates of AChE from Yengijeh (Yg) population increased as the concentration of all three substrates were increased, but were almost constant at concentration of ATC ? 3.98, PTC ? 2.8, and BTC ? 5 mM. The susceptible form of AChE had the most efficient ATC hydrolysis but very low BTC hydrolysis activity. In contrast, AChEs from field populations elicited relatively reduced ATC hydrolysis, but relatively increased BTC hydrolysis. The in vitro inhibition potency of some organophosphates (OPs), on AChEs of the field populations and SS strain was determined. The rank order from the most potent inhibitor to the least as determined by their bimolecular reaction constants (Ki) was ethyl paraoxon > diazoxon > methyl paraoxon for AChE from Aa, B, Dp, and Yg populations, respectively, whereas the rank order for the susceptible strain was methyl paraoxon > ethyl paraoxon > diazoxon.  相似文献   

5.
Organophosphorus pesticides (OPs) are of environmental significance due to their high toxicity to animals. Binding to plasma proteins may effective influence the toxicological properties of xenobiotics. In an attempt to evaluate the affinity of phenthoate (PTA) to bovine serum albumin (BSA) and inhibitory ability of bound PTA to acetylcholinesterase (AChE), we investigated the interactions between phenthoate (PTA) and bovine serum albumin (BSA) using tryptophan fluorescence quenching and subsequent inhibition on AChE activity by PTA. The results showed that PTA caused the fluorescence quenching of BSA because of the formation of a PTA-BSA complex. Quenching constants (Ksv), determined using the Sterns-Volmer equation to provide a measure of the binding affinity between PTA and BSA at 303, 306, 310 and 313 K were (3.4295 ± 0.0763) × 10−4, (3.2446 ± 0.0635) × 10−4, (3.0434 ± 0.0856) × 10−4 and (2.8262 ± 0.0569) × 10−4 M−1, respectively. The thermodynamic parameters, ΔH and ΔS were −25.04 kJ mol−1 and 168.94 J mol−1 K−1, respectively, which indicated that the electrostatic interactions played a major role in PTA-BSA association. The presence of BSA consistently reduced the inhibitory ability of PTA on AChE, with the relative activity being increased from 46.98 to 61.71% for the concentration range of BSA between 0 and 4.0 g L−1.  相似文献   

6.
Rainbow trout (Oncorhynchus mykiss; 116.88 ± 21.69 g) were exposed to sublethal concentrations (25 μg/L) of carbosulfan for 60 days to test if the long term exposure of fish to carbosulfan affects red blood cells acetylcholinesterase (AChE), δ-aminolevulinic acid dehydratase (ALA-D) and paraoxonase (PON) enzyme activity and induces genotoxic and/or mutagenic effects. The exposure resulted in inhibition of AChE and ALA-D activity of rainbow trout when compared to control fish. The activity of PON was not affected by carbosulfan. Interestingly, carbosulfan was found to induce DNA damage in red blood cells (comet assay) and proved to be mutagenic as revealed by the Ames test. Results indicate that blood AChE and ALA-D of rainbow trout may be a sensitive biomarker for assessment of carbosulfan contaminated water bodies. Furthermore, because the Ames test and comet assay were proven successful to detect the genotoxicity of carbosulfan, we proposed that nonlethal techniques such as blood collection from caudal vein of fish should be used to determine potential toxic effects of other pesticides to surrounding environment.  相似文献   

7.
The objective of this study was to investigate the effects of commercial formulation of herbicide 2,4-D on metabolic parameters, acetylcholinesterase (AChE) activity and liver histological evaluation of silver catfish (Rhamdia quelen) exposed for 96 h. AChE activity increased in brain (600 and 700 mg L−1) and decreased in all concentrations tested in muscle tissue. Hepatic glycogen was reduced after 2,4-D exposure ranging from 47.67% (400 mg L−1) until 59.3% (700 mg L−1). Hepatic tissue showed lactate reduction at all 2,4-D concentrations tested and glucose was reduced only at 700 mg L−1. In the highest concentration tested hepatic glycogen and glucose reduced instead plasma glucose levels increased. White muscle tissue showed glycogen reduction in fingerlings exposed to all herbicide concentrations and glucose reduction at 700 mg L−1. Muscle lactate levels increase at all 2,4-D concentrations tested. Vacuolation of hepatocytes and changes in its arrangement cords were observed by histologic analysis in group treated with 700 mg/L of 2,4-D. These results suggest that silver catfish exposed to concentrations of 2,4-D near of CL50 showed metabolic and histological response to compensate some stress caused by herbicide exposure. Taken together parameters measured can be used as biomarkers to monitor herbicide contaminated water.  相似文献   

8.
In vitro inhibition of electric eel acetylcholinesterase (AChE) by single and simultaneous exposure to organophosphorus insecticides diazinon and chlorpyrifos, and their transformation products, formed due to photoinduced degradation, was investigated. Increasing concentrations of diazinon, chlorpyrifos and their oxidation products, diazoxon and chlorpyrifos-oxon, inhibited AChE in a concentration-dependent manner. IC50 (20 min) values, obtained from the inhibition curves, were (in mol/l): (5.1 ± 0.3) × 10−8, (4.3 ± 0.2) × 10−6 and (3.0 ± 0.1) × 10−8 for diazoxon, chlorpyrifos and chlorpyrifos-oxon, respectively, while maximal diazinon concentration was lower than its IC50 (20 min). Calculated KI values, in mol/l, of 7.9 × 10−7, 9.6 × 10−6 and 4.3 × 10−7 were obtained for diazoxon, chlorpyrifos and chlorpyrifos-oxon, respectively. However, 2-isopropyl-4-methyl-6-pyrimidinol (IMP) and 3,5,6-trichloro-2-pyridinol, diazinon and chlorpyrifos hydrolysis products, did not noticeably affect the enzyme activity at all investigated concentrations. Additive inhibition effect was achieved for lower concentrations of the inhibitors (diazinon/diazoxon ?1 × 10−4/1 × 10−8 mol/l i.e., chlorpyrifos/chlorpyrifos-oxon ?2 × 10−6/3 × 10−8 mol/l), while an antagonistic effect was obtained for all higher concentrations of the organophosphates. Inhibitory power of 1 × 10−4 mol/l diazinon irradiated samples can be attributed mostly to the formation of diazoxon, while the presence of non-inhibiting photodegradation product IMP did not affect diazinon and diazoxon inhibitory efficiencies.  相似文献   

9.
The toxicity of organophosphorus pesticides is attributed to their ability to inhibit acetylcholinesterase (AChE), which catalyses the hydrolysis of the neurotransmitting agent acetylcholine (ACh). A simple, reproducible and stable AChE-based amperometric bioelectrode was used, for measuring the enzyme activity before and after the incubation with paraoxon dissolved in organic solvents.The inhibition percentage induced by a paraoxon in organic solvent solutions increases in the following sequence: acetonitrile < water < hexane, suggesting that the paraoxon repartition between the organic solvent and the essential water for enzyme activity plays an important role in establishing the analytical and kinetic parameters of the bioelectrode. Considering the AChE denaturation, induced only by the organic solvent, as a pseudo-inhibition process, the kinetic treatment of the bioelectrode response revealed that it follows a mixed competitive/non-competitive mechanism, while the inhibition due to a non-aqueous solution of paraoxon corresponds to a non-competitive/uncompetitive mechanism. In the last case, the synergetic effect of denaturation and inhibition of enzyme was dominated by the inhibitor presence.  相似文献   

10.
Recovery study was performed at regular intervals to establish the time course of 50% and 100% recovery in neurotransmitter enzyme (acetylcholinesterase, AChE, EC 3.1.1.7) and locomotor behaviour response of mosquito fish, Gambusia affinis exposed to lethal concentration (20.49 mg L−1) of an organophosphorous pesticide, monocrotophos (MCP) for 96 h. In vitro AChE activity studies indicated that MCP could cause 50% inhibition (I50) at 10.2 × 10−5 M. A positive correlation was observed between brain AChE activity and swimming speed during the recovery study. Also, the recovery response of the antioxidant enzymes superoxide dismutase (SOD, EC 1.15.1.1), catalase (CAT, EC 1.11.1.6) and glutathione reductase (GR, EC 1.6.4.2) as well as lipid peroxidation (LPO) as biomarkers of oxidative stress were assessed in viscera of G. affinis. The results showed that the MCP besides its inhibitory effect on target enzyme AChE activity and induction in antioxidant enzyme activities as a characteristic of oxidative stress, which can be used as biomarkers in the pesticide contaminated aquatic streams.  相似文献   

11.
Laboratory tests were conducted to compare the effects of various concentrations of lindane and deltamethrin on mature earthworms (Eisenia fetida) cultured in artificial soil during typical acute (14d) and subchronic (42d) exposure periods. The effects of the two pesticides on earthworm mortality, growth inhibition, and cellulase activity were determined for different exposure durations. The toxicity order for earthworm mortality from the 14-day exposure was lindane > deltamethrin, with median lethal concentrations (LC50) of 162.1 and 432.9 mg kg−1, respectively. Earthworms exposed to deltamethrin showed dose-dependent toxic effects on growth and cellulase activity only from the acute exposures, whereas lindane’s effects on these activities were seen correlated with both the acute and subchronic doses. Also, changes in biomass and cellulase activity during the subchronic exposure period appear to be a more sensitive parameter than the LC50 value in assessing pesticidal injury.  相似文献   

12.
在Fe3O4/Au微粒上固定乙酰胆碱酯酶(AChE),制得磁性复合粒子Fe3O4/Au/AChE。通过磁力将其吸附于涂覆了碳纳米管(CNTs)/纳米ZrO2/普鲁士蓝(PB)/Nafion(Nf)复合膜的丝网印刷碳电极(SPCEs)表面,制得一次性有机磷农药(OPs)酶传感器。采用扫描电镜(SEM)、X射线荧光光谱(XRFS)表征传感器的制备过程,采用循环伏安法(CV)和示差脉冲伏安法(DPV)研究了传感器的电化学性质。利用OPs对AChE的抑制作用,以硫代乙酰胆碱(ATCh)为底物,对乐果进行了检测。在pH=7.5的0.1 mol/L硝酸钾溶液中,乐果浓度的对数与酶电极的抑制率(A)在1.0×10-6~1.0×10-2 mg/L间呈良好的线性关系,检测限为5.6×10-7 mg/L,用于实际样品白菜检测时的添加回收率在88% ~105%之间,与气相色谱法(GC)所得结果一致。该传感器采用复合纳米粒子修饰电极表面,具有较高的比表面活性,响应迅速,检测限低;ZrO2可特异性地富集样品中的OPs,磁性纳米颗粒包被AChE可实现磁场分离和电极表面更新,且具有高灵敏度、低样品量、一次使用可抛弃、便携式等特点,可用于蔬菜等农产品中痕量OPs的快速、简便、准确检测。  相似文献   

13.
The effect of single, binary, and tertiary combination of few plant derived molluscicides alone or in combination with synergist on different enzymes (acetylcholinesterase—AChE, lactic dehydrogenase—LDH, and acid/alkaline phosphatase—ACP/ALP in the nervous tissue of the freshwater snail Lymnaea acuminata were studied. Sublethal in vivo 24 h exposure to 40 and 80% of LC50 of Azadirachta indica oil (AI), oleoresin of Zingiber officinale (OL), Cedrus deodara oil (CD), Allium sativum (AS), and Polianthes tuberosa (PT) bulb powder singly, their binary combination of AI + OL, AS + CD, AS + PT, CD + OL, CD + PT, OL + PT, and tertiary combination of these binary combinations with the synergist piperonyl butoxide (PB) or MGK-264 significantly altered the activity of these enzymes. Tertiary combination with PB or MGK was very effective. Combination of CD + PT + MGK was more effective against AChE whereas, the combination of CD + OL + PB, CD + AS + PB, and CD + PT + PB were more effective against LDH, ACP, and ALP, respectively.  相似文献   

14.
The results showed that survival rates of Biomphalaria alexandrina snails, reproductive potential and hatchability of eggs were evaluated post exposure to Basudin, Selecron and Colchicine. As well, DNA and RNA changes in the cells of ovotestis-digestive gland complex of treated snails were estimated. The current molluscicide Bayluscide was used as a reference compound.The pesticide Selecron proved to be more toxic to B. alexandrina snails than Basudin and Colchicine. Juvenile snails were dead post 3 weeks of exposure to the sublethal concentration LC0 of either Selecron or Basudin, while 26.75% of snails still alive at Bayluscide treatment. In addition, exposure of adult snails to LC0 of Selecron for 24 h/week for 4 weeks markedly reduced their reproductive rate (R0) by 89.9%. Moreover, snails’ eggs failed to hatch post 24 h of exposure to LC90 of either Selecron, Basudin or Bayluscide. Electrophoretic analysis indicated a decrease in the molecular weight of intact DNA in the ovotestis-digestive gland complex of snails treated with 250 ppm of Colchicine and LC25 of Selecron, as it scored 1.2 and 76 bp, respectively, compared to 166.46 bp for control group, while the vice versa was recorded for RNA intensity. It was concluded that the tested pesticides have deleterious effects on snails’ reproductive rate, their eggs and the intensities of DNA and RNA in their ovotestis-digestive gland complex. Therefore, it is expected that reaching of such pesticides to snails’ habitats in water courses during plant pests control could minimize the population density of the snails intermediate hosts of schistosomiasis, hence probably interrupt and reduce the transmission of this parasite.  相似文献   

15.
Nile Tilapia (Oreochromis niloticus) juveniles were exposed to different concentrations of Folidol 600® in static toxicity tests. The 24, 48, 72 and 96 h LC50 values of Folidol 600® to O. niloticus were 17.82, 8.91, 4.00 and 2.70 mg L−1, respectively. The values of hematological parameters increased, and inhibition of cholinesterases activity (AChE, BChE and PChE) in plasma of fish exposed to the higher concentrations of pesticide reached 94%. Furthermore, the exposure of Tilapia to Folidol 600® caused an increase of 4%, 20% and 38.4% in oxygen consumption at 0.1, 0.5 and 1.0 mg L−1, respectively. However, exposure to 2.5, 5.0 and 10 mg L−1 caused a decrease of 33.6%, 35.2% and 42.4% in oxygen consumption relative to the control. The ammonium excretion of fish exposed to 0.0, 0.1, 0.5, 1.0, 2.5, 5.0 and 10.0 mg Folidol 600®/L was 0.12, 0.18, 0.30, 0.33, 0.37, 0.36 and 0.33 μg/g/min, i.e., 50%, 150%, 175%, 208%, 200% and 175% increase, respectively, relative to the control.  相似文献   

16.
The present study was aimed to investigate the effects of purified neem extract azadirachtin on the blood electrolytes of Heteropneustes fossilis for short- and long-term. In short-term exposure the fish were subjected to 0.8 (i.e. 80%) of 96 h LC50 value of azadirachtin (41.89 mg L−1) for 96 h. In long-term exposure the experiment was performed for 28 days by using 0.2 (i.e. 20%) of 96 h LC50 value of azadirachtin (10.47 mg L−1). Fish were sacrificed (anaesthesized with MS 222) from control and experimental (azadirachtin) groups after 24, 48, 72, and 96 h in short-term exposure and after 7, 14, 21, and 28 days in long-term experiment. Blood samples were analyzed for calcium and inorganic phosphate levels. Acute exposure of azadirachtin caused a progressive decrease in the serum calcium levels after 48 h in fish H. fossilis which persists till the close of the experiment (96 h). The serum inorganic phosphate levels remained unaffected till 72 h in the azadirachtin exposed fish. After 96 h the levels exhibit a decrease. Chronic azadirachtin treatment caused a decrease in serum calcium levels at day 14. This decrease continued till the end of the experiment. The serum phosphate level of the azadirachtin treated fish decreased on day 14 and 21. However, on day 28 the levels were close to the normal values.  相似文献   

17.
The inhibitory effects of a recently introduced series of the titled compounds on insect and mammalian acetylcholinesterase (AChE) activity were examined, where the median inhibition concentration (I50) and the inhibition kinetic parameters, bimolecular inhibition rate constant (ki), affinity constant (Ka), and phosphorylation rate constant (kp), were determined for each compound. Results indicated that all examined dioxaphospholenes had less inhibitory effects on mammalian AChE than fenitrothion, a commercial pesticide with moderate mammalian toxicity. The highest selectivity was obtained with compounds containing glutamic and leucine moieties (2.70 and 2.18, respectively) while selectivity of fenitrothion was 0.93. The low inhibitory effects of the examined dioxaphospholenes on mammalian AChE were attributed to their low phosphorylation rates (kp < 2.2 min−1) compared to that of fenitrothion (kp = 4.84 min−1). QSAR equations indicated that the inhibition process is controlled mainly by both the phosphorylation rate (direct effect) and the affinity of compounds toward the enzyme (inverse effect). Although the compounds’ hydrophobicity had no effects on the inhibition process, it affects the compounds’ toxicity since it affects the ability of compounds to penetrate insects to reach the enzyme active site.  相似文献   

18.
The aim of this study was to investigate the impact of carbofuran, a carbamate pesticide on the enzymatic activity of acetylcholinesterase (AChE) in the larvae of the herring nematode parasite Anisakis simplex. A. simplex larvae collected from herring were exposed to carbofuran in vivo at concentrations of 50, 100, 500 and 1000 μg/l, for 24, 48 and 72 h, at a temperature of 4 °C. Generalized Linear Models (GLM) were applied to analyze the relationship between AChE activity and carbofuran concentration, the time of exposure and the biological parameters of the host. The results indicate that A. simplex larvae have a high threshold of sensitivity to carbofuran. The average enzymatic activity was higher in parasites obtained from male hosts, when compared with female hosts. These data suggest that host sex-dependent biological processes may also influence AChE enzymatic activity in parasites.  相似文献   

19.
Agricultural chemicals can induce genetic alterations on aquatic organisms that have been associated with effects on growth, reproduction and population dynamics. The evaluation of DNA damage in fish using the comet assay (CA) frequently involves the utilization of erythrocytes. However, epithelial gill cells (EGC) can be more sensitive, as they are constantly dividing and in direct contact with potentially stressing compounds from the aquatic environment. The aim of the present study was to evaluate (1) the sensitivity and suitability of epithelial gill cells of Prochilodus lineatus in response to different genotoxic agents through the application of the CA, (2) the induction of DNA damage in this cell population after in vivo exposure to cypermethrin. Baseline value of the CA damage index (DI) for EGC of juvenile P. lineatus was 144.68 ± 5.69. Damage increased in a dose-dependent manner after in vitro exposure of EGC to methyl methanesulfonate (MMS) and H2O2, two known genotoxic agents. In vivo exposure of fish to cypermethrin induced a significant increase in DNA DI of EGC at 0.150 μg/l (DI: 239.62 ± 6.21) and 0.300 μg/l (270.63 ± 2.09) compared to control (150.25 ± 4.38) but no effect was observed at 0.075 μg/l (168.50 ± 10.77). This study shows that EGC of this species are sensitive for the application of the CA, demonstrating DNA damage in response to alkylation (MMS), oxidative damage (H2O2), and to the insecticide cypermethryn. These data, together with our previous study on DNA damage induction on erythrocytes of this species, provides useful information for future work involving biomonitoring in regions where P. lineatus is naturally exposed to pesticides and other genotoxic agents.  相似文献   

20.
Experiment was carried out to determine the median lethal concentration (LC50) of fenvalerate to Labeo rohita fingerlings. After determining the LC50 value of fenvalerate, a sub-lethal concentration (1/3rd of LC50) of fenvalerate was exposed for 15 days. Significant alterations in SOD (P < 0.05) activity of liver and gill was observed due to fenvalerate. Catalase activity in gills of fishes was also affected significantly (P < 0.05). WBC, NBT and Hct values were reduced significantly in fenvalerate exposed fishes as compared to control group, whereas blood glucose level showed higher values in fenvalerate exposed group. Serum total protein and albumin were also reduced significantly as a result of fenvalerate exposure. Significant increase in the serum GOT, serum GPT, creatinine, triglyceride and serum ACP was noticed after 15 days of fenvalerate exposure. Results indicated that short term exposure of fenvalerate can induce biochemical and haematological alterations causing stress to L. rohita fingerlings.  相似文献   

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