Combined toxicities of binary mixtures of alachlor, chlorfenvinphos, diuron and isoproturon.

Pesticides are the chemicals of increased concern regarding their adverse environmental effects. In particular, the reports on their joint toxicity effects are scarce in the literature. Therefore, this paper describes the experiments on toxicities of four pesticides: alachlor, chlorfenvinphos, diuron, and isoproturon, toward Vibrio fischeri. In particular, the joint toxicity effects for all possible binary combinations of the pesticides were analyzed. The analysis included the application of concentration addition and independent action models at two toxicity levels: EC10 and EC50. The analysis revealed additive behavior between all pesticide pairs. The only exception was isoproturon and chlorfenvinphos whose combination resulted in synergistic toxic activity. The original form of the logistic function was given preference over the linearized form in describing the response-dose relationships of investigated pesticides.

The effect of two bromfenvinphos impurities: BDCEE and ß-ketophosphonate on oxidative stress induction, acetylcholinesterase activity, and viability of human red blood cells.

Numerous research works have shown that synthesis of pesticides leads to the formation of impurities that may substantially enhance pesticide toxicity. In this study, the effect of manufacturing impurities of pesticide bromfenvinphos (BFVF) such as 1-bromo-2-(2,4-dichlorophenyl)-2-ethoxy ethene (BDCEE) and diethyl [2-(2,4-dichlorophenyl)-2-oxo-ethyl] phosphonate (ß-ketophosphonate) on human erythrocytes, being significantly exposed to xenobiotics has been studied. The cells were treated with the compounds studied in the concentrations ranging from 0.1 µM to 250 µM for 4 h. In order to assess the effect of BDCEE and ß-ketophosphonate on red blood cells hemolytic changes, changes in cell size (FSC parameter) and oxidation of hemoglobin were studied. Moreover, alterations in reactive oxygen species (ROS) formation, reduced glutathione (GSH) level and acetylcholinesterase (AChE) activity were determined. BDCEE induced an increase in ROS level and caused strong oxidation of hemoglobin as well as a slight change in erythrocytes size and hemolysis, while it did not change GSH level and AChE activity. ß-ketophosphonate has not been shown to affect most parameters studied, but it strongly reduced AChE activity. Because changes in the parameters examined were noted at low concentrations of BFVF impurities (5-250 µM), those substances should not negatively affect on red blood cells of humans environmentally exposed to this pesticide.

Comparison and evaluation of pesticide monitoring programs using a process-based mixture model.

A number of European countries run large-scale pesticide monitoring schemes in watersheds aimed at identifying and evaluating the presence of pesticide residues in the environment. These schemes provide national and regional scale assessments of pesticide concentrations within the context of environmental quality assessment, aiming to ensure some degree of ecological protection. The present study is aimed at evaluating the joint effects of the pesticide mixtures detected in monitoring programs, using a process-based mixture model that was parameterized for Daphnia magna. In total, over 15 000 samples containing over 1 million individual measurements were evaluated for effects. It was found that there are only a small number of places where one can expect to have effects on daphnids, based on measured concentrations. The most polluted samples would cause extinction of a daphnid population within only 30 h. The results show that effects are mostly triggered by a limited number of pesticide residues at locations with high emissions. It was also shown that the analytical detection limits are basically too high to exclude mixture effects. So, despite all the effort that is put into chemical monitoring programs, it remains a challenge to make statements on whether or not the environment is protected. Recommendations are offered for a different setup of monitoring programs to improve this situation. Environ Toxicol Chem 2016;35:3113-3123. © 2016 SETAC.

Bromfenvinphos induced suicidal death of human erythrocytes.

The organophosphorus pesticide bromfenvinphos ((E,Z)-O,O-diethyl-O-[1-(2,4-dichlorophenyl)-2-bromovinyl] phosphate) has been shown to decrease hematocrit and hemoglobin levels in blood presumably by triggering oxidative stress of erythrocytes. Oxidative stress is known to activate erythrocytic Ca(2+) permeable unselective cation channels leading to Ca(2+) entry and increase of cytosolic Ca(2+) activity ([Ca(2+)]i), which in turn triggers eryptosis, the suicidal death characterized by cell shrinkage and cell membrane scrambling with phosphatidylserine translocation to the erythrocyte surface. The present study explored, whether and how bromfenvinphos induces eryptosis. To this end, phosphatidylserine exposure at the cell surface was estimated from annexin-V-binding, cell volume from forward scatter, hemolysis from hemoglobin release, [Ca(2+)]i from Fluo3-fluorescence, and ROS formation from DCFDA dependent fluorescence. As a result, a 48hour exposure of human erythrocytes to bromfenvinphos (≥100µM) significantly increased the percentage of annexin-V-binding cells, significantly decreased forward scatter, significantly increased Fluo3-fluorescence, and significantly increased DCFDA fluorescence. The effect of bromfenvinphos on annexin-V-binding and forward scatter was significantly blunted, but not abolished by removal of extracellular Ca(2+). In conclusion, bromfenvinphos triggers cell shrinkage and phospholipid scrambling of the erythrocyte cell membrane, an effect in part due to stimulation of ROS formation and Ca(2+) entry.

Organophosphorous biocides reduce tenacity and cellular viability but not esterase activities in a non-target prosobranch (limpet).

Detecting impacts of organophosphorus biocides (OP) is facilitated by analysing "biomarkers" - biological responses to environmental insults. Understanding is hampered by studying biomarkers in isolation at different levels of biological response and limited work on ecologically-important species. We tested the relevance of esterases as biomarkers of OP-exposure in limpets (Patella vulgata), abundant prosobranchs that structure the assemblages on rocky shores through their grazing. We characterized esterases in haemolymph and tissue, and quantified their dose-dependent inhibition by chlorfenvinphos (0.1-3.0 mM) in vitro. To determine whether esterases are useful biomarkers we exposed limpets to chlorfenvinphos (0-10 µg L(-1)). Despite reduced tenacity (ability to stick to a surface) and haemocyte-viability, esterases remained unaffected. Tenacity was reduced by >50% at 5 µg L(-1) and by 95% at 10 µg L(-1), whilst haemocyte-viability was more sensitive with >40% reductions at concentrations of 0.5 µg L(-1) and above. We discuss results in relation to linking sub-lethal and ecological impacts at contaminated sites.

Oxidative stress in human erythrocytes treated with bromfenvinphos and its impurities.

Bromfenvinphos (BFVF) is an organophosphorus (OP) pesticide which was widely used in agriculture and veterinary practice. During synthesis of this insecticide five main impurities are formed: dihydro-bromfenvinphos, dibromo-bromfenvinphos, 2,4-dichlorophenacyl bromide, 2,4-dichlorophenacylidene bromide and 2,4-dichlorophenacylidyne bromide, which can be present in technical grade bromfenvinphos in amounts from 0.1 to 4%. The aim of this study was to examine the influence of bromfenvinphos and its manufacturing impurities on parameters of oxidative stress, the activity of antioxidative enzymes and the level of reduced glutathione. Human erythrocytes were incubated with bromfenvinphos and its impurities in the concentrations range from 0.5 to 500 µM for 1 h. This study indicated that 2,4-dichlorophenacyl derivatives more strongly oxidized analyzed parameters in human erythrocytes than bromfenvinphos. Investigated compounds caused an increase in lipid peroxidation and oxidation of fluorescent probe DCFH2 - the strongest pro-oxidative changes were provoked by 2,4-dichlorophenacyl bromide. None of the compounds studied in the concentrations from 0.5 to 500 µM changed the activity of SOD and only 2,4-dichlorophenacyl decreased activity of CAT. The level of GSH was only altered by 2,4-dichlorophenacyl derivatives. It was observed that increasing number of bromine atoms in the side chain of those derivatives was associated with decreased GSH level.

Alterations in gills of Lepomis gibbosus, after acute exposure to several xenobiotics (pesticide, detergent and pharmaceuticals): morphometric and biochemical evaluation.

In recent decades, scientific research about the effects of anthropogenic xenobiotics on non-target organisms has increased. Among the likely effects, some studies reported the evaluation of biochemical and morphological changes in specific tissues or organs of fishes, such as gills, which are key organs for the direct action of pollutants in the aquatic environment. This work intended to assess biochemical [oxidative stress/phase II conjugation isoenzymes glutathione S-transferase (GSTs)] and morphological [secondary lamellar length (SLL), secondary lamellar width (SLW), interlamellar distance (ID), basal epithelial thickness (BET) and proportion of the secondary lamellae available for gas exchange (PAGE)] changes in gills, after acute exposure to the pesticide chlorfenvinphos, the detergent sodium dodecylsulphate (SDS) and to the anticholinesterasic pharmaceuticals (neostigmine and pyridostigmine). Our results point to a significant, eventually hormetic, effect in the activity of GSTs following exposure to chlorfenvinphos that significantly increased the activity of GSTs at concentration of 0.2 mg/L. The activity of GSTs increased significantly after exposure to 100 mg/L of neostigmine. Considering the morphometric analysis of the gills, the data obtained showed that chlorfenvinphos exerted mainly minor architectural alterations in gills, with the exception of the highest tested concentration of chlorfenvinphos that produced also a slight decrease of the PAGE. The overall conclusions point to a null or negligible toxicity of the selected toxicants towards L. gibbosus, which may be reverted if exposure is withdrawn.

Combination effects of anticholinesterasics in acetylcholinesterase of a fish species: effects of a metallic compound, an organophosphate pesticide, and a pharmaceutical drug.

The simultaneous presence of distinct compounds in the aquatic environment can be causative of various toxicological interactions. This scenario challenges ecotoxicologists, since the assessment of toxicological effects caused by the simultaneous presence of multiple substances is by far more complicated. An illustrative example can be given by mentioning the anticholinesterasic compounds: by studying the level of cholinesterase impairment of an exposed organism, it is possible to ascertain the level of exposure to all anticholinesterasics (despite their chemical classes and natures) that the organism was subjected to. In this work, we describe the effects of three chemically different, albeit mechanistically, and toxicologically similar compounds (copper, chlorfenvinphos, and pyridostigmine) on cholinesterases of the fish Gambusia holbrooki. The results showed that the combinatorial effects may be of considerable extent, even for levels of exposure that are close to the ones already reported in the wild, for each isolated compound.

Partial protection from organophosphate-induced cholinesterase inhibition by metyrapone treatment.

BACKGROUND: Organophosphates are cholinesterase (ChE) inhibitors with worldwide use as insecticides. Stress response, evidenced by a dramatic and relatively long-lasting (several hours) rise in the plasma glucocorticoid concentration is an integral element of the organophosphate (OP) poisoning symptomatology. In rodents, corticosterone (CORT) is the main glucocorticoid. There are several reports suggesting a relationship between the stressor-induced rise in CORT concentration (the CORT response) and the activity of the cerebral and peripheral ChE. Thus, it seems reasonable to presume that, in OP intoxication, the rise in plasma CORT concentration may somehow affect the magnitude of the OP-induced ChE inhibition. Metyrapone (MET) [2-methyl-1,2-di(pyridin-3-yl)propan-1-one] blocks CORT synthesis by inhibiting steroid 11ß-hydroxylase, thereby preventing the CORT response. Chlorfenvinphos (CVP) [2-chloro-1-(2,4-dichlorophenyl) ethenyl diethyl phosphate] is an organophosphate insecticide still in use in some countries. MATERIAL AND METHODS: The purpose of the present work was to compare the CVP-induced effects - the rise of the plasma CORT concentration and the reduction in ChE activity - in MET-treated and MET-untreated rats. Chlorfenvinphos was administered once at 0.0, 0.5, 1.0 and 3.0 mg/kg i.p. Metyrapone, at 100 mg/kg i.p., was administered five times, at 24-h intervals. The first MET dose was given two hours before CVP. CONCLUSION: The following was observed in the MET-treated rats: i) no rise in plasma CORT concentration after the CVP administration, ii) a reduced inhibition and a faster restitution of blood and brain ChE activities. The results suggest that MET treatment may confer significant protection against at least some effects of OP poisoning. The likely mechanism of the protective MET action has been discussed.

[Serum glucose concentration in subacute intoxication with chlorpyrifos - organophosphate insecticide]. / Stezenie glukozy w surowicy krwi w podostrym zatruciu chlorpyrifosem - insektycydem fosforoorganicznym.

BACKGROUND: Epidemiological studies suggest that exposure to organophosphate insecticides enhances the risk of various diseases, including neurological disorders, e.g. Parkinson's or Alzheimer's disease, arteriosclerosis and diabetes mellitus. For this reason the aim of the presented study was to estimate serum concentration of glucose in subchronic intoxication with low doses of chlorpyrifos, an organophosphate insecticide. MATERIALS AND METHODS: The rats received chlorpyrifos at a daily dose of 0.2, 2 or 5 mg/kg b.w./day for 14 or 28 days. For biochemical determinations of serum glucose in the rats ready-to-use kit was applied. RESULTS: In subacute intoxication with chlorpyrifos the increased serum concentration of glucose was observed after 14 days of intoxication with the highest dose (5 mg/kg b.w.) and after 28 days of intoxication with all dose levels used. CONCLUSIONS: The results of this study showing the increased concentration of serum glucose in subacute intoxication with low doses of chlorpyrifos, as well as the literature data suggest that exposure to organophosphate insecticides can increase the risk of diabetes mellitus. It may thus be concluded that people occupationally exposed to these compounds should be subjected to diagnostic tests for early detection of diabetes.

The effect of exposure to chlorfenvinphos on lipid metabolism and apoptotic and necrotic cells death in the brain of rats.

This study investigated the influence of chlorfenvinphos (0.3 mg/kg bw/24 h corresponding to 0.02 LD50; orally by gastric gavage for 14 and 28 days) on lipid metabolism, and apoptotic and necrotic cells death in the brain of rats as the possible mechanism of neurotoxic action of organophosphate (OP) pesticides at low exposure. Total cholesterol (TCh), triglycerides (TG), phospholipids (PL), and free fatty acids (FFA) were determined and apoptotic, necrotic, and living cells were quantified in the brain. Moreover, the serum and brain acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) were assayed as biomarkers of neurotoxicity. The treatment with chlorfenvinphos increased (duration dependently) the concentrations of TCh and TG and the ratio of TCh/PL, and decreased PL concentration. The prevalence of apoptotic and necrotic cells increased and that of the living brain cells depressed (by 10%) already after 14 days of the exposure. The brain activities of AChE and BChE decreased by 12% and 15%, and by 18% and 25% after 14 and 28 days, respectively, whereas the serum activities of these enzymes were inhibited (by 24% and 18%, respectively) only after the longer treatment. The changes in lipid metabolism and distribution of the living, apoptotic, and necrotic brain cells correlated with AChE and BChE activities in the serum and brain. The results show that chlorfenvinphos may disturb lipid metabolism and induce apoptosis and necrosis in the brain even at the exposure not affecting the serum activities of cholinesterases, and causing only moderate inhibition of their brain activities. Based on the findings it can be concluded that low repeated exposure to OP pesticides may influence the nervous system through disrupting the lipid profile of the nervous tissue and decreasing the number of the nervous cells.

Variation in effects of four OP insecticides on photosynthetic pigment fluorescence of Chlorella vulgaris Beij.

Effects of the insecticides quinalphos, chlorfenvinphos, dimethoate and phorate on photosystem activity of Chlorella vulgaris were investigated by different chlorophyll fluorescence measurements. Exposure to each of the insecticides increased the proportion of inactivated PS II reaction center. Quinalphos and chlorfenvinphos caused OJIP fluorescence reduction at all levels by decreasing the proportion of Q(A)-reducing PS II reaction centers (RCs). The other two insecticides affected OJIP fluorescence rise by hindering the electron transport beyond Q(A). Insecticide treatment resulted in decrease of the density of active RC and performance indices (PI) by enhanced dissipated energy flux per active RC. Antenna size was severely minimized by quinalphos and chlorfenvinphos treatment whereas other two insecticides had no such effect. Each insecticide treatment caused increase of photosystem antenna/core and PS II/PS I fluorescence ratios. Quinalphos and chlorfenvinphos affected the donor sides of photosystems whereas dimethoate and phorate inhibited electron transfer beyond Q(A) (acceptor side).

[Paraoxonase activity and lipid peroxides concentration in serum of rats subchronically intoxicated with chlorpyrifos--organophosphate insecticide]. / Atywnosc paraoksonazy i stezenie nadtlenków lipidów w surowicy krwi szczurów w podostrym zatruciu chlorpyrifosem--insektycydem fosforoorganicznym.

BACKGROUND: Toxicity of organophosphate insecticides is mainly due to the inhibition of acetylcholinesterase (AChE). However, organophosphate insecticides in acute, as well as in chronic and subchronic intoxication may lead to oxidative stress causing the enhancement of lipid peroxidation and changing the activities of antioxidative enzymes. Paraoxonase PON1 is synthesized by the liver. This enzyme hydrolyzes organophosphate compounds, phospholipid hydroperoxides and cholesterol ester hydroperoxides. Its role as an antioxidant has also been suggested. For this reason the aim of the work was to estimate the activity of paraoxonase and the level of serum lipid peroxides in the rats subchronically intoxicated with chlorpyrifos. MATERIALS AND METHODS: The animals received chlorpyrifos at a single daily dose of 0.2, 2 or 5 mg/kg b.w./day for 14 days. For biochemical determinations of paraoxonase activity in serum of rats, Aryloesterase/Paraoksonase Assay Kit (ZeptoMetrix Corporation Buffalo, USA) and for lipid peroxides level, LPO-586 (OXIS International, Foster City, Calif., USA) were used. RESULTS: Chlorpyrifos administration resulted in a decreased activity of paraoxonase in serum. The highest decrease was observed after administration of chlorpyrifos in doses 2 and 5 mg/kg b.w./day. In the intoxication with the highest insecticide dose under study the decreased paraoxonase activity was accompanied by the increased level of lipid peroxides. CONCLUSIONS: In view of the literature data, the finding that low doses of chlorpyrifos lead to statistically significant decrease in paraoxonase activity in serum of rats provides evidence that exposure to organophosphate insecticides induces oxidative stress. It also suggests the need to take into consideration a possible development of arteriosclerosis, hypercholesterolemia and diabetes mellitus in people exposed to these compounds.

[Liver and serum glutathione concentration and liver hydrogen peroxide in rats subchronically intoxicated with chlorfenvinphos--organophosphate insecticide]. / Stezenie glutationu w watrobie i w surowicy krwi oraz nadtlenku wodoru w watrobie szczurów w podostrym zatruciu chlorfenwinfosem--insektycydem fosforoorganicznym.

BACKGROUND: Toxicity of organophosphate insecticides is mainly due to the inhibition of acetylcholinesterase (AChE). However organophosphate insecticides in acute as well as in chronic and subchronic intoxication may lead to oxidative stress causing enhancement of lipid peroxidation and changing the activities of antioxidative enzymes and concentration of non-enzymatic antioxidant. For this reason the aim of the work was to estimate glutathione and hydrogen peroxide levels in the liver, as well as the concentration of total glutathione in serum of rats in subchronic intoxication with chlorfenvinphos. MATERIALS AND METHODS: The animals received chlorfenvinphos, intragastrically with use of a stomach tube, at a one daily dose of 0.3 mg/kg/day for 14 or 28 days. For biochemical determinations BIOXYTECH GSH-400 and BIOXYTECH H2O2-560 Assay kit, OXIS International, Inc., Portland, U.S.A. (reduced glutathione and hydrogen peroxide), and Glutathione Assay Kit, Cayman Chemical Company, U.S.A. (determination of serum total glutathione level) were used. RESULTS: Chlorfenvinphos administration resulted in a decreased level of reduced glutathione in liver accompanied by an increase in liver hydrogen peroxide and serum total glutathione concentrations. The observed changes were more pronounced after 28 days of intoxication. CONCLUSIONS: The common use of organophosphate insecticides results in the environmental pollution, therefore, the decreased liver glutathione level is an additional risk factor for people treated with different medicine (e.g., paracetamol).

Neuroendocrine and behavioral response to amphetamine challenge after exposure to an organophosphorus pesticide.

OBJECTIVES: Exposure to various stressors is known to result in sensitization to psychostimulants, a state related to the psychostimulant dependence and addiction. It has been shown in some studies that the rise in corticosterone (CORT) concentration is indispensable for both the induction and the expression of behavioral sensitization. Therefore, it might be suspected that behavioral hyposensitivity to amphetamine (AMPH) is somehow related to a reduced CORT response to the psychostimulant subsequent to the chlorphenvinphos (CVP) intoxication. MATERIALS AND METHODS: The male adult Wistar rats received single i.p. injections of CVP at the doses 0.5, 1.0 or 3.0 mg/kg b.w., or pure corn oil. CORT concentration was determined in samples of blood drawn from the tail vein before and then 30, 60, 180 min and 24 h after injection. The other rats were divided into two groups and tested, three weeks after the CVP injection for the effect of AMPH (0.5 mg/kg b.w. i.p.) on the serum CORT concentration. In addition, behavioral sensitivity to AMPH was assessed by measuring locomotor activity of the animals in an open-field. RESULTS: 1) The stressor property of CVP was confirmed. The injection resulted in up to tenfold increase in the serum CORT concentration. The magnitude and duration of this response were dose-related. 2) Three weeks after the CVP exposure, the CORT response to AMPH was significantly increased. 3) The behavioral response to the psychostimulant, i.e. augmented locomotion, was significantly reduced compared to the control. CONCLUSIONS: The results confirm that CVP exposure causes behavioral hyposensitivity to AMPH. This effect, however, could not be ascribed to a diminished CORT response.

The effect of bromfenvinphos and its impurities on human erythrocyte.

Bromfenvinphos - (E,Z)-O,O-diethyl-O-[1-(2,4-dichlorophenyl)-2-bromovinyl] phosphate (BFVF) is the insecticide elaborated in Poland, which has been used against Varroa destructor causing honey bees disease called as varroosis. The substances that are formed as a result of bromfenvinphos synthesis are dihydro-bromfenvinphos (O,O-diethyl O-[1-(2,4-dichlorophenyl)vinyl] phosphate); dibromo-bromfenvinphos (O,O-diethyl O-[1-(2,4-dichlorophenyl)-2,2-dibromovinyl] phosphate); 2,4-dichlorophenacyl bromide; 2,4-dichlorophenacylidene bromide and 2,4-dichlorophenacylidyne bromide. In this work, we evaluated the effect of these compounds on hemolysis and hemoglobin oxidation (met-Hb formation) in human erythrocytes. Moreover, the changes in the size (FSC-A) and the shape (SSC-A) of red blood cells were assessed using flow cytometry and phase contrast microscopy. It was proven that bromfenvinphos at concentrations ranging from 0.5 to 250 µM during 1h incubation did not change the parameters examined in human erythrocytes. Similarly, most of bromfenvinphos impurities did not increase hemolysis and methemoglobin level nor changed the size and shape of the erythrocytes. The exception was dibromo-bromfenvinphos, which changed the FSC-A and SSC-A parameters, as well as 2,4-dichlorophenacyl bromide which induced hemolysis, increased the level of met-Hb and changed erythrocytes morphology.

Effects of stress pretreatment on the dynamics of blood cholinesterase activity after exposure to an organophosphorus pesticide in the rat.

A single i.p. administration of 1.0 mg/kg of chlorphenvinphos (CVP), an organophosphorus pesticide, results in an acute stress response, evidenced by a marked (6-7 fold) rise in plasma corticosterone (CORT) concentration, and a diminished behavioural sensitivity to amphetamine (AMPH) three weeks postexposure. Surprisingly, in rats subjected to a single series of inescapable electric footshocks (60 10 msec triplets of 3.0 mA, 2 msec, square pulses during 20 min - IF ) two weeks prior to the CVP exposure, these effects are not observed. It has been assumed that the reduced effectiveness of CVP might be related to some persisting alterations in the functional state of the cholinergic system. The aim of the present work was to discover whether and in what way the IF pretreatment affects i) the cholinesterase activity in blood, and ii) the dynamics of the alterations in the cholinesterase (ChE) activity following the CVP exposure. The experiments were performed on 3 mo. old, male Wistar rats. In the first experiment, the blood samples were taken from the tail vein 15, 60 and 180 min after the IF. In the second experiment, the rats were pretreated with IF and 14 days later given 1.0 mg/kg of CVP i.p. Blood samples were taken 15 min, 60 min, 180 min, 24 h, 7 days, and 14 days after the CVP exposure. In the first experiment no differences in the ChE activity in plasma (pChE) and erythrocytes (rbcChE) were found between the shocked and control rats. In the second experiment, however, in rats pretreated with IF the rbcChE activity of was reduced by CVP less and pChE activity returned to normal faster than in rats not pretreated with IF. The results confirm that exposure to IF, a nonchemical stressor, induces some long-lasting adaptive changes which render the cholinergic system less susceptible to the harmful action of ChE inhibitors. It has been hypothesized that the changes consist in an increase of the antioxidant potential in blood and possibly other tissues.

Assessment of acetylcholinesterase activity in Clarias gariepinus as a biomarker of organophosphate and carbamate exposure.

The objective of this study was to investigate the response of acetylcholinesterase (AChE) activities in Clarias gariepinus in response to Organophosphates (Ops) and carbamate exposure. The AChE activities were determined in plasma, and eye and brain homogenates of unexposed and exposed fish using Ellman's method and 5,5'-dithiobis-2-nitrobenzoic acid (DTNB) chromophore. The baseline AChE activities in plasma, eyes and brain tissues in unexposed fish were comparable between males and females (P > 0.05). Concentrations of pesticides that inhibited 50% (IC(50)) of AChE activities in brain homogenates following in vitro exposures were 0.003, 0.03, 0.15, 190, 0.2, 0.003 and 0.002 microM for carbaryl, chlorfenvinphos, diazinon, dimethoate, fenitrothion, pirimiphosmethyl and profenofos, respectively. The in vivo dose-effect relationships were assessed using chlorfenvinphos and carbaryl at different concentrations that ranged from 0.0003 to 0.06 microM and 0.0005 to 0.05 microM, respectively. Acetylcholinesterase activities were comparable in plasma, and eye and brain homogenates from control and carbaryl-exposed fish. Following exposure of fish to chlorfenvinphos at concentrations above 0.03 microM, a significant inhibition of AChE activities in plasma (84%) and eye homogenate (50%) was observed. The AChE activities in brain homogenate were comparable between chlorfenvinphos-exposed fish and controls. Because carbaryl cause reversible inhibition of AChE activities was found to be more potent than chlorfenvinphos that cause irreversible inhibition following in vitro exposure. Contrary, carbaryl was less potent than chlorfenvinphos after in vivo exposure possibly due to more rapid biotransformation of carbaryl than chlorfenvinphos. Findings from this study have demonstrated that inhibition of AChE activity in C. gariepinus is a useful biomarker in assessing aquatic environment contaminated by anticholinesterases.