Inhibition Of Brain Cholinesterase Activity Research Articles

Novel approaches to mitigating parathion toxicity: targeting cytochrome P450-mediated metabolism with menadione.

Accidental or intentional exposures to parathion, an organophosphorus (OP) pesticide, can cause severe poisoning in humans. Parathion toxicity is dependent on its metabolism by the cytochrome P450 (CYP) system to paraoxon (diethyl 4-nitrophenyl phosphate), a highly poisonous nerve agent and potent inhibitor of acetylcholinesterase. We have been investigating inhibitors of CYP-mediated bioactivation of OPs as a method of preventing or reversing progressive parathion toxicity. It is well recognized that NADPH-cytochrome P450 reductase, an enzyme required for the transfer of electrons to CYPs, mediates chemical redox cycling. In this process, the enzyme diverts electrons from CYPs to support chemical redox cycling, which results in inhibition of CYP-mediated biotransformation. Using menadione as the redox-cycling chemical, we discovered that this enzymatic reaction blocks metabolic activation of parathion in rat and human liver microsomes and in recombinant CYPs important to parathion metabolism, including CYP1A2, CYP2B6, and CYP3A4. Administration of menadione to rats reduces metabolism of parathion, as well as parathion-induced inhibition of brain cholinesterase activity. This resulted in inhibition of parathion neurotoxicity. Menadione has relatively low toxicity and is approved by the Food and Drug Administration for other indications. Its ability to block parathion metabolism makes it an attractive therapeutic candidate to mitigate parathion-induced neurotoxicity.

Vitamin K3 (menadione) redox cycling inhibits cytochrome P450-mediated metabolism and inhibits parathion intoxication

Parathion, a widely used organophosphate insecticide, is considered a high priority chemical threat. Parathion toxicity is dependent on its metabolism by the cytochrome P450 system to paraoxon (diethyl 4-nitrophenyl phosphate), a cytotoxic metabolite. As an effective inhibitor of cholinesterases, paraoxon causes the accumulation of acetylcholine in synapses and overstimulation of nicotinic and muscarinic cholinergic receptors, leading to characteristic signs of organophosphate poisoning. Inhibition of parathion metabolism to paraoxon represents a potential approach to counter parathion toxicity. Herein, we demonstrate that menadione (methyl-1,4-naphthoquinone, vitamin K3) is a potent inhibitor of cytochrome P450-mediated metabolism of parathion. Menadione is active in redox cycling, a reaction mediated by NADPH-cytochrome P450 reductase that preferentially uses electrons from NADPH at the expense of their supply to the P450s. Using human recombinant CYP 1A2, 2B6, 3A4 and human liver microsomes, menadione was found to inhibit the formation of paraoxon from parathion. Administration of menadione bisulfite (40mg/kg, ip) to rats also reduced parathion-induced inhibition of brain cholinesterase activity, as well as parathion-induced tremors and the progression of other signs and symptoms of parathion poisoning. These data suggest that redox cycling compounds, such as menadione, have the potential to effectively mitigate the toxicity of organophosphorus pesticides including parathion which require cytochrome P450-mediated activation.

Prenatal nicotine alters the developmental neurotoxicity of postnatal chlorpyrifos directed toward cholinergic systems: better, worse, or just "different?".

This study examines whether prenatal nicotine exposure sensitizes the developing brain to subsequent developmental neurotoxicity evoked by chlorpyrifos, a commonly-used insecticide. We gave nicotine to pregnant rats throughout gestation at a dose (3mg/kg/day) producing plasma levels typical of smokers; offspring were then given chlorpyrifos on postnatal days 1-4, at a dose (1mg/kg) that produces minimally-detectable inhibition of brain cholinesterase activity. We evaluated indices for acetylcholine (ACh) synaptic function throughout adolescence, young adulthood and later adulthood, in brain regions possessing the majority of ACh projections and cell bodies; we measured nicotinic ACh receptor binding, hemicholinium-3 binding to the presynaptic choline transporter and choline acetyltransferase activity, all known targets for the adverse developmental effects of nicotine and chlorpyrifos given individually. By itself nicotine elicited overall upregulation of the ACh markers, albeit with selective differences by sex, region and age. Likewise, chlorpyrifos alone had highly sex-selective effects. Importantly, all the effects showed temporal progression between adolescence and adulthood, pointing to ongoing synaptic changes rather than just persistence after an initial injury. Prenatal nicotine administration altered the responses to chlorpyrifos in a consistent pattern for all three markers, lowering values relative to those of the individual treatments or to those expected from simple additive effects of nicotine and chlorpyrifos. The combination produced global interference with emergence of the ACh phenotype, an effect not seen with nicotine or chlorpyrifos alone. Given that human exposures to nicotine and chlorpyrifos are widespread, our results point to the creation of a subpopulation with heightened vulnerability.

Effect of ethanolic extract of Acacia auriculiformis leaves on learning and memory in rats.

Background:The effects and benefits of Acacia auriculiformis on health are not well established. This study was planned to evaluate the effect of ethanolic extract of Acacia auriculiformis leaves on learning and memory in rats.Materials and Methods:Learning and memory were evaluated using passive avoidance paradigm and rewarded alternation test (T-maze) after the oral administration of two doses (200mg/kg and 400mg/kg) of ethanolic extract of Acacia auriculiformis with rivastigmine as positive control. Forty eight rats were divided into 4 groups in each study model. Estimation of brain cholinesterase activity was done to substantiate the results of the above mentioned tests. Data was analyzed using one way Analysis of Variance (ANOVA) followed by Tukey's post-hoc test using GraphPad InStat software, version 3.06.Results:The extract produced a dose-dependent improvement in the memory score namely the step through latency in passive avoidance model (P < 0.001) and the percentage of correct responses in rewarded alternation test (P < 0.05). Dose-dependent inhibition of brain cholinesterase activity (P < 0.001) was also noted.Conclusion:The acetylcholinesterase inhibiting property of Acacia auriculiformis contributes to its memory enhancing potential. Further large scale studies are required to elucidate its benefits on cognitive function. This may offer a promising new option for the treatment of dementia and other cognitive deficits.

Prenatal dexamethasone augments the neurobehavioral teratology of chlorpyrifos: Significance for maternal stress and preterm labor

Glucocorticoids are the consensus treatment given in preterm labor and are also elevated by maternal stress; organophosphate exposures are virtually ubiquitous, so human developmental coexposures to these two agents are common. This study explores how prenatal dexamethasone exposure modifies the neurobehavioral teratology of chlorpyrifos, one of the most widely used organophosphates. We administered dexamethasone to pregnant rats on gestational days 17–19 at a standard therapeutic dose (0.2mg/kg); offspring were then given chlorpyrifos on postnatal days 1–4, at a dose (1mg/kg) that produces barely-detectable (<10%) inhibition of brain cholinesterase activity. Dexamethasone did not alter brain chlorpyrifos concentrations, nor did either agent alone or in combination affect brain thyroxine levels. Assessments were carried out from adolescence through adulthood encompassing T-maze alternation, Figure 8 maze (locomotor activity, habituation), novelty-suppressed feeding and novel object recognition tests. For behaviors where chlorpyrifos or dexamethasone individually had small effects, the dual exposure produced larger, significant effects that reflected additivity (locomotor activity, novelty-suppressed feeding, novel object recognition). Where the individual effects were in opposite directions or were restricted to only one agent, we found enhancement of chlorpyrifos' effects by prenatal dexamethasone (habituation). Finally, for behaviors where controls displayed a normal sex difference in performance, the combined treatment either eliminated or reversed the difference (locomotor activity, novel object recognition). Combined exposure to dexamethasone and chlorpyrifos results in a worsened neurobehavioral outcome, providing a proof-of-principle that prenatal glucocorticoids can create a subpopulation with enhanced vulnerability to environmental toxicants.

Prenatal dexamethasone augments the sex-selective developmental neurotoxicity of chlorpyrifos: Implications for vulnerability after pharmacotherapy for preterm labor

Glucocorticoids are routinely given in preterm labor and are also elevated by maternal stress; organophosphate exposures are virtually ubiquitous, so coexposures to these two agents are pervasive. We administered dexamethasone to pregnant rats on gestational days 17–19 at a standard therapeutic dose (0.2mg/kg); offspring were then given chlorpyrifos on postnatal days 1–4, at a dose (1mg/kg) that produces barely-detectable (<10%) inhibition of brain cholinesterase activity. We evaluated indices for acetylcholine (ACh) synaptic function throughout adolescence, young adulthood and later adulthood, in brain regions possessing the majority of ACh projections and cell bodies; we measured nicotinic ACh receptor binding, hemicholinium-3 binding to the presynaptic choline transporter and choline acetyltransferase activity, all known targets for the adverse developmental effects of dexamethasone and chlorpyrifos given individually. Dexamethasone did not enhance the systemic toxicity of chlorpyrifos, as evidenced by weight gain and measurements of cholinesterase inhibition during chlorpyrifos treatment. Nevertheless, it enhanced the loss of presynaptic ACh function selectively in females, who ordinarily show sparing of organophosphate developmental neurotoxicity relative to males. Females receiving the combined treatment showed decrements in choline transporter binding and choline acetyltransferase activity that were unique (not found with either treatment alone), as well as additive decrements in nicotinic receptor binding. On the other hand, males given dexamethasone showed no augmentation of the effects of chlorpyrifos. Our findings indicate that prior dexamethasone exposure could create a subpopulation that is especially vulnerable to the adverse effects of organophosphates or other developmental neurotoxicants.

Depression of the photic after discharge of flash evoked potentials by physostigmine, carbaryl and propoxur, and the relationship to inhibition of brain cholinesterase

The effects of N-methyl carbamate pesticides on the photic after discharge (PhAD) of flash evoked potentials (FEPs) and the relationship between inhibition of brain cholinesterase (ChE) activity and the PhAD were evaluated. FEPs were recorded in Long Evans rats treated with physostigmine (s.c.) 0, 0.05, 0.1, 0.2 or 0.3 mg/kg (free base), in an ascorbic acid/saline vehicle, carbaryl (p.o.) 0, 1, 3, 10, 30, 50 or 75 mg/kg, or propoxur (p.o.) 0, 0.3, 3, 10, 20, 30, or 40 mg/kg in a corn oil vehicle. Physostigmine served as positive control based on literature data. Early (e.g. peak N 36) and late FEP components (peak N 166 and PhAD) are related to the initial retino-geniculate afferent volley and higher cortical processing of visual information, respectively. Compared to controls, the PhAD duration decreased following treatment with 0.1 and 0.3 mg/kg physostigmine, 75 mg/kg carbaryl or 30 mg/kg propoxur. Lesser changes were noted in FEP amplitudes or peak latencies. Treatment with 0.2 or 0.3 mg/kg physostigmine increased peak N 36 latency. Peak N 166 latency increased only following exposure to 40 mg/kg propoxur. None of the compounds altered peak N 36 or N 166 amplitudes. Hypothermia was observed at doses greater than 0.05 mg/kg physostigmine, at 30 or 50 mg/kg carbaryl, and after treatment with 10, 20 or 40 mg/kg propoxur. Inhibition of brain ChE activity occurred at dosages greater than 0.05 mg/kg physostigmine, 1 mg/kg carbaryl, and 0.3 mg/kg propoxur. Linear regression analysis indicated that the decrease in PhAD duration correlated with decrease in brain ChE activity. The results indicate that at 30 min after treatment, inhibition of brain ChE activity did not affect cortical processing of the input from the retino-geniculate volley (evidenced by unaltered peak N 36 amplitude). However, the data suggest that disruption of cortical processing of visual signals related to FEP late components, as indicated by depression of the PhAD, was related to inhibition of brain ChE activity.

Pharmacological Evidence for the Potential of Daucus carota in the Management of Cognitive Dysfunctions

The present study was aimed at investigating the effects of Daucus carota seeds on cognitive functions, total serum cholesterol levels and brain cholinesterase activity in mice. The ethanolic extract of Daucus carota seeds (DCE) was administered orally in three doses (100, 200, 400 mg/kg) for seven successive days to different groups of young and aged mice. Elevated plus maze and passive avoidance apparatus served as the exteroceptive behavioral models for testing memory. Diazepam-, scopolamine- and ageing-induced amnesia served as the interoceptive behavioral models. DCE (200, 400 mg/kg, p.o.) showed significant improvement in memory scores of young and aged mice. The extent of memory improvement evoked by DCE was 23% at the dose of 200 mg/kg and 35% at the dose of 400 mg/kg in young mice using elevated plus maze. Similarly, significant improvements in memory scores were observed using passive avoidance apparatus and aged mice. Furthermore, DCE reversed the amnesia induced by scopolamine (0.4 mg/kg, i.p.) and diazepam (1 mg/kg, i.p.). Daucus carota extract (200, 400 mg/kg, p.o.) reduced significantly the brain acetylcholinesterase activity and cholesterol levels in young and aged mice. The extent of inhibition of brain cholinesterase activity evoked by DCE at the dose of 400 mg/kg was 22% in young and 19% in aged mice. There was a remarkable reduction in total cholesterol level as well, to the extent of 23% in young and 21% in aged animals with this dose of DCE. Therefore, DCE may prove to be a useful remedy for the management of cognitive dysfunctions on account of its multifarious beneficial effects such as, memory improving property, cholesterol lowering property and anticholinesterase activity.

In Vitro and in Vivo Cholinesterase Inhibition in Lacertides by Phosphonate- and Phosphorothioate-Type Organophosphates

The acute toxicity of two organophosphorus (OP) insecticides (trichlorphon and parathion) was assessed in lizards by in vitro and in vivo experiments. Serum and brain cholinesterase (ChE) activities were measured in lizards treated orally with the OPs. Inhibitory effects of trichlorphon were measured 24 h after exposure and those of parathion were determined according to a treatment/sampling schedule designed to investigate the toxic effects at different times of day, since parathion undergoes bioactivation before it inhibits brain ChE. A batch of lizards (batch A) was treated at 10 am and sampled 6 h later; a second batch was exposed to parathion at 4 pm and sampled 24 h later (batch B); a third was treated at 4 pm and ChE measured 6 h later (batch C). In vitro assays showed that brain ChE activity was more sensitive to paraoxon (IC50 = 2.2 · 10−6 M) and dichlorvos (IC50 = 3.2 · 10−5 M) than their parent compounds. Treatment with trichlorphon caused a significant dose-related inhibition of brain and serum ChE activities at doses of 50 and 100 mg/kg. Inhibition of ChEs by parathion depended more on administration time than dose. Significant dose-dependent inhibition of serum ChE was found in lizards of batches A and B. Brain ChE activity was depressed at all treatment doses (> 50% inhibition) in batch A and at the highest dose in batch B (70%). Afternoon treatment and sampling caused dose-related inhibition of serum ChE but no inhibition of brain ChE activity, irrespective of parathion dose. Attempts to correlate the two ChE activities showed a linear relationship in lizards treated with trichlorphon and a nonlinear relationship in lizards treated with parathion. This may be due to bioactivation of parathion to its oxon analogue paraoxon, which inhibits brain ChE activity, and to the buffer effect of serum ChE, which lessens the impact of the organophosphate on the central nervous system. These results show (i) that reptiles have a high resistance to organophosphorus insecticides compared to other vertebrates (e.g., birds and mammals); (ii) that in lacertides the toxic effects of phosphorothioate-type organophosphates may depend strongly on the time of day that exposure occurs; (iii) that a relationship exist between brain and serum ChEs in lizards treated with OP insecticides, suggesting that serum ChE could be used as a surrogate biomarker of effect.

Effectiveness of procyclidine in combination with carbamate prophylactics against diisopropylfluorophosphate poisoning

The protective effect of cholinolytics such as procyclidine and atropine, in combination with carbamate prophylactics, against diisopropylfluorophosphate poisoning was examined in mice. Doses of carbamates were optimized, based on the maximum sign-free dose, the time course of cholinesterase inhibition and the protective potential against diisopropylfluorophosphate poisoning. Centrally-active physostigmine was more toxic than centrally-inactive pyridostigmine and the toxic signs of carbamates appeared to be closely related to the level of inhibition of brain cholinesterase activity. In combination with atropine, physostigmine was more effective than pyridostigmine in protecting mice intoxicated with diisopropylfluorophosphate. Moreover, centrally-active atropine sulfate was a more effective co-antidote to carbamates than centrally-inactive atropine methylnitrate. The most prominent protection was achieved with the combination of carbamates and procyclidine, a centrally-active cholinolytic showing anticonvulsion, which was also observed to prevent diisopropylfluorophosphate-induced convulsions ( Kim et al., 1997). Taken together, it is suggested that procyclidine could be a possible substitute for atropine as an antidote to diisopropylfluorophosphate poisoning.

Subchronic neurotoxicity screening studies with six organophosphate insecticides: an assessment of behavior and morphology relative to cholinesterase inhibition.

Sulprofos, disulfoton, azinphos-methyl, methamidophos, trichlorfon, and tebupirimphos were screened for neurotoxic potential, in accordance with U.S. EPA (FIFRA) requirements. Each organophosphate was administered through the diet for 13 weeks to separate groups of Fischer 344 rats at four dose levels, including a vehicle control. For each study, 12 rats/sex/dietary level were tested using a functional observational battery (FOB), automated measures of activity (figure-8 maze), and detailed clinical observations, with half of the animals perfused at term for microscopic examination of neural and muscle tissues. Separate groups of satellite animals (6/sex/dietary level) were used to measure the effect of each treatment on plasma, erythrocyte (RBC), and brain cholinesterase (ChE) activity. The results show that measures of ChE activity were consistently the most sensitive indices of exposure and assisted in the interpretation of findings. All treatment-related neurobehavioral findings were ascribed to cholinergic toxicity, occurring only at dietary levels that produced more than 20% inhibition of plasma, RBC, and brain ChE activity. Neurobehavioral tests provided no evidence of additional cumulative toxicity after 8 weeks of treatment. The FOB and motor activity findings did not alter the conclusions and generally did not reduce the neurobehavioral no-observed-effect level (NOEL) for any of the six compounds, relative to the results from detailed clinical observations as conducted in these studies. The one exception occurred with tebupirimphos, where the NOEL for motor activity was one dose level lower than the NOEL for the FOB and clinical observations. These results support the value of detailed clinical observations to screen for the neurotoxic potential of organophosphates and a general standard of more than 20% inhibition of brain ChE activity for cholinergic neurotoxicity.

Effects of exposure to carbofuran-sprayed vegetation on the behavior, cholinesterase activity and growth of mallard ducklings(anas platyrhynchos)

Mallard (Anas platyrhynchos) ducklings were led for distances of 50, 150 and 300 m through plots of upland vegetation that had been sprayed with a flowable formulation of carbofuran at rates of 0, 132 and 264 g active ingredient/ha. Duckling behavior was observed and quantified immediately after exposure by using an approach-response apparatus. Brain and plasma cholinesterase (ChE) activities were determined for a subsample of ducklings from each treatment group; the remaining ducklings were monitored for growth effects. No mortality occurred during field exposure, but toxic signs were observed in ducklings moving 150 m, 17 and 100% of the ducklings had difficulty keeping pace due to anti-ChE toxicosis in the low and high spray plots, respectively. Duckling approach-response behavior became significantly slower with increasing spray rate and exposure distance (p = 0.0001). Inhibition of brain ChE activity was directly related to both spray rate and exposure distance; ducklings receiving maximum carbofuran exposure had 29% of the mean ChE activities of controls. Plasma ChE activity was inversely related to spray rate but was not affected by exposure distance and did not correlate well with brain ChE activity (R2 = 0.36). Subsequent growth rates were not affected in a dose-related manner.

Routes of uptake and their relative contribution to the toxicologic response of Northern bobwhite (Colinus virginianus) to an organophosphate pesticide

AbstractIngestion of contaminated food is considered the primary route of exposure in birds to agricultural chemicals. Routes of exposure other than ingestion are not often considered in risk assessments of agricultural chemicals to avian wildlife. However, recent studies demonstrated anorexic or avoidance behaviors in birds exposed to organophosphate (OP) insecticides. These behaviors would tend to limit exposure if ingestion alone were considered. The contribution, if any, of dermal, preening, and respiratory pathways to the exposure of birds to pesticides under field conditions is unknown. In addition, oral exposures are currently assessed in artificial environments that do not reflect real‐life exposure scenarios. To determine the relative contribution of these pathways and to assess exposures under ecological conditions, 270 northern bobwhite (Colinus virginianus) were exposed to simulated aerial crop applications of methyl parathion in an environmentally controlled wind tunnel. The wind tunnel environment consisted of a 25‐cm cotton plant canopy, a 5‐cm‐thick floor of silt‐loam, a temperature of 25°C, 50% RH, UV intensity similar to summer sunlight, and a wind speed of 3.2 km/h. Inhalation, preening, and dermal routes were isolated in groups of birds exposed to each application. Five birds from each group were collected at 1, 4, 8, 24, and 48 h post‐spray to determine cholinesterase (ChE) response to the exposures. Contaminated and uncontaminated darkling beetle (Tenebrio molitar) larvae were presented to free‐ranging sprayed birds in the wind tunnel to assess oral uptake. ChE response was determined at 4, 8, 24, and 48 h postspray. All exposures were replicated. All four routes contributed to the inhibition of brain ChE at different post‐spray periods. Dermal uptake and preening were major contributors to the overall dose and toxic response of birds to methyl parathion. Inhalation was the major route of exposure at 1 h post‐spray. At 4 h post‐spray, uptake through preening caused the greatest inhibition of brain ChE activity. Oral ingestion resulted in less than 20% inhibition of brain ChE during the test. Routes of uptake in order of contribution to toxicologic response from 8 to 48 h post‐spray were dermal &gt; preening ≥ oral &gt; inhalation.

Enhancement of malathion toxicity to the hybrid red-legged partridge following exposure to prochloraz

The agricultural fungicide, prochloraz, which inhibits ergosterol biosynthesis in fungi, was shown to be a potent inducer of liver enzymes in the hybrid red-legged partridge. Pretreatment with a single oral dose of 180 mg/kg resulted in a dramatic potentiation of the toxicity of the organophosphorous insecticide malathion: three out of four prochloraz-pretreated birds died within 10 min from acute poisoning (82–100% inhibition of brain cholinesterase activity) following intraperitoneal administration of 3.3 or 16.7 mg/kg malathion, whereas no significant inhibition of serum cholinesterase activity was seen in control birds 1 hr after administration of the insecticide. Prochloraz pretreatment also resulted in a significant increase in susceptibility to low oral doses of malathion. In vitro metabolism of [ 14C]malathion by microsomes from control birds resulted largely in the formation of the malathion monoacid, whereas the anticholinesterase, malaoxon, was the main metabolite generated by microsomes from prochloraz-pretreated birds. The dramatic amplification of malathion toxicity following induction with prochloraz is attributed to an increased activation of malathion to its active metabolite malaoxon by one or more induced monooxygenase forms; inhibition of “B”-esterase activity by malaoxon may contribute to this amplification of toxicity.

Toxicity of abate® 4E (temephos) in mallard ducklings and the influence of cold

AbstractDiets mixed to contain 0, 0.1, 1.0, 10 and 100 ppm temephos (determined chemically to contain less than 0.5, less than 0.5, 0.89, 6.0 and 59 ppm temephos, respectively) in an Abate® 4E formulation, were fed to mallard (Anas platyrhynchos) ducklings for 7 d. During this period, half of the ducklings in each dietary treatment group were housed in a heated brooder (39 to 41 °C) and half were housed in an unheated brooder (10 to 18°C). Mortality in all dietary groups in the unheated brooder was higher than in the heated brooder. High temephos‐related mortality occurred in the 100 ppm group in the unheated brooder but not in any other diet‐temperature groups. Ingestion of the 100 ppm temephos diet inhibited plasma Cholinesterase (ChE) activity and elevated plasma corticosterone concentration and creatine phosphokinase activity, but other selected plasma chemistries were not affected in a dose‐related manner. Brain ChE activity was depressed only in the 100 ppm dietary groups; maximum inhibition of brain ChE activity was 48%. These findings suggest that diets containing up to 10 ppm temephos do not directly affect duckling survival during the first week of life and that the toxicity of 100 ppm temephos is markedly enhanced by cold.

The effect of methyl parathion on susceptibility of bobwhite quail ( Colinus virginianus) to domestic cat predation

Bobwhite quail (Colinus virginianus) that received either 0, 2, 4, or 8 mg/kg methyl parathion (O,O-dimethyl o-p-nitrophenyl phosphorothioate) treatment were investigated as to their susceptibility to predation by a cat (Felis domesticus) predator. Four hours after receiving methyl parathion (MP), physical activity levels were monitored in quail and included the number of seconds spent still, walking, running, or flying before and after a cat was introduced into an experimental arena. The cholinesterase (ChE) activity for each quail on experiment was determined. Quail that were captured exhibited significantly greater inhibition of brain ChE activity and spent significantly more time being still than noncaptured birds. Birds receiving MP at 8 mg/kg spent more seconds being still than those in other treatment groups and had ChE activity reduced to 42.8% of normal activity. There was a tendency for quail at increasing treatment levels to be more susceptible to capture by the cat predator. The neurological and behavioral effects of methyl parathion may have important ecological ramifications.

The effects of organophosphate-induced cholinergic stimulation on the antibody response to sheep erythrocytes in inbred mice

In a previous study, we demonstrated that parathion suppressed both the primary IgM and IgG response to sheep erythrocytes (SRC) in inbred and outbred mice ( G. P. Casale, S. D. Cohen, and R. A. DiCapua, 1982, Toxicologist 2, 94). Suppression occurred after a dosage which produced cholinergic effects but was absent after a lower dosage which did not produce cholinergic signs. This information suggested that immunosuppression might be mediated indirectly as a result of toxic chemical stress. The present study evaluated the relationship between the anticholinesterase action of parathion, malathion, and dichlorvos (DDVP) and their effects on the primary humoral response to SRC. Male C57B1 6 mice were given a single dose of parathion (16 mg/kg, po), malathion (720 mg/kg, po), or DDVP (120 mg/kg, po) 2 days after immunization with SRC. Two days later, tissues were removed for cholinesterase (CHE) assay and enumeration of splenic antibody-forming cells (PFC). All three compounds produced moderate to severe cholinergic poisoning. DDVP produced cholinergic signs beginning 1 2 hr after dosing and lasting 1 2 to 1 hr. This profile was associated with a rapid but transient inhibition of brain CHE activity. In contrast, malathion and parathion produced prolonged cholinergic poisoning (4 to 7 hr) and prolonged suppression of brain CHE activity. All three compounds suppressed the primary IgM response. However, when they were given as multiple lower doses, none of the compounds suppressed the primary IgG response. These latter treatments produced no cholinergic signs. The cholinomimetic agent, arecoline (65 mg/kg, ip) produced a short-lived cholinergic crisis but no IgM suppression. Sustained-release arecoline produced prolonged cholinergic poisoning (3 to 5 hr) and reduced the number of IgM PFC to 50% of control. These results demonstrated that organophosphate-induced immunosuppression was associated with severe cholinergic stimulation. The immunosuppression may result from direct action of acetylcholine upon the immune system or it may be secondary to the toxic chemical stress associated with cholinergic poisoning.

Inhibitory effect of leptophos on carboxylesterase (isocarboxazid amidase) in rat liver

The organophosphate insecticide, leptophos, inhibited rat liver isocarboxazid amidase (ISOCase) activity to 20% of control at 5.0 mg/kg 1 h after administration, but at this dose brain cholinesterase (ChE) activity was not affected. The activity of ISOCase decreased to 29 and 0% of control 24 h after treatment with leptophos at doses of 2.5 and 5.0 mg/kg, respectively. With repeated administration of leptophos at a dose of 1 mg/kg for 10 days, ISOCase activity decreased to 34% of control on day 1 and the inhibition increased to 85% on day 10 without inhibition of brain ChE activity. After cessation of three successive daily doses (1 mg/kg/day), the ISOCase activity was gradually restored near to control levels in 8 days. Pretreatment with carboxylesterase inhibitors, triorthocresylphosphate (TOCP) and bis- p-nitrophenylphosphate (BNPP), potentiated the inhibition of brain ChE by leptophos, suggesting that ISOCase might take a role in leptophos detoxification.

Diagnostic criteria for carbaryl poisoning in sheep.

Sheep were used to study the effect of carbaryl in liver, heart, and brain and on cholinesterase activity. Carbaryl residues greater than or equal to 0.01 ppm in the brain were present in all sheep dying after dosing. Sheep dying acutely had higher levels of carbaryl and greater than or equal to 50% inhibition of brain cholinesterase activity, while sheep with prolonged death had lower carbaryl levels and less cholinesterase inhibition. Prolonged deaths were associated with pulmonary embarrassment, enteritis, hyperthermia, and metabolic acidosis. Carbaryl was rapidly degraded in stored blood samples but was stable in dead brain tissue.

Recovery of brain and plasma cholinesterase activities in ducklings exposed to organophosphorus pesticides

Brain and plasma cholinesterase (ChE) activities were determined for mallard ducklings (Anas platyrhynchos) exposed to dicrotophos and fenthion. Recovery rates of brain ChE did not differ between ducklings administered a single oral dose vs. a 2-week dietary dose of these organophosphates. Exposure to the organophosphates, followed by recovery of brain ChE, did not significantly affect the degree of brain ChE inhibition or the recovery of ChE activity at a subsequent exposure. Recovery of brain ChE activity followed the general model Y = a + b(logX) with rapid recovery to about 50% of normal, followed by a slower rate of recovery until normal ChE activity levels were attained. Fenthion and dicrotophos-inhibited brain ChE were only slightly reactivated in vitro by pyridine-2-aldoxime methiodide, which suggested that spontaneous reactivation was not a primary method of recovery of ChE activity. Recovery of brain ChE activity can be modeled for interpretation of sublethal inhibition of brain ChE activities in wild birds following environmental applications of organophosphates. Plasma ChE activity is inferior to brain ChE activity for environmental monitoring, because of its rapid recovery and large degree of variation among individuals.