Whole Blood Cholinesterase Activity Research Articles

Plasma cholinesterase activity is influenced by interactive effect between omethoate exposure and CYP2E1 polymorphisms

The aim of this study was to explore the association between metabolizing enzyme gene polymorphisms and the decrease in cholinesterase activity induced by omethoate exposure. A total of 180 workers exposed to omethoate over an extended period were recruited along with 115 healthy controls. Cholinesterase activity in whole blood, erythrocyte, and plasma was detected using acetylthiocholine and the dithio-bis-(nitrobenzoic acid) method. Six polymorphic loci of GSTT1(+/-), GSTM1(+/-), GSTP1 rs1695, CYP2E1 rs6413432, CYP2E1 rs3813867, and PON2 rs12026 were detected by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP). The gene-environment interactions were analyzed using the generalized linear model method. The cholinesterase activity of erythrocyte and plasma in the exposure group was significantly lower than that in the control group (P < 0.001) in general. The plasma cholinesterase activity in the TT + AT genotype in CYP2E1 rs6413432 was lower than that in the AA genotype in the exposure group (P = 0.016). Interaction between the AA genotype in CYP2E1 rs6413432 and omethoate exposure had a significant effect on plasma cholinesterase activity (P = 0.079). The decrease in plasma cholinesterase activity was associated with interaction between the AA genotypes in rs6413432 and omethoate exposure.

Decrease in Cellular Nanovesicles Concentration in Blood of Athletes More Than 15 Hours After Marathon.

IntroductionCellular nanovesicles (CNVs), that are shed from cells, have been recognized as promising indicators of health status. We analyzed the effect of long-distance running on concentration of CNVs, along with some standard blood parameters, in 27 athletes two days before and >15 hours after physical effort.MethodsCNVs were isolated by repetitive centrifugation and washing of samples, and assessed by flow cytometry. Cholinesterase (ChE) and glutathione S-transferase (GST) activity were measured spectrophotometrically. Interleukin 6 (IL-6) and tumor necrosis factor-α (TNF-α) concentrations were measured using enzyme-linked immunosorbent assay (ELISA). C-reactive protein (CRP) was measured with immunoturbidimetric determination and lipidogram parameters were measured by enzymatic colorimetric assay. Flow cytometry was used for blood cell count and mean platelet volume (MPV) measurement.ResultsMore than 15 hours after physical effort a decrease was found in CNVs’ concentration in isolates from blood (46%; p<0.05), in ChE activity in whole blood (47%; p<0.001), in plasma (34%; p<0.01), and in erythrocyte suspension (54%; p<0.001), as well as in GST activity in erythrocyte suspension (16%; p<0.01) and in IL-6 concentration in plasma (63%; p<0.05). We found no change in GST activity in plasma and in TNF-α concentration in plasma. Correlations (>0.8; p<0.001) between CNVs’ concentration and ChE activity, and GST activity, respectively, in erythrocyte suspension were found.ConclusionWe found that >15 hours post-physical effort, CNVs’ concentration was below the initial value, concomitant with other measured parameters: ChE and GST activity as well as IL-6 concentration, indicating a favorable effect of physical effort on health status. CNVs’ concentration and ChE activity in isolates from peripheral blood proved to have potential as indicators of the response of the human body to inflammation after physical effort. Physical activity should be considered as an important factor in preparation of subjects for blood sampling in procedures focusing on CNV-containing diagnostic and therapeutic compounds.

Determining a threshold sub-acute dose leading to minimal physiological alterations following prolonged exposure to the nerve agent VX in rats.

VX, a potent inhibitor of cholinesterase (ChE), is considered as one of the most toxic, persistent and least volatile nerve agents. VX is absorbed in various environmental surfaces and is gradually released long after its initial dispersal. Its toxicity is mainly caused by disrupting central and peripheral cholinergic nervous system activity, leading to potential long-term detrimental effects on health. The primary objective of the present study was to assess the threshold VX dose leading to minimal physiological alterations following prolonged VX exposure. Characterization of such a threshold is crucial for dealing with unresolved operative dilemmas such as when it is safe enough to resettle a population that has been evacuated from a VX-contaminated area. Rats, continuously exposed to various doses of VX (0.225-45µg/kg/day) for 4weeks via implanted mini-osmotic pumps, showed a dose-dependent and continuous decrease in ChE activity in whole blood, brain and muscles, ranging between 20 and 100%. Exposure to 13.5µg/kg/day led to a stable low ChE activity level (~20%), accompanied by transient and negligible electrocorticogram spectral power transformations, especially in the theta and alpha brain wave frequencies, and a significant decrease in total brain M2 receptor density. These changes were neither accompanied by observable signs of intoxication nor by changes in motor function, circadian rhythm or TSPO level (a reliable marker of brain damage). Following exposure to lower doses of 2.25 and 0.225µg/kg/day, the only change measured was a reduction in ChE activity of 60 and 20%, respectively. Based on these results, we delineate ChE inhibition as the physiological measure most susceptible to alterations following prolonged VX exposure, and determine for the first time the threshold sub-acute VX dose for minimal physiological effects (up to 20% reduction in ChE activity) in the rat as 0.225µg/kg/day.

Cholinergic enzymes and inflammatory markers in rats infected by Sporothrix schenckii

The aim of this study was to evaluate the cholinesterase activity in serum, whole blood, and lymphocytes, as well as to verify its relation to immune response in rats experimentally infected by Sporothrix schenckii. For this study, 63 Wistar rats (Rattus norvegicus), male, adult were divided into three groups: the negative control group (GC: n = 21), the group infected subcutaneously (GSC: n = 21), and the group infected intraperitoneally (GIP: n = 21). The groups were divided into subgroups and the following variables were evaluated at 15, 30, and 40 days post-infection (PI): acetylcholinesterase (AChE) activity in lymphocytes and whole blood, butyrylcholinesterase (BChE) activity in serum, cytokines levels (IL-1, IL-6, TNFα, and INF-γ), immunoglobulins levels (IgA, IgG, IgM, and IgE), and protein profile by electrophoresis. Both infected groups showed increased levels of inflammatory parameters (P < 0.05) in tissue and inflammatory infiltrates. The activities of AChE in lymphocytes and BChE in serum increased (P < 0.05) significantly in animals from the GSC group on day 40 PI compared to the GC group. Regarding the GIP, there was a marked increase in the AChE activity in lymphocytes on days 30 and 40 PI, and in whole blood on days 15, 30, and 40 PI compared to GC. Furthermore, IL-10, an anti-inflammatory cytokine, was also present in high levels during chronic systemic S. schenckii infections in animals. Therefore, it is concluded that cholinesterase has an important modulatory role in the immune response during granulomatous infection by S. schenckii.

Development and validation of a simple assay for the determination of cholinesterase activity in whole blood of laboratory animals

Current methods for measuring acetylcholinesterase (AChE) activities in whole blood use butyrylcholinesterase (BChE)-selective inhibitors. However, the poor selectivity of these inhibitors results in the inhibition of AChE activity to some degree, leading to errors in reported values. The goal of this study was to develop and validate a simple assay for measuring AChE and BChE activities in whole blood from humans as well as experimental animals. Blood was fractionated into plasma and erythrocytes, and cholinesterase activities were titrated against ethopropazine and (-)-huperzine A to determine the lowest concentration of ethopropazine that inhibited BChE completely without affecting AChE activity and the lowest concentration of (-)-huperzine A that inhibited AChE completely without interfering with BChE activity. Results indicate that 20 µm ethopropazine can be successfully used for the accurate measurement of AChE activity in blood from humans as well as animals. Use of (-)-huperzine A is not required for measuring BChE activity in normal or 'exposed' blood samples. The method was validated for blood from several animal species, including mice, rats, guinea pigs, dogs, minipigs, and African green, cynomolgus and rhesus monkeys. This method is superior to all reported methods, does not require the separation of erythrocyte and plasma fractions, and is suitable for measuring cholinesterase activities in fresh or frozen blood from animals that were exposed to nerve agents or those that were administered high doses of BChE. The method is simple, direct, reproducible, and reliable and can easily be adapted for high-throughput screening of blood samples. Published 2012. This article is a US Government work and is in the public domain in the USA.

Pharmacokinetics and Toxicodynamics of Pralidoxime Effects on Paraoxon-Induced Respiratory Toxicity

Empirical studies suggest that the antidotal effect of pralidoxime depends on plasma concentrations with therapeutic effects associated with concentrations above 4 mg/l. The purpose of this study was to determine the pharmacokinetic-toxicodynamic (PK-TD) relationships for the antidotal effect of pralidoxime on paraoxon-induced toxicity in rats. Diethylparaoxon inactivation of whole-blood cholinesterase activity was studied both in vitro and in male Sprague-Dawley rats. Toxin-induced respiratory effects were measured via whole-body plethysmography in control and pralidoxime-treated animals (50 mg/kg im injection). In the in vitro analysis, cholinesterase reactivation by pralidoxime in blood-poisoned diethylparaoxon (10nM) was proportional to the logarithm of drug concentrations. A mechanism-based TD model was developed, which well described the inhibition of cholinesterases by diethylparaoxon and reactivation with pralidoxime. The in vitro pralidoxime EC(50) was estimated to be 4.67 mg/l. Animals exposed to diethylparaoxon exhibited a decrease in respiratory rate and an increase in expiratory time, and pralidoxime treatment resulted in a rapid complete but transient (< 30 min) correction in respiratory toxicity. In contrast, there was a fast and total reactivation of blood cholinesterase activity over the 210-min study period. The in vitro TD model was extended to capture the time-course of in vivo pralidoxime antidotal effects, which explained the complex relationship between drug exposure and pharmacological response profile. This study provides insights into the role of oxime-rescue of paraoxon-induced toxicity, and the final PK-TD model might prove useful in optimizing the design and development of such therapy.

Occupational Exposure to Pesticides: Blood Cholinesterase Activity in a Farming Community in Ghana

A survey was undertaken to establish the extent of pesticide exposure in a farming community. Cholinesterase (ChE) activity in whole blood was used as a marker for assessing exposure to pesticides. Complete data were gathered for 63 farmers at Akumadan (exposed) and 58 control subjects at Tono, both prominent vegetable-farming communities in Ghana, by means of a questionnaire and blood cholinesterase analyses (acetylcholine assay). Although whole-blood ChE was significantly lower in the exposed than the control participants, it was not significantly correlated with either confounders of age, sex, body weight, and height or high-risks practices. The high-risks practices revealed during the survey included lack of use of personal protective clothing, short reentry intervals, and wrong direction of spraying of pesticides by hand or knapsack sprayer. About 97% of exposed participants had experienced symptoms attributable to pesticide exposure. The frequent symptoms were reported as weakness and headache. There is the need to review safety precautions in the use and application of pesticides in Ghana.

Comparison of methods used for the determination of cholinesterase activity in whole blood

Cholinesterases (ChEs) are classified as either acetylcholinesterase (AChE) or butyrylcholinesterase (BChE) based on their substrate and inhibitor specificity. Organophosphate and carbamate compounds commonly represented by herbicides, pesticides, and nerve gases irreversibly inhibit ChEs. Therefore, exposure to organophosphates and carbamates is normally assessed by measuring ChE activity in blood. There are two approaches for measuring AChE and BChE activity present in whole blood: (1) separating blood into erythrocytes, which contain only AChE, and plasma which contains only BChE, to measure their activity individually, or (2) use a BChE-specific inhibitor to measure the activity of AChE in whole blood. A number of studies have reported the use of different inhibitors for the simultaneous measurement of AChE and BChE activities. However, the inhibitors used for completely inhibiting BChE activity also inhibited AChE activity leading to errors in reported values. The goal of this study was to find the most accurate and simple method for the simultaneous determination of AChE and BChE activity in animal whole blood. Solutions containing human AChE and BChE in various proportions were prepared and AChE and BChE activities were measured using three reported methods. Results demonstrate that ethopropazine and (−) huperzine A appear to be the most specific ChE inhibitors. Preliminary results with human and animal whole blood suggest that 20 μM ethopropazine and 500 nM (−) huperzine A can be used for measuring AChE and BChE activities across species.

Acute Toxic Effects of Inhaled Dichlorvos Vapor on Respiratory Mechanics and Blood Cholinesterase Activity in Guinea Pigs

Using a modified noninvasive volume-displacement plethysmography system, we investigated the effects of inhaled dichlorvos (2,2-dimethyl dichlorovinyl phosphate, or DDVP) vapor on the respiratory mechanics and blood cholinesterase activity of guinea pigs. Data revealed significant dose-dependent changes in several pulmonary parameters. Animals exposed to a DDVP concentration of 35 mg/m3 did not show any significant changes in frequency, tidal volume, or minute ventilation. However, animals exposed to 55 mg/m3 DDVP showed significantly decreased respiratory frequency and significantly increased tidal volume with no significant changes in minute ventilation. Similarly, animals exposed to 75 mg/m3 DDVP showed significantly decreased respiratory frequency along with significantly increased tidal volume. The decreased respiratory frequency was large enough in the high exposure group to offset the increased tidal volume. This effect resulted in significantly decreased minute ventilation by the end of exposure, which remained attenuated 10 min after exposure. An analysis of whole-blood cholinesterase activity revealed significantly decreased activity for both acetylcholinesterase (AChE) and butylcholinesterase (BChE). Peak inhibition occurred for both enzymes at the end of exposure for all three concentrations and rapidly recovered within several minutes of exposure. Analysis of blood samples using gas chromatography–mass spectroscopy (GC-MS) revealed that minute ventilation may only play a minimal role in the dosimetry of inhaled DDVP vapor.

Effects of oral tetrachlorvinphos fly control (Equitrol®) administration in horses: Physiological and behavioural findings

Highly reactive horses may pose risks to humans involved in equestrian activities. Among the factors that may affect horses' reactivity to external stimuli are pesticides used for fly control in equine facilities. The organophosphorus (OP) insecticide tetrachlorvinphos (TCVP) is used as a feed-through larvicide to prevent completion of the fly larval life cycle in horse manure. TCVP exerts its effect by inhibiting the enzyme cholinesterase (ChE) leading to the accumulation of the neurotransmitter acetylcholine (AChE) in synapses of the central and peripheral nervous systems. The aim of the present study was to investigate alterations of whole-blood ChE levels associated with feeding a commercially available product (Equitrol, Farnam Companies, Inc.) to horses for fly control. A second aim was to report neurological, physiological and behavioural findings in addition to profiles of selected immune markers (IFN-gamma, IL-12p40 and COX-2) and serum thyroid hormones during and after a 30-day treatment period of TCVP feeding. The results indicated significant decreases in whole-blood ChE activity and concomitant behavioural alterations, manifested as increased reactivity and decreased controllability in treated horses. No changes were detected in physiological or neurological parameters, immune markers or thyroid hormones in treated (n=6) or control (n=4) horses during the course of the study.

Whole-Blood and Plasma Cholinesterase Levels in Normal Koi (Cyprinus Carpio)

Organophosphate and carbamate insecticides produce their toxic effects by binding with cholinesterase enzymes, resulting in the inhibition of acetylcholine catabolism. Excessive acetylcholine levels cause excessive synaptic neurotransmitter activity. In most animal species, whole blood is the preferred sample for evaluation of cholinesterase activity. Previous reports of cholinesterase activity in carp have involved sampling of serum, plasma, brain, liver, muscle, and heart. Whole-blood samples were obtained from 12 healthy koi (Cyprinus carpio). Whole-blood cholinesterase activity was measured in all 12 samples. Plasma cholinesterase activity was also measured in 4 samples. The overall average of the whole-blood cholinesterases was 0.08 micromol/ml/minute (SD = 0.024). The results for the plasma cholinesterases were similar, with an average of 0.08 micromol/ml/minute (SD = 0.012). Because these reference values are below the minimum detection limit for the method of analysis used, results from blood samples from koi suspected to be suffering from anticholinesterase toxicosis may be uninterpretable.

ACETYLCHOLINESTERASE CONCENTRATIONS IN HEPARINIZED BLOOD OF HISPANIOLAN AMAZON PARROTS (AMAZONA VENTRALIS)

Organophosphate and carbamate pesticides inhibit acetylcholinesterase (AChE) at nerve synapses. Blood samples from 22 Hispaniolan Amazon parrots (Amazona ventralis) were assayed for cholinesterase levels by two different techniques. Using the modified Michel method, the whole-blood cholinesterase activity levels ranged from 0.082 to 0.616 deltapH/hr with a mean value of 0.35 deltapH/hr. A reference range (0.08-0.62 deltapH/hr) for cholinesterase was established in birds. The modified Ellman spectrophotometric method was used to measure AChE activity by adding acetylthiocholine or pseudocholinesterase (plasma cholinesterase) activity by adding butyrylthiocholine. The reference range of the AChE activity using the modified Ellman spectrophotometric method was 0-1.12 micromol/ml/min with a mean of 0.48 micromol/ml/min, and for pseudocholinesterase the range was 0.09-0.98 micromol/ml/min with a mean of 0.53 micromol/ml/min.

Determination of whole blood cholinesterase in different animal species using specific substrates

Whole blood cholinesterase was measured using acetyl-, butyryl- and propionylthiocholine as substrates in 10 healthy adult dogs, cats, horses, pigs, goats, sheep and cows, in order to determine and characterise the cholinesterase activity in whole blood of the main domestic animals. An in vitro exposure test with two anticholinesterase compounds, the organophosphate insecticide coumaphos and the carbamate insecticide imidocarb, was also performed.In whole blood of ruminants and pigs, acetylthiocholine yielded the highest cholinesterase activity and other substrates were poorly hydrolysed; in dogs and cats, although acetylthiocholine showed the highest cholinesterase activity, butyryl- and propionylthiocholine also produced high cholinesterase values; in horses, propionylthiocholine was the substrate that yielded the highest cholinesterase activity, closely followed by butyrylthiocholine. All within- and between-run coefficients of variation observed in whole blood samples were less than 5 and 7 per cent, respectively, except when butyrylthiocholine was used as substrate in ruminant blood samples. Butyryl- and propionylthiocholine were the substrates that yielded higher inhibitions after coumaphos exposure, whereas the use of acetylthiocholine showed the highest cholinesterase inhibition after imidocarb exposure.The use of at least two substrates (acetyl and butyrylthiocholine) is recommended for whole blood cholinesterase analyses in domestic animals since it will allow monitoring of both acetyl- and butyrylcholinesterase activities, respectively, and a more accurate detection of exposure to anticholinesterase compounds. However, acetylthiocholine could be used as a unique substrate for whole blood cholinesterase determination in porcine and ruminant samples since butyrylcholinesterase activity is very low in these species. Additionally, propionylthiocholine could be used as an alternative substrate to butyrylthiocholine in horse whole blood samples.

Automated Spectrophotometric Method Using 2,2’-Dithiodipyridine Acid for Determination of Cholinesterase in Whole Blood

An automated method using 2,2'-dithiodipyridine (2-PDS) as chromophore for determination of whole-blood cholinesterase activity was developed. Assay procedures, optimal concentrations of chromophore and substrate, detection limit, precision, backgrounds, and sensitivity of the method were compared with those of an earlier automated method based on the Ellman method and using 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) as chromophore. The new method has the advantages of automation (resulting in increase throughput rate and decrease in amount of reagents used) and good precision and sensitivity. Sample dilutions also are reduced in the new method because hemoglobin interference is less.

Effect of Time and Storage Temperature on Cholinesterase Activity in Blood from Normal and Organophosphorus Insecticide-Treated Horses

Delays between time of sampling and time of testing are common; therefore, the length of time that blood can be stored at various temperatures was evaluated for effects on cholinesterase activity. Six horses were treated with 16 g of trichlorfon per os, 6 horses were treated with 15 g of dichlorvos per os, and 10 horses were untreated controls. The cholinesterase activity in whole blood from each horse was measured using an adaptation of the Ellman colorimetric method. The blood from each horse was then divided into 3 groups and stored at 5 C (refrigerated), 20 C (room temperature), or 38 C (incubated). Subsequent cholinesterase activities were measured daily and then at weekly intervals. The cholinesterase activities did not significantly increase or were measured daily and then at weekly intervals. The cholinesterase activities did not significantly increase or decrease (P > 0.05) in the blood from the untreated horses until after 1 week for any of the 3 temperature groups. The cholinesterase activities did not significantly increase or decrease (P > 0.05) in the stored blood from the trichlorfon-treated horses for 4 weeks in all 3 temperature groups. The cholinesterase activities significantly increased (P < 0.05) in the stored blood from the dichlorvos-treated horses after 1 week when the blood was refrigerated and by 24 hours when the blood was stored at room temperature or incubated. Therefore, blood from normal or organophosphate-treated horses can be used for cholinesterase evaluation for up to 1 week when stored at 5 C.

PH Method for Determination of Cholinesterase in Whole Blood: Collaborative Study

A collaborative study of an indirect method for determination of cholinesterase activity was conducted with cattle blood. Ten collaborators analyzed 8 samples of varying degrees of cholinesterase inhibition. The sample is diluted with a buffer, and the pH change that results from the hydrolysis of acetylcholine is monitored. Repeatability relative standard deviations (RSDr) ranged from 6.6 to 8.6%, and reproducibility relative standard deviations (RSDR) ranged from 10.7 to 17.3% for mean activities of 0.60-0.40 delta pH/h. All samples that were completely inhibited were detected. The method was adopted first action by AOAC International for determination of cholinesterase activity in whole bovine blood.

Rapid Whole-Blood Cholinesterase Assay with Potential Use for Biological Monitoring During Chemical Weapons Disposal

Abstract A protocol Is described to determine whole-blood cholinesterase activity using an automated highspeed centrifugal analyzer. Throughput rate was increased 10-fold (from 6 to 60 samples/h), and precision, as measured by a reduction In Intra-allquot variation, was significantly Improved (from 4.7 to 3.0%) by using an automated system rather than a manual system. Similar cholinesterase activity was determined for samples analyzed by both the automated and manual protocols, and Interlaboratory comparison Indicated no change In accuracy. The lower limit of detection was estimated to be 0.10 μmol/mL/mln. The 10-fold Increase In throughput facilitates use of the automated protocol when many analyses must be performed in a limited time.

Aldicarb Toxicosis in a Flock of Sheep

Aldicarb toxicosis was diagnosed in 200 sheep that died suddenly. Carbamate insecticide toxicosis was suspected based on observed clinical signs (hypersalivation, diarrhea, urination, paddling, seizures, miosis, and deaths occurring within 1 hour). Tissue samples were submitted from 4 Columbian ewes for pathologic and analytical evaluation. Severe diffuse pulmonary edema was observed on gross and histologic examination. Inhibition of cholinesterase activity in retina (21.2-68.1% of normal activity, n = 3), brain (40.6-45.6% of normal activity, n = 3), and whole blood (27% of normal activity, n = 1) supported a diagnosis of carbamate toxicosis. Reversal of brain and whole blood cholinesterase activities (reactivation factor greater than 1.4) following an in vitro 1 hour incubation at 37 C was also consistent with carbamate poisoning. Aldicarb toxicosis was confirmed following its detection in rumen contents at 1.5, 5.5, and 334 ppm using both high-pressure liquid chromatography with UV detection and gas chromatography with nitrogen/phosphorus detection.

Enzymatic-Spectrophotometric Method for Determination of Cholinesterase Activity in Whole Blood: Collaborative Study

Ten collaborating laboratories assayed 4 blind duplicate pairs of whole bovine blood for cholinesterase activity. The 4 sample pairs ranged from normal (100%) to severely organo-phosphorus-inhibited (less than 10%) activity. Collaborators also received commercially available human lyophillized serum as an external control and a chromate solution to evaluate spectrophotometer performance. The Ellman kinetic assay was performed on a 1:1000 dilution of the whole blood in pH 8.0 phosphate buffer. The method monitors the increase in absorbance at 412 nm caused by formation of 5-thio-2-nitrobenzoic acid (yellow reaction product). Repeatability standard deviations (RSDr) ranged from 4.30 to 14.2%; reproducibility standard deviations (RSDR) ranged from 6.99 to 19.3%. The lower limit of detection was estimated to be 0.10 mumole/mL/min. The method has been approved interim official first action by AOAC.

Effect of Dursban in the drinking water of chicks.

Dursban®, ( O, O -diethyl O -(3,5,6-trichloro-2-pyridyl)phosphorothioate), was added to the drinking water of young Leghorn cockerels at 8 levels ranging from 0.08 to 1280 ppm active ingredient. Experiments were begun with chicks 3 or 5 days old and were concluded after 3 and 4 weeks. Mortality at 80 ppm was 20%, and mortality at 320 and 1280 ppm was 100%. No adverse effects increased were apparent at lower levels. Solvent and inert ingredients of Dursban emulsifiable concentrate had no adverse effect on the chicks up to 717.7 ppm. Cholinesterase activity in whole blood was monitored at all treatment levels, and the activity decreased at 80 ppm Dursban and above. At lower levels there was no significant difference from the controls. Weight gain per day diminished with increased levels of Dursban. Starvation of treated chicks did not affect blood-cholinesterase activity.