Research Article Experimental Measurements for the Effect...

Research ArticleExperimental Measurements for the Effect ofDilution Procedure in Blood Esterases as AnimalsBiomarker for Exposure to OP Compounds

Kasim Sakran Abass

Department of Basic Nursing Sciences College of Nursing University of Kirkuk Kirkuk Iraq

Correspondence should be addressed to Kasim Sakran Abass kasimabassusacom

Received 11 February 2014 Revised 2 March 2014 Accepted 2 March 2014 Published 22 April 2014

Academic Editor Miroslav Pohanka

Copyright copy 2014 Kasim Sakran AbassThis is an open access article distributed under the Creative Commons Attribution Licensewhich permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

Organophosphate compounds can bind to carboxylesterase which may lower the concentration of organophosphate pesticidesat the target site enzyme cholinesterase It is unclear from the literature whether it is the carboxylesterase affinity for theorganophosphate andor the number of carboxylesterase molecules that is the dominant factor in determining the protectivepotential of carboxylesterase The fundamental dilutions and kinetic effects of esterase enzyme are still poorly understood Thisstudy aims to confirm and extend our current knowledge about the effects of dilutions on esterases activities in the blood forbirds with respect to protecting the enzyme from organophosphate inhibition There was significantly higher esterases activitiesin dilution 1 10 in the all blood samples from quail duck and chick compared to other dilutions (1 5 1 15 1 20 and 1 25) inall cases Furthermore our results also pointed to the importance of estimating different dilutions effects prior to using in birdsas biomarker tools of environmental exposure Concentration-inhibition curves were determined for the inhibitor in the presenceof dilutions 1 5 1 10 plus 1 15 (to stimulate carboxylesterase) Point estimates (concentrations calculated to produce 20 50 and80 inhibition) were compared across conditions and served as a measure of esterase-mediated detoxification Results with well-known inhibitors (malathion) were in agreement with the literature serving to support the use of this assay Among the thiol-estersdilution 1 5 was observed to have the highest specificity constant (119896cat119870119898) and the119870119898 and 119896cat values were 176 120583M and 16765 sminus1respectively for S-phenyl thioacetate ester while detected in dilution 1 15 was the lowest specificity constant (119896cat119870119898) and the119870119898and 119896cat values were 943 120583M and 1154 sminus1 respectively for acetylthiocholine iodide ester

1 Introduction

An esterase is a hydrolase enzyme that splits esters into anacid and an alcohol in a chemical reaction with water calledhydrolysis [1 2] Carboxylesterase (CbE)cholinesterase(ChE) family members are under esterase enzyme and areresponsible for controlling the nerve impulse detoxificationand various developmental functions and are a major targetof pesticides and chemical warfare agents [2ndash4] Comparativestructural analysis of these enzymes is thus important [25 6] Organophosphates (OP) bind strongly and largelyirreversibly to both ChE in cholinergic synapses and to CbEwhich is found in blood and tissues [7ndash9] The reactionwith the enzyme is the key to the considerable number ofpharmacological actions possessed by OP although otherbiochemical properties of these compounds have recently

been recognised as important [10] OP compounds are agroup of artificially synthesized substances used in birds tocontrol pests and to enhance agricultural production [1112] However OP are used as insecticides acaricides andchemical agents and share a common neurotoxic mechanismof action [13] The biochemical alterations leading to manyof the deleterious effects have been studied in neuronal celllines however nonneuronal toxic effects of OP are far lesswell characterized in vitro [12 14] At present many hundredsof thousands of tons of OP compounds are used in thisway throughout the world Apart from the interest in OP asmilitary weapons the past thirty years have seen an unprece-dented growth in their use as insecticides stimulating theproduction of certain compounds such as malathion whichare degraded by higher organisms while remaining toxicto arthropods [12] Generally the cause of death in severe

Hindawi Publishing CorporationBioMed Research InternationalVolume 2014 Article ID 451982 13 pageshttpdxdoiorg1011552014451982

2 BioMed Research International

0 20 40 60 80 100 120 140 160 180 200

A

B

B

B

a

b

b

ab

b

AB1 5

1 10

1 15

1 20

1 25

X-d

ilutio

n

CbE ChE

Esterase activity (nmol minminus1 mLminus1)

(a)

0 50 100 150 200 250 300 350

A

B

B

B

B1 5

1 10

1 15

1 20

1 25

X-d

ilutio

nCbE ChE


(b)

0 100 200 300 400 500 600 700

A

A

AB

AB

B

a

b

b

b

b

1 5

1 10

1 15

1 20

1 25

X-d

ilutio

n

CbE ChE


(c)

Figure 1 Esterase enzyme activities in plasma diluted for quail (a) duck (b) and chick (c) A Bdenote significantly different between meanChE with different dilutions (analysis of variance (ANOVA) 119875 lt 005) a bdenote significantly different between mean CbE with differentdilutions (analysis of variance (ANOVA) 119875 lt 005) Each experiment is performed in duplicate (119899 = 5 in each bird)

OP intoxication is a combination of several factors whichare (a) circulatory effects by decreased cardiac output withbradycardia and peripheral vascular effects and (b) centralnervous system by generalised cortical activity causing con-vulsions which aggravate the cardiovascular system [15 16]Consequently more efficient processes such as chemical

oxidation are needed to remove OP during consumption offood production [6 17 18] Malathion is an organophosphateparasympathomimetic that binds irreversibly to esteraseenzyme It is converted to malaoxon by the multifunctionoxidases in mammals and insects or undergoes hydrolysis ofone of the CH

3OndashP bonds [19ndash21] Deactivation may also

BioMed Research International 3

0 200 400 600 800 1000 1200

A

A

B

B

B

a

b

b

b

b

1 5

1 10

1 15

1 20

1 25

X-d

ilutio

n

CbE ChE


(a)

0 50 100 150 200 250 300 350

A

B

B

B

B

a

a

b

b

b1 5

1 10

1 15

1 20

1 25

X-d

ilutio

nCbE ChE


(b)

0 200 400 600 800 1000

A

AB

B

B

B

a

b

b

b

b1 5

1 10

1 15

1 20

1 25

X-d

ilutio

n

CbE ChE


(c)

Figure 2 Esterase enzyme activities in erythrocyte diluted for quail (a) duck (b) and chick (c) Key to the figures is listed under Figure 1

occur by hydrolysis of the PndashS linkage However very fewstudies are focused on the dilution of malathion in the birdsaccordingly this paper was aimed to investigate the effectsof dilution on CbE and ChE activities in the blood frombirds used for human consumption A further aim was toindicate the value of maximal inhibitory concentrations anddifferent kinetic effects ofmalathion in different dilutions as abiochemical biomarker of exposure to pesticides toxicology

2 Materials and Methods

21 Chemicals CbE substrate S-phenyl thioacetate (PSA)98 purity ChE substrate acetylthiocholine iodide(AcTChI) 98 purity and 5 51015840-dithiobis(2-nitrobenzoicacid) (DTNB) were supplied by the Sigma ChemicalCompany Malathion [S-12-bis(ethoxycarbonyl) ethylOO-dimethyl phosphorodithioate] was obtained from GL


0 100 200 300 400

A

A

B

a

B

b

AB

b

b

b1 5

1 10

1 15

1 20

1 25

X-d

ilutio

n

CbE ChE


(a)

0 100 200 300 400 500

Aa

B

b

b

Aa

B

Bb1 5

1 10

1 15

1 20

1 25

X-d

ilutio

nCbE ChE


(b)

0 200 400 600 800 1000

A

B

B

B

B

a

b

b

b

b

CbE ChE


1 5

1 10

1 15

1 20

1 25

X-d

ilutio

n

(c)

Figure 3 Esterase enzyme activities in serum for quail (a) duck (b) and chick (c) Key to the figures is listed under Figure 1

Industries (E) Ltd Guwahati India All other reagents andsolvents used in this paper were of analytical grade

22 Sample Collection Blood samples from healthy quailduck and chick from local markets in Kirkuk were used inthis paper They were maintained in batches of 3ndash15 birds incages with dimensions of 75 times 75 times 75 cm in a room with

constant lighting at a temperature of 25ndash35∘C and relativehumidity was between (45ndash50) which was controlled byelectric heaters The floor litter consisted of wood shavingswater and feed were available ad libitum Blood sampleswere from male and female of white and brown Japanesequail Pekin duck Anas platyrhynchos and chicks (Ross 308Gallus gallus) ranging in age from 2 to 4 weeksThe ScientificCommittee of the College of Nursing at the University of


0 5 10 15 20 25 30 35 40 45 50

0

200

400

600

800

1000

0

200

400

600

800

1000

1200

Mean

Upper specification

Lower specificationminus200

minus400

minus200

minus400

CbE

activ

ity (n

mol

min

minus1

mLminus

1)

ChE

activ

ity (n

mol

min

minus1

mLminus

1)

CbEChE

X data

Figure 4 Distribution of the individual values of CbE and ChEactivities in all tested birds (quail duck and chick) and bloodsamples (plasma serum and erythrocyte) in dilutions (1 5 1 101 15 1 20 and 1 25)

Kirkuk (Iraq) has approved the present study Experimentsdetected with institutional regulations addressing animal useand good attention and care 6 were given to the birds used inthis work

23 Sample Preparation To obtain plasma 1mL bloodsamples were added to anticoagulant (EDTA 72mg finalconcentration 018) in 5mL centrifuge tubes Plasma wasseparated by centrifugation at 5000 g for 10min [2] To obtainserum blood samples were allowed to clot for at least 2 hat 25∘C after which they were centrifuged at 5000 g for10min The erythrocytes were washed three times with twovolumes of phosphate buffer (01M pH 74) centrifuged asdescribed above between washes Next the packed erythro-cytes diluted in 20 volumes of hypotonic sodium phosphatebuffer (67mM pH 74) to facilitate haemolysis followedby centrifugation at 5000 g for 10min The supernatant wasremoved and the pellet resuspended in hypotonic phosphatebuffer

24 Enzyme Measurement Esterase enzyme activity wasdetermined at room temperature 25∘C by the Ellmanmethod[22] using thioacetate (PSA substrate) for measuring CbEactivity or by using thiocholine (AcTChI substrate) formeasuring ChE activity [2 23] Subsequent combinationof thioacetate or thiocholine derivatives with DTNB formsthe yellow anion 5-thio-2-nitrobenzoic acid which absorbsstrongly at 410 nm [24] Substrate solutions (2mM for PSAwhile 1mM for AcTChI) were prepared and used on the sameday and kept on ice during use The blood samples werediluted with phosphate buffer (01M pH 8) with a 1 5 10 1520 and 25 ratios (wv) of parts of buffer All measurementsin this paper were carried out in duplicate Specific inhibitor

incubation of the samples with 01mM final concentrationof bis(p-nitrophenyl)phosphate in case of measuring of CbEwhile samples incubated with 4mM final concentration oftetraisopropyl pyrophosphoramide in case of measuring ofChE

25 In Vitro Exposure to OP Compound For the measure-ment of maximal inhibitory concentrations (IC

20IC50

andIC80) blood samples were inhibited for 30min at room

temperature 25∘C with appropriate concentration malathioncompound depending on preliminary range finding tests[2 25] Controls were incubated with phosphate buffer pH80 included Then the enzyme activity was determined asdescribed in above section Then the data were fitted withnonlinear regression analysis using a single exponential decayby SigmaPlot 11 (Systat software Inc)TheMichaelis constant(119870119898) and turnover number (119896cat) were determined according

to Abass [2]

26 Statistical Analysis The statistical analysis was per-formedusing SigmaPlot version 11Differences among groupswere determined by one-way analysis of variance (ANOVA)followed by Fisherrsquos LSD test All results are presented asmean plusmn SE and significance is accepted at 119875 lt 005

3 Results

31 Determination of Blood Esterase in Birds by the Enzyme-Dilution Method The effect of dilutions on CbE and ChEactivities was determined in serum plasma and erythrocytefor quail duck and chick as described in above section ofMaterials andMethods (Figures 1ndash3) CbE and ChE activitiesin dilution 1 10 observed highest activity in the plasmaserum and erythrocyte for quail duck and chick (Figures1ndash3) It was found that plasma CbE was significant (119875 lt005) in dilution 1 25 among other dilutions for quail andchick (Figures 1(a) and 1(c)) Blood plasma from the ChEactivity in blood from the plasma was significantly different(119875 lt 005) in dilution 1 25 among other dilutions for quailand duck (Figures 1(a) and 1(b)) and in dilution 1 10 amongother dilutions for chick (Figure 1(c)) The plasma activitiesof CbE ranged between 701 and 1781 nmolminminus1mLminus1 forquail between 999 and 1967 nmolminminus1mLminus1 for duck andbetween 996 and 3875 nmolminminus1mLminus1 for chick samplesacross different dilutions while for plasma ChE rangedbetween 833 and 1723 nmolminminus1mLminus1 for quail between1197 and 3053 nmolminminus1mLminus1 for duck and between3077 and 5907 nmolminminus1mLminus1 for chick samples acrossdifferent dilutions (Figures 1(a)ndash1(c)) There was significance(119875 lt 005) in serum CbE within dilution 1 25 among otherdilutions used for quail and chick (Figures 2(a) and 2(c))while serum CbE was not significant (119875 gt 005) betweendilution 1 20 and dilution 1 25 for duck (Figure 2(b)) ChEactivity was a significantly different (119875 lt 005) between dilu-tion 1 10 and dilution 1 15 with other dilutions used for quail(Figure 2(a)) while in duck it was seen as not significant (119875 gt005) between dilution 1 20 and dilution 1 25 (Figure 2(b))whereas for chick was significant (119875 lt 005) between dilution


Quail

0 200 400 600 800 10000

200

400

600

800

1000

ChE activity (nmol minminus1 mLminus1)

CbE

activ

ity (n

mol

min

minus1

mLminus

1)

R2 = 0114

P lt 0218

f = 3478 + 0232lowastx

(a)

Duck

0 200 400 600 800 1000 12000

200

400

600

800

1000

1200


CbE

activ

ity (n

mol

min

minus1

mLminus

1)

f = 3404 + 135lowastx

R2 = 0641

P lt 00003

(b)

Chick

0 100 200 300 400 5000

100

200

300

400

500


CbE

activ

ity (n

mol

min

minus1

mLminus

1)

R2 = 0767

P lt 00001

f = 6667 + 0843lowastx

(c)

Figure 5 Regression analyses between CbE and ChE activities in all tested blood contents from quail duck and chick across differentdilutions (1 5 1 10 1 15 1 20 and 1 25)

1 25 among other dilutions (Figure 2(c)) The serum activi-ties of CbE ranged between 802 and 1806 nmolminminus1mLminus1for quail between 581 and 3876 nmolminminus1mLminus1 for duckand between 629 and 4631 nmolminminus1mLminus1 for chick sam-ples across different dilutions whereas for serumChE rangedbetween 2111 and 4203 nmolminminus1mLminus1 for quail between823 and 3502 nmolminminus1mLminus1 for duck and between2973 and 7084 nmolminminus1mLminus1 for chick samples acrossdifferent dilutions (Figures 2(a)ndash2(c))The enzyme activity inthe serum blood at the higher dilution ratio declined fasterthan that of samples at lower dilution ratio

Blood from erythrocyte was seen significant (119875 lt 005)within dilution 1 25 among other dilutions for quail CbEand chick ChE (Figures 3(a) and 3(c)) and was seen notsignificant (119875 gt 005) between dilution 1 20 and dilution1 20 for duck (Figure 3(b)) while there were significant(119875 lt 005) differences within dilution 1 25 among otherdilutions used for chick CbE and ChE (Figure 3) ChE wasseen not significant (119875 gt 005) between dilution 1 20 anddilution 1 25 for quail (Figure 3(a)) erythrocyte activitiesof CbE ranged between 501 and 8506 nmolminminus1mLminus1 forquail between 684 and 3188 nmolminminus1mLminus1 for duck


Plasma

0 100 200 300 4000

200

400

600


CbE

activ

ity (n

mol

min

minus1

mLminus

1)

f = minus851 + 153 lowastx

R2 = 0812

P lt 00001

(a)

Serum

0 200 400 600 8000

200

400

600

800


CbE

activ

ity (n

mol

min

minus1

mLminus

1)

R2 = 0694

P lt 00001

f = 14831 + 0876lowastx

(b)

Erythrocyte

0 200 400 600 800 1000 12000

200

400

600

800

1000

1200


CbE

activ

ity (n

mol

min

minus1

mLminus

1)

R2 = 0391

P lt 00126

f = 7427 + 0616lowastx

(c)

Figure 6 Regression analyses between CbE and ChE activities in all tested birds for plasma serum and erythrocyte across different dilutions(1 5 1 10 1 15 1 20 and 1 25)

and between 1842 and 9047 nmolminminus1mLminus1 for chicksamples across different dilutions whereas for erythrocyteChE ranged between 353 and 9964 nmolminminus1mLminus1 forquail between 423 and 2935 nmolminminus1mLminus1 for duck andbetween 654 and 5164 nmolminminus1mLminus1 for chick samplesacross different dilutions (Figures 3(a)ndash3(c)) Overall studythe dilution 1 25 gave a lowest enzymatic activity in bothCbE and ChE (Figures 1ndash3) The distribution of the meanindividuals values of esterase enzyme activity in all birdsand blood contents in different dilutions was detected thehighest esterase activities in ChE (3187 nmolminminus1mLminus1)

compared to CbE (2144 nmolminminus1mLminus1) (Figure 4) In allcases (quail duck and chick used five dilutions) the linearregression found between CbE and ChE was seen to be R2

= 0114 119875 lt 0218 for quail R2 = 0641 119875 lt 00003 forduck and R2 = 0767 119875 lt 00007 for chick (Figure 5) Inaddition the author found a linear regression between alltested blood samples (1198772 = 0812 119875 lt 00001 for plasma1198772

= 0694 119875 lt 00001 for serum and 1198772 = 0391 119875 lt00126 for erythrocyte) (Figure 6) The author found blooddilution only with erythrocyte detects in higher activities ofCbE and ChE than plasma and serum (Figure 7) Pearson


Esterases activity0 200 400 600 800 1000 1200

Plas

ma

Seru

mEr

ythr

ocyt

e

Figure 7 Box plots of the range esterase enzyme activity reportedto be taken in different dilutions (1 5 1 10 1 15 1 20 and 1 25)

correlation coefficient (r) calculated to measure the degreeof relationship between CbE and ChE in the plasma serumand erythrocyte for testing birds was observed significantlyin plasma serum and erythrocyte with dilution 1 10 for quailand chick in addition to a significant effect with dilution 1 20for serum of quail and duck (Table 1)

32 Kinetic Dilution Methodology for the OP CompoundConcentration-inhibition curves for malathion in the serumfor quail in presence of dilution 1 5 dilution 1 10 anddilution 1 15 were found to be the highest reaction rates indilution 1 10 for long time (4 h) whereas reducing reaction10 rates in dilution 1 10 for short time (1 h) (Figure 8) Thesubstrate specificity of the diluted esterases was investigatedusing two different kinds of substrates PSA for the CbEactivity and AcTChI for the ChE activity Point estimatesare reported in Table 2 along with the ratios of the differentdilutions values The somewhat smaller ratios at the higherconcentrations (less than half at the IC

80compared to IC

20)

with dilution 1 10 but not dilution 1 15 suggest some satu-ration of the CbE in that range of the curve Esterase activityof the enzyme was determined using PSA and AcTChI-esters containing a various thiol side chain length As shownin Table 3 the highest 119896cat value was observed with PSA-substrate (dilution 1 5) whereas AcTChI-substrate (dilution1 15) showed the lowest 119896cat valueHowever the119870119898 valuewasthe highest with PSA-substrate (dilution 1 15) and the lowestwith AcTChI-substrate (dilution 1 10) Among the thiol-esters dilution 1 5was observed to have the highest specificityconstant (119896cat119870119898) and the 119870

119898and 119896cat values were 176 120583M

and 16 765 sminus1 respectively for PSA ester while detected indilution 1 15 the lowest specificity constant (119896cat119870119898) and the119870119898and 119896cat values were 943 120583M and 1154 sminus1 respectively for

AcTChI ester

4 Discussion

41 The Evaluation of Esterase Enzyme Dilution Assays Twoforms of esterase enzyme activities have been identified inmammalian blood These are distinguished according totheir substrate specificity [2] In fact CbE prefers PSA assubstrate (at high concentrations) but ChE prefers AcTChIas substrate (at low concentrations) [26ndash29] In this paperthe author showed five dilutions to measure the activities ofCbE and ChE in plasma serum and erythrocyte for quailduck and chick to reduce the turbidity of the blood samplesand to ensure that the optical signal falls within the linearrange of detection throughout the esterase enzyme activitymeasurement and the enzymedilution slows down the rate ofspending of substrate so providing an extended timewindowfor statement of steady-state esterase enzyme kinetics [1830] The results from this study show that dilution 1 10had high esterases enzyme activity over different dilutionsused and ranged between 1723 and 5903 nmolminminus1mLminus1for plasma between 2934 and 9964 nmolminminus1mLminus1 forerythrocyte and between 1806 and 7635 nmolminminus1mLminus1for serum (Figures 1ndash3) A similar observation has beenmade by Quinn et al [31] reporting that dilution 1 10 forplasma and erythrocyte had higher activity than dilution1 25 These interesting results are also similar with findingof [32] where they found it higher in the blood for ChEactivity with dilution 1 10The reason for this is probably dueto adsorption changes occurring faster at dilution 1 10 thanother dilutions [33] Esterase activity in the blood was at thehigher dilution ratio and declined faster than that of samplesat lower dilution ratio Linear regression of between CbE andChE activities was observed in five dilutions samples for allbirds tested with blood with exception of quail (Figure 5)

42 A Kinetic Model for Substrate and Dilution of Blood Sam-ples under OP-Sufficient Conditions The results of kineticstudies using thiol side esters for the esterase enzyme showthat PSA for the CbE activity and AcTChI for the ChEactivity (dilution 1 10) are the most suitable substrate forthe enzyme kinetic The somewhat smaller ratios at thehigher concentrations (less than half at the IC

80compared to

IC20) with dilution 1 10 but not dilution 1 15 suggest some

saturation of the CbE in that range of the curve (Table 2)Thisis in agreement with the previous study [34] In summarythis study used an in vitro assay to measure detoxificationby CbE and ChE of a malathion compound (Table 3) Thisscreening method could be useful for predicting the degreeof esterase detoxification and this information could beuseful in assessment of animal variability due to differencesin esterase activities [2] Dilution 1 5 was observed to havethe highest specificity constant (119896cat119870119898) and the 119870

119898and

119896cat values were 176 120583M and 16765 sminus1 respectively for PSAester while dilution 1 15 had the lowest specificity constant(119896cat119870119898) and the 119870

119898and 119896cat values were 943 120583M and

1154 sminus1 respectively for AcTChI ester For many of thisinhibitor these are the first data available for informingesterase enzyme mediated detoxification Our findings indi-cate that detoxification patterns are chemical-specific and


Table1Pearsoncorrelationcoeffi

cientand119875values

betweenCb

EandCh

Ein

theb

lood

forb

irdsa

tdifferentd

ilutio

ns

Dilu

tion

Plasma

Erythrocyte

Serum

Quail

Duck

Chick

Quail

Duck

Chick

Quail

Duck

Chick

15

0219

minus0111

minus0402

minus064

8minus0063

0112

0411

0102

0071

(022)

(052

)(022)

(021)

(082)

(021)

(022)

(071)

(010

)

110

minus0337

0081

0123

0253

minus0580

minus0326

0026

0209

0138

(010

)(090)

(004)

(013

)(013

)(002)

(093)

(052

)(006)

115

0311

minus0179

minus0083

0152

minus0451

minus0224

0544

minus0091

0417

(031

)(032

)(021)

(054)

(012

)(074

)(004)

(092)

(009)

120

minus0245

0162

0242

0352

minus0324

0157

044

6minus0216

0429

(074

)(013

)(012

)(043)

(024)

(055)

(004)

(002)

(018

)

125

0221

0205

0134

minus0078

minus0121

minus0251

0559

minus0075

0317

(011)

(089)

(043)

(013

)(042)

(065)

(032

)(008)

(012

)Va

lues

inthetablearethe119903(119875

value)


Table2Malathion

pointestim

ates(IC 2

0IC

50and

IC80120583M)from

concentration-inhibitio

ncurvesin

thep

resenceo

fdilu

tions

15110and

115Twopreincub

ationcond

ition

swereu

sed

long

(4h)

andshort(1h

)

Preincub

ationcond

ition

Slop

IC20(120583M)

Slop

IC50(120583M)

Slop

IC80(120583M)

Long

(4h)

Dilu

tion(15)

022plusmn0655a

315plusmn0991a

067plusmn0126

11plusmn0432a

066plusmn0718

11plusmn0121

Dilu

tion(110)

14plusmn0122

050plusmn0654

089plusmn0117

078plusmn0121

064plusmn0611

12plusmn0211

Dilu

tion(115)

011plusmn0418b

63plusmn12

b071plusmn0112

098plusmn0212

084plusmn0801

083plusmn0221

Short(1h

)Dilu

tion(15)

034plusmn0372a

21plusmn

0876a

029plusmn0501a

24plusmn0311a

037plusmn048

a19plusmn0421

Dilu

tion(110)

066plusmn0931

11plusmn0765

027plusmn0145

26plusmn0911

029plusmn018

24plusmn0987

Dilu

tion(115)

012plusmn0583b

58plusmn11

b053plusmn0196b

131plusmn

0876

024plusmn041

b29plusmn0766

Values

inthetablearethemeanplusmnSE

ofthreeindividu

alserum

esterase

(CbE

)for

quailFo

rthe

measuremento

fIC 2

0IC

50and

IC80este

rase

enzymewas

inhibitedfor3

0min

atroom

temperature

25∘

Cwith

approp

riate

concentrations

form

alathion

compo

und

Then

results

were

fittedwith

anexpo

nentiald

ecay

usingSigm

aPlot11

abdeno

tess

ignificantd

ifference

(analysis

ofvaria

nce(A

NOVA

)119875lt005)b

etween

dilutio

n15anddilutio

n115


00 05 10 15 200

20

40

60

80

100

120C

ontro

l act

ivity

()

Concentration (120583M)

Dilution (1 5)Dilution (1 10)Dilution (1 15)

(a)

00 05 10 15 20

Con

trol a

ctiv

ity (

)

0

20

40

60

80

100

120

140

160

180

200



(b)

Figure 8 Concentration-inhibition curves for malathion in serum for quail in the presence of dilution 1 5 dilution 1 10 and dilution 1 15Three preincubation conditions were used long (4 h) (a) and short (1 h) (b) Experimental data (symbols) and model output (lines) are fittedby single rectangular equation of a hyperbola using a SigmaPlot 11

Table 3 Kinetic parameters for hydrolysis of esterase in various dilutions

Preincubation condition CbE ChE119870119898(120583M) 119896cat (s

minus1) 119896cat119870119898 (sminus1

120583Mminus1) 119870119898(120583M) 119896cat (s

minus1) 119896cat119870119898 (sminus1

120583Mminus1)Dilution (1 5) 176 plusmn 153a 16765 plusmn 865a 953a 265 plusmn 234a 2023 plusmn 232a 76a

Dilution (1 10) 543 plusmn 213 12765 plusmn 932 235 234 plusmn 324 17165 plusmn 654 734Dilution (1 15) 753 plusmn 284 7653 plusmn 121b 102 943 plusmn 712 1154 plusmn 111 12Values in the table are the mean plusmn SE of three individual serum esterase (CbE and ChE) for quail For the measurement of 119870

119898and 119896cat esterase enzyme was

inhibited for 30min at room temperature 25∘C with appropriate concentrations for malathion compound Then results were fitted with an exponential decayusing SigmaPlot 11 abdenotes significant difference (analysis of variance (ANOVA) 119875 lt 005) between dilution 1 5 and other dilutions

that the influence of known susceptibility factors such as CbEand ChE should not be generalized across this inhibitor [35]

5 Conclusions

In conclusion this is the first paper that provided originaldata concerning an enzymological dilution characterizationin the blood samples from birds used for human consump-tionThis paper succeed in establishing a gentle dilution in allthree tested birds (quail duck and chick) and blood (plasmaserum and erythrocyte) the dilution 1 10 recorded higheresterase enzyme activity than other dilutions (1 5 1 15 1 20and 1 25) Furthermore our results also pointed at the impor-tance of estimating different kinetic dilutions effects prior tousing in birds as biomarker tools of environmental exposureto anti-CbE or anti-ChE pesticides intoxication Finally in

spite of this paper further studies are required under differentlaboratories and different OP pesticide compounds in orderto improve and to increase our knowledge about this veryinteresting enzyme as a potential biochemical marker forpesticide compounds

Conflict of Interests

Theauthor declares that there is no conflict of interests and hehas no financial or personal relationships with other peopleor organizations that can inappropriately influence his workthere is no professional or other personal interest of anynature or kind in any product service andor company thatcould be construed as influencing the position presented inthis paper


Acknowledgments

The author wishes to thank the farmers who participated inthis study and the animal health assistants and laboratorytechnicians for their expert work in collecting the samplesand for their useful assistance The author is also thankful tomanagement and principal of College of Nursing Kirkuk forthe providing facilities for present work

References

[1] R Vanholme I Cesarino K Rataj et al ldquoCaffeoyl shikimateesterase (CSE) is an enzyme in the lignin biosynthetic pathwayin Arabidopsisrdquo Science vol 341 no 6150 pp 1103ndash1106 2013

[2] K S Abass ldquoAmethod for fast assessment ofOPCB exposure inthe Japanese quail (Coturnix coturnix japonica) using combinedesterases enzyme activity as biomarkersrdquo Enzyme Research vol2014 Article ID 812302 15 pages 2014

[3] P R Rivadeneira M Agrelo S Otero and G Kristoff ldquoDif-ferent effects of subchronic exposure to low concentrations ofthe organophosphate insecticide chlorpyrifos in a freshwatergastropodrdquo Ecotoxicology and Environmental Safety vol 90 pp82ndash88 2013

[4] R C Lajmanovich A M Attademo P M Peltzer C M Jungesand M C Cabagna ldquoToxicity of four herbicide formulationswith glyphosate on Rhinella arenarum (Anura Bufonidae)tadpoles B-esterases and glutathione S-transferase inhibitorsrdquoArchives of Environmental Contamination and Toxicology vol60 no 4 pp 681ndash689 2011

[5] C J Jackson J-W Liu P D Carr et al ldquoStructure and functionof an insect 120572-carboxylesterase (120572Esterase7) associated withinsecticide resistancerdquo Proceedings of the National Academy ofSciences of the United States of America vol 110 no 25 pp10177ndash10182 2013

[6] H Chan ldquoSeparation of pollutants from restaurant effluents asanimal feed fertilizer and renewable energy to produce highwater quality in a compact areardquoWater Resources and Industryvol 3 pp 35ndash47 2013

[7] K Kanavouras M N Tzatzarakis V Mastorodemos APlaitakis and A M Tsatsakis ldquoA case report of motor neurondisease in a patient showing significant level of DDTs HCHsand organophosphate metabolites in hair as well as levels ofhexane and toluene in bloodrdquo Toxicology and Applied Pharma-cology vol 256 no 3 pp 399ndash404 2011

[8] S Jain R S Ahmed V K Arora and B D Banerjee ldquoBio-chemical and histopathological studies to assess chronic toxicityof triazophos in blood liver and brain tissue of ratsrdquo PesticideBiochemistry and Physiology vol 100 no 2 pp 182ndash186 2011

[9] H Kara and A Ak ldquoEffects of CoQ10 on the erythrocyte andheart tissue cholinesterase nitric oxide and malondialdehydelevels in acute organophosphate toxicityrdquo Critical Care vol 15supplement 1 article P367 2010

[10] A C Agra-Neto T H Napoleao E V Pontual et al ldquoEffectofMoringa oleifera lectins on survival and enzyme activities ofAedes aegypti larvae susceptible and resistant to organophos-phaterdquo Parasitology Research vol 113 no 1 pp 175ndash184 2014

[11] R C Gupta ldquoOrganophosphates and carbamatesrdquo inVeterinaryToxicology pp 477ndash488 Academic Press Oxford UK 2007

[12] C M Timothy ldquoOrganophosphates history chemistry phar-macologyrdquo in Organophosphates and Health pp 1ndash36 ImperialCollege Press London UK 2001

[13] N van den Eede A C Dirtu H Neels and A Covaci ldquoAna-lytical developments and preliminary assessment of humanexposure to organophosphate flame retardants from indoordustrdquo Environment International vol 37 no 2 pp 454ndash4612011

[14] G Ding S Han P Wang et al ldquoIncreased levels of 8-hydroxy-21015840-deoxyguanosine are attributable to organophosphate pesti-cide exposure among young childrenrdquo Environmental Pollutionvol 167 pp 110ndash114 2012

[15] J L Carey C Dunn and R J Gaspari ldquoCentral respiratoryfailure during acute organophosphate poisoningrdquo RespiratoryPhysiology and Neurobiology vol 189 no 2 pp 403ndash410 2013

[16] G Lemercier P Carpentier H Sentenac Roumanou andP Morelis ldquoHistological and histochemical changes in thecentral nervous system of the rat poisoned by an irreversibleanticholinesterase organophosphorus compoundrdquo Acta Neu-ropathologica vol 61 no 2 pp 123ndash129 1983

[17] C OrsquoDonnell B K Tiwari P J Cullen and R G Rice ldquoOzonefor water treatment and its potential for process water reuse inthe food industryrdquo Ozone in Food Processing pp 177ndash199 2012

[18] K A Askar A C Kudi and A J Moody ldquoSpontaneous reac-tivation and aging kinetics of acetylcholinesterase inhibited bydichlorvos and diazinonrdquo The Journal of Toxicological Sciencesvol 36 no 2 pp 237ndash241 2011

[19] W N Aldridge J W Miles D L Mount and R D VerschoyleldquoThe toxicological properties of impurities in malathionrdquoArchives of Toxicology vol 42 no 2 pp 95ndash106 1979

[20] W ECotham Jr andT F Bidleman ldquoDegradation ofmalathionendosulfan and fenvalerate in seawater and seawatersedimentmicrocosmsrdquo Journal of Agricultural and Food Chemistry vol37 no 3 pp 824ndash828 1989

[21] J P Haliscak and RW Beeman ldquoStatus of malathion resistancein five genera of beetles infesting farm-stored corn wheat andoats in the United Statesrdquo Journal of Economic Entomology vol76 no 4 pp 717ndash722 1983

[22] G L Ellman K D Courtney V Andres Jr and R MFeatherstone ldquoA new and rapid colorimetric determination ofacetylcholinesterase activityrdquo Biochemical Pharmacology vol 7no 2 pp 88ndash95 1961

[23] J Nilin M Monteiro I Domingues S Loureiro L V Costa-Lotufo and A M Soares ldquoBivalve esterases as biomarkeridentification and characterization in European cockles (Ceras-toderma edule)rdquo Bulletin of Environmental Contamination andToxicology vol 88 no 5 pp 707ndash711 2012

[24] J L Trimm G Salama and J J Abramson ldquoSulfhydryl oxida-tion induces rapid calcium release from sarcoplasmic reticulumvesiclesrdquoThe Journal of Biological Chemistry vol 261 no 34 pp16092ndash16098 1986

[25] K A Askar A C Kudi and A J Moody ldquoComparison of twostorage methods for the analysis of cholinesterase activities infood animalsrdquo Enzyme Research vol 2010 Article ID 90424911 pages 2010

[26] F Tecles and J J Ceron ldquoDetermination of whole bloodcholinesterase in different animal species using specific sub-stratesrdquo Research in Veterinary Science vol 70 no 3 pp 233ndash238 2001

[27] B W Wilson and W Philip ldquoCholinesterase inhibitionrdquo inEncyclopaedia of Toxicology pp 588ndash599 Elsevier New YorkNY USA 2005

[28] E K Tarkiainen J T Backman M Neuvonen P J NeuvonenM Schwab and M Niemi ldquoCarboxylesterase 1 polymorphism


impairs oseltamivir bioactivation in humansrdquo Clinical Pharma-cology andTherapeutics vol 92 no 1 pp 68ndash71 2012

[29] S E Pratt S Durland-Busbice R L Shepard K Heinz-TahenyP W Iversen and A H Dantzig ldquoHuman carboxylesterase-2hydrolyzes the prodrug of gemcitabine (LY2334737) and confersprodrug sensitivity to cancer cellsrdquo Clinical Cancer Researchvol 19 no 5 pp 1159ndash1168 2013

[30] L T-H Kao and M Gratzl ldquoSerum cholinesterase assay usinga reagent-free micro pH-statrdquo Analytical Biochemistry vol 389no 2 pp 93ndash96 2009

[31] R P Quinn P de Miranda L Gerald and S S Good ldquoA sensi-tive radioimmunoassay for the antiviral agent BW248U [9-(2-hydroxyethoxymethyl)guanine]rdquo Analytical Biochemistry vol98 no 2 pp 319ndash328 1979

[32] K A Askar A C Kudi and A J Moody ldquoComparative analysisof cholinesterase activities in food animals using modifiedEllman and Michel assaysrdquo Canadian Journal of VeterinaryResearch vol 75 no 4 pp 261ndash270 2011

[33] H Soreq and S Seidman ldquoAcetylcholinesterasemdashnew roles foran old actorrdquoNature ReviewsNeuroscience vol 2 no 4 pp 294ndash302 2001

[34] V C Moser and S Padilla ldquoEsterase metabolism ofcholinesterase inhibitors using rat liver in vitrordquo Toxicologyvol 281 no 1ndash3 pp 56ndash62 2011

[35] K A Askar and A C Kudi ldquoIn vitro kinetic characterizationof inhibition of acetylcholinesterase by organophosphate andcarbamate compounds in food animalsrdquo Toxicological andEnvironmental Chemistry vol 94 no 3 pp 596ndash604 2012

Submit your manuscripts athttpwwwhindawicom

PainResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom

Volume 2014

ToxinsJournal of

VaccinesJournal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

AntibioticsInternational Journal of

ToxicologyJournal of


StrokeResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Drug DeliveryJournal of



Advances in Pharmacological Sciences

Tropical MedicineJournal of


Medicinal ChemistryInternational Journal of


AddictionJournal of



BioMed Research International

Emergency Medicine InternationalHindawi Publishing Corporationhttpwwwhindawicom Volume 2014


Autoimmune Diseases


Anesthesiology Research and Practice

ScientificaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of


Pharmaceutics


MEDIATORSINFLAMMATION

of


0 20 40 60 80 100 120 140 160 180 200

A

B

B

B

a

b

b

ab

b

AB1 5

1 10

1 15

1 20

1 25

X-d

ilutio

n

CbE ChE


(a)

0 50 100 150 200 250 300 350

A

B

B

B

B1 5

1 10

1 15

1 20

1 25

X-d

ilutio

nCbE ChE


(b)

0 100 200 300 400 500 600 700

A

A

AB

AB

B

a

b

b

b

b

1 5

1 10

1 15

1 20

1 25

X-d

ilutio

n

CbE ChE


(c)

Figure 1 Esterase enzyme activities in plasma diluted for quail (a) duck (b) and chick (c) A Bdenote significantly different between meanChE with different dilutions (analysis of variance (ANOVA) 119875 lt 005) a bdenote significantly different between mean CbE with differentdilutions (analysis of variance (ANOVA) 119875 lt 005) Each experiment is performed in duplicate (119899 = 5 in each bird)

OP intoxication is a combination of several factors whichare (a) circulatory effects by decreased cardiac output withbradycardia and peripheral vascular effects and (b) centralnervous system by generalised cortical activity causing con-vulsions which aggravate the cardiovascular system [15 16]Consequently more efficient processes such as chemical

oxidation are needed to remove OP during consumption offood production [6 17 18] Malathion is an organophosphateparasympathomimetic that binds irreversibly to esteraseenzyme It is converted to malaoxon by the multifunctionoxidases in mammals and insects or undergoes hydrolysis ofone of the CH

3OndashP bonds [19ndash21] Deactivation may also


0 200 400 600 800 1000 1200

A

A

B

B

B

a

b

b

b

b

1 5

1 10

1 15

1 20

1 25

X-d

ilutio

n

CbE ChE


(a)

0 50 100 150 200 250 300 350

A

B

B

B

B

a

a

b

b

b1 5

1 10

1 15

1 20

1 25

X-d

ilutio

nCbE ChE


(b)

0 200 400 600 800 1000

A

AB

B

B

B

a

b

b

b

b1 5

1 10

1 15

1 20

1 25

X-d

ilutio

n

CbE ChE


(c)






0 100 200 300 400

A

A

B

a

B

b

AB

b

b

b1 5

1 10

1 15

1 20

1 25

X-d

ilutio

n

CbE ChE


(a)

0 100 200 300 400 500

Aa

B

b

b

Aa

B

Bb1 5

1 10

1 15

1 20

1 25

X-d

ilutio

nCbE ChE


(b)

0 200 400 600 800 1000

A

B

B

B

B

a

b

b

b

b

CbE ChE


1 5

1 10

1 15

1 20

1 25

X-d

ilutio

n

(c)






0 5 10 15 20 25 30 35 40 45 50

0

200

400

600

800

1000

0

200

400

600

800

1000

1200

Mean

Upper specification


minus400

minus200

minus400

CbE

activ

ity (n

mol

min

minus1

mLminus

1)

ChE

activ

ity (n

mol

min

minus1

mLminus

1)

CbEChE

X data







20IC50



to Abass [2]


3 Results



Quail

0 200 400 600 800 10000

200

400

600

800

1000


CbE

activ

ity (n

mol

min

minus1

mLminus

1)

R2 = 0114

P lt 0218

f = 3478 + 0232lowastx

(a)

Duck

0 200 400 600 800 1000 12000

200

400

600

800

1000

1200


CbE

activ

ity (n

mol

min

minus1

mLminus

1)

f = 3404 + 135lowastx

R2 = 0641

P lt 00003

(b)

Chick

0 100 200 300 400 5000

100

200

300

400

500


CbE

activ

ity (n

mol

min

minus1

mLminus

1)

R2 = 0767

P lt 00001

f = 6667 + 0843lowastx

(c)





Plasma

0 100 200 300 4000

200

400

600


CbE

activ

ity (n

mol

min

minus1

mLminus

1)


R2 = 0812

P lt 00001

(a)

Serum

0 200 400 600 8000

200

400

600

800


CbE

activ

ity (n

mol

min

minus1

mLminus

1)

R2 = 0694

P lt 00001

f = 14831 + 0876lowastx

(b)

Erythrocyte

0 200 400 600 800 1000 12000

200

400

600

800

1000

1200


CbE

activ

ity (n

mol

min

minus1

mLminus

1)

R2 = 0391

P lt 00126

f = 7427 + 0616lowastx

(c)








Plas

ma

Seru

mEr

ythr

ocyt

e




80compared to IC

20)




AcTChI ester

4 Discussion



80compared to



119898and







betweenCb

EandCh

Ein

theb

lood

forb

irdsa

tdifferentd

ilutio

ns

Dilu

tion

Plasma

Erythrocyte

Serum

Quail

Duck

Chick

Quail

Duck

Chick

Quail

Duck

Chick

15

0219

minus0111

minus0402

minus064

8minus0063

0112

0411

0102

0071

(022)

(052

)(022)

(021)

(082)

(021)

(022)

(071)

(010

)

110

minus0337

0081

0123

0253

minus0580

minus0326

0026

0209

0138

(010

)(090)

(004)

(013

)(013

)(002)

(093)

(052

)(006)

115

0311

minus0179

minus0083

0152

minus0451

minus0224

0544

minus0091

0417

(031

)(032

)(021)

(054)

(012

)(074

)(004)

(092)

(009)

120

minus0245

0162

0242

0352

minus0324

0157

044

6minus0216

0429

(074

)(013

)(012

)(043)

(024)

(055)

(004)

(002)

(018

)

125

0221

0205

0134

minus0078

minus0121

minus0251

0559

minus0075

0317

(011)

(089)

(043)

(013

)(042)

(065)

(032

)(008)

(012

)Va

lues


value)


Table2Malathion

pointestim

ates(IC 2

0IC

50and

IC80120583M)from


ncurvesin

thep

resenceo

fdilu

tions

15110and

115Twopreincub

ationcond

ition

swereu

sed

long

(4h)

andshort(1h

)

Preincub

ationcond

ition

Slop

IC20(120583M)

Slop

IC50(120583M)

Slop

IC80(120583M)

Long

(4h)

Dilu

tion(15)

022plusmn0655a

315plusmn0991a

067plusmn0126

11plusmn0432a

066plusmn0718

11plusmn0121

Dilu

tion(110)

14plusmn0122

050plusmn0654

089plusmn0117

078plusmn0121

064plusmn0611

12plusmn0211

Dilu

tion(115)

011plusmn0418b

63plusmn12

b071plusmn0112

098plusmn0212

084plusmn0801

083plusmn0221

Short(1h

)Dilu

tion(15)

034plusmn0372a

21plusmn

0876a

029plusmn0501a

24plusmn0311a

037plusmn048

a19plusmn0421

Dilu

tion(110)

066plusmn0931

11plusmn0765

027plusmn0145

26plusmn0911

029plusmn018

24plusmn0987

Dilu

tion(115)

012plusmn0583b

58plusmn11

b053plusmn0196b

131plusmn

0876

024plusmn041

b29plusmn0766

Values


ofthreeindividu

alserum

esterase

(CbE

)for

quailFo

rthe

measuremento

fIC 2

0IC

50and

IC80este

rase

enzymewas

inhibitedfor3

0min

atroom

temperature

25∘

Cwith

approp

riate

concentrations

form

alathion

compo

und

Then

results

were

fittedwith

anexpo

nentiald

ecay

usingSigm

aPlot11

abdeno

tess

ignificantd

ifference

(analysis

ofvaria

nce(A

NOVA

)119875lt005)b

etween

dilutio

n15anddilutio

n115


00 05 10 15 200

20

40

60

80

100

120C

ontro

l act

ivity

()



(a)

00 05 10 15 20

Con

trol a

ctiv

ity (

)

0

20

40

60

80

100

120

140

160

180

200



(b)




minus1) 119896cat119870119898 (sminus1

120583Mminus1) 119870119898(120583M) 119896cat (s

minus1) 119896cat119870119898 (sminus1






5 Conclusions






Acknowledgments


References










































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of


0 200 400 600 800 1000 1200

A

A

B

B

B

a

b

b

b

b

1 5

1 10

1 15

1 20

1 25

X-d

ilutio

n

CbE ChE


(a)

0 50 100 150 200 250 300 350

A

B

B

B

B

a

a

b

b

b1 5

1 10

1 15

1 20

1 25

X-d

ilutio

nCbE ChE


(b)

0 200 400 600 800 1000

A

AB

B

B

B

a

b

b

b

b1 5

1 10

1 15

1 20

1 25

X-d

ilutio

n

CbE ChE


(c)






0 100 200 300 400

A

A

B

a

B

b

AB

b

b

b1 5

1 10

1 15

1 20

1 25

X-d

ilutio

n

CbE ChE


(a)

0 100 200 300 400 500

Aa

B

b

b

Aa

B

Bb1 5

1 10

1 15

1 20

1 25

X-d

ilutio

nCbE ChE


(b)

0 200 400 600 800 1000

A

B

B

B

B

a

b

b

b

b

CbE ChE


1 5

1 10

1 15

1 20

1 25

X-d

ilutio

n

(c)






0 5 10 15 20 25 30 35 40 45 50

0

200

400

600

800

1000

0

200

400

600

800

1000

1200

Mean

Upper specification


minus400

minus200

minus400

CbE

activ

ity (n

mol

min

minus1

mLminus

1)

ChE

activ

ity (n

mol

min

minus1

mLminus

1)

CbEChE

X data







20IC50



to Abass [2]


3 Results



Quail

0 200 400 600 800 10000

200

400

600

800

1000


CbE

activ

ity (n

mol

min

minus1

mLminus

1)

R2 = 0114

P lt 0218

f = 3478 + 0232lowastx

(a)

Duck

0 200 400 600 800 1000 12000

200

400

600

800

1000

1200


CbE

activ

ity (n

mol

min

minus1

mLminus

1)

f = 3404 + 135lowastx

R2 = 0641

P lt 00003

(b)

Chick

0 100 200 300 400 5000

100

200

300

400

500


CbE

activ

ity (n

mol

min

minus1

mLminus

1)

R2 = 0767

P lt 00001

f = 6667 + 0843lowastx

(c)





Plasma

0 100 200 300 4000

200

400

600


CbE

activ

ity (n

mol

min

minus1

mLminus

1)


R2 = 0812

P lt 00001

(a)

Serum

0 200 400 600 8000

200

400

600

800


CbE

activ

ity (n

mol

min

minus1

mLminus

1)

R2 = 0694

P lt 00001

f = 14831 + 0876lowastx

(b)

Erythrocyte

0 200 400 600 800 1000 12000

200

400

600

800

1000

1200


CbE

activ

ity (n

mol

min

minus1

mLminus

1)

R2 = 0391

P lt 00126

f = 7427 + 0616lowastx

(c)








Plas

ma

Seru

mEr

ythr

ocyt

e




80compared to IC

20)




AcTChI ester

4 Discussion



80compared to



119898and







betweenCb

EandCh

Ein

theb

lood

forb

irdsa

tdifferentd

ilutio

ns

Dilu

tion

Plasma

Erythrocyte

Serum

Quail

Duck

Chick

Quail

Duck

Chick

Quail

Duck

Chick

15

0219

minus0111

minus0402

minus064

8minus0063

0112

0411

0102

0071

(022)

(052

)(022)

(021)

(082)

(021)

(022)

(071)

(010

)

110

minus0337

0081

0123

0253

minus0580

minus0326

0026

0209

0138

(010

)(090)

(004)

(013

)(013

)(002)

(093)

(052

)(006)

115

0311

minus0179

minus0083

0152

minus0451

minus0224

0544

minus0091

0417

(031

)(032

)(021)

(054)

(012

)(074

)(004)

(092)

(009)

120

minus0245

0162

0242

0352

minus0324

0157

044

6minus0216

0429

(074

)(013

)(012

)(043)

(024)

(055)

(004)

(002)

(018

)

125

0221

0205

0134

minus0078

minus0121

minus0251

0559

minus0075

0317

(011)

(089)

(043)

(013

)(042)

(065)

(032

)(008)

(012

)Va

lues


value)


Table2Malathion

pointestim

ates(IC 2

0IC

50and

IC80120583M)from


ncurvesin

thep

resenceo

fdilu

tions

15110and

115Twopreincub

ationcond

ition

swereu

sed

long

(4h)

andshort(1h

)

Preincub

ationcond

ition

Slop

IC20(120583M)

Slop

IC50(120583M)

Slop

IC80(120583M)

Long

(4h)

Dilu

tion(15)

022plusmn0655a

315plusmn0991a

067plusmn0126

11plusmn0432a

066plusmn0718

11plusmn0121

Dilu

tion(110)

14plusmn0122

050plusmn0654

089plusmn0117

078plusmn0121

064plusmn0611

12plusmn0211

Dilu

tion(115)

011plusmn0418b

63plusmn12

b071plusmn0112

098plusmn0212

084plusmn0801

083plusmn0221

Short(1h

)Dilu

tion(15)

034plusmn0372a

21plusmn

0876a

029plusmn0501a

24plusmn0311a

037plusmn048

a19plusmn0421

Dilu

tion(110)

066plusmn0931

11plusmn0765

027plusmn0145

26plusmn0911

029plusmn018

24plusmn0987

Dilu

tion(115)

012plusmn0583b

58plusmn11

b053plusmn0196b

131plusmn

0876

024plusmn041

b29plusmn0766

Values


ofthreeindividu

alserum

esterase

(CbE

)for

quailFo

rthe

measuremento

fIC 2

0IC

50and

IC80este

rase

enzymewas

inhibitedfor3

0min

atroom

temperature

25∘

Cwith

approp

riate

concentrations

form

alathion

compo

und

Then

results

were

fittedwith

anexpo

nentiald

ecay

usingSigm

aPlot11

abdeno

tess

ignificantd

ifference

(analysis

ofvaria

nce(A

NOVA

)119875lt005)b

etween

dilutio

n15anddilutio

n115


00 05 10 15 200

20

40

60

80

100

120C

ontro

l act

ivity

()



(a)

00 05 10 15 20

Con

trol a

ctiv

ity (

)

0

20

40

60

80

100

120

140

160

180

200



(b)




minus1) 119896cat119870119898 (sminus1

120583Mminus1) 119870119898(120583M) 119896cat (s

minus1) 119896cat119870119898 (sminus1






5 Conclusions






Acknowledgments


References










































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of


0 100 200 300 400

A

A

B

a

B

b

AB

b

b

b1 5

1 10

1 15

1 20

1 25

X-d

ilutio

n

CbE ChE


(a)

0 100 200 300 400 500

Aa

B

b

b

Aa

B

Bb1 5

1 10

1 15

1 20

1 25

X-d

ilutio

nCbE ChE


(b)

0 200 400 600 800 1000

A

B

B

B

B

a

b

b

b

b

CbE ChE


1 5

1 10

1 15

1 20

1 25

X-d

ilutio

n

(c)






0 5 10 15 20 25 30 35 40 45 50

0

200

400

600

800

1000

0

200

400

600

800

1000

1200

Mean

Upper specification


minus400

minus200

minus400

CbE

activ

ity (n

mol

min

minus1

mLminus

1)

ChE

activ

ity (n

mol

min

minus1

mLminus

1)

CbEChE

X data







20IC50



to Abass [2]


3 Results



Quail

0 200 400 600 800 10000

200

400

600

800

1000


CbE

activ

ity (n

mol

min

minus1

mLminus

1)

R2 = 0114

P lt 0218

f = 3478 + 0232lowastx

(a)

Duck

0 200 400 600 800 1000 12000

200

400

600

800

1000

1200


CbE

activ

ity (n

mol

min

minus1

mLminus

1)

f = 3404 + 135lowastx

R2 = 0641

P lt 00003

(b)

Chick

0 100 200 300 400 5000

100

200

300

400

500


CbE

activ

ity (n

mol

min

minus1

mLminus

1)

R2 = 0767

P lt 00001

f = 6667 + 0843lowastx

(c)





Plasma

0 100 200 300 4000

200

400

600


CbE

activ

ity (n

mol

min

minus1

mLminus

1)


R2 = 0812

P lt 00001

(a)

Serum

0 200 400 600 8000

200

400

600

800


CbE

activ

ity (n

mol

min

minus1

mLminus

1)

R2 = 0694

P lt 00001

f = 14831 + 0876lowastx

(b)

Erythrocyte

0 200 400 600 800 1000 12000

200

400

600

800

1000

1200


CbE

activ

ity (n

mol

min

minus1

mLminus

1)

R2 = 0391

P lt 00126

f = 7427 + 0616lowastx

(c)








Plas

ma

Seru

mEr

ythr

ocyt

e




80compared to IC

20)




AcTChI ester

4 Discussion



80compared to



119898and







betweenCb

EandCh

Ein

theb

lood

forb

irdsa

tdifferentd

ilutio

ns

Dilu

tion

Plasma

Erythrocyte

Serum

Quail

Duck

Chick

Quail

Duck

Chick

Quail

Duck

Chick

15

0219

minus0111

minus0402

minus064

8minus0063

0112

0411

0102

0071

(022)

(052

)(022)

(021)

(082)

(021)

(022)

(071)

(010

)

110

minus0337

0081

0123

0253

minus0580

minus0326

0026

0209

0138

(010

)(090)

(004)

(013

)(013

)(002)

(093)

(052

)(006)

115

0311

minus0179

minus0083

0152

minus0451

minus0224

0544

minus0091

0417

(031

)(032

)(021)

(054)

(012

)(074

)(004)

(092)

(009)

120

minus0245

0162

0242

0352

minus0324

0157

044

6minus0216

0429

(074

)(013

)(012

)(043)

(024)

(055)

(004)

(002)

(018

)

125

0221

0205

0134

minus0078

minus0121

minus0251

0559

minus0075

0317

(011)

(089)

(043)

(013

)(042)

(065)

(032

)(008)

(012

)Va

lues


value)


Table2Malathion

pointestim

ates(IC 2

0IC

50and

IC80120583M)from


ncurvesin

thep

resenceo

fdilu

tions

15110and

115Twopreincub

ationcond

ition

swereu

sed

long

(4h)

andshort(1h

)

Preincub

ationcond

ition

Slop

IC20(120583M)

Slop

IC50(120583M)

Slop

IC80(120583M)

Long

(4h)

Dilu

tion(15)

022plusmn0655a

315plusmn0991a

067plusmn0126

11plusmn0432a

066plusmn0718

11plusmn0121

Dilu

tion(110)

14plusmn0122

050plusmn0654

089plusmn0117

078plusmn0121

064plusmn0611

12plusmn0211

Dilu

tion(115)

011plusmn0418b

63plusmn12

b071plusmn0112

098plusmn0212

084plusmn0801

083plusmn0221

Short(1h

)Dilu

tion(15)

034plusmn0372a

21plusmn

0876a

029plusmn0501a

24plusmn0311a

037plusmn048

a19plusmn0421

Dilu

tion(110)

066plusmn0931

11plusmn0765

027plusmn0145

26plusmn0911

029plusmn018

24plusmn0987

Dilu

tion(115)

012plusmn0583b

58plusmn11

b053plusmn0196b

131plusmn

0876

024plusmn041

b29plusmn0766

Values


ofthreeindividu

alserum

esterase

(CbE

)for

quailFo

rthe

measuremento

fIC 2

0IC

50and

IC80este

rase

enzymewas

inhibitedfor3

0min

atroom

temperature

25∘

Cwith

approp

riate

concentrations

form

alathion

compo

und

Then

results

were

fittedwith

anexpo

nentiald

ecay

usingSigm

aPlot11

abdeno

tess

ignificantd

ifference

(analysis

ofvaria

nce(A

NOVA

)119875lt005)b

etween

dilutio

n15anddilutio

n115


00 05 10 15 200

20

40

60

80

100

120C

ontro

l act

ivity

()



(a)

00 05 10 15 20

Con

trol a

ctiv

ity (

)

0

20

40

60

80

100

120

140

160

180

200



(b)




minus1) 119896cat119870119898 (sminus1

120583Mminus1) 119870119898(120583M) 119896cat (s

minus1) 119896cat119870119898 (sminus1






5 Conclusions






Acknowledgments


References










































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of


0 5 10 15 20 25 30 35 40 45 50

0

200

400

600

800

1000

0

200

400

600

800

1000

1200

Mean

Upper specification


minus400

minus200

minus400

CbE

activ

ity (n

mol

min

minus1

mLminus

1)

ChE

activ

ity (n

mol

min

minus1

mLminus

1)

CbEChE

X data







20IC50



to Abass [2]


3 Results



Quail

0 200 400 600 800 10000

200

400

600

800

1000


CbE

activ

ity (n

mol

min

minus1

mLminus

1)

R2 = 0114

P lt 0218

f = 3478 + 0232lowastx

(a)

Duck

0 200 400 600 800 1000 12000

200

400

600

800

1000

1200


CbE

activ

ity (n

mol

min

minus1

mLminus

1)

f = 3404 + 135lowastx

R2 = 0641

P lt 00003

(b)

Chick

0 100 200 300 400 5000

100

200

300

400

500


CbE

activ

ity (n

mol

min

minus1

mLminus

1)

R2 = 0767

P lt 00001

f = 6667 + 0843lowastx

(c)





Plasma

0 100 200 300 4000

200

400

600


CbE

activ

ity (n

mol

min

minus1

mLminus

1)


R2 = 0812

P lt 00001

(a)

Serum

0 200 400 600 8000

200

400

600

800


CbE

activ

ity (n

mol

min

minus1

mLminus

1)

R2 = 0694

P lt 00001

f = 14831 + 0876lowastx

(b)

Erythrocyte

0 200 400 600 800 1000 12000

200

400

600

800

1000

1200


CbE

activ

ity (n

mol

min

minus1

mLminus

1)

R2 = 0391

P lt 00126

f = 7427 + 0616lowastx

(c)








Plas

ma

Seru

mEr

ythr

ocyt

e




80compared to IC

20)




AcTChI ester

4 Discussion



80compared to



119898and







betweenCb

EandCh

Ein

theb

lood

forb

irdsa

tdifferentd

ilutio

ns

Dilu

tion

Plasma

Erythrocyte

Serum

Quail

Duck

Chick

Quail

Duck

Chick

Quail

Duck

Chick

15

0219

minus0111

minus0402

minus064

8minus0063

0112

0411

0102

0071

(022)

(052

)(022)

(021)

(082)

(021)

(022)

(071)

(010

)

110

minus0337

0081

0123

0253

minus0580

minus0326

0026

0209

0138

(010

)(090)

(004)

(013

)(013

)(002)

(093)

(052

)(006)

115

0311

minus0179

minus0083

0152

minus0451

minus0224

0544

minus0091

0417

(031

)(032

)(021)

(054)

(012

)(074

)(004)

(092)

(009)

120

minus0245

0162

0242

0352

minus0324

0157

044

6minus0216

0429

(074

)(013

)(012

)(043)

(024)

(055)

(004)

(002)

(018

)

125

0221

0205

0134

minus0078

minus0121

minus0251

0559

minus0075

0317

(011)

(089)

(043)

(013

)(042)

(065)

(032

)(008)

(012

)Va

lues


value)


Table2Malathion

pointestim

ates(IC 2

0IC

50and

IC80120583M)from


ncurvesin

thep

resenceo

fdilu

tions

15110and

115Twopreincub

ationcond

ition

swereu

sed

long

(4h)

andshort(1h

)

Preincub

ationcond

ition

Slop

IC20(120583M)

Slop

IC50(120583M)

Slop

IC80(120583M)

Long

(4h)

Dilu

tion(15)

022plusmn0655a

315plusmn0991a

067plusmn0126

11plusmn0432a

066plusmn0718

11plusmn0121

Dilu

tion(110)

14plusmn0122

050plusmn0654

089plusmn0117

078plusmn0121

064plusmn0611

12plusmn0211

Dilu

tion(115)

011plusmn0418b

63plusmn12

b071plusmn0112

098plusmn0212

084plusmn0801

083plusmn0221

Short(1h

)Dilu

tion(15)

034plusmn0372a

21plusmn

0876a

029plusmn0501a

24plusmn0311a

037plusmn048

a19plusmn0421

Dilu

tion(110)

066plusmn0931

11plusmn0765

027plusmn0145

26plusmn0911

029plusmn018

24plusmn0987

Dilu

tion(115)

012plusmn0583b

58plusmn11

b053plusmn0196b

131plusmn

0876

024plusmn041

b29plusmn0766

Values


ofthreeindividu

alserum

esterase

(CbE

)for

quailFo

rthe

measuremento

fIC 2

0IC

50and

IC80este

rase

enzymewas

inhibitedfor3

0min

atroom

temperature

25∘

Cwith

approp

riate

concentrations

form

alathion

compo

und

Then

results

were

fittedwith

anexpo

nentiald

ecay

usingSigm

aPlot11

abdeno

tess

ignificantd

ifference

(analysis

ofvaria

nce(A

NOVA

)119875lt005)b

etween

dilutio

n15anddilutio

n115


00 05 10 15 200

20

40

60

80

100

120C

ontro

l act

ivity

()



(a)

00 05 10 15 20

Con

trol a

ctiv

ity (

)

0

20

40

60

80

100

120

140

160

180

200



(b)




minus1) 119896cat119870119898 (sminus1

120583Mminus1) 119870119898(120583M) 119896cat (s

minus1) 119896cat119870119898 (sminus1






5 Conclusions






Acknowledgments


References










































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of


Quail

0 200 400 600 800 10000

200

400

600

800

1000


CbE

activ

ity (n

mol

min

minus1

mLminus

1)

R2 = 0114

P lt 0218

f = 3478 + 0232lowastx

(a)

Duck

0 200 400 600 800 1000 12000

200

400

600

800

1000

1200


CbE

activ

ity (n

mol

min

minus1

mLminus

1)

f = 3404 + 135lowastx

R2 = 0641

P lt 00003

(b)

Chick

0 100 200 300 400 5000

100

200

300

400

500


CbE

activ

ity (n

mol

min

minus1

mLminus

1)

R2 = 0767

P lt 00001

f = 6667 + 0843lowastx

(c)





Plasma

0 100 200 300 4000

200

400

600


CbE

activ

ity (n

mol

min

minus1

mLminus

1)


R2 = 0812

P lt 00001

(a)

Serum

0 200 400 600 8000

200

400

600

800


CbE

activ

ity (n

mol

min

minus1

mLminus

1)

R2 = 0694

P lt 00001

f = 14831 + 0876lowastx

(b)

Erythrocyte

0 200 400 600 800 1000 12000

200

400

600

800

1000

1200


CbE

activ

ity (n

mol

min

minus1

mLminus

1)

R2 = 0391

P lt 00126

f = 7427 + 0616lowastx

(c)








Plas

ma

Seru

mEr

ythr

ocyt

e




80compared to IC

20)




AcTChI ester

4 Discussion



80compared to



119898and







betweenCb

EandCh

Ein

theb

lood

forb

irdsa

tdifferentd

ilutio

ns

Dilu

tion

Plasma

Erythrocyte

Serum

Quail

Duck

Chick

Quail

Duck

Chick

Quail

Duck

Chick

15

0219

minus0111

minus0402

minus064

8minus0063

0112

0411

0102

0071

(022)

(052

)(022)

(021)

(082)

(021)

(022)

(071)

(010

)

110

minus0337

0081

0123

0253

minus0580

minus0326

0026

0209

0138

(010

)(090)

(004)

(013

)(013

)(002)

(093)

(052

)(006)

115

0311

minus0179

minus0083

0152

minus0451

minus0224

0544

minus0091

0417

(031

)(032

)(021)

(054)

(012

)(074

)(004)

(092)

(009)

120

minus0245

0162

0242

0352

minus0324

0157

044

6minus0216

0429

(074

)(013

)(012

)(043)

(024)

(055)

(004)

(002)

(018

)

125

0221

0205

0134

minus0078

minus0121

minus0251

0559

minus0075

0317

(011)

(089)

(043)

(013

)(042)

(065)

(032

)(008)

(012

)Va

lues


value)


Table2Malathion

pointestim

ates(IC 2

0IC

50and

IC80120583M)from


ncurvesin

thep

resenceo

fdilu

tions

15110and

115Twopreincub

ationcond

ition

swereu

sed

long

(4h)

andshort(1h

)

Preincub

ationcond

ition

Slop

IC20(120583M)

Slop

IC50(120583M)

Slop

IC80(120583M)

Long

(4h)

Dilu

tion(15)

022plusmn0655a

315plusmn0991a

067plusmn0126

11plusmn0432a

066plusmn0718

11plusmn0121

Dilu

tion(110)

14plusmn0122

050plusmn0654

089plusmn0117

078plusmn0121

064plusmn0611

12plusmn0211

Dilu

tion(115)

011plusmn0418b

63plusmn12

b071plusmn0112

098plusmn0212

084plusmn0801

083plusmn0221

Short(1h

)Dilu

tion(15)

034plusmn0372a

21plusmn

0876a

029plusmn0501a

24plusmn0311a

037plusmn048

a19plusmn0421

Dilu

tion(110)

066plusmn0931

11plusmn0765

027plusmn0145

26plusmn0911

029plusmn018

24plusmn0987

Dilu

tion(115)

012plusmn0583b

58plusmn11

b053plusmn0196b

131plusmn

0876

024plusmn041

b29plusmn0766

Values


ofthreeindividu

alserum

esterase

(CbE

)for

quailFo

rthe

measuremento

fIC 2

0IC

50and

IC80este

rase

enzymewas

inhibitedfor3

0min

atroom

temperature

25∘

Cwith

approp

riate

concentrations

form

alathion

compo

und

Then

results

were

fittedwith

anexpo

nentiald

ecay

usingSigm

aPlot11

abdeno

tess

ignificantd

ifference

(analysis

ofvaria

nce(A

NOVA

)119875lt005)b

etween

dilutio

n15anddilutio

n115


00 05 10 15 200

20

40

60

80

100

120C

ontro

l act

ivity

()



(a)

00 05 10 15 20

Con

trol a

ctiv

ity (

)

0

20

40

60

80

100

120

140

160

180

200



(b)




minus1) 119896cat119870119898 (sminus1

120583Mminus1) 119870119898(120583M) 119896cat (s

minus1) 119896cat119870119898 (sminus1






5 Conclusions






Acknowledgments


References










































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of


Plasma

0 100 200 300 4000

200

400

600


CbE

activ

ity (n

mol

min

minus1

mLminus

1)


R2 = 0812

P lt 00001

(a)

Serum

0 200 400 600 8000

200

400

600

800


CbE

activ

ity (n

mol

min

minus1

mLminus

1)

R2 = 0694

P lt 00001

f = 14831 + 0876lowastx

(b)

Erythrocyte

0 200 400 600 800 1000 12000

200

400

600

800

1000

1200


CbE

activ

ity (n

mol

min

minus1

mLminus

1)

R2 = 0391

P lt 00126

f = 7427 + 0616lowastx

(c)








Plas

ma

Seru

mEr

ythr

ocyt

e




80compared to IC

20)




AcTChI ester

4 Discussion



80compared to



119898and







betweenCb

EandCh

Ein

theb

lood

forb

irdsa

tdifferentd

ilutio

ns

Dilu

tion

Plasma

Erythrocyte

Serum

Quail

Duck

Chick

Quail

Duck

Chick

Quail

Duck

Chick

15

0219

minus0111

minus0402

minus064

8minus0063

0112

0411

0102

0071

(022)

(052

)(022)

(021)

(082)

(021)

(022)

(071)

(010

)

110

minus0337

0081

0123

0253

minus0580

minus0326

0026

0209

0138

(010

)(090)

(004)

(013

)(013

)(002)

(093)

(052

)(006)

115

0311

minus0179

minus0083

0152

minus0451

minus0224

0544

minus0091

0417

(031

)(032

)(021)

(054)

(012

)(074

)(004)

(092)

(009)

120

minus0245

0162

0242

0352

minus0324

0157

044

6minus0216

0429

(074

)(013

)(012

)(043)

(024)

(055)

(004)

(002)

(018

)

125

0221

0205

0134

minus0078

minus0121

minus0251

0559

minus0075

0317

(011)

(089)

(043)

(013

)(042)

(065)

(032

)(008)

(012

)Va

lues


value)


Table2Malathion

pointestim

ates(IC 2

0IC

50and

IC80120583M)from


ncurvesin

thep

resenceo

fdilu

tions

15110and

115Twopreincub

ationcond

ition

swereu

sed

long

(4h)

andshort(1h

)

Preincub

ationcond

ition

Slop

IC20(120583M)

Slop

IC50(120583M)

Slop

IC80(120583M)

Long

(4h)

Dilu

tion(15)

022plusmn0655a

315plusmn0991a

067plusmn0126

11plusmn0432a

066plusmn0718

11plusmn0121

Dilu

tion(110)

14plusmn0122

050plusmn0654

089plusmn0117

078plusmn0121

064plusmn0611

12plusmn0211

Dilu

tion(115)

011plusmn0418b

63plusmn12

b071plusmn0112

098plusmn0212

084plusmn0801

083plusmn0221

Short(1h

)Dilu

tion(15)

034plusmn0372a

21plusmn

0876a

029plusmn0501a

24plusmn0311a

037plusmn048

a19plusmn0421

Dilu

tion(110)

066plusmn0931

11plusmn0765

027plusmn0145

26plusmn0911

029plusmn018

24plusmn0987

Dilu

tion(115)

012plusmn0583b

58plusmn11

b053plusmn0196b

131plusmn

0876

024plusmn041

b29plusmn0766

Values


ofthreeindividu

alserum

esterase

(CbE

)for

quailFo

rthe

measuremento

fIC 2

0IC

50and

IC80este

rase

enzymewas

inhibitedfor3

0min

atroom

temperature

25∘

Cwith

approp

riate

concentrations

form

alathion

compo

und

Then

results

were

fittedwith

anexpo

nentiald

ecay

usingSigm

aPlot11

abdeno

tess

ignificantd

ifference

(analysis

ofvaria

nce(A

NOVA

)119875lt005)b

etween

dilutio

n15anddilutio

n115


00 05 10 15 200

20

40

60

80

100

120C

ontro

l act

ivity

()



(a)

00 05 10 15 20

Con

trol a

ctiv

ity (

)

0

20

40

60

80

100

120

140

160

180

200



(b)




minus1) 119896cat119870119898 (sminus1

120583Mminus1) 119870119898(120583M) 119896cat (s

minus1) 119896cat119870119898 (sminus1






5 Conclusions






Acknowledgments


References










































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of



Plas

ma

Seru

mEr

ythr

ocyt

e




80compared to IC

20)




AcTChI ester

4 Discussion



80compared to



119898and







betweenCb

EandCh

Ein

theb

lood

forb

irdsa

tdifferentd

ilutio

ns

Dilu

tion

Plasma

Erythrocyte

Serum

Quail

Duck

Chick

Quail

Duck

Chick

Quail

Duck

Chick

15

0219

minus0111

minus0402

minus064

8minus0063

0112

0411

0102

0071

(022)

(052

)(022)

(021)

(082)

(021)

(022)

(071)

(010

)

110

minus0337

0081

0123

0253

minus0580

minus0326

0026

0209

0138

(010

)(090)

(004)

(013

)(013

)(002)

(093)

(052

)(006)

115

0311

minus0179

minus0083

0152

minus0451

minus0224

0544

minus0091

0417

(031

)(032

)(021)

(054)

(012

)(074

)(004)

(092)

(009)

120

minus0245

0162

0242

0352

minus0324

0157

044

6minus0216

0429

(074

)(013

)(012

)(043)

(024)

(055)

(004)

(002)

(018

)

125

0221

0205

0134

minus0078

minus0121

minus0251

0559

minus0075

0317

(011)

(089)

(043)

(013

)(042)

(065)

(032

)(008)

(012

)Va

lues


value)


Table2Malathion

pointestim

ates(IC 2

0IC

50and

IC80120583M)from


ncurvesin

thep

resenceo

fdilu

tions

15110and

115Twopreincub

ationcond

ition

swereu

sed

long

(4h)

andshort(1h

)

Preincub

ationcond

ition

Slop

IC20(120583M)

Slop

IC50(120583M)

Slop

IC80(120583M)

Long

(4h)

Dilu

tion(15)

022plusmn0655a

315plusmn0991a

067plusmn0126

11plusmn0432a

066plusmn0718

11plusmn0121

Dilu

tion(110)

14plusmn0122

050plusmn0654

089plusmn0117

078plusmn0121

064plusmn0611

12plusmn0211

Dilu

tion(115)

011plusmn0418b

63plusmn12

b071plusmn0112

098plusmn0212

084plusmn0801

083plusmn0221

Short(1h

)Dilu

tion(15)

034plusmn0372a

21plusmn

0876a

029plusmn0501a

24plusmn0311a

037plusmn048

a19plusmn0421

Dilu

tion(110)

066plusmn0931

11plusmn0765

027plusmn0145

26plusmn0911

029plusmn018

24plusmn0987

Dilu

tion(115)

012plusmn0583b

58plusmn11

b053plusmn0196b

131plusmn

0876

024plusmn041

b29plusmn0766

Values


ofthreeindividu

alserum

esterase

(CbE

)for

quailFo

rthe

measuremento

fIC 2

0IC

50and

IC80este

rase

enzymewas

inhibitedfor3

0min

atroom

temperature

25∘

Cwith

approp

riate

concentrations

form

alathion

compo

und

Then

results

were

fittedwith

anexpo

nentiald

ecay

usingSigm

aPlot11

abdeno

tess

ignificantd

ifference

(analysis

ofvaria

nce(A

NOVA

)119875lt005)b

etween

dilutio

n15anddilutio

n115


00 05 10 15 200

20

40

60

80

100

120C

ontro

l act

ivity

()



(a)

00 05 10 15 20

Con

trol a

ctiv

ity (

)

0

20

40

60

80

100

120

140

160

180

200



(b)




minus1) 119896cat119870119898 (sminus1

120583Mminus1) 119870119898(120583M) 119896cat (s

minus1) 119896cat119870119898 (sminus1






5 Conclusions






Acknowledgments


References










































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of




betweenCb

EandCh

Ein

theb

lood

forb

irdsa

tdifferentd

ilutio

ns

Dilu

tion

Plasma

Erythrocyte

Serum

Quail

Duck

Chick

Quail

Duck

Chick

Quail

Duck

Chick

15

0219

minus0111

minus0402

minus064

8minus0063

0112

0411

0102

0071

(022)

(052

)(022)

(021)

(082)

(021)

(022)

(071)

(010

)

110

minus0337

0081

0123

0253

minus0580

minus0326

0026

0209

0138

(010

)(090)

(004)

(013

)(013

)(002)

(093)

(052

)(006)

115

0311

minus0179

minus0083

0152

minus0451

minus0224

0544

minus0091

0417

(031

)(032

)(021)

(054)

(012

)(074

)(004)

(092)

(009)

120

minus0245

0162

0242

0352

minus0324

0157

044

6minus0216

0429

(074

)(013

)(012

)(043)

(024)

(055)

(004)

(002)

(018

)

125

0221

0205

0134

minus0078

minus0121

minus0251

0559

minus0075

0317

(011)

(089)

(043)

(013

)(042)

(065)

(032

)(008)

(012

)Va

lues


value)


Table2Malathion

pointestim

ates(IC 2

0IC

50and

IC80120583M)from


ncurvesin

thep

resenceo

fdilu

tions

15110and

115Twopreincub

ationcond

ition

swereu

sed

long

(4h)

andshort(1h

)

Preincub

ationcond

ition

Slop

IC20(120583M)

Slop

IC50(120583M)

Slop

IC80(120583M)

Long

(4h)

Dilu

tion(15)

022plusmn0655a

315plusmn0991a

067plusmn0126

11plusmn0432a

066plusmn0718

11plusmn0121

Dilu

tion(110)

14plusmn0122

050plusmn0654

089plusmn0117

078plusmn0121

064plusmn0611

12plusmn0211

Dilu

tion(115)

011plusmn0418b

63plusmn12

b071plusmn0112

098plusmn0212

084plusmn0801

083plusmn0221

Short(1h

)Dilu

tion(15)

034plusmn0372a

21plusmn

0876a

029plusmn0501a

24plusmn0311a

037plusmn048

a19plusmn0421

Dilu

tion(110)

066plusmn0931

11plusmn0765

027plusmn0145

26plusmn0911

029plusmn018

24plusmn0987

Dilu

tion(115)

012plusmn0583b

58plusmn11

b053plusmn0196b

131plusmn

0876

024plusmn041

b29plusmn0766

Values


ofthreeindividu

alserum

esterase

(CbE

)for

quailFo

rthe

measuremento

fIC 2

0IC

50and

IC80este

rase

enzymewas

inhibitedfor3

0min

atroom

temperature

25∘

Cwith

approp

riate

concentrations

form

alathion

compo

und

Then

results

were

fittedwith

anexpo

nentiald

ecay

usingSigm

aPlot11

abdeno

tess

ignificantd

ifference

(analysis

ofvaria

nce(A

NOVA

)119875lt005)b

etween

dilutio

n15anddilutio

n115


00 05 10 15 200

20

40

60

80

100

120C

ontro

l act

ivity

()



(a)

00 05 10 15 20

Con

trol a

ctiv

ity (

)

0

20

40

60

80

100

120

140

160

180

200



(b)




minus1) 119896cat119870119898 (sminus1

120583Mminus1) 119870119898(120583M) 119896cat (s

minus1) 119896cat119870119898 (sminus1






5 Conclusions






Acknowledgments


References










































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of


Table2Malathion

pointestim

ates(IC 2

0IC

50and

IC80120583M)from


ncurvesin

thep

resenceo

fdilu

tions

15110and

115Twopreincub

ationcond

ition

swereu

sed

long

(4h)

andshort(1h

)

Preincub

ationcond

ition

Slop

IC20(120583M)

Slop

IC50(120583M)

Slop

IC80(120583M)

Long

(4h)

Dilu

tion(15)

022plusmn0655a

315plusmn0991a

067plusmn0126

11plusmn0432a

066plusmn0718

11plusmn0121

Dilu

tion(110)

14plusmn0122

050plusmn0654

089plusmn0117

078plusmn0121

064plusmn0611

12plusmn0211

Dilu

tion(115)

011plusmn0418b

63plusmn12

b071plusmn0112

098plusmn0212

084plusmn0801

083plusmn0221

Short(1h

)Dilu

tion(15)

034plusmn0372a

21plusmn

0876a

029plusmn0501a

24plusmn0311a

037plusmn048

a19plusmn0421

Dilu

tion(110)

066plusmn0931

11plusmn0765

027plusmn0145

26plusmn0911

029plusmn018

24plusmn0987

Dilu

tion(115)

012plusmn0583b

58plusmn11

b053plusmn0196b

131plusmn

0876

024plusmn041

b29plusmn0766

Values


ofthreeindividu

alserum

esterase

(CbE

)for

quailFo

rthe

measuremento

fIC 2

0IC

50and

IC80este

rase

enzymewas

inhibitedfor3

0min

atroom

temperature

25∘

Cwith

approp

riate

concentrations

form

alathion

compo

und

Then

results

were

fittedwith

anexpo

nentiald

ecay

usingSigm

aPlot11

abdeno

tess

ignificantd

ifference

(analysis

ofvaria

nce(A

NOVA

)119875lt005)b

etween

dilutio

n15anddilutio

n115


00 05 10 15 200

20

40

60

80

100

120C

ontro

l act

ivity

()



(a)

00 05 10 15 20

Con

trol a

ctiv

ity (

)

0

20

40

60

80

100

120

140

160

180

200



(b)




minus1) 119896cat119870119898 (sminus1

120583Mminus1) 119870119898(120583M) 119896cat (s

minus1) 119896cat119870119898 (sminus1






5 Conclusions






Acknowledgments


References










































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of


00 05 10 15 200

20

40

60

80

100

120C

ontro

l act

ivity

()



(a)

00 05 10 15 20

Con

trol a

ctiv

ity (

)

0

20

40

60

80

100

120

140

160

180

200



(b)




minus1) 119896cat119870119898 (sminus1

120583Mminus1) 119870119898(120583M) 119896cat (s

minus1) 119896cat119870119898 (sminus1






5 Conclusions






Acknowledgments


References










































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of


Acknowledgments


References










































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of














Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of

Research Article Experimental Measurements for the Effect...

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