Journal of Scientifi c & Industri al Research Vol. 60, A ugust 200 I. pp 668-674

Toxicity Evaluation of Zectran on Certain Blood Biochemical Parameters in Passer domesticus (Linneaus)

Atri Gupta , Ravi Kant Upad hyay ' and P N Saxena'il' Department of Zoo logy, lnstitute of Bas ic Sciences ,

Kh andari Campus. Agra 282 002. UP, India

Received: 06 December 2000: accepted: 15 M ay 200 I

The tox icit y of zectran has been eva luated on the basis of blood biochemical changes, especiall y concerning sublethal dose>

for acute and subchronic treatment. The LD"' of zectra n in house sparrows has been observed to be 37.3 45 mg/kg body weight and shows dose dependent mortalit y in Pas.l"l' r do111esticus, zectran also formidably shows effec t on glucose and l ipid metabo­li sm. which is noticed in hyperli pemia in both acute and subchronic treatments. A reducti on in serum cholesterol and an increase in the blood glucose level has been observed after acute and subchro nic treatment in the blood after orai administ rati on of

subleth al doses of zect ran to Passer domesticus.

Introduction Like all the other anima ls, specia lly plants , man is in

con. tan t competiti on with numerou s bi ological spec ies that ranges from bacteria, viruses , in sects, and other para­sites of domestic animals. To combat with these agents , man has ex tensively used pes ti c ides which are causing

threats to non-targeting organi sms and creating seri ous problem leading to many pathological diseases. These harmful chemicals put ad verse effect on the body of ani ­mals and inhibit cho linestrase activity' ·'. O rganophos­

phate pesticides genera ll y induce bioc he mical a nd behavioural changes in animals after dietary consump­ti onn . These pest ic ides effec t the metabol ic acti vities of the animalsx and show lethal effects. In the present investi gation biochemical changes, i.e., serum choles­tero l, serum total lipids and blood glucose are eva luated after chronic and subchron ic zec tran treatment in Passer domesticus.

Materials and Methods

The adult hou se sparrows (Passer domesticus) were procured from the local an imal catchers. the birds were kept in laboratory conditions for about a week be fore starting the experiment. The birds were housed in iron

* Author for correspondence ~ D epartment o f Zoo logy, D een Daya l Upad hyay G orak hpur University, Gorakhpur 223 009

mesh cages measurin g 43 x 29 em between 25 11 ± 511 C and re lative humidit y 60±5 per cent. The birds were pro­vided 12 h/d photoperiod The cages were c leaned regu­larl y to avo id foul sme ll and ma intain hyg iene.

Th e bird s we re fe el o n seed s o f pear l mill e t (Penn isetum typhoides) family g ramminae The test com­

pound zectran was procured from the authorized insec­ticide deale r. The LDSO was clete rmined'1 by prov iding test compound for at least 96 h. The sparrows were di­vided into four g roups each consist ing of four ind ividu­als . Each group was treated with d ifferent doses of mexacarbate by dissolving it in wate r. Therea fte r. four dos es I 0 , 20, 40 and 60 mg, res pe c ti ve ly, we re adm inistred orally per kg body weight . T he data were analyzed stati sti ca ll y by log dose/probi t regress ion line method. For acute treatment birds were subjected to sub­lethal dose of zectran for I d and for sub-chroni c treat­ment birds were treated for 5, I 0 and 15 d.

The sparrows of experimenta l sets were observed for the changes in blood biochemical parameters afte r acute ( lei ) and sub-chroni c (5 , 10 and 15 dl. The effec t of zect ran was estimated in serum tota ll ipids 111

• serum cho­leste rol11 , and for g lucose 12

• Blood was dep roteini zecl by usin g 0.4 mL of sod ium tungstate and 0.4 mL of 2/'J M sulphuric acid.

The statistical calculations were completed by using stati stical methods g iven by Fi sher and Yates 11 in bio-

GUPTA eta/.: TOXI CITY EVALUATION OF ZECTRAN 669

Table !- -Survival numbers and percentage of Passerdonresticus (Linnaeus) on treatment with zec tan (A l=Mexacarhate)

Sl No. Dose (mg/kg bw) 0. or indi vidual treated Exposure time (h ) Survival number Survival percentage

I. IOmg/ kg 4

2. 20 mg I kg 4

3. 30 mg /kg 4

4. 40 mg I kg 4

logical statistics. The calculations done were mean S.D. , SE, df and t value were signified by the Fischer's for­mula. The probability was determined as at P>0.05, P= 0.05. 0.02, 0.0 I, and. 00 I , which indicates not-signifi­cant, significant, highly significant, and very highly sig­nificant values . The working probit value was calculated by using following formula . Weighting coefficient for each point are obtained statistically~ .Each coefficient is multiplied by the number of sparrows used . The value of 'b' for regress ion equation has been obtained by the following formula :

b= SWxy- X Swy Swx2- X Swx

The value of y corresponding to the original values of x has been calculated from the following formula, y=Y + b (X-X) .

The X chi squared (.¥1 ) values has been determined by the following formula ,

)(2 = (S'H-Y2- YSWY) - b (S WXy- X S""Y).

The value was compared with the tabulated value for n-2 degree of freedom (where n is the number of experi­

ments) for 5 per cent level of significance.

Results and Discussion

Determination of LD10

The birds of different experimental sets have been ex­posed to different doses of mexacarbate and four doses I 0 mg/kg. 20 mg/kg, 40 mg/kg and 60 mg/kg, respec­ti vely, body weights were selected and the survival num­

ber and survival percentage of birds of each dose were

noted after 96 h (Table I ) . The survival percentage

showed a corresponding decrease with the increase

96 0

96 3 75

96 2 50

96 25

dose of mexacarbate. LD50

was calcu lated from log­

dose/probit mortality method 14. The tes t doses were

converted to their logrithms . (Table 2). Emperical

probit values corresponding to the per cent mortal­

ity were obtained and plotted against log doses

(Figure I). Thereafter, the provis ional line fittin g to

the point was drawn and the expected probit va lue

was noted (Figure I ). The calculated value of LD,0

was focused to be 37.345 mg/kg for Pa sser domesticus (Table3).

Assessment of Zectran Induced Biochemical Changes

The following biochemical changes were observed

in the blood of Passer_domesticus at I d of acute, and

5, I 0 and 15 d of subchronic treatment. All three

parameters serum total lipids, serum cholesterol , and

blood glucose were observed .

(A) Serum Total Lipids

The semm total lipids decrease after I d of acute treat­ment . The decrease in the serum total lipid has been ob­served to be highly significant (p < 0.0 I ) (Table 4, Figure 2). Inversely, the serum total lipids decrease after 5 d of sub-chronic treatment..

The semm total lipids decrease after I 0 d sub-chro ni c treatment. The decrease in the serum total lipid has been observed to be very highly s igniicant (p < 0.00 I )

(Table 4, Figure 2). The serum total lipids decrease after sub-chronic treatment.

(B) Semm Cholesterol

The serum cholestero l decreases after I d of acute

treatment. The decrease in the serum cholesterol has

been observed to be highly s ignificant (p < 0 .0 I )

(Table 5 , Figure3) .

Table 2 - Determination of LD50 by log-dose probit regress ion analysis after oral adminsitration of different doses of zectran to Passer domesticus (Linnaeus)

Sl No. Dose level No. of Per cent Log Empirical Expected Working Weighting Weight of zectran individual mortal ity dose pro bit pro bit probit coefficient (mg/kg BW)

X y w wx WY WXY

10 4 - 1.000 - 3.6 3.06 0.302 1.208 1.028 3.696 3.696

2 20 4 25 1.30 1 4.33 4.3 4.33 0.532 2. 128 2.768 9.2 14 11 .988

3 40 4 50 1.602 5.00 5.0 5.00 0.637 2.548 4.082 12.740 20.409

4 60 4 75 1.778 5.67 5.4 5.66 0.61 2.404 4.274 13 .607 24.139

SW SWX SWY SWXY

8.288 12.332 39.257 60.286

WX 2

1.208

3.602

6.539

7.600

SWX2

18.948

wyz

11.3110

39.8980

63.7000

77.0135

SWY 2

191.9224

0\ -l 0

._ V> Q

z 0 :::t' tT1 V>

< 0 r 0\ 0

~ c: 0 c: V> ~J

N

8

GUPTA et a/.: TOXICITY EVALUATION OF ZECTRAN 671

514

~ 4B2

E 0 S? 450 -­a>

S 41B 111 Ill 0 u ~ 3B6 l')

'0

Treatments

0 Cont rol

~Acute

E3 Sub-chronic

~:::~L-~~----~~7-----~~:-Figure !-Shows empirical probit values corresponding to

the per cent mortality in Passer domesticus

(C) Blood Glucose

The blood glucose increases after I d of acute treat­ment. The increase in the blood glucose has been observed to be highly significant (p < 0 .0 I) (Table 6, Figure 4) .

The blood glucose increases after I 0 d of sub­chronic treatment. The blood glucose increases after

15 d of sub-chronic treatment.

Discussion

The toxicity of zectran has been observed to be dose dependent as it has been found to increase with the increase in the dose level. the LD50 determined for zectran for house sparrows is 37.345 mg/kg body weight (Table I) and variance calculated 0 .0131 .

The mortality in the house sparrow is an outcome of inhibition of cholinestrase enzyme (ChE). that

plays an important role in the nervous transmission breaks down acetylcholine at cholinergic site and is responsible for acetytcholine accumlation at the syn­apses J 5- 17.

The fall in serum total lipids, cholesterol may be because of lipo-protein modifying ability of liver and accumlation and its into bile acids 1R. Simulta­neous increase in blood glucose causes hyperglyce­mia after both the acute and subchronic (5d, I Od,

325

'E Joo 0 0

]'m 0

~ 2 so u; " 0 £ 22 5

E ~

;;; 200 V>

175

Sd 10d 15d Trootment in days

Tr~o t mc:-nts

Q con rol

~ Acute

EJ Sub-c hronrc

Figure 2- Shows seru m total lipids of Passer dom esticus afte r acute and subc hroni c treatment s with Zectran. P< 0.0 I**­Hi gh ly significant. P< 0.001 ***- Very highly significant

and 15d) treatment show significant effect on the blood physiologyl 9

-21 which may be due to exces­

sive gluconeogenesis in the live r, and resulting in

excessive discharge of glucose in the blood stream.

Alterations in the biochemical compostion of blood on exposure to zectran sugges t th at the me­

tabolism of carbohydrate a nd fat is c losely intenlinked and is interconvertible as is ev ident by the decline in the serum cholesterol and an increase in blood glucose.

The fall in serum lipids is accompanied with a fall in serum cholesterol , which is derived partly from the diet and partly synthesized in certain body ti ssues. Further, cholesterol is a major constituent of lipid . A decrease in cholesterol is noticed all along with the decrease in se­rum total lipids.

Since the lipoprotein is associated with the choles­terol ester and to the free cholesterol , it is possible that the liver might have regulated the plasma choles terol. Further, it is established fact that li ver suppli es endog­enous cholesterol and cholesterol ester to pl as ma in fair sense is concerned with cholesterol homeostasis22 .

Cholesterol has a significant effect on toxicity assess­ment as it plays an indirect role in defence agai nst the xenobiotics and an elevation shows the act ivity of ACTH secretion23 ·24 . Again the reduction in serum cholesterol may probably correlates with its simultaneous accumlation in the liver25

. Simultaneous increase in blood glucose after both the acute (ID) and subchronic ( I 0 and 15 d) has been observed after malathion treatment. A

672 J SCI I 0 RES VOL 60 AUGUST 2001

Tab le 3-Tox icity eva luati on of zectran (AI-mexacarbate) to Passer dom esticus spec ify ing Fiducial limits

Expe rimental birds

House sparrows Passer domesti cus

Compound

Zectran (A I =Mexacarbate)

Regress ion equat ion

Y=4.737+3. 123 (X-1 .468)

Variance Fiducial limits

37.332 0.0 13 1 1111

= Log 1.541·1 (35 . 156) '· 1

M2

= Log 1.5991+1 (39.539) Table 4-Serum total lipids of Passer domesticus on treatment with zec tran (AI = mexaca rbate)

Sl No. Treatment# Dose Ex posure Serum total li pids Mean± S.Em S igni fica nt leve l (mglkg BW) (in d) (mg. I 100 mL) range

I Control x 11 20- 1247.27 11 70.9 ± 31.74 ** p <().01

2 Acute 10 mg I kg 9 16.36- 10 18. 18 975.75 ± 24 .9 3 Control x 5 1094 - 11 45.45 11 20 ± 11.99 *** p < 0.001 4 Sub-chronic 1.5 mg I kg 5 763.63- 7R9.09 793.33± 15.09 5 Control x 10 1094 - 1145.45 11 20 ± 11.99 "'** p < 0.00 I 6 Sub-chronic 1.5 mg I kg 10 509 .09- 636.36 568.48 ± 30. 19 7 Control x 15 11 20 - 1247.27 11 70.9 ± 31.74 ''** p < 0 .00 I 8 Sub-chronic 1.5 mg I kg 15 330.90-407.27 373 .32 ± 18.33

x =Con trols were given in the same quantit y of diluent (Distill ed water), S Em= Standard Error of mean , P =Probability. # =The no. of sparrows for all treatment s were kept three, ** = Highl y Signifi can t,*** =Very highl y signifi cant

Table 5-Serum cholesterol o f Passer domesticus on treatment with zectran (AI= mexacarbate)

Sl No Treatment# Dose Exposure Serum cholesterol Me::t n ± S.Em Sig ni ficant level (mglkg BW) (in d) (mg I I 00 mL) range

I Contro l x I 308 .86 - 326.67 3 15.54 + 4.9 ** p <0.01 2 Acute 10 mg I kg I 27 1. 11 - 277.7S 27 I.S5 + 2.6

3 Control x 5 306.67 - 324.44 3 13.70 + 4.45 *** p <O.OO I 4 Sub-chronic 1.5 mg I kg 5 220.00 - 233 .33 227.40 + 3. 1 5 Control x 10 306.67 - 324.44 3 13.70 + 4.45 *** p < 0.001 6 Sub-chronic 1.5 mg I kg 10 202.22 - 2 1 I. II 206.67 + 2.09 7 Control x 15 308.86 - 326.67 3 15.54 + 4.5 *** p < 0 .001 8 Sub-chronic 1.5 mg I kg 15 186.67 - 194.44 190.00 + 1.88

x =Controls were given in the same quantity of diluen t (Distill ed wate r), S Em= Standard error o f mean . P =Probabi lity. # =Then no. of sparrows for all treatments were kept three. ** = Highly signifi cant ,*** = Very highl y s igni fica nt Table 6--Biood glucose of Passer dom esticus on treatment with zectran (AI= mexaca rbate)

Sl No. Tretreatment # Doe Ex posure Blood g lucose Mean± S.Em S ignifi cant leve l (mglkg BW) (in d) (mg. I I 00 mL) range Control x I 32 1.17 -335.77 328.46 ± 3.4 **p<O.O I

2 Acute 10 mg I kg I 350.36- 370.80 36 1.06 ± 4 .8

3 Control x 5 3 18.28 - 327.007 322. 14 ± 2. 1 *** p < 0 .001 4 Sub-chronic 1.5 mg I kg 5 408.76 - 4 17.5 1 4 15.56 ± 2.8 5 Control x 10 3 18.25 - 327.007 322. 14 ± 2 .1 *** p < 0.00 1 6 Sub-chronic 1.5 mg I kg 10 445.25- 459.85 452.55 ± 3.4

7 Contro l x 15 32 1.1 7-335.77 328.46 ± 3.4 *** p < 0 .001 8 Sub-chronic 1.5 mg I kg 15 474.45 - 487.59 481.26±3 . 1

x =Controls were given in the same quantity o f diluent (Distilled water),~ EM= Standard error o f mean. P =Probabi lity.# =The no. o f sparrows for all treatments were kept three,** = Highl y s igni f icant ,** * =Very highl y s ignifican t

------

GUPTA el a/.: TOXICITY EVALUATION OF ZECTRA 673

1120

E 96 0 0 0 -"' 600 5. i 640

.§ 460

.8 E 2 320 "' <f)

160

0 ;;::

~ ~ ~ t/:

% ~

~

~

t/ 1d

F'

-~-

~ Sd 10d 15d

Treutment in days

Tr eatments

Control 0

~ Acute

~ Sub-chronic

Figure 3- Shows serum cholesterol or Passer domeslicus after acute and subchron ic treatments wi th Zectran . P< 0.0 1 **- Hi gh ly significant. P< 0.00 1 ***- Very hi ghl y signifi cant

0

a ~------------------~~~~--~ y=4.737+3.123 (x-1.488)

u ·;: ·a. 4

E w

2 l-~J---1J.0-0----~,~.2~0-----71.~4~o----~,~.G~o~--~,~.s~o Log Oose (x)

Figure 4--Shows blood glucose of Passer domeslicus after acute and subchronic treatments with Zectran. P< 0.0 I**- Hi ghl y sig­nillcant. P< 0.00 I*** - Very highly significant

combined effect of heptachl or and dimethoate simulta­neous decrease in serum cholesteerol and it is inferred that decrease in bile formation in the rats gets intox i­cated and result s in retenti on of cholesterol in liver"6

. It is due to fact th at dose dependent hypocho leste rolemia prevails in the blood of Passer domesticus in the present inves ti gation and is in contradi c ti on to Harsan and Santolucito27 who hypothes ized that increase in ACTH acti vity is responsible for enhancement of plasma corti­costerone and cholesterol level in liver2x

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