Prallethrin induced serum biochemical changes in ...

Accepted Manuscript Prallethrin induced serum biochemical changes in Wistar rats A. Akhtar, A.A. Deshmukh, C.G. Raut, A.P. Somkuwar, S.S. Bhagat PII: DOI: Reference:

S0048-3575(11)00202-1 10.1016/j.pestbp.2011.12.009 YPEST 3420

To appear in:

Pesticide Biochemistry and Physiology

Received Date: Accepted Date:

18 May 2011 26 December 2011

Please cite this article as: A. Akhtar, A.A. Deshmukh, C.G. Raut, A.P. Somkuwar, S.S. Bhagat, Prallethrin induced serum biochemical changes in Wistar rats, Pesticide Biochemistry and Physiology (2012), doi: 10.1016/j.pestbp. 2011.12.009

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Title: PRALLETHRIN INDUCED SERUM BIOCHEMICAL CHANGES IN WISTAR RATS A. AKHTARa, A.A.DESHMUKHa, C.G.RAUT b, A. P. SOMKUWARa AND S. S. BHAGATc • Author names and affiliations: 1. AZHAR AKHTAR a Department of Pharmacology and Toxicology, College of Veterinary and Animal Sciences, Udgir, Maharashtra, India; Email:[email protected] 2. ANANT A. DESHMUKH a Department of Pharmacology and Toxicology, College of Veterinary and Animal Sciences, Udgir, Maharashtra, India; Email: [email protected] 3. CHANDRASHEKHAR G. RAUT b

Division of Laboratory Animals, National Institute of Virology, Pune, Maharashtra, India;Email: [email protected] 4. ARJU P. SOMKUWAR a

Department of Pharmacology and Toxicology, College of Veterinary and Animal Sciences, Udgir, Maharashtra, India;Email: [email protected] 5. SUSHIL S. BHAGAT c

Department of Veterinary Pathology, College of veterinary and Animal Sciences, Udgir, Maharashtra, India;Email: [email protected]

• Corresponding author. AZHAR AKHTAR Email:[email protected] Clintox Bioservices Pvt ltd Plot 4B, Phase I, S.P. Biotech Park Turkapally, Shameerpet, Hyderabad, Andhra Pradesh, India, 500 078 Primary Phone:

+918004279256

Secondary Phone: +918960025777 Fax Number +914023480667

Abstract The present study was designed as a repeated dose 28-day oral toxicity study in rodent. All the rats were randomly divided into five groups (C1, C2, T1, T2 and T3) each containing 10 Wistar rats (5 male and 5 female). Group C1 served as control as no treatment was administered. Group C2 was administered groundnut oil (1ml/100g b.wt) and served as vehicle control. Group T1 was put on high dose 153.33 mg/kg b.wt (LD50/3), while Group T2 received intermediate dose of 92 mg/kg b.wt (LD50/5), and Group T3 was administered low dose of 46 mg/kg b.wt (LD50/10) of Prallethrin suspended in 1ml/100 g b.wt of groundnut oil. Blood samples were collected from all groups on the 7th,14th, 21st and 28th day of the experiment for measurement of serum glucose, serum urea, serum triglyceride, serum cholesterol, serum aspartate aminotransferase (AST), serum alanine aminotransferase (ALT) and serum alkaline phosphatase (ALP). According to data obtained on the 7th day of the study, no statistically significant change in any of the treatment groups was observed as compared to the control group. On the 14th day of the study, in comparison to the control group, triglyceride level and ALP activity were found to be significantly increased in the Group T1 female and Group T1 male rats respectively. On the 21stday of the study, compared to the controls, significant increase in cholesterol and ALP levels were present in both T1 and T2 females and in addition to this total protein and triglycerides levels were also significantly increased in Group T1 female rats. In Group T1 male total protein, triglycerides, ALT and ALP activity was found to be increased significantly as compared to healthy control group. On the 28th day, all the recorded biochemical parameters were found to be significantly increased, except BUN and AST in group T1 female rats. In Group T2 female rats, significantly increased cholesterol, ALT and ALP levels were observed. In Group T3 female rats, none of the parameters were found to be significantly affected. Among male rats, only total protein level was found to be increased in Groups T2 and T3.Whereas, total protein, triglyceride, ALT and ALP were significantly elevated in Group T1 male rats at the end of the study. In conclusion, the results of this study demonstrate that subacute oral administration of prallethrin; at dose levels of 1/3 LD50 and 1/5 LD50for 28 days induces moderate toxic effects on different biochemical parameters.

Keywords: Subacute Toxicity, Prallethrin, Biochemical Parameters

1. INTRODUCTION Synthetic pyrethroids are synthetic chemical analogs and derivatives of pyrethrins and represent the third largest class of chemical insecticides after organophosphates and chloronicotinyl insecticides [1].Pyrethroids are much more effective against a wide spectrum of economically important pests than the organochlorine, organophosphate and carbamate insecticides. In majority of species thus far investigated, high-level exposures to pyrethroids induce toxic signs that are characteristic of a strong excitatory action on the nervous system. The pyrethroids have been divided into two classes (Type I and Type II) on the basis of their chemical structure and toxic manifestations. Type I pyrethroids do not include a cyano group, and signs of toxicity include whole body fine muscular tremors. Type II pyrethroids include a cyano group and are characterized by more complex toxic manifestations such as choreoathetosis (writhing, shaking) and salivation [2, 3.] The development of permethrin, the first potent and photostable pyrethroid by Elliott and Janes (1978) [4] catalyzed intensive research and development of this class of compound globally. As a result many photostable pyrethroids were synthesized that proved extremely effective agricultural insecticides and then replaced earlier pyrethroids and insecticides of other classes in a variety of nonagricultural uses. It is worth mentioning that, despite the development of latest congeners of the early chemical pesticides, the target species selectivity of pesticides is not as well developed as might be hoped for, and non-target species are frequently affected because they possess physiological and/or biochemical systems similar to those of the target organisms. In the synthetic pyrethroid armory, Prallethrin {CAS No: 23031-36-9 (racemic)} is the most popular Type I synthetic pyrethroid which produces a rapid knock-down in household insect pests such as mosquitoes, houseflies and cockroaches [5].At present, Prallethrin has prevalent household presence in the form of mosquito repellant mats, coils, liquid vaporizers etc. and therefore there could be direct and indirect exposure in pets and humans through accidental continued contamination of feed/food and water. Moreover, there is no data available on repeated oral exposure of prallethrin at subacute doses. Hence, the present investigation was undertaken to assess the effects of Prallethrin on biochemical parameters in male and female Wistar rats at repeated doses of for a period of 28 days. In a separate study, the oral LD50 of prallethrin was determined to be 460mg/kg b.wt. and 640 mg/kg b.wt. in female and male rats respectively [6]. In the present study, 460mg/kg b.wt dose was used as the approximate median lethal dose for both the sexes of rats for the calculation of three subacute doses.

i.e.153.33mg/kg/b.wt/day

(1/3LD50),

92mg/kg/b.wt/day

(1/5LD50)

and

46mg/kg/b.wt/day

(1/10LD50) 2. MATERIALS AND METHODS Prallethrin Technical (95.53 %) was obtained from M/S Shogun Organics Ltd., Thane, India. Fifty post weaning Wistar rats (25 male and 25 female), aged 6-8 weeks with body weight of 145–200 g were used in the study. The temperature in the experimental animal room was maintained at 22°C (± 3°C) with artificial lighting, the sequence being 12 hours light, and 12 hours dark. The animals were provided with pellet food (M/S Amrut feeds Ltd., Pune, India) and drinking water ad libitum. The experimental animals were housed in conventional polypropylene cages in small groups (five each) of same sex. The rats were randomly assigned to control and treatment groups. The animals were allowed to acclimatize to the laboratory conditions for five days prior to the start of study. The present study was designed as a repeated dose 28-day oral toxicity study in rodent [7]. The experimental animals (five groups of males and five groups of females) were administered with Prallethrin at concentrations of 153.33 mg/kg b.wt. (Group T1: 1/3 LD50), 92 mg/kg b.wt (Group T2 1/5 LD50) and 46 mg/kg b.wt (Group T3 1/5 LD50). Prallethrin (1mL/100g b.wt.) suspended in an emulsion of groundnut oil, was administered intragastrically, once a day, for a period of 28 days. Two control groups were used for parallel studies. No treatment was administered in Control Group C1 and was designated as healthy control. The vehicle control Group C2, was administered intragastrically the groundnut oil used for suspending the Prallethrin. At 7th, 14th, 21st and 28th day of the treatment, blood samples were collected from orbital plexus of all the rats to perform biochemical (serum glucose, serum urea, serum triglyceride, serum cholesterol, serum aspartate aminotransferase (AST), serum alanine aminotransferase (ALT) and serum alkaline phosphatase (ALP)) estimations by standard kits obtained from Crest Biosystems on Evolution 3000, Tulip Diagnostics autoanalyzer. All the rats were sacrificed by ether anesthesia with cervical dislocation on 29th day of the experiment in a confined disinfected area and were subjected to post mortem examination for gross alterations. Suitable pieces of organs were collected and kept in 10 % formalin for histopathological examinations. The formalin fixed tissues were processed by paraffin wax embedding method of tissue sectioning as per method described by Culling [8]. Sections were cut at 4-5 microns thickness with rotatatory microtome and were stained with Haematoxylin and

Eosin (H & E) stains [9]. The H & E stained slides were observed under microscope and lesions were recorded. One-way ANOVA with Dunnett’s post test was performed using GraphPad Prism version 5.00 for Windows, GraphPad Software, San Diego California USA, to compare the effects of treatment of various doses of Prallethrin on biochemical variables to that of control group (C1).

3. RESULTS 3.1 Serum Biochemistry 3.1.1

Serum glucose

Subacute poisoning with Prallethrin in female rats did not result in any significant change in mean serum glucose levels of all the treatment groups when compared to the control group (C1) up to the end of third week. However, at the end of fourth week, group T1 female rats (high prallethrin dose) exhibited a statistically significant rise (p < 0.05) in mean serum glucose level when compared to the control group (C1). Moreover, mean serum glucose levels in all the treatment groups of male rats did not reveal any statistical significant difference when compared with the control group (C1), during the entire period of study (Fig. 1). 3.1.2

Blood urea nitrogen (BUN)

Despite per os administration of Prallethrin at various dose levels for 28 consecutive days, Prallethrin did not produce any statistically significant change in the mean serum urea levels in rats of both sexes of all the treatment groups as compared to the control group (C1) (Fig. 2). 3.1.3

Serum total protein

Prallethrin treated female rats of Group T1 showed a statistically significant increase (p < 0.05) in mean total protein level from third week onwards and continued till the end of fourth week, when compared to the control Group (C1) at respective periods. On the contrary to the female rats of Group T1, no statistically significant difference was detected in groups T2 and T3 when compared to the control group (C1) female rats throughout the study. Male rats of all treatment groups exhibited a rise in the mean total serum protein level from third week onwards until the end of fourth week. However, a statistically significant increase (p < 0.05) was found in only male rats of Group T1 at the end of third week and in male rats of all treatment groups at the end of fourth week as compared to male rats of the Control Group (C1) at respective periods (Fig. 3).

3.1.4

Serum cholesterol

Female rats of all the treatment groups exhibited an increase in mean values of serum cholesterol from the end of first week onwards until the end of fourth week. However, the female rats of groups T1 and T2 showed a statistically significant increase (p < 0.05) in mean values of serum cholesterol at the end of third and fourth week, compared to the control group (C1) female rats at respective periods. On the contrary to the female rats, male rats of only treatment group T1 exhibited slight statistically non-significant increased mean values of serum cholesterol from the end of first week onwards until the end of fourth week when compared to the control group (C1) during the entire period of study (Fig. 4). 3.1.5

Serum triglycerides

In response to daily administration of Prallethrin, female rats of treatment group T1 and T2 exhibited a dose dependent increase in mean serum triglyceride values from the end of second week onwards until the end of fourth week of study. However, the female rats of only group T1 exhibited a statistically significant (p < 0.05) increase in mean serum triglyceride values at the end of second, third and fourth week, when compared to control group (C1) at respective periods. Similarly, the male rats of only treatment group T1 exhibited a statistically significant increase (p < 0.05) in mean serum triglyceride values from the end of third week onwards until the end of fourth week, compared to control group (C1) at respective intervals (Fig. 5). 3.1.6

Serum aspartate aminotransferase (AST)

Administration of Prallethrin in the treatment groups T1 and T2 (both sexes) produced an increase in the mean serum AST activity at different stages of the study. However, the results were statistically non-significant when compared with the control group (C1) throughout the study (Fig. 6). 3.1.7

Serum alanine aminotransferase (ALT)

Female rats of all the treatment groups exhibited a dose dependent increase in mean serum ALT activity from the end of first week onwards until the end of fourth week. However, the female rats of groups T1 and T2 showed a statistically significant increase in mean serum ALT activity at the end of fourth week, compared to control group (C1) female rats (Fig. 7). Similarly, male rats of all the treatment groups exhibited a marked increase in mean serum ALT activity from the end of third week onwards until the end of study. However, the male rats of only group T1 showed a

statistically significant increase in mean serum ALT activity at the end of third and fourth week, compared to the control group (C1) male rats at respective periods (Fig. 7). 3.1.8

Serum alkaline phosphatase (ALP)

Female rats of all the treatment groups exhibited a dose dependent increase in the mean serum ALP activity from the end of second week onwards until the end of study. However, female rats of groups T1 and T2 showed a statistically significant increase in the mean serum ALP activity from the end of third week onwards until the end of fourth week, compared to the control group (C1) female rats at respective periods (Fig. 8). Male rats of all treatment groups exhibited increased mean serum ALP activities from the end of first week onwards until the termination of study. However, the male rats of treatment group T1 exhibited a statistically significant increase in the mean ALP activity at the end of second, third and fourth week, as compared to the control group (C1) male rats at respective periods (Fig. 8). 3.1.9

Histopathological Observations

Histopathological changes were observed in all the major organs of prallethrin treated animals as compared to the control group. In contrast to the normal histological examination of liver tissue of controls, hemorrhages, congestion, dilatation of sinusoidal spaces (fig.9) and other necrobiotic changes were observed in liver of all prallethrin treated rats. Similarly, diffuse hemorrhages with oedema (fig.10) and emphysema in alveoli were seen in the lung tissue of prallethrin-treated rats as compared to the normal histological examination of lung tissue in control rats. The kidneys exhibited cellular swelling, cystic degeneration, and mononuclear infiltration (fig.11). In testis section, there was intertubular oedema (fig. 12) with vacuolation within tubules.

DISCUSSION In toxicity studies, a variety of biochemical parameters are measured to evaluate a broad range of physiological and metabolic functions (e.g. serum glucose for carbohydrate metabolism, and serum triglycerides for lipid metabolism.), affected target organ identification and tissue injury assessment. Moreover, a combination of some common biochemical parameters provide better information from pattern recognition, e.g. plasma enzymes like AST, ALT and ALP for hepatotoxicity and urea and creatinine for glomerular function[10]. The result of the present study showed that per os administration of Prallethrin increased the level of serum glucose in female rats at highest dose (Group T1) (Fig. 1). This might be due to

stress caused by Prallethrin exposure. Stress causes decreased insulin synthesis and increased adrenocorticotropic hormones and glucagons synthesis which converts hepatic glycogen into glucose [11]. Among earlier similar studies carried out on other synthetic pyrethroids in various animal species Manna et al., [12] reported a significant rise in serum glucose levels in Wistar rats of both the sexes after administration of -cypermethrin for 30 days. Blood urea nitrogen estimates blood - filtering capacity of the kidneys and does not become significantly elevated until kidney function is reduced 60 – 75 % [13]. In this study, Prallethrin did not produce any statistically significant change in the mean serum urea levels in rats of both sexes of all the treatment groups (Fig. 2) as compared to the control group because the kidney might not be damaged to such an extent that it affected BUN levels. Contrary to the present findings, it is reported that various synthetic pyrethroids intoxication result in increase in urea levels [14]. Serum total protein (Fig. 3) was found to be significantly increased in both sexes in dose dependent manner at various time points in this study. The values are raised mainly due to rise in its constitutive fractions like albumin, globulins etc. and a simple hyperproteinemia without change in albumin: globulin ratio occurs due to dehydration [15]. Dehydration might have played a role in causing hyperproteinemia in the present study as Prallethrin treated rats were dehydrated after prolonged duration of tremors. Disparate to the present findings, it is reported that intoxication with other pyrethroids causes either a decrease [16] or no change [17] in total protein levels. Carlson and Kalmodin Hedman [18] reported that the accumulation of pesticides in liver was associated with the disturbance of lipid metabolism and an elevation of serum cholesterol. In this study, Prallethrin significantly increased serum cholesterol only in female rats of both high (Group T1) and medium dose (Group T2) groups. Prallethrin induced increase in serum cholesterol can be attributed to the effects of pesticides on the permeability of liver cell membrane [19] and /or liver dysfunction [20] as confirmed by the increment of ALT and ALP levels. Perusal to the available literature reveals that Al-Qarawi and Adam [21] studied the effects of oral administration of the combination of Malathion and superphosphate or urea on Najdi sheep. Similarly, Yousef et al. [14] reported an increase in serum triglycerides values in male rabbits fed with 24 mg/kg b.wt of cypermethrin at the end of sixth week. Therefore, increase in serum

triglyceride level in the present study may indicate impairment of lipid metabolism in both sexes of Wistar rats. Serum enzymes like AST, ALT and ALP represent the functional status of the liver [22]. AST is also present in skeletal muscle and heart and most commonly associated with damage to these. Therefore, the present insignificant increased values of AST (Fig. 6) may indicate that subacute exposure of Prallethrin to Wistar rats (both sexes) might not have adverse effect on skeletal muscles and heart but indicate evidenced liver damage as AST escapes from the disrupted hepatic parenchyma cells with necrosis. Evaluation of ALT is usually associated with hepatic damage or disease and a significant increase in serum ALT were also reported with other pyrethroids such as fenvalerate [23] in buffalo calves and permethrin [24] and fluvalinate [25] in rats and, so the elevation of ALT enzyme activity (Fig. 7) in this study suggests probable liver tissue damage due to Prallethrin as evidenced by the histopathological lesions in liver of prallethrin treated rats. Liver being the organ of biotransformation of synthetic pyrethroids, early pathological changes like congestion, haemorraghes and other necrobiotic changes in the liver are probably associated due to decreased free radical (O-2) scavenger formation and most prominent lesions produced by them include vacuolar degeneration, degeneration of hepatic cords and hepatocytes, focal to extensive necrosis, enlargement and dilation of sinusoids [26]. The higher base value of ALP is an indicator of growth in Wistar rats, as bone alkaline phosphate tends to be higher in young animals. Moreover, serum ALP activity increases in case of damage to the hepatic cells and obstruction of the bile ducts [14]. Therefore, the present increase in ALP activity (Fig. 8) is probably the consequence of Prallethrin induced pathological changes in the liver. Similar to the present findings, Manna et al., [11] found a rise in serum ALP activity in both the sexes of Wistar rats after administration of -cypermethrin for 30 days. In the present study, high concentration of Prallethrin in lungs may have caused inflammation leading to progressive oedema. In testis, there were vacuole formation and edematous fluid accumulation, which might be due to testicular degeneration induced by Prallethrin. All these observations suggest that biochemical alterations induced by prallethrin toxicity are species, dose and sex dependent and there is scope for further investigation in context with the sex hormones affecting the degree of toxic signs as females exhibited more toxic manifestations than males at the same dose levels.

4. CONCLUSIONS

The results of this study presents strong evidence of moderate toxic effect of the Prallethrin at dose levels of 1/3 LD50 and 1/5 LD50 for 28 days on biochemical functions which correlate well with the histopathological changes in various vital organs. Although the data cannot be extrapolated directly from rats to human being, but it may be concluded that prallethrin use may cause hazardous effects in various levels to nontarget species.

5.

ACKNOWLEDGMENT

The authors acknowledge the gift of technical grade Prallethrin by M/S Shogun Organics Ltd., Thane, India, to carry out the research work.

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2. J.R. Coats, Mechanisms of toxic action and structure-activity relationships for organochlorine and synthetic pyrethroid insecticides. Environ Health Perspect. 87 (1990) 255-262. 3. R.D. Verschoyle, W.N. Aldridge, Structure-activity relationships of some pyrethroids in rats. Arch Toxicol. 45 (1980) 325-329. 4. M. Elliot, N.F. Janes, Synthetic pyrethroids - a new class of insecticides. Chem. Soc. Revs., 7 (1978) 473. 5.

T. Matsunaga,M. Makita, A. Higo, I. Nishibe, K. Dohara, G.Shinjo,, Studies on prallethrin, a new synthetic pyrethroid for indoor applications. The insecticidal activities of prallethrin. Jpn. J. Sanit. Zool. 38 (1987) 219-223.

6. The WHO recommended Specifications and Evaluations for Public Health Pesticides Prallethrin, WHO, Geneva, 2004, pp.1-17. 7. OECD (Organization for Economic Cooperation and Development) Repeated dose 28-day oral toxicity study in rodents. In Guideline for the Testing of Chemicals, 1995, pp. 407. 8. C. F. A. Culling, A Hand Book of Histopathological and Histochemical Techniques. 3rd ed. Butter worth and Co. Ltd. 1974,pp- 29-22. 9. L.G Luna, Manual of histologic staining methods of the Armed Forces Institute of Pathology. 3rd ed. McGraw Hill Book Co., New York.1968. 10. C.O. Evans,General Introduction, in: G.O. Evans. (Ed.), Animal Clinical Chemistry - A Primer for Toxicologists. USA Taylor & Francis Inc., Frost Road, Suite 101, Bristol,1996, pp. 1- 9

11. G. Eraslan, A. Bilgili, D. Essiz M. Akdogan,F. Saindokuyucu, The effects of deltamethrin on some serum biochemical parameters in mice. Pesticide Biochemistry and Physiology. 87,( 2007), 123-130. 12. S. Manna, D. Bhattacharyya, S. Das, T.K. Mandal, Repeated dose toxicity of

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cypermethrin in rats. J. Vet. Sci. 5 ,(2004),241–245. 13. S.C. Gad, Repeat dose toxicity studies, in: S.C. Gad (Ed.), Preclinical Development Handbook. John Wiley &Sons, Inc., Hoboken, New Jersey, 2008, 221-225 14. M.I. Yousef, F.M. El-Demerdash, K.I. Kamel, K.S. Al-Salhen, Changes in some hematological and biochemical indices of rabbits induced by isoflavones and cypermethrin. Toxicology. 189(2003)223-234. 15. J.J. Kaneko, Serum Proteins and the Dysproteinemias, in: J.J. Kaneko, J.W. Harvey,M.L Bruss, (Eds),Clinical Biochemistry of Domestic Animals. Harcourt Brace and Company Asia PTE Ltd. Singapore,1997, pp134 16. F.M. El-demerdash, M.I. Yousef , K.S. Al-Salhen,Protective effects of isoflavone on some biochemical parameters affected by cypermethrin in male rabbits. J. Environ. Sci. Health B. 189, (2003) 365- 378. 17. F. Sayim, N.U.K. Yavasoglu, Y. Uyanikgil, H.Aktug, A. Yavasoglu, M. Turgut, Neurotoxic Effects

of

Cypermethrin

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Wistar

Rats:

a

Hematological,

Biochemical

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Histopathological Study. Journal of Health science, 51 (2005) 300-307. 18. L.A. Carlson, B. Kolmodin-Hedman, Hyper alpha lipoprotenemia in men exposed to chlorinated hydrocarbon pesticides. Acta. Med. Cand. 192 (1972) 29-32. 19. K.G. Adham, A. Khairalla, M. Abu-Shaban, N. Addel-Mguid, A.A. El-Mmoneim, Environmental stress in Lake Maryut and physiological response of Tilapia zilliGerv. J. Environ. Sci. Health 32 (1997) 2585-2598. 20. S.Kalender, A. Ogutcu, M. Uzunhisarcikli, F.Acikgoz, D. Durak, Y. Ulusoy, Y. Kalender,Diazinon-induced hepatotoxicity and protective effect of vitamin E on some biochemical indices and ultrastructural changes. Toxicology. 211(2005)197–206. 21. A.A. Al-Qarawi, S.E. Adam, Effects of malathion plus superphosphate or urea on Najdi sheep. Vet. Hum. Toxicol. 45 (2003) 3-6. 22. S. Manna, D. Bhattacharyya, T.K. Mandal, S. Das,Sub-Chronic Toxicity Study of AlfaCypermethrin in Rats. Iranian Journal of Pharmacology & Therapeutics. 5 (2006) 163-166. 23. G. Singh, L.D. Sharma, A.H. Ahmad, S.P. Singh, Fenvalerate induced dermal toxicity in buffalo calves. Journal of Applied Animal Research. 16 (1999) 205-210.

24. M.A. Shah, P.K. Gupta, Subacute toxicity studies on permethrin. Indian J. Toxicol. 8 (2001) 61-67. 25. U.K.Garg, A.K. Pal, G.J. Jha, S.B. Jadhao, Haemato-biochemical and immunopathophysiological effects of chronic toxicity with synthetic pyrethroid organophosphate and chlorinated pesticides in broiler chicks. International Immunopharmacology. 4 (2004)1709–1722. 26. A.Yavasoglu, F. Sayim, Y. Uynikgil, M. Turgut, N. U. karabay- Yavasoglu, The pyrethroid cypermethrin induced biochemical and histological alterations in rat liver. journal of health sciences. 52 (2006) 774-780.

. Control

Vehicle

46 mg/kg

92 mg/kg

153.33 mg/kg

100.00

*

90.00 80.00

Glucose (mg/dl)

70.00 60.00 50.00 40.00 30.00 20.00 10.00 0.00 7

14 Female

21

28

7

14 Male

21

28

Experimental Days

Figure 1: Effect of repeated oral administration of Prallethrin on Serum Glucose of Wistar rats *-

Significant (P < 0.05), **- Very significant (P < 0.01), ***- Extremely significant (P < 0.001)

Control

Vehicle

46 mg/kg

92 mg/kg

153.33 mg/kg

60.00

50.00

BUN (mg/dl)

40.00

30.00

20.00

10.00

0.00 7

14

21

Female

28

7

14

21

Male

28

Experimental Days

Figure 2: Effect of repeated oral administration of Prallethrin on BUN of Wistar rats *-


Control

Vehicle

46 mg/kg

92 mg/kg

153.33 mg/kg

20.00 18.00 16.00

Total Protein ( g/dl )

* 14.00 12.00

***

* *

10.00

** *

8.00 6.00 4.00 2.00 0.00 7

14

21 Female

28

7

14

21 Male

28

Experimental Days

Figure3: *-

Effect of repeated oral administration of Prallethrin on Serum Total Proteins of Wistar rats


Control

Vehicle

46 mg/kg

92 mg/kg

153.33 mg/kg

100.00 90.00 80.00

** *

Cholesterol (mg/dl)

70.00

** *

60.00 50.00 40.00 30.00 20.00 10.00 0.00 7

14

21

Female

28

7

14

21

Male

28

Experimental Days Figure 4: Effect of repeated oral administration of Prallethrin on Serum Cholesterol of Wistar rats ,*-


Control

Vehicle

46 mg/kg

92 mg/kg

153.33 mg/kg

140.00

120.00 **

Triglyceride (mg/dl)

100.00

**

*

** ***

80.00

60.00

40.00

20.00

0.00 7

14

21

Female

28

7

14

21

Male

28

Experimental Days

Figure 5: Effect of repeated oral administration of Prallethrin on Serum Triglycerides of Wistar rats ,*-


Control

Vehicle

46 mg/kg

92 mg/kg

153.33 mg/kg

200.00 180.00 160.00 140.00

AST (IU/L)

120.00 100.00 80.00 60.00 40.00 20.00 0.00 7

14

21

Female

28

7

14

21

Male

28

Experimental Days Figure 6: ,*-

Effect of repeated oral administration of Prallethrin on AST of Wistar rats


Control

Vehicle

46 mg/kg

92 mg/kg

153.33 mg/kg

90.00 80.00

**

**

** **

70.00

ALT (IU/L)

60.00 50.00 40.00 30.00 20.00 10.00 0.00 7

14

21

Female

28

7

14

21

Male

28


Effect of repeated oral administration of Prallethrin on ALT of Wistar rats


Control

Vehicle

46 mg/kg

92 mg/kg

153.33 mg/kg

500.00 *

450.00 * 400.00

**

* * **

350.00

*

ALP (U/L)

300.00 250.00 200.00 150.00 100.00 50.00 0.00 7

14

21

Female

28

7

14

21

Male

28


Effect of repeated oral administration of Prallethrin on ALP of Wistar rats Significant (P < 0.05), **- Very significant (P < 0.01), ***- Extremely significant (P < 0.001)

Figure 9: Photomicrograph of prallethrin treated rat liver tissue showing congestion and disruption of sinusoids on 29th day (H & E X 200).

Figure10: Photomicrograph of prallethrin treated rat lung showing severe congestion, emphysema and oedema on 29th day (H & E X 200).

Figure 11:Photomicrograph of prallethrin treated rat kidney showing cellular swelling and mononuclear infiltration on 29th day (H & E X 200).

Figure 12: Photomicrograph of prallethrin treated rat testis showing oedema in intertubular spaces on 29th day (H & E X 200).

EXPERIMENTAL TIMEPOINTS

HIGH DOSE GROUP (153.33 mg/kg b.wt/day)

PRALLETHRIN TECHNICAL

ORAL ADMINISTRATION OF PRALLETHRIN IN WISTAR RAT

MEDIUM DOSE GROUP (92 mg/kg b.wt/day) LOW DOSE GROUP (46 mg/kg b.wt/day)

th

th

st

14 DAY

7 DAY

th

21 DAY



TRIGLYCERIDES

   

TOTAL PROTEIN CHOLESTEROL TRIGLYCERIDES ALP



ALP

     

TOTAL PROTEIN TRIGLYCERIDES ALT ALP CHOLESTEROL ALP

FEMALE

________

MALE

________

FEMALE

________

________

MALE

________

________

________

FEMALE

________

________

________

MALE

________

________

________

28 DAY              

GLUCOSE TOTAL PROTEIN CHOLESTEROL TRIGLYCERIDES ALT ALP TOTAL PROTEIN TRIGLYCERIDES ALT ALP CHOLESTEROL ALT ALP TOTAL PROTEIN

________  TOTAL PROTEIN

EFFECT OF PRALLETHRIN ADMINISTRATION ON SERUM BIOCHEMICAL PARAMETERS IN WISTAR RATS 

Statistically significant increase in comparison to healthy control group ;

------- No statistically significant change in comparison to healthy control group

HIGHLIGHTS • Female rats were more susceptible to toxic effects of Prallethrin than male rats, at different dose levels. • Majority of the studied parameters got significantly elevated at the end of fourth week. • Elevated biochemical parameters correlated well with the histopathological changes in various vital organs.