Oxidative stress and cytotoxic potential of anticholinesterase insecticide, malathion in reproductive toxicology of male adolescent mice after acute exposure

Document Type : Original Article


Laboratory of Animal Physiology, Department of Biology, Faculty of Sciences, University Tunis El Manar, Tunis, Tunisia


Objective(s):The present study was undertaken to determine the effects of acute exposure to malathion on oxidative stress and cytotoxic potential of anticholinesterase insecticide, malathion in reproductive toxicology of adolescent male mice.
Materials and Methods: Thirty two adolescent male mice at pubertal age were treated with 500 mg/kg body weight (BW) of malathion for three days. After exposure, biochemical markers and sperm analysis were evaluated and finally histological modifications of testis and sperm were assessed.
Results: Our data showed that treatment of male mice with malathion (500 mg/kg, BW) could lead to oxidative stress. Induced oxidative stress status can be assessed due to increased malondialdhyde (MDA) content, decreased thiol group content, as well as increased antioxidant enzyme activities. On the other hand, exposure to malathion at the pubertal age led to alteration of semen parameters; sperm production and percentage of motile sperm were decreased in the treated groups compared to the control.  Furthermore, exposure of male mice to malathion led to a decrease of testosterone level, inhibition of acetylcholinesterase, and decrease of the reproductive performance of male mice after three days of treatment at the age of puberty.
Conclusion: The importance to carry out in vitro reproductive toxicology assays lies on the need of knowing the alterations these insecticides may cause at cellular level, since they are endocrine disruptors that interfere with reproductive functions.


1. Kamath V, Rajini PS. Altered glucose homeostasis and oxidative impairment in pancreas of rats subjected to dimethoate intoxication. Toxicology 2007; 231:137–146.
3. Selmi S, El-Fazaa S, Gharbi N. Oxidative stress and alteration of biochemical markers in liver and kidney by malathion in rat pups. Toxicol Ind Health 2013; 1-7.
4. Fortunato JJ, Feier G, Vitali AM, Petronilho FC, Dal-Pizzol F, Quevedo J. Malathion-induced oxidative stress in rat brain regions. Neurochem Res 2006; 31:671–678.
5. Debnath D, Mandal TK. Study of quinalphos (an environmental oestrogenic insecticide) formulation (Ekalux 25 E.C.)-induced damage of the testicular tissues and antioxidant defense systems in Sprague–Dawley albino rats. J Appl Toxicol 2000; 20:197–204.
6. Padungtod C, Savitz DA, Overstreet JW, Christiani DC, Ryan LM, Xu X. Occupational pesticide exposure and semen quality among Chinese workers. J Occup Environ Med 2000; 42:982–992.
7. Ramezani A, Goudarzi I, Lashkarboluki T, Ghorbanian MT, Abrari K, Elahdadi Salmani M. Role of oxidative stress in ethanol-induced neurotoxicity in the developing cerebellum. Iran J Basic Med Sci 2012; 15:965-974.
8. Khastar H, Kadkhodaee M, Sadeghipour HR, Seifi B, Hadjati J, Najafi A, et al. Liver oxidative stress after renal ischemia-reperfusion injury is leukocyte dependent in inbred mice. Iran J Basic Med Sci 2011; 14:534-539.
9. da Silva AP, Meotti FC. Lactational exposure to malathion inhibits brain acetylcholinesterase in mice. Neurotoxicology 2006; 27:1101–1105.
10. Lasram MM, Alya BA, El Elj N, Selmi S, El Fazaa S, Gharbi N. Effect of short-time malathion administration on glucose homeostasis in Wistar rat. Pestic Biochem Physiol 2008; 92:114–119.
11.  Videira RA, Antunes-Madeira MC. Changes induced by malathion methylparathion and parathion on membrane lipid physicochemical properties correlate with their toxicity. Biochem Biophys Acta 2001; 1511:360–368.
12.  Petrelli G, Mantovani A.  Environmental risk factors and male fertility and reproduction. Contraception 2002; 65:297–300.
13.  Pina-Guzman B, Solıs-Heredia MJ Quintanilla-Vega B. Diazinon alters sperm chromatin structure in mice by phosphorylating nuclear protamines. Toxicol Appl Pharmacol 2005; 202: 189–198.
14. Ellman GA. New and rapid colorimetric determination of acetylcholinesterase activity. Biochem Pharmacol 1961; 7:88-95.
15. Vega SG, Guzman P, Garcia L, Espinosa J, De Nava CC. Sperm shape abnormality and urine mutagenicity in mice treated with niclosamide. Mutat Res 1988; 204:269–276.
16. Tardif S, Laforest JP, Comier N, Bailey JL. The importance of porcine sperm parameters on fertility in vivo. Theriogenology 1999; 52:447–459.
17.  Lobet JM, Colomina MT, Sivent JJ. Reproductive toxicology of aluminium in male mice. Fund Appl Toxicol 1995; 25:45-51.
18. Kakinuma K, Yamaguchi T, Kaneda M, Shimada K, Tomita Y, Chance B. determination of H2O2 release by the treatment of human blood polymorphonuclear leucocytes with myristate. J Biochem1979; 86:87-95.
19. Aebi H. Catalase in vitro. Methods Enzymol 1984; 105:121–126. 
20. Bradford MM. A rapid and sensitive method for the quantitation of µg quantities of protein utilizing the principal of protein-dye binding. Anal Biochem 1976; 72:248–254.
21. Rubin C, Esteban E, Kieszak S, Hill Jr, RH, Dunlop B, Yacovac R, et al. Assessment of human exposure and human health effects after indoor application of methyl parathion in Lorain County, Ohio, 1995–1996. Environ Health Perspect 2002; 110:1047–1051.
22. Pope CN. Organophosphorus pesticides: do they all have the same mechanism of toxicity? J Toxicol Environ Health B Crit Rev 1999; 2:161–181.
23. Maroni M, Colosio C, Feridi A, Fait A. Organophosphorus pesticides. Toxicology 2000; 143:9–37.
24. Sharma Y, Somia B, Irshadb M, Datta G, Dograa TD. Effects of acute dimethoate administration on antioxidant status of liver and brain of experimental rats. Toxicology 2005; 206:49–57.
25. Kidd H, James DR. The agrochemicals handbook. 3rd ed. Cambridge, UK: Royal Society of Chemistry Information Services; 1991.
26.  Uzunhisarcikli M, Kalender Y, Dirican K, Kalender S, Ogutcu A, Buyukkomurcu F. Acute, subacute and subchronic administration of methyl parathion- induced testicular damage in male rats and protective role of vitamins C and E. Pestic Biochem Physiol 2007; 87: 115–122.
27. Farag AT, Ewediah MH, El-Okazy AM. Reproductive toxicology of acephate in male mice. Reprod Toxicol 2000; 14:457–462.
28. Khan IA, Reddy BV, Mahboob M, Rahman MF, Jamil K. Effects of phosphorothionate on the reproductive system of male rats. J Environ Sci Health B 2001; 36:445–456.
29. Xu L, Zhan N, Liu R, Song L, Wang X. Joint action of phoxim and fenvalerate on reproduction in male rats. Asian J Androl 2004; 6:337–341.
30.  Burruel VR, Raabe OG, Overstreet JW, Wilson BM, Wiley LM. Paternal effects from methamidophos administration in mice. Toxicol Appl Pharm 2000; 165:148–157.
31.  Contreras HR, Bustos-Obregón E. Morphological alterations in mouse testis by a single dose of malathion. J Exp Zool 1999; 284:355–359.
32. Griswold MD. The central role of Sertoli cells in spermatogenesis, Semin. Cell Dev Biol 1998; 9:411–416.
33. Roohbakhsh A, Moghaddam AH, Delfan KM. Anxiolytic-like effect of testosterone in male rats: GABAC receptors are not involved. Iran J Basic Med Sci 2011; 14:376-382.
34. Lafuente A, Cabaleiro T, Caride A, Esquifino AI. Toxic effects of methoxychlor administered subcutaneously on the hypothalamic-pituitary-testicular axis in adult rats. Food Chem Toxicol 2008; 46:1570–1575.
35. Durak D, Uzun FG, Kalender S, Ogutcu A, Uzunhisarcikli M, Kalender Y. Malathion-induced testicular toxicity in male rats and the protective effect of vitamins C and E. Food Chem Toxicol 2009; 47:1903–1908.
36. Piña-Guzmán B, Solís-Heredia MJ, Rojas-García AE, Urióstegui-Acosta M, Quintanilla-Vega B. Genetic damage caused bymethyl-parathion inmouse spermatozoa is related to oxidative stress. Toxicol Appl Pharmacol 2006; 216:216–224.
37. Kianifard D, Sadrkhanlou RA, Hasanzadeh S.The ultrastructural changes of the sertoli and leydig cells following streptozotocin induced diabetes. Iran J Basic Med Sci 2012; 15:623-635.
38. Rezg R, Mornagui B, El-Fazaa S, Gharbi N. Biochemical evaluation of hepatic damage in subchronic exposure to malathion in rats: effect on superoxide dismutase and catalase activities using native PAGE. C R Biol 2008; 331:655–662.
39. Rezg R, Mornagui B, El-Fazaa S, Gharbi N. Caffeic acid attenuates malathion induced metabolic disruption in rat liver, involvement of acetylcholinesterase activity. Toxicology 2008; 250:27–31.