Environmental Science and Pollution Research

, Volume 25, Issue 16, pp 15616–15629 | Cite as

Assessment of neurohepatic DNA damage in male Sprague–Dawley rats exposed to organophosphates and pyrethroid insecticides

  • Doha Yahia
  • Marwa F. Ali
Research Article


The current work was undertaken to test the genotoxic potential of chlorpyrifos (CPF), dimethoate, and lambda cyhalothrin (LCT) insecticides in rat brain and liver using the single cell gel electrophoresis (comet assay). Three groups of adult male Sprague–Dawley rats were exposed orally to one third LD50of CPF, dimethoate, or LCT for 24 and 48 h while the control group received corn oil. Serum samples were collected for estimation of malondialdehyde (MDA) and glutathione peroxidase (GPx); the brain and liver samples were used for comet assay and for histopathological examination. Results showed that signs of neurotoxicity appeared clinically as backward stretching of hind limb and splayed gait in dimethoate and LCT groups, respectively. CPF, LCT, and dimethoate induced oxidative stress indicated by increased MDA and decreased GPx levels. CPF and LCT caused severe DNA damage in the brain and liver at 24 and 48 h indicated by increased percentage of DNA in tail, tail length, tail moment, and olive tail moment. Dimethoate induced mild DNA damage in the brain and liver at 48 h. Histopathological changes were observed in the cerebrum, cerebellum, and liver of exposed rats. The results concluded that CPF, LCT, and dimethoate insecticides induced oxidative stress and DNA damage associated with histological changes in the brain and liver of exposed rats.


CPF Dimethoate LCT DNA damage Comet Oxidative stress Brain Liver 


Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All procedures involving animals were done in accordance with the ethical standards of Assiut University. All rats were handled according to the standard guidelines for care and use of experimental animals.


  1. Abd Elkawy M, Oliman G, Abd-El Rehim E (2013) Effect of Solanum nigrum Linn against lambda cyhalothrin-induced toxicity in rats. IOSR J Pharm Bio Sci 5:55–62Google Scholar
  2. Abd Ellah MR, Nishimori K, Goryo M, Okada K, Yasuda J (2004) Glutathione peroxidase and glucose 6 phosphate dehydrogenase activities in bovine blood and liver. J Vet Med Sci 66(10):1219–1221CrossRefGoogle Scholar
  3. Abdallah F, Fetoui H, Fakhfakh F, Keskes L (2012) Caffeic acid and quercetin protect erythrocytes against the oxidative stress and the genotoxic effects of lambda-cyhalothrin in vitro. Hum Exp Toxicol 31(1):92–100CrossRefGoogle Scholar
  4. Abdel-Mobdy YE, Abdel-Rahim EA (2015) Toxicological influences of lambda cyhalothrin and evaluation of the toxicity ameliorative effect of pomegranate in albino rats, global. Veterinaria 14(6):913–921Google Scholar
  5. Abdollahi M, Ranjbar A, Shadnia S, Nokfar S, Rezaie A (2004) Pesticides and oxidative stress: a review. Med SciMonit.10: 141147Google Scholar
  6. Al-Awthan YS, Mohamed A, Gamal HE, Esam MA (2012) Dimethoate-induced oxidative stress and morphological changes in the liver of Guinea pig and the protective effect of vitamin C and E. Asian J Biol Sci 5:9–19CrossRefGoogle Scholar
  7. Arce ZR, Rojas-García AE, Benitez-Trinidad A, Herrera-Moreno JF, Medina-Díaz IM, Barrón-Vivanco BS, Villegas GP, Hernández-Ochoa I, Sólis Heredia MJ, Bernal-Hernández YY (2017) Oxidative stress and genetic damage among workers exposed primarily to organophosphate and pyrethroid pesticides. Environ Toxicol 32:1754–1764CrossRefGoogle Scholar
  8. Astiz M, de Alaniz MJ, Marra CA (2009) Effect of pesticides on cell survival in liver and brain rat tissues. Ecotoxicol Environ Saf 72:2025–2032CrossRefGoogle Scholar
  9. Ayala A, Munoz MF (2014) Arguelles S. Lipid peroxidation production, metabolism and signaling mechanisms of malondialdehyde and 4-hydroxy-2-nonenal. Oxidative Med Cell Longevity 1–31Google Scholar
  10. Bagchi D, Balmoori J, Bagchi M, Ye X, Williams CB, Stohs SJ (2002) Comparative effects of TCDD, endrin, naphthalene and chromium (IV) on oxidative stress and tissue damage in the liver and brain tissues of mice. Toxicology 175:73–82CrossRefGoogle Scholar
  11. Ballesteros ML, Durando PE, Nores ML, Diaz MP, Bistoni MA, Wunderlin DA (2009) Endosulfan induces changes in spontaneous swimming activity and acetylcholinesterase activity of Jenynsia multidentata (Anablepidae, Cyprinodontiformes). Environ Pollut 157:1573–1580CrossRefGoogle Scholar
  12. Bancroft D, Stevens A, Turmer R (1996) Theory and practice of histological technique, 4thed, Churchill Living Stone, Edinburgh, London, Melbourne. pp. 47-67Google Scholar
  13. Bebe FN, Panemangalore M (2003) Exposure to low doses of endosulfan and chlorpyrifos modifies endogenous antioxidants in tissue of rats. J Environ Sci Heal B 38:349–363CrossRefGoogle Scholar
  14. Bertram C, Hass R (2008) Cellular responses to reactive oxygen species induced DNA damage and aging. BiolChem 389:211–220Google Scholar
  15. Bolognesi C (2003) Genotoxicity of pesticides: a review of human biomonitoring studies. Mutat Res/Rev Mutat Res 543:251–272CrossRefGoogle Scholar
  16. Braun R, Schoneich J, Weissflog L, Debek W (1982) Activity of organophosphorus insecticides in bacterial tests for mutagenicity and DNA repair-direct alkylation vs metabolic activation and breakdown. I: butanoate, vinylbutonate, dichlorvos, demethyldichlorovos and demethyl vinylbutonate. Chem Biol Interact 39:339–350CrossRefGoogle Scholar
  17. Celik A, Mazmanci BC, Amlica Y, Askin AC, Omelekoglu U (2003) Cytogenitic effects of lambda-cyhalothrin on wistar rat bone marrow. Mutat Res 539:91–97CrossRefGoogle Scholar
  18. Celik A, Mazmanci BC, Amlica YC, Omelekoglu U, Askin A (2005) Evaluation of cytogenetic effects of lambda-cyhalothrin on Wistar rat bone marrow by gavage administration. Ecotoxicol Environ Saf 61:128–133CrossRefGoogle Scholar
  19. Collins AR, Dobson VL, Dusinská M, Kennedy G, Stĕtina R (1997) The comet assay: what can it really tell us? Mutat Res 375(2):183–193CrossRefGoogle Scholar
  20. Deeba F, Raza I, Muhammad N (2016) Chlorpyrifos and lambda cyhalothrin-induced oxidative stress in human erythrocytes: in vitro studies. Toxicol Ind Health:1–11Google Scholar
  21. DeLeve LD, Kaplowitz N (1991) Glutathione metabolism and its role in hepatotoxicity. Pharmacol Ther 52:287–305CrossRefGoogle Scholar
  22. Dogan D, Can C, Kocyigit A, Dikilitas M, Taskin A, Bilinc H (2011) Dimethoate-induced oxidative stress and DNA damage in Oncorhynchus mykiss. Chemosphere 84:39–46CrossRefGoogle Scholar
  23. Ducolomb Y, Casas E, Valdez A, Gonzalez G, Altamirano- Lozano M, Betancourt M (2009) In vitro effect of malathion and diazinon on oocytes fertilization and embryo development in porcine. Cell Biol Toxicol 25:623–633CrossRefGoogle Scholar
  24. El-Damaty EM, Farrag AH, Rowayshed G, Fahmy HM (2012) Biochemical and Histopathological effects of systemic pesticides on some functional organs of male albino rats. J Appl Sci Res 8(11):5459–5469Google Scholar
  25. El-Demerdash F (2007) Lambda-cyhalothrin-induced changes in oxidative stress biomarkers in rabbit erythrocytes and alleviation effect of some antioxidants. Toxicol in Vitro 21(3):392–397CrossRefGoogle Scholar
  26. Elsharkawy EE, Yahia D, El-Nisr N (2013) Sub-chronic exposure to chlorpyrifos induces hematological, metabolic disorders and oxidative stress in rat: attenuation by glutathione. Environ Toxicol Pharmacol 35:218–227CrossRefGoogle Scholar
  27. Ezzi L, Salah IB, Haouas Z, Sakly A, Grissa I (2016) Histopathological and genotoxic effects of chlorpyrifos in rats. Environ SciPollut Res 23:4859–4867CrossRefGoogle Scholar
  28. Fahmy AM, Abdalla EF (2001) Cytogenetic effects by the natural pyrethrin and the synthetic lambda-cyhalothrin in mice in vivo. Cytologia 66:139–149CrossRefGoogle Scholar
  29. Felton GW, Summers CB (1995) Antioxidant systems in insects. Arch Insect Biochem Physiol 29:187–197CrossRefGoogle Scholar
  30. Fetoui H, Garoui M, Makni-Ayadi F, Zeghal N (2008) Oxidative stress induced by lambda-cyhalothrin (LTC) in rat erythrocytes and brain: attenuation by vitamin C. Environ Toxicol Pharmacol 26(2):225–231CrossRefGoogle Scholar
  31. Fetoui H, Garoui MZ, eghal N (2009) Lambda-cyhalothrin-induced biochemical and histopathological changes in the liver of rats: ameliorative effect of ascorbic acid. Exp Toxicol Pathol 61:189–196CrossRefGoogle Scholar
  32. Fetoui H, Makni M, Garoui M, Zeghal N (2010) Toxic effects of lambda-cyhalothrin, a synthetic pyrethroid pesticide, on the rat kidney: involvement of oxidative stress and protective role of ascorbic acid. Exp Toxicol Pathol 62(6):593–599CrossRefGoogle Scholar
  33. Glickman AH, Lech JJ (1982) Differential toxicity of trans-permethrin in rainbow trout and mice. II. Role of target organ sensitivity. Toxicol Appl Pharmacol 66:162–171CrossRefGoogle Scholar
  34. Goel A, Chanuhan DP, Dhawan DK (2000) Protective effects of zinc in chlorpyrifos induced hepatotoxicity; a biochemical and trace element study. Bio Trace Elemen Res 74:171–183CrossRefGoogle Scholar
  35. Gokcimen A, Gulle K, Demirin H, Bayram D, Kocak A, Altuntas I (2007) Effects of diazinon at different doses on rat liver and pancreas tissues. Pestic Biochem Physiol 87:103–108CrossRefGoogle Scholar
  36. Grotto D, Maria LS, Valentini J, Paniz C, Schmitt G, Garcia SC, Pomblum VJ, Rocha JB, Farina M (2009) Importance of the lipid peroxidation biomarkers and methodological aspects FOR malondialdehyde quantification. Química Nova 32(1):169–174CrossRefGoogle Scholar
  37. He L, Troiano J, Wang A, Goh K (2008) Environmental chemistry, ecotoxicity, and fate of lambda cyhalothrin. Rev Environ Contam Toxicol 195:71–91Google Scholar
  38. Heikal TM, Mossa AH, Nawwar GA, MEl-sherbiny M, Ghanem HZ (2012) Protective effect of a synthetic antioxidant “acetyl gallate derivative” against dimethoate induced DNA damage and oxidant antioxidant status in male rats. Environ Anal Toxicol 2:155Google Scholar
  39. Hussain LA (2010) Role of oxidative stress in organophosphate insecticide toxicity—short review. Pest Biochem Physiol 98:145–150CrossRefGoogle Scholar
  40. Kale M, Rathore N, John S, Bhatnagar D (1999) Lipid peroxidative damage on pyrethroid exposure and alterations in antioxidant status in rat erythrocytes: a possible involvement of reactive oxygen species. Toxicol Lett 105:197–205CrossRefGoogle Scholar
  41. Kammon AM, Brar RS, Banga HS, Sodhi S (2010) Patho-biochemical studies on hepatotoxicity and nephrotoxicity on exposure to chlorpyrifos and imidacloprid in layer chickens. Vet Arhi v 80:663–672Google Scholar
  42. Kassie F, Parzefall W, Knasmuller S (2000) Single cell gel electrophoresis assay: a new technique for human biomonitoring studies. Mutat Res 463:13–31CrossRefGoogle Scholar
  43. Kavitha P, Rao JV (2008) Toxic effects of chlorpyrifos on antioxidant enzymes and target enzyme acetylcholinesterase interaction in mosquito fish, Gambusia affinis. Environ Toxicol Pharm 26:192–198CrossRefGoogle Scholar
  44. Kidd H, James DR (1991) The agrochemicals handbook, 3rd edn. Royal Society of Chemistry Information Services, Cambridge, pp 27–33Google Scholar
  45. Knopper LD, Mineau P, McNamee JP, Lean DR (2005) Use of comet and micronucleus assays to measure genotoxicity in meadow voles (Microtus pennsylvanicus) living in golf course ecosystems exposed to pesticides. Ecotoxicology 14(3):323–335CrossRefGoogle Scholar
  46. Lee KM, Park SY, Lee K, Oh S, Ko S (2017) Pesticide metabolite and oxidative stress in male farmers exposed to pesticide. Ann Occup Environ Med 29:5CrossRefGoogle Scholar
  47. Linn S (1998) DNA damage by iron and hydrogen peroxide in vitro and in vivo. Drug Metab Rev 30:313–326CrossRefGoogle Scholar
  48. Livingstone DR (2001) Contaminant-stimulated reactive oxygen species production and oxidative damage in aquatic organisms. Mar Pollut Bull 42:656–666CrossRefGoogle Scholar
  49. Lone MY, Baba BA, Raj P, Shrivastava VK, Bhide M (2013) Haematological and hepatopathological changes induced by dimethoate in Rattus rattus. Indo Am J Pharm Res 3:4360–4365Google Scholar
  50. Lowe J, Lennox G, Leigh PN (1997) Disorders of movement and system degenerations. In: Graham DI, Lantos PL (eds) Greenfield’s neuropathology, vol 2, 6th edn. Arnold, London, pp 312–314Google Scholar
  51. Mani VM, Ahmed AN, Ali AL, Gokulakrishnan A, Vinoth K, Hussain AA (2016) Hepatoprotective effect of quercetin on lambda-cyhalothrin induced hepatotoxicity in male Wistar rats. Int J Sci Humanit 2:401–416Google Scholar
  52. Mansour SA, Mossa AT (2010) Oxidative damage, biochemical and histopathological alterations in rats exposed to chlorpyrifos and the antioxidant role of zinc. Pestic Biochem Physiol 96:14–23CrossRefGoogle Scholar
  53. Meister A (1988) Glutathione metabolism and its selective modification. J Biol Chem 263(33):17205–17208Google Scholar
  54. Mitchelmore CL, Chipman JK (1998) DNA strand breakage in aquatic organisms and the potential value of the comet assay in environmental monitoring. Mutat Res 399:135–147CrossRefGoogle Scholar
  55. Modesto KA, Martinez CB (2010) Effects of roundup transorb on fish: hematology, antioxidant defenses and acetylcholinesterase activity. Chemosphere 81:781–787CrossRefGoogle Scholar
  56. Monteiro DA, Almeida JA, Rantin FT, Kalinin AL (2006) Oxidative stress biomarkers in the freshwater characid fish, Brycon cephalus, exposed to organophosphorus insecticide Folisuper 600 (methyl parathion). Comp Biochem Phys 143:141–149CrossRefGoogle Scholar
  57. Mostafalou S, Abdollahi M (2013) Pesticides and human chronic diseases: evidences, mechanisms and perspectives. Toxicol Appl Pharmacol 268:157–177CrossRefGoogle Scholar
  58. Narahashi T, Carter DB, Frey J, Ginsburg K, Hamilton BJ, Nagata K, Roy ML, Song JH, Tatebayashi H (1995) Sodium channels and GABAA receptor-channel complex as targets of environmental toxicants. Toxicol Lett 82-83:239–245CrossRefGoogle Scholar
  59. Naravaneni R, Jamil K (2005) Evaluation of cytogenetic effects of lambda-cyhalothrin on human lymphocytes. J Biochem Mol Toxicol 19(5):304–310CrossRefGoogle Scholar
  60. Nasuti C, Cantalamessa F, Falcioni G, Gabbianelli R (2003) Different effects of type I and type II pyrethroids on erythrocyte plasma membrane properties and enzymatic activity in rats. Toxicology 191:233–244CrossRefGoogle Scholar
  61. Ogut S, Gultekin F, Kisioglu AN, Kucukoner E (2011) Oxidative stress in the blood of farm workers following intensive pesticide exposure. Toxicol Ind Health 27:820–825CrossRefGoogle Scholar
  62. Oppenheimer DR, Esiri MM (1992) Diseases of the basal ganglia, cerebellum and motor neurons. In: Adams JH, Duchen LW (eds) Greenfield’s neuropathology, 5th edn. Arnold, London, pp 988–1045Google Scholar
  63. Oral B, Guney M, Demirin H (2006) Endometrial damage and apoptosis in rats induced by dichlorvos and ameliorating effect of antioxidant vitamin E and C. Reprod Toxicol 22:783–790CrossRefGoogle Scholar
  64. Piperakis SM, Kontogianni K, Siffel C, Piperakis MM (2005) Measuring of effects of pesticides on occupationally exposed humans with the comet assay. Environ Toxicol 21:355–359CrossRefGoogle Scholar
  65. Prakasam AS, Sethupathy SL (2001) Plasma and RBCs antioxidant status in occupational male pesticide sprayers. Clin Chim Acta 310:107–112CrossRefGoogle Scholar
  66. Prasanthi K, MuralidharaRajini PS (2005) Morphological and biochemical perturbations in rat erythrocytes following in vitro exposure to Fenvalerate and its metabolite. Toxicol in Vitro 19:449–456CrossRefGoogle Scholar
  67. Radad K, Hassanein K, Moldzio R, Rausch WD (2013) Vascular damage mediates neuronal and non-neuronal pathology following short and long-term rotenone administration in Sprague-Dawley rats. Exp Toxicol Pathol 65:41–47CrossRefGoogle Scholar
  68. Ray DE, Fry JR (2006) A reassessment of the neurotoxicity of pyrethroid insecticides. Pharmacol Ther 111:174–193CrossRefGoogle Scholar
  69. Saafi EB, Louedi M, Elfeki A, Zakhama A, Najjar MF, Hammami M, Achour L (2011) Protective effect of date palm fruit extract (Phoenix dactylifera L.) on dimethoate induced-oxidative stress in rat liver. Exp Toxicol Pathol 63:433–441CrossRefGoogle Scholar
  70. Sakonlaya D, Apisarnthanarak A, Yamada N, Tomtitchong P (2014) Modified toluidine blue: an alternative stain for Helicobacter pylori detection in routine diagnostic use and post-eradication confirmation for gastric cancer prevention. Asian Pac J Cancer Prev 15:6983–6987CrossRefGoogle Scholar
  71. Sasaki YF, Izumiyama F, Nishidate E, Matsusaka N, Tsuda S (1997) Detection of rodent liver carcinogen genotoxicity by the alkaline single cell gel electrophoresis (Comet) assay in multiple mouse organs (liver, lung, spleen, kidney and bone marrow). Mutat Res 391:201–214CrossRefGoogle Scholar
  72. Sharma Y, Bashir S, Irshad M, Gupta SD, Dogra TD (2005) Effects of acute dimethoate administration on antioxidant status of liver and brain of experimental rats. Toxicology 206:49–57CrossRefGoogle Scholar
  73. Sharma RK, Upadhyay G, Siddiqi NJ, Sharma B (2013) Pesticides-induced biochemical alterations in occupational North Indian suburban population. Hum Exp Toxicol 32:1213–1227CrossRefGoogle Scholar
  74. Singh M, Sandhir R, Kiran R (2006) Erythrocyte antioxidant enzymes in toxicological evaluation of commonly used organophosphate pesticides. Indian J Exp Biol 44:580–583Google Scholar
  75. Soderlund DM, Clarc GM, Sheets LP, Mullin LS, Piccirillo VJ (2002) Mechanisms of pyrethroid neurotoxicity: implications for cumulative risk assessment. Toxicology 171:3–59CrossRefGoogle Scholar
  76. Soltaninejad K, Abdollahi M (2009) Current opinion of the science of organophosphate pesticides and toxic stress: a systematic review. Med Sci Monit 15:75–90Google Scholar
  77. Southwood J (1985) Acute oral toxicity studies. Unpublished report no. CTL/P/1102 from Central Toxicology Laboratory, Macclesfield, England. Submitted to WHO by Syngenta Crop Protection AG. PP321Google Scholar
  78. Sparling DW, Fellers G (2007) Comparative toxicity of chlorpyrifos, diazinon, malathion and their oxon derivatives to larval Rana boylii. Environ Pollut 147:535–539CrossRefGoogle Scholar
  79. Tice RR, Agurell E, Anderson D, Burlinson B, Hartmann A, Kobayashi H, Miyamae Y, Rojas E, Ryu JC, Sasaki YF (2000) Single cell gel/comet assay: guidelines for in vitro and in vivo genetic toxicology testing. Environ Mol Mutagen 35(3):206–221CrossRefGoogle Scholar
  80. Tsuda S, Kosaka Y, Matsusaka N, Sasaki YF (1998) Detection of pyrimethamine-induced DNA damage in mouse embryo and maternal organs by the modified alkaline single cell gel electrophoresis assay. Mutat Res 415:69–77CrossRefGoogle Scholar
  81. Tuzmen N, Candan N, Kaya E, Demiryas N (2008) Biochemical effects of chlorpyrifos and deltamethrin on altered antioxidative defence mechanisms and lipid peroxidation in rat liver. Cell Biochem Funct 26:119–124CrossRefGoogle Scholar
  82. US EPA (1988) Fact sheet number 171: karate (PP321). Washington, DCGoogle Scholar
  83. Vijverberg HP, van den Bercken J (1990) Neurotoxicological effect and the mode of action of pyrethroid insecticides. Crit Rev Toxicol 21:105–126CrossRefGoogle Scholar
  84. Wang WX (2002) Interaction of trace metals and different marine food chains. Mar Ecol Prog Ser 243:295–309CrossRefGoogle Scholar
  85. Wang X, Xing H, Li X, Xu S, Wang X (2011) Effects of atrazine and chlorpyrifos on the mRNA levels of IL-1 and IFN-γ 2b in immune organs of common carp. Fish Shellfish Immunol 31:126–133CrossRefGoogle Scholar
  86. WHO (1990) Cyhalothrin, environmental health criteria, 99; Geneva, SwitzerlandGoogle Scholar
  87. Wiener SW, Hoffman RS (2004) Nerve agents: a comprehensive review. J Intensive Care Med 19(1):22–37CrossRefGoogle Scholar
  88. Worthing CR, Walker SB (1987) The Pesticide manual: a world compendium. 8th ed, Thornton Heath, [Surrey, England]: British Crop Protection CouncilGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Forensic Medicine and Toxicology, Faculty of Veterinary MedicineAssiut UniversityAssiutEgypt
  2. 2.Department of Veterinary Pathology and Clinical Pathology, Faculty of Veterinary MedicineAssiut UniversityAssiutEgypt

Personalised recommendations