Shireen Abd El-satar, Nasr Nasr, Khaled Khailo, Hossam Sayour


This study was conducted to evaluate hemotoxicity and genotoxicity induced by lead acetate and chlorpyrifos in catfish (Clarias gariepinus) as a model for checking genotoxic pollutants in aquatic surroundings. Lead acetate was added by a dose of 24.4 mg/L (20% of the LC50) daily, chlorpyrifos was added by a dose of 1.65 mg/L (1/10 LC50) daily. Fish were kept in standard condition in which water temperature (25°C± 2); pH (7.6 ± 0.4) and dissolved oxygen (5.4±0.4 mg/L). Blood and liver were sampled after 4 weeks. The result revealed a significant reduction of RBCs count, Hb, hematocrit in fish exposed to lead and chlorpyrifos. Furthermore, MDA level and catalase activity showed significant increase and decrease, respectively in groups exposed to lead acetate and chlorpyrifos in comparison with the control fish. Toxic effect of lead acetate and chlorpyrifos confirmed by histopathological changes in liver sections which showed marked hepatic vacuolation and parenchymal hemorrhage. DNA damage detected by comet assay also revealed a significance increase in tail length, tail DNA% and tail moment at sub-lethal concentration of lead acetate and chlorpyrifos. This study concluded that lead acetate and chlorpyrifos have hemotoxic and genotoxic effect probably through, at least in part, induction of reactive oxygen species and chlorpyrifos has more hemotoxic and genotoxic effect than lead acetate.    

Key words: catfish; lead acetate; chlorpyrifos; DNA damage

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McGlashan D, Hughes J. Genetic evidence for historical continuity between populations of the Australian freshwater fish Craterocephalus stercusmuscarum (Atherinidae) east and west of the Great Dividing Range, Journal of Fish Biology 2001; 59: 55–67.

Ali F K, El-Shafai S A, Samhan F A, Khalil W K. Effect of water pollution on expression of immune response genes of Solea aegyptiaca in Lake Qarun, African Journal of Biotechnology 2008; 7(10).

Mahfouz M, Hegazi M, El-Magd M, Kasem E. Metabolic and molecular responses in Nile tilapia, Oreochromis niloticus during short and prolonged hypoxia, Marine and Freshwater Behaviour and Physiology 2015; 48(5): 319–40.

Ayadi I, Monteiro S M, Regaya I, Coimbra A, Fernandes F, Oliveira M M, Peixoto F, Mnif W. Biochemical and histological changes in the liver and gills of Nile tilapia Oreochromis niloticus exposed to Red 195 dye, RSC Advances 2015; 5(106): 87168–78.

Pandey S, Parvez S, Sayeed I, Haque R, Bin-Hafeez B, Raisuddin S. Biomarkers of oxidative stress: a comparative study of river Yamuna fish Wallago attu (Bl. & Schn.), Science of the Total environment 2003; 309(1-3): 105–15.

Gilliom R J, Hamilton P A, Pesticides in the nation's streams and ground water, 1992-2001-a summary, 2006.

Monroy M, Maceda-Veiga A, de Sostoa A. Metal concentration in water, sediment and four fish species from Lake Titicaca reveals a large-scale environmental concern, Science of the Total environment 2014; 487: 233–44.

Authman M M, Zaki M S, Khallaf E A, Abbas H H. Use of fish as bio-indicator of the effects of heavy metals pollution, Journal of Aquaculture Research & Development 2015; 6(4): 1–13.

Patra R, Swarup D, Dwivedi S. Antioxidant effects of α tocopherol, ascorbic acid and L-methionine on lead induced oxidative stress to the liver, kidney and brain in rats, Toxicology 2001; 162(2): 81–8.

Gurer H, Ercal N. Can antioxidants be beneficial in the treatment of lead poisoning?, Free Radical Biology and Medicine 2000; 29(10): 927–45.

El-Magd M A, Kahilo K A, Nasr N E, Kamal T, Shukry M, Saleh A A. A potential mechanism associated with lead-induced testicular toxicity in rats, Andrologia 2016; 49(9): e12750.

Dai W, Liu S, Fu L, Du H, Xu Z. Lead (Pb) accumulation, oxidative stress and DNA damage induced by dietary Pb in tilapia (Oreochromis niloticus), Aquaculture Research 2012; 43(2): 208–14.

Abdallah G M, El-Sayed E-S M, Abo-Salem O M. Effect of lead toxicity on coenzyme Q levels in rat tissues, Food and Chemical Toxicology 2010; 48(6): 1753–6.

Nordberg , Fowler , Nordberg , Friberg. Handbook on the Toxicology of Metals. 3rd Edn. Academic press, Amesterdam., ISBN 2007; -10: 0123694132, pp: 10–24.

Makino S, Tsuruta H, Takata T. Relationship between blood lead level and urinary ALA level in workers exposed to very low levels of lead, Industrial health 2000; 38(1): 95–8.

Flora S J, Pande M, Mehta A. Beneficial effect of combined administration of some naturally occurring antioxidants (vitamins) and thiol chelators in the treatment of chronic lead intoxication, Chemico-Biological Interactions 2003; 145(3): 267–80.

Ademuyiwa O, Ugbaja R, Ojo D, Owoigbe A, Adeokun S. Reversal of aminolevulinic acid dehydratase (ALAD) inhibition and reduction of erythrocyte protoporphyrin levels by vitamin C in occupational lead exposure in Abeokuta, Nigeria, Environmental Toxicology and Pharmacology 2005; 20(3): 404–11.

Sabra F S, Mehana E. Pesticides toxicity in fish with particular reference to insecticides, Asian Journal of Agriculture and Food Sciences (ISSN: 2321–1571) 2015; 3(01).

Lari S Z, Khan N A, Gandhi K N, Meshram T S, Thacker N P. Comparison of pesticide residues in surface water and ground water of agriculture intensive areas, Journal of Environmental Health Science and Engineering 2014; 12(1): 11.

Banaee M, Mirvagefei A, Rafei G, Majazi Amiri B. Effect of sub-lethal diazinon concentrations on blood plasma biochemistry, International Journal of Environmental Research 2008; 2(2): 189–8.

Wang Y, Qiu Y, Fei Y, Li L, Zhu Z, Zhao J, Yao E, Yao Y. Measurement and preliminary human health risk assessment of representative organochlorines in farmed Mandarin fish, Huan jing ke xue= Huanjing kexue 2011; 32(8): 2385–90.

Rao J V, Begum G, Pallela R, Usman P, Rao R N. Changes in behavior and brain acetylcholinesterase activity in mosquito fish, Gambusia affinis in response to the sub-lethal exposure to chlorpyrifos, International Journal of Environmental Research and public health 2005; 2(3): 478–83.

Akhgari M, Abdollahi M, Kebryaeezadeh A, Hosseini R, Sabzevari O. Biochemical evidence for free radicalinduced lipid peroxidation as a mechanism for subchronic toxicity of malathion in blood and liver of rats, Human & experimental toxicology 2003; 22(4): 205–11.

Cicchetti R, Argentin G. The role of oxidative stress in the in vitro induction of micronuclei by pesticides in mouse lung fibroblasts, Mutagenesis 2003; 18(2): 127–32.

Orbea A, Ortiz-Zarragoitia M, Solé M, Porte C, Cajaraville M P. Antioxidant enzymes and peroxisome proliferation in relation to contaminant body burdens of PAHs and PCBs in bivalve molluscs, crabs and fish from the Urdaibai and Plentzia estuaries (Bay of Biscay), Aquatic Toxicology 2002; 58(1-2): 75–8.

Sharawy Z Z, Thiele R, Abbas E M, El-Magd M A, Hassaan M S, Peter C, Schmidt J, Saborowski R, Goda A M A-S, Slater M J. Antioxidant response, body composition of whiteleg shrimp Litopenaeus vannamei co-cultured with Nile tilapia Oreochromis niloticus in recirculating aquaculture, Aquaculture Environment Interactions 2017; 9: 257–68.

Ahmad I, Hamid T, Fatima M, Chand H S, Jain S K, Athar M, Raisuddin S. Induction of hepatic antioxidants in freshwater catfish (Channa punctatus Bloch) is a biomarker of paper mill effluent exposure, Biochimica et Biophysica Acta (BBA)-General Subjects 2000; 1523(1): 37–48.

Narra M R, Regatte R R, Kodimyala R. Effects of chlorpyrifos on enzymes as biomarkers of toxicity in Fresh water field crab Barytelphusa guerini, International Journal of Environmental Sciences 2012; 2(4): 2015.

Karami A, Christianus A, Ishak Z, Courtenay S, Syed M, Azlina M N, Noorshinah H. Effect of triploidization on juvenile African catfish (Clarias gariepinus), Aquaculture international 2010; 18(5): 851–58.

Hassanain M, Abdel-Rahman E H, Abo-Hegab S, Tawfik M, Abbas W T. Induction of cytochrome P450 1A1 as a biomarker of Benzo-a-pyrene pollution in Egyptian fresh water fish, Pakistan journal of biological sciences: PJBS 2007; 10(8): 1161–9.

Mekkawy I A, Mahmoud U M, Osman A G, Sayed A E-D H. Effects of ultraviolet A on the activity of two metabolic enzymes, DNA damage and lipid peroxidation during early developmental stages of the African catfish, Clarias gariepinus (Burchell, 1822), Fish physiology and biochemistry 2010; 36(3): 605–26.

Sayed A E-D H, Hamed H S. Induction of apoptosis and DNA damage by 4-nonylphenol in African catfish (Clarias gariepinus) and the antioxidant role of Cydonia oblonga, Ecotoxicology and environmental safety 2017; 139: 97–101.

Alkahemal-Balawi H F, Ahmad Z, Al-Akel A S, Al-Misned F, Suliman E-A M, Al-Ghanim K A. Toxicity bioassay of lead acetate and effects of its sub-lethal exposure on growth, haematological parameters and reproduction in Clarias gariepinus, African Journal of Biotechnology 2011; 10(53): 11039-–47.

Narra M R, Rajender K, Reddy R R, Rao J V, Begum G. The role of vitamin C as antioxidant in protection of biochemical and haematological stress induced by chlorpyrifos in freshwater fish Clarias batrachus, Chemosphere 2015; 132: 172–8.

Natt M P, Herrick C A. A new blood diluent for counting the erythrocytes and leucocytes of the chicken, Poultry Science 1952; 31(4): 735-738.

Hesser E F. Methods for routine fish hematology, The Progressive Fish‐Culturist 1960; 22(4): 164-–71.

Nwani C D, Somdare P O, Ogueji E O, Nwani J C, Ukonze J A, Nwadinigwe A O. Genotoxicity assessment and oxidative stress responses in freshwater African catfish Clarias gariepinus exposed to fenthion formulations, Drug and chemical toxicology 2017; 40(3): 273–80.

Ohkawa H, Ohishi N, Yagi K. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction, Analytical biochemistry 1979; 95(2): 351–8.

Aebi H, [13] Catalase in vitro, Methods in enzymology, Elsevier1984, pp. 121–6.

Bancroft J D, Cook H C, Manual of histological techniques and their diagnostic application, Churchill Livingstone1994.

Singh N P, McCoy M T, Tice R R, Schneider E L. A simple technique for quantitation of low levels of DNA damage in individual cells, Experimental cell research 1988; 175(1): 184–91.

Badawy A A, El-Magd M A, AlSadrah S A. Therapeutic Effect of Camel Milk and Its Exosomes on MCF7 Cells In Vitro and In Vivo, Integrative Cancer Therapies 2018; 7(4): 1235-–46.

El-Adawy M, El-Aziz M A, El-Shazly K, Ali N G, El-Magd M A. Dietary propionic acid enhances antibacterial and immunomodulatory effects of oxytetracycline on Nile tilapia, Oreochromis niloticus, Environmental Science and Pollution Research. DOI:10.1007/s11356-018-3206-5 2018.

Hayati A, Pratiwi H, Khoiriyah I, Winarni D, Sugiharto, Histopathological assessment of cadmium effect on testicles and kidney of Oreochromis niloticus in different salinity, AIP Conference Proceedings, AIP Publishing, 2017, p. 020014.

Osman A G, Mekkawy I A, Verreth J, Wuertz S, Kloas W, Kirschbaum F. Monitoring of DNA breakage in embryonic stages of the African catfish Clarias gariepinus (Burchell, 1822) after exposure to lead nitrate using alkaline comet assay, Environmental Toxicology: An International Journal 2008; 23(6): 679–87.

Nguyen L, Janssen C. Embryo-larval toxicity tests with the African catfish (Clarias gariepinus): comparative sensitivity of endpoints, Archives of environmental contamination and toxicology 2002; 42(2): 256–62.

Adeyemo O K. Haematological profile of Clarias gariepinus (Burchell, 1822) exposed to lead, Turkish Journal of Fisheries and Aquatic Sciences 2007; 7(2).

Shah S L. Hematological parameters in tench Tinca tinca after short term exposure to lead, Journal of Applied Toxicology: An International Journal 2006; 26(3): 223–8.

Jenkins F, Smith J, Rajanna B, Shameem U, Umadevi K, Sandhya V, Madhavi R. Effect of sub-lethal concentrations of endosulfan on hematological and serum biochemical parameters in the carp Cyprinus carpio, Bulletin of Environmental Contamination and Toxicology 2003; 70(5): 0993–7.

Toni C, Ferreira D, Kreutz L C, Loro V L, Barcellos L J G. Assessment of oxidative stress and metabolic changes in common carp (Cyprinus carpio) acutely exposed to different concentrations of the fungicide tebuconazole, Chemosphere 2011; 83(4): 579–84.

Xing H, Wang X, Sun G, Gao X, Xu S, Wang X. Effects of atrazine and chlorpyrifos on activity and transcription of glutathione S-transferase in common carp (Cyprinus carpio L.), Environmental Toxicology and Pharmacology 2012; 33(2): 233–44.

Abdelhady D, El-Abasy M, Abou-Asa S, Elbialy Z, Shukry M, Hussein A, Saleh A, El-Magd M. The ameliorative effect of Aspergillus awamori on aflatoxin B1-induced hepatic damage in rabbits, World Mycotoxin Journal 2017; 10(4): 363–73.

Sharbidre A A, Metkari V, Patode P. Effect of methyl parathion and chlorpyrifos on certain biomarkers in various tissues of guppy fish, Poecilia reticulata, Pesticide Biochemistry and Physiology 2011; 101(2): 132–41.

Leonard S S, Vallyathan V, Castranova V, Shi X, Generation of reactive oxygen species in the enzymatic reduction of PbCrO 4 and related DNA damage, Oxygen/Nitrogen Radicals: Cell Injury and Disease, Springer2002, pp. 309–15.

El-Sokkary G H, Kamel E S, Reiter R J. Prophylactic effect of melatonin in reducing lead-induced neurotoxicity in the rat, Cellular & molecular biology letters 2003; 8(2): 461–70.

Kaur M, Jindal R. Oxidative Stress Response in Liver, Kidney and Gills of Ctenopharyngodon Idellus (Cuvier & Valenciennes) Exposed To Chlorpyrifos, MOJ Biol Med 2017; 1(4): 00021.

Abdelhady D H, El-Magd M A, Elbialy Z I, Saleh A A. Bromuconazole-induced hepatotoxicity is accompanied by upregulation of PXR/CYP3A1 and downregulation of CAR/CYP2B1 gene expression, Toxicol Mech Methods 2017; 27(7): 544–50.

Rubio R, Tineo P, Torreblanca A, Del Ramo J, Mayans J D. Histological and electron microscopical observations on the effects of lead on gills and midgut gland of Procambarus clarkii, Toxicological & Environmental Chemistry 1991; 31(1): 347–52.

Deb N, Das S. Chlorpyrifos toxicity in fish: A Review, Curr. World Environ 2013; 8(1): 77-84.

Ballatori N. Transport of toxic metals by molecular mimicry, Environmental health perspe-ctives 2002; 110 (Suppl 5): 689.

61. Danadevi K, Rozati R, Banu B S, Rao P H, Grover P. DNA damage in workers exposed to lead using comet assay, Toxicology 2003; 187(2-3): 183–93.

Celik A, Öğenler O, Çömelekoğlu Ü. The evaluation of micronucleus frequency by acridine orange fluorescent staining in peripheral blood of rats treated with lead acetate, Mutagenesis 2005; 20(6): 411–5.

Fracasso M E, Perbellini L, Soldà S, Talamini G, Franceschetti P. Lead induced DNA strand breaks in lymphocytes of exposed workers:

role of reactive oxygen species and protein kinase C, Mutation Research/Genetic Toxicology and Environmental Mutagenesis 2002; 515(1): 159–9.

Yin X H, Li S N, Zhang L, Zhu G N, Zhuang H S. Evaluation of DNA damage in Chinese toad (Bufo bufo gargarizans) after in vivo exposure to sublethal concentrations of four herbicides using the comet assay, Ecotoxicology 2008; 17(4): 280–6.

DOI: http://dx.doi.org/10.26873/SVR-807-2019


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