• Maher M Khadairi Assistant professor, Director of CBRNSSD , University of Babylon
  • Moayed Jy Al-amari Department of Biology, College of Science, University of Babylon, Hillah, Iraq
  • Ayad Mj Al-mamoori Department of Biology, College of Science, University of Babylon, Hillah, Iraq



Cyanobacteria, Purification, Microcystin-leucine arginine, Biochemical markers, DNA damage markers



 Objective: This study determined the effect of purified microcystin-leucine arginine (MC-LR) on biochemical and DNA damage parameters in rats.

Methods: Utilization of preparative high-performance liquid chromatography in analysis, purification and collection of MC-LR, then intraperitoneally injection of purified MC-LR to rats. At the end of exposure, animals were sacrificed, and liver cell was isolated to measure the biochemical markers such as superoxide dismutase (SOD), catalase (CAT) and glutathione (GSH) as well as measured malondialdehyde (MDA), reactive oxygen species (ROS) and cytochrome P450 (Cyt P450), and DNA damage markers such as comet length, tail length, and tail moment were measured with the single cell gel electrophoresis also called comet assay.

Results: The present results showed significantly increased activities of SOD as well as concentration of MDA, ROS with increasing concentration of MC-LR but the activities of CAT and GSH, as well as Cyt P450, were significantly decreased with increasing MC-LR dose while makers of DNA damage such as comet length, tail length, and tail moment also significantly increased with increasing MC-LR dose.

Conclusion: This study demonstrated that chronic exposure to MC-LR toxin can induce alteration of biochemical and DNA damage markers.

Author Biography

Maher M Khadairi, Assistant professor, Director of CBRNSSD , University of Babylon

Assistant professor, Biology Dept.


Amado LL, Monserrat JM. Oxidative stress generation by microcystins in aquatic animals: Why and how. Environ Int 2010;36(2):226-35.

Atencio L, Moreno I, Jos A, Pichardo S, Moyano R, Blanco A, et al. Dose-dependent antioxidant responses and pathological changes in tenca (Tinca tinca) after acute oral exposure to Microcystis under laboratory conditions. Toxicon 2008;52(1):1-12.

WHO. Blue-Green Algae in Inland Waters: Assessment and Control of Risks to Public Health. Geneva: World Health Organization (WHO) Document, Annex G; 1998.

Kumar M. Harvesting of valuable eno-and exo-metabolites from cyanobacteria: A potential source. Asian Pharm Clin Res 2014;2:974-2441.

Naseri A, Karami M, Nadoushan MJ. Failing of information transmission by dorsal hippocampus due to microinjection of colchicine in rats cortical area 1. Asian Pharm Clin Res 2015;4:974-2441.

Campos A, Vasconcelos V. Molecular mechanisms of microcystin toxicity in animal cells. Int J Mol Sci 2010;11(1):268-87.

Zegura B, Sedmak B, Filipic M. Microcystin-LR induces oxidative DNA damage in human hepatoma cell line HepG2. Toxicon 2003;41(1):41-8.

Pinho GL, da Rosa CM, Maciel FE, Bianchini A, Yunes JS, Proença LA, et al. Antioxidant responses and oxidative stress after microcystin exposure in the hepatopancreas of an estuarine crab species. Ecotoxicol Environ Saf 2005;61(3):353-60.

Livingstone DR. Contaminant-stimulated reactive oxygen species production and oxidative damage in aquatic organisms. Mar Pollut Bull 2001;42(8):656-66.

Bouaicha N, Maatouk I, Plessis MJ, Perin F. Genotoxic potential of microcystin-LR and nodularin in vitro in primary cultured rat hepatocytes and in vivo in rat liver. Environ Toxicol 2005;20(3):341-7.

Tredici MR. Mass production of microalgae: Photobioreactors. In: Richmond A, editor. Handbook of Microalgae Culture: Biotechnology and Applied Phycology. Oxford: Blackwell Science; 2004.

Namikoshi M, Choi BW, Sun F, Rinehart KL, Evans WR, Carmichael WW. Chemical characterization and toxicity of dihydro derivatives of nodularin and microcystin-LR, potent cyanobacterial cyclic peptide hepatotoxins. Chem Res Toxicol 1993;6(2):151-8.

Lawton LA, Edwards C. Purification of microcystins. J Chromatogr A 2001;912(2):191-209.

Qiu T, Xie P, Liu Y, Li G, Xiong Q, Hao L, et al. The profound effects of microcystin on cardiac antioxidant enzymes, mitochondrial function and cardiac toxicity in rat. Toxicology 2009;257(1-2):86-94.

Marklund S, Marklund G. Involvement of the superoxide anion radical in the autoxidation of pyrogallol and a convenient assay for superoxide dismutase. Eur J Biochem 1974;47(3):469-74.

Claiborne A. Catalase activity. In: Greenwald RA, editor. CRC Handbook of Methods for Oxygen Radical Research. Boca Raton, FL: CRC Press; 1985. p. 283-4.

Moreno I, Pichardo S, Jos A, Gómez-Amores L, Mate A, Vazquez CM, et al. Antioxidant enzyme activity and lipid peroxidation in liver and kidney of rats exposed to microcystin-LR administered intraperitoneally. Toxicon 2005;45(4):395-402.

Aust SD. Lipid peroxidation. In: Greenwald RA, editor. CRC Handbook of Methods for Oxygen Radical Research. Boca Raton, FL: CRC Press; 1985. p. 203-7.

Burtis CA, Ashwood ER. Tietz Textbook of Clinical Chemistry. 3rd ed. Philadelphia, PA: W.B. Saunders Co.; 1999.

Singh NP, McCoy MT, Tice RR, Schneider EL. A single technique for quantification of low levels of DNA damage in individual cells. Exp Cell Res 1988;175:184-91.

Conners DE. Biomarkes of Oxidative Stress in Fresh Water Clam (Corbicula fluminea) as Mechanistic Tool to Evaluate the Impairment of Stream Ecosystem Health by Lawn Care Pesticides. PhD Thesis. USA: The University of Georgia; 2004.

Chen T, Cui J, Liang Y, Xin X, Owen Young D, Chen C, et al. Identification of human liver mitochondrial aldehyde dehydrogenase as a potential target for microcystin-LR. Toxicology 2006;220(1):71-80.

da Silva CA, Oba ET, Ramsdorf WA, Magalhães VF, Cestari MM, Oliveira Ribeiro CA, et al. First report about saxitoxins in freshwater fish Hoplias malabaricus through trophic exposure. Toxicon 2011;57(1):141-7.

Harada KI. Chemistry and detection of microcystins. In: Watanabe MF, Harada KI, Carmichael WW, Fujiki H, editors. Toxic Microcystis. Boca Raton, FL: Chemical Rubber Company, CRC Press; 1996. p. 103-48.

Sicinska P, Bukowska B, Michalowicz J, Duda W. Damage of cell membrane and antioxidative system in human erythrocytes incubated with microcystin-LR in vitro. Toxicon 2006;47(4):387-97.

Mitrovic SM, Pflugmacher S, James KJ, Furey A. Anatoxin-a elicits an increase in peroxidase and glutathione S-transferase activity in aquatic plants. Aquat Toxicol 2004;68(2):185-92.

Ozturk O, Gumus S. Changes in glucose-6phosphate dehydrogenase, copper, zinc-superoxide dismutase and catalase activities, glutathione and its metabolizing enzymes, and lipid peroxidation in rat erythrocytes with age. Exp Gerontol 2004;39:211-6.

van Bladeren PJ. Glutathione conjugation as a bioactivation reaction. Chem Biol Interact 2000;129(1-2):61-76.

Campolo J, De Maria R, Caruso R, Accinni R, Turazza F, Parolini M, et al. Blood glutathione as independent marker of lipid peroxidation in heart failure. Int J Cardiol 2007;117:45-50.

Li ZH, Zlabek V, Grabic R, Li P, Randak T. Modulation of glutathione-related antioxidant defense system of fish chronically treated by the fungicide propiconazole. Comp Biochem Physiol C Toxicol Pharmacol 2010;152(3):392-8.

Tripathy A. Oxidative stress, reactive oxygen species (ROS) and antioxidative defense system. Int J Curr Res Biosci Plant Biol 2016;3(10):79-89.

Cazenave J, Bistoni Mde L, Pesce SF, Wunderlin DA. Differential detoxification and antioxidant response in diverse organs of Corydoras paleatus experimentally exposed to microcystin-RR. Aquat Toxicol 2006;76(1):1-12.

Galal A, Souich PD. 21-aminosteroids prevent the down-regulation of hepatic cytochrome P450 induced by hypoxia and inflammation in conscious rabbits. Br J Pharmacol 1999;128(2):374-9.

Moore MJ, Mitrofanov IV, Valentini SS, Volkov VV, Kurbskiy AV, Zhimbey EN, Eglinton LB, Stegeman JJ. Cytochrome P4501A expression, chemical contaminants and histopathology in roach, goby sturgeon and chemical contaminants in sediments from the Caspian Sea, Lake Balkhash and the Ily River Delta, Kazakhstan. Mar Pollut Bull 2013;46:107-119.

Zhang B, Liu Y, Li X. Alteration in the expression of cytochrome P450s (CYP1A1, CYP2E1, and CYP3A11) in the liver of mouse induced by microcystin-LR. Toxins (Basel) 2015;7(4):1102-15.

Brooks WP, Codd GA. Immunological and toxicological studies on Microcystis aeruginosa peptide toxin. Br Phycol J 2007;87:22-301.

Hudder A, Song W, O’Shea KE, Walsh PJ. Toxicogenomic evaluation of microcystin-LR treated with ultrasonic irradiation. Toxicol Appl Pharmacol 2007;220(3):357-64.

Fawell JK, Mitchell RE, Everett DJ, Hill RE. The toxicity of cyanobacterial toxins in the mouse: I microcystin-LR. Hum Exp Toxicol 1999;18(3):162-7.

Li X, Ma J, Fang Q, Li Y. Transcription alterations of microRNAs, cytochrome P4501A1 and 3A65, and AhR and PXR in the liver of zebrafish exposed to crude microcystins. Toxicon 2013;73:17-22.

Zegura B, Volcic M, Lah TT, Filipic M. Different sensitivities of human colon adenocarcinoma (CaCo-2), astrocytoma (IPDDC-A2) and lymphoblastoid (NCNC) cell lines to microcystin-LR induced reactive oxygen species and DNA damage. Toxicon 2008;52(3):518-25.

Nong Q, Komatsu M, IzumoK. Involvement of reactive oxygen species in Microcystin-LR-induced cytogenotoxicity. Free Radic Res 2007;41(12):1326-37.

Lankoff A, Banasik A, Obe G. Effect of microcystin LR and cyanobacterial extract from polish reservoir of drinking water on cell cycle progression, mitotic spindle, and apoptosis in CHO-K1 cells. Toxicol Appl Pharmacol 2004;189(3):204-13.

Dias E, Louro H, Pinto M, Santos T, Antunes S, Pereira P, et al. Genotoxicity of microcystin-LR in in vitro and in vivo experimental models. Biomed Res Int 2014;2014:949521.

Georg O, Amaeze NH, Soghanmu TO, Otitoloju AA. Biomarkers responses in Tympanotous fuscatus Var. Radula (L) inhibiting an oil-impacted and fire-ravaged mangrove ecosystem, current advance in environmental science. Aman V King Sci Publ 2014;2:101-11.

Alodeani EA. Botulinum toxin Type A: An effective, safe and minimally invasive treatment option of axillary and palmar hyperhidrosis. Int J Pharm Pharm Sci 2016;7:975-1491.

Ramesh S, Dilipan E, Mayavu P. Effects of drugs against antioxidant and cytotoxic (HEp2 cell line) activity compounds from marine animals conusamadis venom (GMELIN, J.F, 1791). Int J Pharm Pharm Sci 2014;7:975-1491.



How to Cite

Khadairi, M. M., M. J. Al-amari, and A. M. Al-mamoori. “THE BIOCHEMICAL ALTERATION AND DNA DAMAGE IN RATS (RATTUS RATTUS) AFTER CHRONIC INTRAPERITONEALLY INJECTION TO PURIFIED MICROCYSTIN-LR FROM ANABAENA CIRCINALIS”. Asian Journal of Pharmaceutical and Clinical Research, vol. 10, no. 11, Nov. 2017, pp. 277-83, doi:10.22159/ajpcr.2017.v10i11.20565.



Original Article(s)