Evaluation of Micronucleus Assay and Genotoxic Effect in <I>Oreochromis mossambicus</I> Exposed to a Fumaronitrile


  • PG and Research Centre in Biotechnology, MGR College, Hosur – 635130, Tamil Nadu
  • PG and Research Centre in Biotechnology, MGR College, Hosur – 635130, Tamil Nadu
  • Centre for Research and PG studies in Botany, Thiagarajar College, Madurai - 625009, Tamil Nadu




Comet Assay, DNA Damage, Fumaronitrile, <I>Oreochromis mossambicus</I>


Chemical contaminants of major concern in a polluted environment include persistent organic chemical, pesticides, oils, nutrients, heavy metals, etc. Some of these chemicals are known genotoxicants and can cause damage to the genetic material of the exposed organism. Apart from the enzymatic biomarkers, other parameters, such as DNA damage, should be evaluated for assessing the consequences of exposure of chemical. Damaged DNA may potentiate subsequent deleterious cellular events such as disease (e.g. cancer) and reduced reproductive competence. DNA damage in a variety of aquatic animals has been associated with reduced growth, abnormal development and reduced survival of embryos, larvae and adults. The DNA damage which is induced by fumaronitrile exposure could be evaluated by using the comet assay. The comet assay [alkaline single-cell gel electrophoresis] is a widely used technique to detect DNA damage due to environmental stress. The toxicological effect of fumaronitrile originates from its oxidizing action. Once entered in to the cell, in the presence of cellular reluctant, it induces DNA fragmentation, which is considered to be premutagenic change. The kind of the nuclear alterations observed in erythrocytes of Oreochromis mossambicus exposed to the fumaronitrile is binucleated cells, cells with bleb bed and lobed nuclei.


Download data is not yet available.


Ritter L, Solomon KR, Forget J, Stemeroff M, Leary CO.Persistent organic pollutants. United Nations Environ Prog.Retrieved. 2007; 9–16.

Loganthan, BG, Kannan K. Global organochlorine contamination trends: An overview. Ambio. 1994; 23(3):187–91.

Van der Oost R, Beyer JNPE. Vermeulen fish bioaccumulation and biomarkers in environmental risk assessment: A review. Environ Toxicol Pharmacology. 2003; 13(2):57–149.https://doi.org/10.1016/S1382-6689(02)00126-6

Brausch JM, Connors KA, Brooks BW, Rand GM. Human pharmaceuticals in the aquatic environment: A review of recent toxicological studies and considerations for toxicity testing. Rev Environ Contam Toxicol. 2012; 218:1–99. PMid: 22488604. https://doi.org/10.1007/978-1-4614-3137-4_1

Bolognesi C, Hayashi M. Micronucleus assay in aquatic animals. Mutagenesis. 2011; 26:205–13. PMid: 21164204.https://doi.org/10.1093/mutage/geq073

Mir MI, Khan S, Bhat SA, Ahmad Arif Reshi, Shah FA, Balki MH, Rusheeba Manzoor A. Scenario of genotoxicity in fishes and its impact on fish industry. J Environ Sci Toxico Food Tech. 2014; 8(6):65–76. https://doi.org/10.9790/2402 -08626576

Villela IV, OliveiraIM, Silva J, Henriques JAP. DNA damage and repair in haemolynph cells of golden mussel (Limnoperna fortunei) exposed to environmental contaminants.Mutat Res. 2006; 605(12):78–86. PMid: 16697250.https://doi.org/10.1016/j.mrgentox.2006.02.006

Pawar DH. River water pollution, an environmental crisis a case study of Panchaganga river of Kolhapur city. Inter J Ecol Develop Sum. 2012; 9(1):131–3.

Kawanishi S, Ohnishi S, Ma N, Hiraku Y, Oikaw S, Murata M. Nitrative and oxidative DNA damage in infectionrelated carcinogenesis in relation to cancer stem cells.Genes Environ. 2017; 38:26. PMid: 28050219 PMCid: PMC5203929. https://doi.org/10.1186/s41021-016-0055-7

American Public Health Association. Standard methods for examination of water and waste water. 17th Ed. New York: APHA, AWWA, WPCF; 1989.

Fenech M, Chang WP, Kirsch-Volders M, Holland N, Bonassi S, Zeiger E. HUMN project: Detailed description of the scoring criteria for the cytokines is-block micronucleus assay using isolated human lymphocytes cultures. Mutat Res. 2003; 53 (2):65–75. https://doi.org/10.1016/S13835718(02)00249-8

Zhong LI, Hong-Wei L, Gui-Wei-ZA. A rapid PCR quality DNA extraction method in fish. Acta Hydrobiologica Sinica. 2012; 36(2):365–7.

Ramaswamy BR, Shanmugam G, Velu G, Rengarajan B, Larsson DG. GC-MS analysis and ecotoxicological risk assessment of triclosan, carbamazepine and parabens in Indian rivers. J Hazard Mater. 2011; 186(3):1586–93. PMid: 21216531. https://doi.org/10.1016/j.jhazmat.2010.12.037

Shanmugam G, Sampath S, Selvaraj K, Larsson D, Ramaswamy B. Nonsteroidal anti-inflammatory drugs in Indian rivers. Environ Sci Pollut. 2013; 21:921–31. PMid: 23832803. https://doi.org/10.1007/s11356-013-1957-6

Babu Rajendran R, Govindaraj S, Geetha V, Bhuvaneshwari R, Joakim Larsson D. GC-MS analysis and ecotoxicological risk assessment o triclosan, carbamazepine and parabens in Indian rivers. J of Hazardous Materials. 2011; 186:1586–93. PMid: 21216531. https://doi.org/10.1016/j .jhazmat.2010.12.037

Livingstone DR. Review biotechnology and pollution monitoring: Use of molecular biomarkers in the aquatic environment. J Chem Tech Biotech. 1993; 57:195–211.https://doi.org/10.1002/jctb.280570302

Hickey EJ, Raje RR, Reid VE, Gross SM, Ray SD. Diclofenac induced in vivo nephrotoxicity may involve oxidative stress-mediated massive genomic DNA fragmentation and apoptotic cell death. Free Radic Biol Med. 2001; 31:139–52.https://doi.org/10.1016/S0891-5849(01)00560-3

Kammann U, Bunke M, Steinhart H, Theobald N. A permanent fish cell line (EPC) for genotoxicity testing of marine sediments with the comet assay. Mutat Res. 2001; 498:61–77.https://doi.org/10.1016/S1383-5718(01)00268-6

Frenzilli G, Bosco E, Barale R.Validation of single gel assay in human leukocytes with 18 reference compounds.Mutat Res. 2000; 35:206–21. https://doi.org/10.1016/ S1383-5718(00)00042-5

Banu BS, Danadevi K, Rahman MF, Ahuja YR, Kaiser J.Genotoxic effect of monocrotophos to sentinel species using the comet assay. Food Chem Toxicol. 2001; 39:361–6.https://doi.org/10.1016/S0278-6915(00)00141-1

Triebskorn R, Casper H, Heyd A, Eikemper R, Kohler HR.Toxic effects of the non-steroidal anti-inflammatory drug diclofenac part II. Cytological effects in liver, kidney, gills and intestine of rainbow trout (Oncorhynchus mykiss).Aquat Toxicol. 2004; 68:151–66. PMid: 15145225. https:// doi.org/10.1016/j.aquatox.2004.03.015

Haap T, Triebskorn R, Kohler HR. Acute effects of diclofenac and DMSO to Daphnia magna: Immobilisation and hsp70induction. Chemosp. 2008; 73:353–9. PMid: 18649920. https://doi.org/10.1016/j.chemosphere.2008.05.062

Schnetzer A, Miller PE, Schaffner RA, Stauffer BA, Jones BH, Weisberg SB, DiGiacomo PM, Berelson WM, Caron DA. Blooms of pseudo-nitzschia and domoic acid in the san pedro channel and Los Angeles harbor areas of the Southern California Bight 2003-2004. Harmful Algae.2007; 6:372–87. https://doi.org/10.1016/j.hal.2006.11.004

Carvalho Pinto-Silva CR, Moukha S, Matias WG, Creppy EE. Domoic acid induces direct DNA damage and apoptosis in Caco-2 cells: Recent advances. Environ Toxicol. 2008; 23(6):657–63. PMid: 18293405. https://doi.org/10.1002/ tox.20361

Ahamed M, Karns M, Goodson M, Rowe J, Hussain SM, Schlager JJ, Hong Y. DNA damage response to different surface chemistry of silver nanoparticles in mammalian cells.Toxicol Appl Pharm. 2008; 233:404–10. PMid: 18930072.https://doi.org/10.1016/j.taap.2008.09.015

Vevers WF, Jha AN. Genotoxic and cytotoxic potential of titanium dioxide (TiO2) nanoparticles on fish cells in vitro.Ecotoxi. 2008; 17:410–20. PMid: 18491228. https://doi .org/10.1007/s10646-008-0226-9

Reeves JF, Davies SJ, Dodd NJF, Jha AN. Hydroxyl radicals (OH) are associated with titanium dioxide (TiO2) nanoparticleinduced cytotoxicity and oxidative DNA damage in fish cells. Mutat Res. 2008; 640:113–22. PMid: 18258270.https://doi.org/10.1016/j.mrfmmm.2007.12.010

Singh M, Kaur P, Sandhir R, Kiran R. Protective effects of vitamin E against atrazine-induced genotoxicity in rats.Mut Res. 2008; 654:145–9. PMid: 18582598. https://doi .org/10.1016/j.mrgentox.2008.05.010

Ramesh R, Kavitha P, Kanipandian N, Arun S, Thirumurugan R, Subramanian P. Alteration of antioxidant enzymes and impairment of DNA in the SiO2 nanoparticles exposed zebra fish (Danio rerio). Environ Monit Assess. 2013; 185:5873–81. PMid: 23196406. https://doi .org/10.1007/s10661-012-2991-4

Russo C, Russo L, Morescalchi MA, Stingo V. Assessment of environmental stress by the micronucleus test and the comet assay on the genome of teleost population from two natural environments. Ecotox Environ Saf. 2004; 57:168– 74. https://doi.org/10.1016/S0147-6513(03)00027-7

Schultz N, Norrgren L, Grawe J, Johannison A, Medhage O. Micronucleus frequency in circulating erythrocytes from rainbow trout (Oncorhynchus mykiss) subjected to radiation, an image analysis and flow cytometric study.Comp. Biochem Physiol.1993; 105:207–11. https://doi .org/10.1016/0742-8413(93)90196-R

Gustavino B, Scornajenghi KA, Minissi S, Ciccotti E. Micronuclei induced in erythrocytes of Cyprinus carpio (Telostei, Pisces) by X-rays and colchicine. Mutat Res. 2001; 494:151–9. https://doi.org/10.1016/S13835718(01)00191-7

Sawhney AK, Johal MS. Erythrocyte alterations induced by malathion in Channa punctatus (Bloch). Bull Environ Contam Toxicol. 2000; 64: 398–405. PMid: 10757665.https://doi.org/10.1007/s001280000014

Campos-Ventura B, De-Angelis DF, Marin-Morales MA.Mutagenic and genotoxic effects of the atrazine herbicide in Oreochromis niloticus (Perciformes, Cichlidae) detected by the micronuclei test and the comet assay. Pestic Biochem Physiol. 2008; 90:42–51. https://doi.org/10.1016/j.pestbp.2007.07.009

Cavas T. In vivo genotoxicity evaluation of atrazine and atrazine based herbicide on fish Carassius auratus using the micronucleus test and the comet assay. Food and Chem Toxic. 2011; 49:1431–5. PMid: 21443921. https://doi .org/10.1016/j.fct.2011.03.038

Grisolia CK. A comparison between mouse and fish micronucleus test using cyclophosphamide, mitomycin C andvarious pesticides. Mutat Res. 2002; 518:145–50. https:// doi.org/10.1016/S1383-5718(02)00086-4

Cavas T, Ergene-Gozukara S. Micronuclei, nuclear lesions and interphase silver-stained nucleolar organizer regions (AgNORs) as cytogenotoxicity indicators in Oreochromis niloticus exposed to textile mill effluent. Mutat Res.2003; 538(1-2):81–91. https://doi.org/10.1016/S1383 -5718(03)00091-3

Barsiene J. Genotoxic impacts in Klaipeda marine Port and Butinge oil terminal areas (Baltic Sea). Mar Environ Res. 2004; 54:475–9. https://doi.org/10.1016/S0141 -1136(02)00160-5



How to Cite

Chinnadurai, K., Balaji, P., & Eyini, M. (2020). Evaluation of Micronucleus Assay and Genotoxic Effect in <I>Oreochromis mossambicus</I> Exposed to a Fumaronitrile. Toxicology International, 26(3&amp;4), 70–79. https://doi.org/10.18311/ti/2019/v26i3&4/24267