Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Alkaline Comet Assay for Genotoxic Effect Detection in Neotropical Fish Prochilodus lineatus (Pisces, Curimatidae)

  • 251 Accesses

  • 27 Citations


Toxicants on fish may induce genetic alterations that can be used as genotoxic markers. We evaluated DNA damage using alkaline comet assay applied on erythrocytes after in vivo exposure of Prochilodus lineatus to different concentrations of Cypermethrin (0.300, 0.150, 0.075 and 0.000 μg/L) as a probable chemical mutagen. The results revealed a significantly higher level of DNA damage at all concentrations of Cypermethrin tested compared to control and background level (p < 0.05). We have standardized the technique for one of the most common native fish species that will be useful for biomonitoring genotoxicity in polluted waters of the region.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2


  1. Ali D, Nagpure NS, Kumar S, Kumar R, Kushwaha B (2008) Genotoxicity assessment of acute exposure of chlorpyrifos to freshwater fish Channa punctatus (Bloch) using micronucleus assay and alkaline single-cell gel electrophoresis. Chemosphere 71:1823–1831. doi:10.1016/j.chemosphere.2008.02.007

  2. Cavalcante DG, Martínez CB, Sofia SH (2008) Genotoxic effects of Roundup® on the fish Prochilodus lineatus. Mutat Res 655:41–46

  3. de Andrade VM, de Freitas T, da Silva J (2004) Comet assay using mullet. (Mugil sp.) and sea catfish (Netuma sp.) erytrocites for the detection of genotoxic pollutants in aquatic environment. Mutat Res 560:57–67

  4. de Campos Ventura B, de Angelis D, Marin-Morales MA (2008) 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 90:42–51. doi:10.1016/j.pestbp.2007.07.009

  5. Frenzilli G, Nigro M, Lyons BP (2009) The comet assay for the evaluation of genotoxic impact in aquatic environments. Mutat Res 681:80–92. doi:10.1016/j.mrrev.2008.03.001

  6. Houston AH (1990) Blood and Circulation. In: Schreck CB, Moyle PB (eds) Methods for fish biology. Am Fish Soc Meryland, USA, p 684

  7. Jha AN (2008) Ecotoxicological applications and significance of the comet assay. Mutagenesis 23:207–221. doi:10.1093/mutage/gen014

  8. Markert BA, Breure AM, Zechmeister HG (2003) Bioindicators & Biomonitoring. Principles, Concepts and Applications. Elsevier, Oxford, UK

  9. Matsumoto ST, Mantovani MS, Malagutti MIA, Dias AL, Fonseca IC, Marin-Morales MA (2006) Genotoxicity and mutagenicity of water contaminated with tannery effluents, as evaluated by the micronucleus test and comet assay using the fish Oreochromis niloticus and chromosome aberrations in onion root-tips. Genet Mol Biol 29:148–158. doi:10.1590/S1415-47572006000100028

  10. Nacci DE, Cayula S, Jackim E (1996) Detection of DNA damage in individual cells from marine organisms using the single cells gel assay. Aquat Toxicol 35:197–210. doi:10.1016/0166-445X(96)00016-1

  11. Nehls S, Segner H (2001) Detection of DNA damage in two cell lines from rainbow trout, RTG-W1, using the comet assay. Environ Toxicol 16:321–329. doi:10.1002/tox.1039

  12. Parma MJ, Loteste A, Campana M, Baccheta C (2007) Changes of hematological parameters in Prochilodus lineatus (Pisces, Prochilodontidae) exposed to sublethal concentration of cypermethrin. J Environ Biol 28(1):147–149

  13. Rodriguez Ferreiro G, Cancino Badías L, Lopez-Nigro M, Palermo A, Mudry M, González Elio P, Carballo M (2002) DNA single strand breaks in peripheral blood lymphocytes induced by three nitroimidazole derivatives. Toxicol Lett 132:109–115. doi:10.1016/S0378-4274(02)00039-5

  14. Rodriguez-Cea A, Ayllon F, Garcia Vazquez E (2003) Micronucleous test in freshwater fish: an evaluation of its sensitivity for application in field surveys. Ecotoxicol Environ Saf 56:442–448. doi:10.1016/S0147-6513(03)00073-3

  15. Singh NP, McCoy MT, Tice RR, Schneider EL (1988) A simple technique for quantitation of low levels of DNA damage in individuals cells. Exp Cell Res 175:184–191. doi:10.1016/0014-4827(88)90265-0

  16. Vanzella TP, Cólus Martinez CB, MS I (2007) Genotoxic and mutagenic effects of diesel oil water soluble fraction on a neotropical fish species. Mutat Res 631:36–43

Download references


The work was supported by research grant PI CAI + D N° 21/123 2006 from Universidad Nacional del Litoral, Argentina.

Author information

Correspondence to M. J. Parma.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Simoniello, M.F., Gigena, F., Poletta, G. et al. Alkaline Comet Assay for Genotoxic Effect Detection in Neotropical Fish Prochilodus lineatus (Pisces, Curimatidae). Bull Environ Contam Toxicol 83, 155–158 (2009). https://doi.org/10.1007/s00128-009-9771-z

Download citation


  • Comet assay
  • Prochilodus lineatus
  • Genotoxic effects
  • Cypermethrin