Genotoxic effects of the water-soluble fraction of heavy oil in the brackish/freshwater amphipod Quadrivisio aff. lutzi (Gammaridea) as assessed using the comet assay
Amphipod crustaceans have been widely used as invertebrate models in ecotoxicology due to their importance in the food chain. However, few studies have evaluated the genotoxic effects of pollutants in this model using the comet assay. The main obstacle to using amphipods in the comet assay is the difficulty in obtaining enough blood cells from a single individual. In this study, we evaluated the genotoxic effects of the water-soluble fraction (WSF) of heavy oil on the brackish/freshwater amphipod Quadrivisio aff. lutzi, which is common in the coastal lagoons of southeastern Brazil, using hemocytes obtained from single amphipods (without pooling) after optimizing hemolymph extraction. The comet assay revealed significantly higher DNA damage levels (2- to 6-fold higher) in treated amphipods compared to untreated ones with a sublethal concentration of 17.6 % of the WSF within 72 h of treatment. Two independent experiments confirmed an “up and down” pattern of DNA damage, measured as the % of DNA contained in the tail of the comets. Elevations in DNA damage levels were observed at the 6 and 48 h time points, while very low levels of DNA damage were observed at the 24 and 72 h time points. Furthermore, the comet assay revealed gender variability in the levels of DNA damage after short-term exposure.
KeywordsEcotoxicology Comet assay Petroleum Invertebrate Amphipoda Gender
The authors are grateful for the financial support that was provided by FINEP/PETROBRAS S/A, Research Fund No. 01.06.1191.00, and to FUNEMAC for three IC Grants. We are thankful to Cristiana Serejo for the taxonomic identification of the amphipods and to Lisia Gestinari for the taxonomic identification of the algae to which the amphipods were associated. We are also particularly grateful to two anonymous reviewers for their valuable suggestions and comments which have improved the manuscript considerably.
Conflicts of interest
The experiments outlined in the present work comply with current Brazilian laws. The authors declare that they have no conflicts of interest.
- ANP (2011) Anuário Estatístico 2011—Anuário Estatístico Brasileiro de Petróleo, Gas Natural e Biocombustíveis. http://www.anp.gov.br/?pg=56346#Se_01. Accessed 21 Feb 2012
- Browne M (2009) Imaging and image analysis in the comet assay. In: Dhawan A, Anderson D (eds) The comet assay in toxicology. RSC Publishing, Cambridge, pp 343–390Google Scholar
- Finney DJ (1964) Probit analysis: a statistical treatment of the sigmoid response curve, 2nd edn. Cambridge University Press, LondonGoogle Scholar
- Ingersoll CG, MacDonald DD, Brumbaugh WG, Johnson BT, Kemble NE, Kunz JL, May TW, Wang N, Smith JR, Sparks DW, Ireland DS (2002) Toxicity assessment of sediments from the Grand Calumet River and Indiana Harbor Canal in Northwestern Indianan, USA. Arch Environ Contam Toxicol 43:156–167CrossRefGoogle Scholar
- Katsumiti A, Domingos FXV, Azevedo M, da Silva MD, Damian RC, Almeida MIM, Silva de Assis HC, Cestari MM, Randi MAF, Oliveira Ribeiro CA, Freire CA (2009) An assessment of acute biomarker responses in the demersal catfish Cathorops spixii after the Vicuña oil spill in a harbour estuarine area in Southern Brazil. Environ Monit Assess 152:209–222CrossRefGoogle Scholar
- Pellegrini M, Colodette G, Martinez I, Neves L (2008) Jubarte field production enhanced with wellbore ESP. Offshore 68, Brazil, 6 pp. http://www.offshore-mag.com/index/article-display/334453/articles/offshore/volume-68/issue-7/brazil/jubarte-field-production-enhanced-with-wellbore-esp.html. Accessed 2 Aug 2011
- Pérez-Cadahía B, Laffon B, Pásaro E, Méndez J (2004) Evaluation of PAH bioaccumulation and DNA damage in mussels (Mytilus galloprovincialis) exposed to spilled Prestige crude oil. Comp Biochem Physiol C 138:453–460Google Scholar
- Petroleum (2009) Information on petroleum and crude oil. http://www.petroleum.co.uk/. Accessed 26 July 2011
- Shoemaker CR (1933) Amphipoda from Florida and the West Indies. Am Mus Novit 598:1–24Google Scholar
- Stebbing TRR (1907) The Fauna of brackish ponds at port canning, lower Bengal. Part V. Definition of a new genus of Amphipoda, and description of the typical species. Rec Indian Mus 1:159–162Google Scholar
- Trevisan OV, França FA, Clarissa AO, Lisboa AC (2009) Elements of a heavy oil technology development program. Terrae 6:21–29Google Scholar