Abstract
The present study applied the comet assay to erythrocytes of Oreochromis niloticus with the aim of improving protocols to detect DNA damage in these cells, by using two distinct pHs (pH = 12.1 and pH > 13) and evaluating whether there is a correspondence between silver and ethidium bromide staining. Comets were visually examined and, the frequency of cells with and without damage was obtained, as well as the distribution of classes and scores. By using the Kruskal-Wallis test, our results revealed that pH 12.1 is more effective, although both pHs can be used. Our findings also suggest that silver staining can substitute ethidium bromide, an expensive and highly toxic stain that requires specific equipment for examination.
Highlights
The development of new methodologies and the application of more sensitive assays to detect genotoxicity in different samples have been the subject of several scientific studies on environmental monitoring (Al-Sabti and Metcalfe, 1995; Lemos et al, 2005)
Compared with other genotoxicity tests, the advantages of the comet assay are the detection of slight DNA damage, the low number of cells required, low cost, precision, ease of application, reproducibility, and short period of time to conduct the experiment (Belpaeme et al, 1998; Tice et al, 2000; Bücker et al, 2006)
This technique is the result of studies undertaken by Östling and Johanson, who developed the methodology of DNA electrophoresis in micro-gel, and those by Singh et al (1988), who improved the technique
Summary
Application of the comet assay in erythrocytes of Oreochromis niloticus (Pisces): A methodological comparison. Fontanetti1 1Departamento de Biologia, Instituto de Biociências, Universidade Estadual Paulista “Júlio de Mesquita Filho”, Rio Claro, SP, Brazil. 2Departamento de Genética, Instituto de Biociências, Universidade Federal do Pará, Belém, PA, Brazil
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