An alkaline single-cell gel electrophoresis (comet) assay for environmental biomonitoring with native rodents

Abstract

The main advantages of single-cell gel electrophoresis (SCG) are its applicability to any eukaryotic organism and cell type, its low cost and the short time required to obtain results. These properties make the SCG assay particularly useful in screening for environmental genotoxicity. The present study describes a modified version of this technique for use in field work with native rodents and examines some factors which influence the outcome of the assay. Wild rodents (Ctenomys torquatus, "tuco-tuco") from a region close to a strip coal mine and from a region with no coal mines were used. Animals from the coal mining region had significantly more DNA damage than those from the control area. The use of this SCG technique for direct sampling in the field should facilitate environmental genotoxicity studies with natural populations, without the need to remove the animals from their habitat or to sacrifice them.

Potential Role of Mosquito Larvae <i>Culex Pipiens</i> as a Biological Indicator of Environmental Water Pollution in Egypt

“…Our results for C. pipiens indicate that pollution-induced DNA damage is detectable by the SCG assay, and confirm the viability of this method in the environmental biomonitoring. The study of animals in their habitat and without the need to sacrifice them provides an ideal approach for environmental evaluation (Silva et al, 2000). Van et al (2003) showed that the assay is easy, generally modest and simple to perform.…”

“…Eukaryotic organism and cell types have also been tested with this assay (Petras et al, 1995;Tice, 1995;Verschaeve and Gilles, 1995;Sasaki et al, 1997a, b). Some studies used small mammalian species, living in or close to polluted areas for detection of hazardous pollution (Farbairn et al, 1995;Petras et al, 1995;Tice, 1995;Baker et al, 1996;Salagovic et al, 1996;Ralph et al, 1997;Silva et al, 2000). Also, comet assay has been applied to cells of insects, including Shistocerca gregaria, Drosophila melanogaster, Curculio sikkimensis, mosquito larvae, Ephestia kuehniella and in grasshoppers Chorthippus brunneus (Siddique et al, 2005;Todoriki et al, 2006;Augustyniak et al, 2006;Yousef et al;, Abdel-Gawad et al, 2011Isabel and Maria, 2014;Pandir and Guven, 2014).…”

Cellular Responses to Tritium Exposure in Rainbow Trout: Hto- And Obt-Spiked Feed Exposure Experiments

“…Finally, a cover slip was mounted over each slide and at least 100 cells were scored at 400× magnification using a fluorescent microscope. The scoring criteria (derived from da Silva et al [14]) are presented in Table 3.…”

DNA damage in children and adolescents with cardiovascular disease risk factors

“…Cells were visually examined to determine DNA damage scores ranging from class 0 (no migration) to class 4 (maximal migration) based on tail intensity (size and shape). Two DNA damage parameters were calculated accordingly: the damage index (DI), which summed the scores of all 100 cells and ranged from 0 (cells with no migration) to 400 (cells with maximal migration), and the damage frequency (DF), which is the percentage of damaged cells (Da Silva et al 2000). For the CBMN assay, an aliquot of blood (0.5 mL) was added to 5 mL of RPMI 1640 medium supplemented with 20% fetal calf serum and 0.2% phytohemagglutinin.…”