Genotoxicity of cadmium in marine diatom Chaetoceros tenuissimus using the alkaline Comet assay
Genotoxic effects of cadmium on phytoplankton Chaetoceros tenuissimus have been evaluated using DNA damage by Comet assay. Cadmium concentrations ranging from 2.4 to 10 mg/l were used to evaluate the effects. Results showed that as the concentration of Cd increased growth of the diatom decreased. Alkaline single-cell gel electrophoresis (Comet assay) method, which is highly sensitive in detection of DNA damage in eukaryotic cells, was used to observe genomic changes in marine diatom cells. DNA damage was measured as percent number of comets and normal cells. 65% cells were found to be damaged at 10 mg/l concentration of Cd as compared to 23% in 2.4 mg/l and only 5% in controls. More than 50% apoptic cells were observed on 8th day at 10 mg/l and 12th day at 7.5 mg/l concentrations. At lower Cd concentrations (4.5 mg/l and below) the damage was below 30% till the last day. This suggested that higher Cd levels have early damaging effects on cell nuclear material and that % injury increases with advancement of exposure period. One advantage of use of C. tenuissimus is the ease with which it can be cultured in a defined medium. C. tenuissimus diatom can be used as an in vivo model for ecogenotoxicity assessment using the Comet assay.
KeywordsGrowth Chaetoceros tenuissimus Comet assay Single-cell gel electrophoresis DNA damage
The authors are grateful to Dr. S. R. Shetye, Director, NIO for encouragement. Thanks are also due to Dr. S. G. Dalal for his help in statistical analysis. This work was carried out with funding (GAP 0323) from the Department of Ocean Development, New Delhi. This is NIO contribution No. 4119.
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