Detection of DNA damage in yolk-sac larvae of the Japanese Medaka, Oryzias latipes, by the comet assay.

This study was set up to determine the suitability of the early life stage (ELS) alkaline comet assay for the detection of DNA strand breaks induced by genotoxicants in whole organism. This assay was performed on cells of medaka 2 days posthatch (dph). An efficient procedure for cell dissociation using enzymatic and mechanical digestion was developed. This protocol ensures 80% viability of cells and low DNA damage background. Cells from 2 dph medaka larvae were exposed in vitro to model genotoxicants, hydrogen peroxide, cadmium, and fluoranthene, followed by comet assay analysis. Results show a significant increase in the percentage of DNA damage of dissociated cells by all the tested compounds when compared to controls. The assay was also performed in vivo on medaka larvae (2 dph) exposed for 24 h to waterborne cadmium or fluoranthene. Significant induction of DNA damage levels were observed following larvae exposure to cadmium and fluoranthene at concentrations of 0.1 and 50 µM, respectively. This study demonstrates that cells of embryo life stage medaka respond to known DNA damaging agents and that the ELS comet assay may be a useful biomarker to detect DNA strand breakage in whole body of pluricellular organism induced by a range of agents. This technique may provide a sensitive, nonspecific endpoint of genotoxicity as part of ELS toxicity test.

Occurrence and concentrations of benzotriazole UV stabilizers in marine organisms and sediments from the Ariake Sea, Japan.

The benzotriazole UV stabilizers, which are used in a variety of plastic products, were analyzed in marine organisms and sediments collected from the Ariake Sea, Japan. The UV stabilizers, such as UV-320, UV-326, UV-327, and UV-328 were detected in all of the samples analyzed, suggesting the production and use of these compounds in Japan. High concentrations of UV stabilizers were found in clams, oysters, and gastropods collected from the tidal flat at concentrations on the order of several hundreds of ng/g on a lipid weight (wt.) basis. The higher trophic species, such as hammerhead sharks and coastal birds, accumulated UV stabilizers, with mean concentrations of 190 ng/g and 74 ng/g (lipid wt.), respectively. These results indicate that benzotriazole UV stabilizers are persistent and bioaccumulative in the marine food-chains. The benzotriazole UV stabilizers were also detected in coastal and river sediments around the Ariake Sea, at concentrations in the range of 7.9-720 ng/g (dry weight basis). Significant correlations were found between concentrations of UV stabilizers and organic carbon content in sediments, implying adsorption of these compounds to organic matter. To our knowledge, this is the first report of ubiquitous contamination and bioaccumulation of benzotriazole UV stabilizers in the marine environment.