Use of DNA Strand Damage (Comet Assay) and Embryo Hatching Effects to Assess Contaminant Exposure in Blue Crab (Callinectes sapidus) Embryos

摘要

After fertilization, blue crab eggs are embedded in a "sponge" which is attached to the female abdomen during embryo deveolpment. Embryos after 9 stages in the egg sac hatch into a swimming zoea stage (stage 10). We have developed a bioassay where embryo development is monitored in culture plates with and without toxicants in the water. Toxicant effects are based on determining the percentage of embryos which hatch to zoea. Hatching EC_50 (toxicant concentration at which 50percent of the embryos fail to hatch) for a number of pesticides, organometallics and metals were determined. The test takes from 2 to 6 days depending on the embryo stage selected for the study. In addition to embryo development effects the prevalence of DNA single-strand breaks in individual embryo cells were determined using the single cell gel electrophoresis method (Comet assay). A good correlation between DNA strand breakage and embryo defects was found after exposure to genotoxic contaminants. Thus, the bioassay linking DNA damage to embryo hatching effects is rapid, sensitive and mechanistically relevant.