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Interspecific differences in oxidative DNA damage after hydrogen peroxide exposure of sea urchin coelomocytes.
Liu F
,
Last KS
,
Henry TB
,
Reinardy HC
.
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Interspecific comparison of DNA damage can provide information on the relative vulnerability of marine organisms to toxicants that induce oxidative genotoxicity. Hydrogen peroxide (H2O2) is an oxidative toxicant that causes DNA strand breaks and nucleotide oxidation and is used in multiple industries including Atlantic salmon aquaculture to treat infestations of ectoparasitic sea lice. H2O2 (up to 100 mM) can be released into the water after sea lice treatment, with potential consequences of exposure in nontarget marine organisms. The objective of the current study was to measure and compare differences in levels of H2O2-induced oxidative DNA damage in coelomocytes from Scottish sea urchins Echinus esculentus, Paracentrotus lividus, and Psammechinus miliaris. Coelomocytes were exposed to H2O2 (0-50 mM) for 10 min, cell concentration and viability were quantified, and DNA damage was measured by the fast micromethod, an alkaline unwinding DNA method, and the modified fast micromethod with nucleotide-specific enzymes. Cell viability was >92% in all exposures and did not differ from controls. Psammechinus miliaris coelomocytes had the highest oxidative DNA damage with 0.07 ± 0.01, 0.08 ± 0.01, and 0.07 ± 0.01 strand scission factors (mean ± SD) after incubation with phosphate-buffered saline, formamidopyrimidine-DNA glycosylase, and endonuclease-III, respectively, at 50 mM H2O2. Exposures to 0.5 mM H2O2 (100-fold dilution from recommended lice treatment concentration) induced oxidative DNA damage in all three species of sea urchins, suggesting interspecific differences in vulnerabilities to DNA damage and/or DNA repair mechanisms. Understanding impacts of environmental genotoxicants requires understanding species-specific susceptibilities to DNA damage, which can impact long-term stability in sea urchin populations in proximity to aquaculture farms.
Figure 1. H2O2 concentration-dependent DNA damage (SSF, fast micromethod) of coelomocytes in three species of sea urchins, E. esculentus (■, n = 8), P. lividus (▲, n = 4), and P. miliaris (●, n = 4). Data were modelled with three-parameter logistic regression and data points are means ± SD (n = 4–8).). *Significant concentration-dependent increase in DNA damage (P < .05, ANOVA). **Significant species difference in overall level of SSF (P < .05, ANOVA).
Figure 2. DNA damage (SSF) in E. esculentus (n = 6, A), P. lividus (n = 6, B), and P. miliaris (n = 6, C) coelomocytes exposed to H₂O₂. Fast micromethod modified to include either no enzyme (white bars), incubation with FPG (grey bars), or incubated with Endo III (black bars). Data are means ± SD, *Significant difference in SSF between no enzyme, FPG, and Endo III (P < .05, GLM). DNA damage SSF was significantly affected by H₂O₂ concentrations for all treatments (P < .05, GLM).
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