Marine pollutants such as heavy metals (HMs) are considered among the most copious oxidative stress (OS) inducers in marine organisms which leads to reactive oxygen species (ROS) formation. Complementary to our previous bioassays studies, the present research focuses on Catalase (CAT), Glutathione S-transferase (GST) and Malondialdehyde (MDA) as oxidative stress biomarkers and the integrated biomarker response (IBR) indexes (IBR1 and IBR2) as an ecotoxicological assessment tool in Mytilus galloprovincialis using central composite face centered (CCF) design. The oxidative stress biomarkers were measured in adult mussels (45-55 mm) on 3 days-exposed under different sub-lethal concentrations of cadmium (Cd), zinc (Zn), and copper (Cu). Using multiple regressions, ANOVA analysis revealed that experimental data fitted to second-order (quadratic) polynomial equations. The results showed that types, concentrations and metals combinations has a direct effect in CAT and GST activities, MDA level and IBR indexes. Additionally, metal-metal interactions were found synergistic (supra-additive), antagonistic (infra-additive) or zero interaction in the toxicological effect. As necessary, the optimization of the experimental results was done in order to determine the optimal conditions for the oxidative stress responses and IBR indexes. It was demonstrated that the CCF design combined with the multi-biomarker approach and IBR index can be used as an appropriate tool in ecotoxicological modulation and prediction of oxidative stress and antioxidant status by heavy metals in the mussels Mytilus galloprovincialis.
Keywords: CCF design; Heavy metals; LC50; Modeling optimization; Mytilus galloprovincialis; Oxidative stress.
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