2. Environmental Monitoring Methods SOURCES Concentrations in media (external dose) Emission rates SOURCES Concentrations in media (external dose) Emission rates INDICATORS OF EXPOSURE Bioaccumulation markers Biomarkers of exposure INDICATORS OF EXPOSURE Bioaccumulation markers Biomarkers of exposure INDICATORS OF EFFECTS Biomarkers of effects Injury Population & Community parameters INDICATORS OF EFFECTS Biomarkers of effects Injury Population & Community parameters Chemical monitoring Bioaccumulation & Biological effect monitoring Biological effect, Health & ecosystem monitoring Environmental factors modifying exposure Environmental factors modifying exposure Environmental factors modifying susceptibility Environmental factors modifying susceptibility Other causes

3. Major Processes in Pollutant Impact Contaminant Mixtures Particulates Speciation, Binding & Transport Toxicity & Cell injury Physicochemical Behavior, Activation & Detoxication Exposure Biomarkers Pathological Endpoints Prognostic Biomarkers Ecological Impact Mortality, Reduced Fecundity Increased Susceptibility to Other Stresses & Loss of Diversity Human Health Impact Toxicity & Diseases via food chain UV & Visible Radiation UV & Visible Radiation Uptake Import by adsorptive Endocytosis & Phagocytosis Uptake Trans-Membrane Diffusion

4. Biological Effects & Ecological Relevance Physical factors & Biological Agents Pollutant Chemicals & Radionuclides Molecular & Cellular TissueOrgansIndividual animal SHORT TERMLONG TERM Biological Complexity, Response Time & Biological Significance EARLY BIOMARKER SIGNALS LATER EFFECTS

5. Environmental Pollutants Lethal effects Sublethal effects Changes in enzyme activities Damage to DNA, proteins & membrane lipids & membrane lipids Effects on behavioral, reproductive reproductive & immunological activities Environmental Pollutants & Effects to Organisms

6. Mussels: Bioindicators for Aquatic Pollution Freshwater Ecosystems Marine Ecosystems Sentinel Species Bioindicators Biomonitoring Wide geographical distribution Easy to collect Filter-feeding organisms Bioconcentrate xenobiotics Easily transferred from control to contaminated areas

7. Chemical Pollutants & Oxidative Stress ROS Organic & Inorganic Pollutants Organic & Inorganic Pollutants Aquatic Organisms Aquatic Organisms Polyclic Aromatic Hydrocarbons Organochlorine & Organophoshate pesticides Polychlorinated biphenyls Aromatic diols Quinones Nitroaromatics Aromatic hydroxylamines Dioxins Bipyridyls Transition metal chelates Redox Potential Molecular Biomarkers Of Oxidative Stress Molecular Biomarkers Of Oxidative Stress Lipid Peroxidation Enzyme Activities DNA Damage Protein Degradation * ROS: Reactive Oxygen Species

8. ROS & biochemical & physiological responses in cells of aquatic organisms LipidPeroxidation Anthropogeni c activities Environmental factors MT CA T PUFAs SO D Organic pollutants Solar radiatio n Temperature Salinity Μ n+ Metal transport Metal complexation Ions diffusion Ο2─Ο2─ Metal competition Metals Μ n+ Oxidative DNA damage Proteindamage Changes in Enzyme activities ΗΟ  Η2Ο2Η2Ο2 Ο2─Ο2─ Proteins Η2Ο2Η2Ο2

9. Superoxide dismutase (SOD) SOD

10. Catalase (CAT) CAT

11. Lipid Peroxidation (LPO) Initiation Propagation Termination Inhibition Antioxidants OxidizedAntioxidants Degradation of Lipid Peroxides Aldehydes and other products Biomarkers of Oxidative stress LH: lipid substrate L: lipid radical LOOH: lipid hydroperoxide LOO: lipid peroxyl radical NRP: nonradical product

12. Oxidative Stress in Mussels from the Saronikos gulf (Greece) 3 polluted sites (1, 2, 3) in the Saronikos gulf Reference site (4) “Clean” marine farm PAHs Metals (Fe,Cd,Cu,Cr,Ni,Pb) Biomarkers of Oxidative Stress Sampling at 4 seasons Superoxide dismutase (SOD) activity Catalase (CAT) activity Lipid Peroxidation (LPO) levels Measurements of PAHs and Metal concentrations in mussel tissues revealed elevated levels of chemical pollution at Sites 1 & 2 Measurements of PAHs and Metal concentrations in mussel tissues revealed elevated levels of chemical pollution at Sites 1 & 2

13. Representative chart for PAHs concentrations in mussel tissues

14. Results for SOD activity

15. Results for CAT activity

16. Results for Lipid Peroxidation levels

17. Conclusions Pollution in coastal waters of Saronikos gulf Pollution in coastal waters of Saronikos gulf causes moderate levels of oxidative stress causes moderate levels of oxidative stress in tissues of mussels in tissues of mussels The biomarkers of SOD, CAT & LPO are suitable The biomarkers of SOD, CAT & LPO are suitable for appraising the seawater quality for appraising the seawater quality and can be used as cost effective test system for Environmental Risk Assessment

18. Factors that affect biomarker responses of oxidative stress in aquatic organisms Biomarker Responses Biomarker Responses Health condition Sex Age Nutritional status Metabolic activities Migratory behavior Reproductive & Developmental status Population density Season Ambient temperature Heterogeneity of the Environmental Pollution