1. Biological effects related to the Prestige oil spill in demersal fish from the northern Iberian shelf Martínez-Gómez C., Campillo J.A., Benedicto J., Fernández B., Valdés J., García I. & Sánchez F*. Spanish Institute of Oceanography. Oceanographic Centre of Murcia (Marine Environment Protection Area) Oceanographic Centre of Santander (Fisheries Area) Spanish Institute of Oceanography

2. European Space Agency. December 2002 www10.uniovi.es/prestige/ fotos/fotosSatelite.htm Spanish Institute of Oceanography

3. The potencial toxicity of the fuel should not be evaluated on the only basis of the 16 EPA PAH content in the fuel (less than 0.1% --- 63 Tons). The fuel contains more than 3% GC-resolved alkylated PAH and total aromatic content is around 50%. Spanish Institute of Oceanography It may have potentially carcinogenic, mutagenic and teratogenigc effects for the marine organisms and humans. They may act also as endocrine disruptors. Prestige fuel oil (J. Oudot. http://www.le-cedre.fr/fr/prestige ) Heavy metal contamination in the Northeast Atlantic Ocean (Prego & Cobelo-García., 2004)

4. Biological effects of the pollution: Sublethal effects Pollution contributes to the stress in marine organisms Marine organisms are provided with physiologial and biochemical mechanisms to survive in degraded habitats Great part of the biological changes are due to low exposition levels of pollutants for long time periods. Spanish Institute of Oceanography Chronic Pollution

5. MAIN OBJECTIVE: HANDICAP: Spanish Institute of Oceanography To carry out a medium term biomonitoring programe, using biochemical biomarkers related with the PAHs exposure in demersal fish from the Galician-Cantabrian shelf...... In order to evaluate the possible biological effects related with the Prestige oil spill Until the Prestige sank, no studies using biochemical biomarkers in fish species had been made on the Galician-Cantabrian shelf.

6. (Bayne et al., 1985; Moore & Simpson, 1992; Moore et al., 1994; Moore, 2002a). Spanish Institute of Oceanography

8. Lepidorhombus boscii : Muddy bottoms Depths of 120 to 350 m Pietrapiana et al., 2002 Callyonimus lyra : sandy bottoms depths less than 150 m Burgeot and Galgani, 1994 Spanish Institute of Oceanography Sánchez et al., 2002

9. SELECTED BIOMARKERS EROD activity in microsomal fractions (Burke & Mayer, 1974). Glutation-S-Transferase activity in citosolic fractions (Habig & Jakoby, 1981) Glutation Reductasa activity in citosolic fractions (Karacoh et al., 1998). Catalase activity in citosolic fractions (Greenwald, 1985) Evaluation of genotoxic effects using the alkaline filter elution method recommended by MED POL (UNEP/RAMOGE, 1999). Spanish Institute of Oceanography

10. “An assessment of the impact of the Prestige oil spill on the ecosystems on the continental shelf and its resources. Spring period”. Marzo 2003-Sept 2003 “Distribution, evolution and effects of the fuel-oil in the coast affected by the Prestige Oil Spill. Integrated study. Jan 2004- Dec 2006

11. Spanish Institute of Oceanography Survey carried out in April 2003 71-120 m (n=12) size range [18-20] cm (age class 2) 121-200 m (n=12) 201-500 m (n=6) size range [18-20] cm (age class 2)

12. 9.008.007.006.005.004.003.002.00 9.008.007.006.005.004.003.002.00 42.00 43.00 44.00 42.00 43.00 44.00 10 20 40 60 80 MF FE EPPAAB EROD activity L. boscii EROD activity levels (pmol*min-1*mg prot.microsomal-1) Spanish Institute of Oceanography RESULTS APRIL 2003

13. GST activity rates (nmol*min -1 *mg prot. cit.) 9.008.007.006.005.004.003.002.00 9.008.007.006.005.004.003.002.00 42.00 43.00 44.00 42.00 43.00 44.00 75 100 125 150 200 MF FE EPPA AB GST activity L. boscii Spanish Institute of Oceanography RESULTS APRIL 2003

14. 9.008.007.006.005.004.003.00 2.00 9.008.007.006.005.004.003.002.00 42.00 43.00 44.00 42.00 43.00 44.00 5 10 15 20 25 MF FE EPPAAB GR activity L. boscii Spanish Institute of Oceanography RESULTS APRIL 2003 GR activity rates (mmol*min -1 *mg prot. cit.)

15. Spanish Institute of Oceanography RESULTS APRIL 2003

16. Spanish Institute of Oceanography RESULTS APRIL 2003 Catalase (mmol*min -1 *mg prot. cit.)

17. Spanish Institute of Oceanography RESULTS APRIL 2003 Genotoxicity: Elution rate constant

18. 9.008.007.006.005.004.003.00 2.00 9.008.007.006.005.004.003.002.00 42.00 43.00 44.00 42.00 43.00 44.00 50 100 200 300 400 MF FE EPPAAB EROD activity C. lyra Spanish Institute of Oceanography EROD activity levels (pmol*min-1*mg prot.microsomal-1) RESULTS APRIL 2003

19. Spanish Institute of Oceanography RESULTS APRIL 2003 GST activity rates (nmol*min -1 *mg prot. cit.) 9.008.007.006.005.004.003.002.00 9.008.007.006.005.004.003.002.00 42.00 43.00 44.00 42.00 43.00 44.00 100 125 150 200 250 MF FE EPPA AB GST Activity C. lyra

20. Spanish Institute of Oceanography RESULTS APRIL 2003 GR activity rates (mmol*min -1 *mg prot. cit.) 9.008.007.006.005.004.003.002.00 9.008.007.006.005.004.003.002.00 42.00 43.00 44.00 42.00 43.00 44.00 15 18 21 24 27 MF FE EPPAAB GR activity C. lyra

21. Spanish Institute of Oceanography RESULTS APRIL 2003 Catalase activity rates (mmol*min -1 *mg prot. cit.)

22. Spanish Institute of Oceanography RESULTS APRIL 2003 Genotoxicity: Elution rate constant 9.008.007.006.005.004.003.002.00 9.008.007.006.005.004.003.002.00 42.00 43.00 44.00 42.00 43.00 44.00 30 60 90 120 150 MF FE EPPA AB (K x 1000) Genotoxic damages C. lyra

23. EROD activity and fuel (p=0.024) Elution rate “k” and tar aggregates p=0.025 Sánchez et al., 2002 González et al., 2003 Rodríguez et al., 2003

24. 9.008.007.006.005.004.003.002.00 9.008.007.006.005.004.003.002.00 42.00 43.00 44.00 42.00 43.00 44.00 10 12 15 17 20 MF FE EPPA AB PMS content mg PMS/ml MF C. lyra 9.008.007.006.005.004.003.002.00 9.008.007.006.005.004.003.002.00 42.00 43.00 44.00 42.00 43.00 44.00 5 10 15 20 25 MF FE EPPAAB PMS content mg PMS/ml MF L. boscii Spanish Institute of Oceanography

25. size range [19-21] cm (age class 2) Spanish Institute of Oceanography Survey carried out in October 2003

26. González et al., 2004)

27. Spanish Institute of Oceanography Survey carried out in October 2003

28. 4 times minimum value

29. 5 Spanish Institute of Oceanography

30. 3-4

31. Spanish Institute of Oceanography

32. The simultaneous use of both species allowed us to obtain information from different shelf environments since C. lyra is representative of the inner shelf (70-120 m), whereas L. boscii represents the middle and external shelf (120-350 m). Systematically, the biomarker results indicate that the fish populations from Finisterre, Galicia N, Asturias E and Cantabria areas are exposed to compounds that affect the detoxification process and, in the case of Galicia N these compounds may also to have a genotoxic potential, at least in the coastal shelf environment. We can conclude that....

33. The results obtained in Asturias W and the Galicia S offshore point to a lower degree of exposure to organic xenobiotic compounds than those observed in another areas studied. In the Basque Country area the results are not conclusive. However, the GR and catalase values measured in April (in L. boscii collected in the Basque Country) pointed to the existence of an oxidative stress comparable to that observed in Galicia N, although EROD and GST levels were lower. The presence of a certain type of metal pollution and another organic compounds may be interacting with the biotransformation enzymes ADAPTATIVE PROCESSES ¿? We can conclude that....

34. The fact that in a systematic way, the pollutant concentrations in sediments were slightly higher in the Cantabrian than in the Galician zones contrasts with the observation that highest levels of EROD induction in C. lyra was observed in Finisterre and that DNA integrity was significantly lower in Galicia N. These results, therefore, may be indicating a possible relation between cause and effect but …. We can conclude that....

35. Significantly high induction values of GST, GR and catalase found in L. boscii in the areas of Finisterre and Galicia N cannot only be explained by taking into account the low pollutant concentrations measured in offshore sediments from the mid and external continental shelf (OSPAR, 2000; Soriano et al., 2003; IEO, 2004), which again make us think that the POS is the cause of the impact… In any case, the biomarkers are reflecting in both species the situation prevailing in the environmental bottom shelf. Taking into account the results obtained up to date with L. boscii, we consider appropriate their use as target species in the Northern Iberian Continental shelf. The use of C. lyra depends on its abundance in the catches. We can conclude that....

36. Spanish Institute of Oceanography FURTHER WORKS.... Responses using separated genders Evolution along time

37. Spanish Institute of Oceanography... To those who doesn´t appear in the picture and to the crew of the R/V “Vizconde de Eza” and “Cornide de Saavedra”................ Thank you...

38. Spanish Institute of Oceanography The work was carried out partly with the help of a Support Personnel Research Training fellowship to Valdés, J. from the SIO This study was performed within the frameworks of research projects ECOPREST and DEEP II, both funded by the Spanish Science and Technology Ministry through an Strategic Action on accidental marine dumpings