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ECOSYSTEM HEALTH ASSESSMENT BECPELAG (2001-2002) Water Column Survey (2003-2004) Impacts of oil and gas extraction in the North Sea Oceanographic vessel Mytilus galloprovincialis Mullus barbatus Mytilus edulis Gadus morhua BEEP (2001-2004) Sampling areas in Europe for biomonitoring of biological effects of pollutants in sentinel species PRESTEPSE (2003-2006) IMPRES (2003-2007) PRESTIMES (2008) K-EGOKITZEN (2007-2009) Gorliz Arriluze URDAIBAI (1991-1994) TRANSPLANT STUDY (1998) DERBIUR (2006-2008) OKATUR (2007-2008) MARINE AND COASTAL BIOMONITORING Miren P. Cajaraville (PR), Ionan Marigómez, Eduardo Angulo, Mari Carmen Barbero, Manu Soto, Teresa Serrano, Ibon Cancio, Amaia Orbea, Maren Ortiz-Zarragoitia, Beñat Zaldibar, Izaskun Zorita, Eider Bilbao, Urtzi Izagirre, Damien Raingeard, Larraitz Garmendia, Vega Asensio, Amaia Rodríguez, AneMiren Castilla, Pamela Ruiz, Eunate Puy, Aitzpea Errasti, Oihane Díaz de Cerio, Alberto Katsumiti, Unai Vicario, María Múgica. Department of Zoology and Cell Biology, Laboratory of Cell Biology and Histology, Faculty of Science and Technology, University of the Basque Country. CELL BIOLOGY IN ENVIRONMENTAL TOXICOLOGY (CBET) RESEARCH GROUP RESEARCH ACTIVITIES: RESEARCH ACTIVITIES: RESEARCH ACTIVITIES: RESEARCH ACTIVITIES: • ECOSYSTEM HEALTH ASSESSMENT • ENDOCRINE DISRUPTION • TOXICOGENOMICS • CHEMICAL CARCINOGENESIS • NANOPARTICLE TOXICITY SET OF BIOMARKERS USED FOR ECOSYSTEM HEALTH ASSESSMENT • Cell viability • Endocytic/phagocytic ability • ROS production • Cytoskeleton integrity General and specific toxicity in vitro Immunotoxicity • Peroxisome proliferation • Phase I (EROD), II and III metabolism of xenobiotics Organic chemical exposure • Metallothionein levels • MT20 gene expression • Intralysosomal metal levels Metal exposure Exposure to specific pollutants Alterations in reproduction • Vitellogenin (Vtg) and Vtg-like protein levels • Gonad index • Estrogen receptor gene reporter assays • Steroid hormone metabolism-aromatase • OECD reproduction test in Eisenia Endocrine disruption Alterations in reproduction • Micronuclei frequency • 7,8 oxo dG intensity • Comet assay DNA damage Genotoxicity • Lipid peroxidation • Anti-oxidant enzyme measurements Oxidative damage • Lysosomal membrane stability • Lysosomal structural changes • Lipofuscin levels • Neutral lipid accumulation Lysosomal alterations • Cell-type composition • Digestive epithelial thickness • Parasite infestation • Inflammation, necrosis, neoplasia and other lesions Histopathology • Stress on stress Mortality General stress MEASUREMENT ENDPOINT ISSUE • Research and teaching activities within the fields of Cell Biology and Environmental Toxicology (both aquatic and terrestrial ecosystems) since 1983. • Recognized as Consolidated Research Group since 2001. • Lab facilities for histology, light, fluorescence and electron microscopy, image analysis, cell fractionation and biochemistry, cell culture and molecular biology. Experimental aquarium service • Main research fields: - Development and application of early warning cellular and molecular biomarkers of pollution exposure and effects and in vitro toxicity assays - Understanding of the cellular and molecular mechanisms of toxicity of chemical pollutants • Participation in bilateral projects with French, Portuguese, Italian, German, Norwegian, and Swedish groups and in EU projects XENOFISH (IV FP), BEEP (V FP), PRAGMA (VI FP) and NANORETOX (VII FP). • Research Masters: Contaminación y Toxicología Ambientales; European MSc in Marine Environment & Resources. Advanced & International Postgraduate Courses. Since the early 80’s, CBET has been investigating the biological effects of contaminants in both terrestrial and aquatic ecosystems with a common conceptual and methological approach (biomarkers + bioassays + bioanalytical chemistry) applied to target sentinel species that are representative of the corresponding ecosystem in terms of health. Marine ecosystem health assessment. Pioneering monitoring programmes were started in the Abra estuary (1991-93) and in Urdaibai (1991-94). Since 1998, ecosystem health assessment has been carried out in North Sea and Mediterranean Sea within the framework of international projects/contracts (XENOFISH, 1998-99; BECPELAG, 2001-02; WCM, 2004; BEEP, 2001-04). Since 2000, the biological impact of the refinery sewage discharges outside the Abra estuary is being annually assessed (Petronor contract). After the Prestige oil spill, its biological impact was assessed in 22 localities from Portugal to the Basque Country (2003-06) and in laboratory experiments (IMPRES; PRESTEPSE). This investigation is being completed with lab experiments (PRAGMA; 2006-07) and a running field research (PRESTIMES; 2008). Further on, monitoring campaigns have been continued (IMPRES II; 2006-07) in the Basque coast in order to obtain baseline values suitable as reference values for future accidental spills. Soil ecosystem health assessment. Laboratory studies on the pollution effects in soil organisms (slugs) started in early 80’s. Later, mining areas in Wales and volcanic areas in Açores were used as experimental fields, in 1994 (PRAXIS XXI, 1996-99). Currently, within the framework of the BERRILUR strategic project (2003-ongoing) and PACARI (2006-09), which included slugs, earthworms and amoebas as sentinels/experimental models this field is of increasing activity. Most efforts are being addressed to harmonize scientifically based data with regulatory policies (Law for Soil protection) and to understand the consequences of chronic pollution for soil ecosystem health and its assessment. Ecosystem health and climate crisis. Due to the increasing concern on the risks posed by climate crisis, we are currently involved in the investigation of the interactions between ecosystem health, chemical pollution and (global and local) threats resulting from anthropic changes in climatic conditions (K-EGOKITZEN, 2007-09). List of 5 most relevant publications of the CBET Group: CAJARAVILLE, MP; BEBIANNO, M; BLASCO, J; PORTE, C; SARASQUETE C; VIARENGO, A. The use of biomarkers to assess the impact of pollution in coastal environments of the Iberian peninsula. A practical approach. Science of the Total Environment 247: 295-311 (2000). HYLLAND K, TOLLEFSEN KE, RUUS A, JONSSON G, SUNDT RC, SANNI S, UTVIK TIR, JOHNSEN S, NILSSEN I, PINTURIER L, BALK L, BARSIENE J, MARIGOMEZ I, FEIST SW, BORSETH JF. Water column monitoring near oil installations in the North Sea 2001-2004. Marine Pollution Bulletin, 56: 414-429 (2008) MARIGÓMEZ, I; SOTO, M; ORBEA, A; CANCIO, I; CAJARAVILLE, MP. Chapter 14: Biomonitoring of environmental pollution in the Basque coast using molecular, cellular and tissue-level biomarkers: an integrative approach. In: Oceanography and marine environment of the Basque Country. A BORJA & M COLLINS (eds). Elsevier Oceanography Series nº 70, Elsevier, Amsterdam, pp 335-364 (2004). MARIGÓMEZ, I; SOTO, M; CANCIO, I; ORBEA, A; GARMENDIA, L; CAJARAVILLE, MP. Cell and tissue biomarkers in mussel, and histopathology in hake and anchovy from Bay of Biscay after the Prestige oil spill (Monitoring campaign 2003). Marine Pollution Bulletin, 53: 287-304 (2006). ZALDIBAR B, CANCIO I, SOTO M, MARIGÓMEZ I. Changes in cell-type composition in digestive gland of slugs and its influence in biomarkers following transplantation between a relatively unpolluted and a chronically metal-polluted site. Environmental Pollution, in press (2008) ZORITA, I; APRAIZ, I; ORTIZ-ZARRAGOITIA, M; ORBEA, A; CANCIO, I; SOTO, M; MARIGÓMEZ, I; CAJARAVILLE, MP. Assessment of biological effects of environmental pollution along the NW Mediterranean Sea using mussels as sentinel organisms. Environmental Pollution, 148: 236-250 (2007). Assessment of biological effects of environmental pollution along the North Iberian coast Biomonitoring activities carried out in the North Sea and the Mediterranean Sea Impact of the Prestige oil spill Caging experiment in the Basque coast Locations of the North Iberian coast sampled after the Prestige oil spill classified by the expert system (Dagnino et al., 2007), an index that integrates the response of different biomarkers to assess ecosystem health. Decision makers can easily identify the degree of affection represented in the colour grading bar. Evaluation of biological effects of environmental pollution in the estuaries of Bizkaia ABRA ESTUARY (1991-1993) PETRONOR (2000-ongoing) Mytilus galloprovincialis Chelon labrosus Crassostrea gigas Carcinus maenas Urdaibai BERRILUR (2003-ongoing) PACARI (2006-2009) Eisenia fetida Delika Fe mine Derio Mungia Field laboratories used to investigate the impact of pollutants in soil ecosystem health. Abandoned mining areas (Fe and Zn mines) and the sorroundings of a highway are the model experimental fields. Reference site is Delika. Micrographs of the digestive gland of slugs after H/E staining. (E) Slug from a clean area. (F) Slug from a Zn mine. 3 6 April 2003 April 2006 9 SUANCES 10 PEDREÑA 11 LAREDO 12 MUSKIZ 13 ARRIGUNAGA 14 GORLIZ 15 MUNDAKA 16 ORIO 17 HONDARRIBIA 1 OIA 2 AGUIÑO 3 CALDEBARCOS 4 CAMELLE 5 SEGAÑO 6 ESTACA 7 LLANES 8 SAN VICENTE Healthy Low stress Medium stress High stress Pathological 3 3 3 3 3 3 3 3 3 3 3 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 3 3 3 3 3 3 6 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 Merlucius merlucius Engraulius encrasilocus Mytilus galloprovincialis Dyctiostelum discoideum 50 100 150 200 μmol/μg soil 20 40 60 80 100 120 % Control Delika Fe mine Run Off Zn mine Zn mine A B (A) Total metal load in studied areas. (B) Cell viability of E. fetida coelomocytes after 3 days exposure to reference and contaminated soils. Zn mine www.ehu.es/GrupoBCTA Ionan Marigómez (PR), Miren P. Cajaraville, Manu Soto, Eduardo Angulo, Mari Carmen Barbero, Teresa Serrano, Ibon Cancio, Amaia Orbea, Maren Ortiz-Zarragoitia, Beñat Zaldibar, Izaskun Zorita, Eider Bilbao, Urtzi Izagirre, Damien Raingeard, Larraitz Garmendia, Vega Asensio, Amaia Rodríguez, AneMiren Castilla, Pamela Ruiz, Eunate Puy, Aitzpea Errasti, Oihane Díaz de Cerio, Alberto Katsumiti, Unai Vicario, María Múgica. CBET Research Group (September 2006) SOIL MONITORING B C D Oxidative stress A A B C D E F Arion ater Abra
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