POLLUTION BY XENOBIOTICS : BIOMARKERS FOR EARLY DETECTION OF POLLUTION EFFECTS Ibon Cancio EUSKALHERRIKOUNIBERTSITATEA UNIVERSITY OF THE BASQUE COUNTRY emantazabalzazu DEPARTMENT OF ZOOLOGY AND ANIMAL CELL DYNAMICS

POLLUTION Nutrients Light Noise Smell Toxic compounds XENOBIOTICS Alien species

CHEMICAL ANALYSIS OF THE WATER - Knowledge of the chemical products in the environment - Drawbacks: * Requires selection of substances to be tested, it is difficult and expensive * It has little biological meaning: toxic? Synergistic, antagonistic effects? Legal action against polluting factory

BIOMONITORING - Chemical analysis on biota * Mussel watch, slug watch... * Bioaccumulation: fraction of pollutants that is accumulated in biological tissues - Drawbacks: * Still need to select substances * Pollutants may not accumulate (be metabolised) * Biological meaning? (does accumulation relate to a possible impairment of animal performance?) Slug Watch

BIOMARKERS: LOOKING FOR BIOLOGICAL ANSWERS - THE MAIN AIM OF ENVIRONMENTAL TOXICOLOGY IS TO KNOW THE EFFECT OF POLLUTANTS ON POPULATIONS AND ECOSYSTEMS: Can we measure biologically exposure to chemicals and the effects caused by them?

* Noise * Ecological relevance Population Organism Physisological Cellular Molecular Community Ecosystem * Specificity * Speed Biomarkers at different levels of organization

BIOACCUMULATION: low water pollutant concentrations may be concentrated in biological structures Cd Cl Biota Cells Cellular structures

MOLECULAR AND CELLULAR BIOMARKERS - Quick responses that precede answers at higher levels of biological organization - Quantitative and qualitative relationships between exposure and biomarker response * DEFINITION: alterations at the molecular or cellular level that indicate exposure to contaminants and/or injuries induced by them (McCarthy & Shugart, 1990)

BIOMARKERS ARE APPLIED ON SENTINEL SPECIES

CELLS AND TISSUES UNDER STUDY Those showing high exposure to pollutants, accumulation rates and responsiveness to pollutants Liver Gills Dig. gland Gonads

SAMPLING STRATEGIES Shipyard River coming from Gernika city * * * * * Sampling points

STRATEGIES: ANIMAL TRANSPLANT

ANALYSIS OF PROTEINS THAT SCAVENGE SPECIFIC POLLUTANTS OR THAT CHANGE THEIR ACTIVITY UNDER EXPOSURE * Organophosphorous pesticides inhibit brain Acetylcholinesterase * Organic pollutants induce enzymes of the detoxification system (CYP450) * Metals induce Metallothionein synthesis in cells to sequester them

ST MT C 1h 1d 7d det Cod Cod Turbot 14.4 kDa MT-level (a.u.) h1 d7 dDetox. Control 0.01 ppm Cd 0.1 ppm Cd QUANTIFICATION OF METALOTHIONEIN LEVELS

* ORGANOTINS inhibit aromatase activity IMPOSEX, INTERSEX Aromatase Testosterone Estradiol Antifouling paints TBT

IMPOSEX IN THE DOGWELK Progression of vas deferens and penis growth in a female dogwelk

POLLUTANTS ACCUMULATE IN LYSOSOMES * Induce changes in the structure of lysosomes

LSC - FIELD STUDY Aegean Sea / Biscay Gulf LIMANI MUSKIZ OLYMPIADAPLENTZIA Polluted site Reference site

PEROXISOME PROLIFERATION Normal rat liver Treated with PP

PEROXISOME PROLIFERATORS Hypolipidemic drugs and other therapeutic agents Herbicides Plasticizers Steroids Solvents and lubricants Food flavors Natural products Crude oils and petroleum products

ENVIRONMENTAL ESTROGENS Chemical substances that mimic female sexual hormones * Alter sex ratios of wild populations * Impair reproductive performance Normal testis Intersex testis

VITELLOGENIN IN FISH a biomarker of endocrine disruption Male fish exposed to environmental estrogens express Vtg Liver Female gonad E2 Vtg 2 1