Environmental Monitoring MethodsEnvironmental 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 monitoringBioaccumulation

& Biological effect monitoring

Biological effect,Health & ecosystem

monitoring

Environmental factorsmodifying exposure

Environmental factorsmodifying exposure

Environmental factorsmodifying susceptibilityEnvironmental factors

modifying susceptibility

Other causesOther causes

Major Processes in Pollutant ImpactMajor Processes in Pollutant Impact

ContaminantMixtures

ParticulatesSpeciation,

Binding & Transport

Toxicity & Cell injuryPhysicochemical Behavior,Activation & Detoxication

Exposure Biomarkers

Pathological EndpointsPrognostic Biomarkers

Ecological ImpactMortality, Reduced Fecundity

Increased Susceptibility to Other Stresses & Loss of Diversity

Human Health ImpactToxicity & Diseases

via food chain

UV & Visible Radiation

UV & Visible Radiation

UptakeImport by adsorptive

Endocytosis & Phagocytosis

UptakeTrans-Membrane

Diffusion

Biological Effects & Ecological Biological Effects & Ecological RelevanceRelevance

Physical factors&

Biological Agents

Pollutant Chemicals&

Radionuclides

Molecular&

CellularTissue Organs Individual animal

SHORT TERM LONG TERMBiological Complexity, Response

Time & Biological Significance

EARLY BIOMARKER SIGNALS LATER EFFECTS

Environmental Pollutants

Lethal effectsLethal effects

Sublethal effectsSublethal effects

Changes in enzyme activitiesChanges in enzyme activitiesDamage to DNA, proteinsDamage to DNA, proteins

& membrane lipids& membrane lipidsEffects on behavioral,Effects on behavioral,

reproductivereproductive& immunological activities& immunological activities

Environmental Pollutants & Effects to Environmental Pollutants & Effects to OrganismsOrganisms

Mussels: Bioindicators for Aquatic Mussels: Bioindicators for Aquatic PollutionPollution

Freshwater Ecosystems

Marine Ecosystems

Sentinel SpeciesBioindicators

Biomonitoring

Wide geographical distributionEasy to collect

Filter-feeding organismsBioconcentrate xenobiotics

Easily transferred from control to contaminated areas

Chemical Pollutants & Oxidative StressChemical Pollutants & Oxidative Stress

ROSOrganic & Inorganic

PollutantsOrganic & Inorganic

Pollutants

AquaticOrganismsAquatic

Organisms

Polyclic Aromatic HydrocarbonsOrganochlorine

& Organophoshate pesticidesPolychlorinated biphenyls

Aromatic diolsQuinones

NitroaromaticsAromatic hydroxylamines

DioxinsBipyridyls

Transition metal chelates

Redox

Potential

Molecular BiomarkersOf

Oxidative Stress

Molecular BiomarkersOf

Oxidative Stress

Lipid PeroxidationEnzyme Activities

DNA DamageProtein Degradation

* ROS: Reactive Oxygen Species

ROS & biochemical & physiological responses ROS & biochemical & physiological responses in cells of aquatic organismsin cells of aquatic organisms

LipidLipidPeroxidationPeroxidation

LipidLipidPeroxidationPeroxidation

Anthropogenic activities

Environmental

factors

MT

CAT

PUFAsPUFAs

SOD

Organicpollutants

Solar radiatio

n

Temperature

Salinity

Μn

+Metal Metal transporttransport

Metalcomplexation

Ions Ions diffusiondiffusion

Ο2─

Metal Metal competitiocompetitio

nn

Metals

Μn

+Oxidative DNA Oxidative DNA

damagedamage

Oxidative DNA Oxidative DNA damagedamage

Protein Protein damagedamage

Protein Protein damagedamage

Changes in Changes in Enzyme activitiesEnzyme activities

Changes in Changes in Enzyme activitiesEnzyme activities

ΗΟ

Η2Ο

2 Ο2─

Proteins

Η2Ο

2

Superoxide dismutase (SOD)Superoxide dismutase (SOD)

2O2 + 2H H2O2 + O2SOD

SODSOD

Catalase (CAT)Catalase (CAT)

2H2O2 2H2O + O2CAT

CATCATCATCAT

Lipid Peroxidation (LPO)Lipid Peroxidation (LPO)LH

L

R

RH

LOO

NRP

LOO

LH

LOOH

O2

InitiationInitiation

PropagationPropagation

TerminationTermination

InhibitionInhibition

AntioxidantsAntioxidants

OxidizedOxidizedAntioxidantsAntioxidants

DegradationDegradation of Lipid Peroxides of Lipid Peroxides

AldehydesAldehydesand and otherother products products

Biomarkers ofBiomarkers ofOxidative stressOxidative stress

LH: lipid substrateL•: lipid radicalLOOH: lipid hydroperoxideLOO•: lipid peroxyl radicalNRP: nonradical product

Oxidative Stress in Mussels from the Saronikos gulf Oxidative Stress in Mussels from the Saronikos gulf (Greece)(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) activityCatalase (CAT) activity

Lipid Peroxidation (LPO) levels

Measurements of PAHs and Metal concentrations in mussel tissuesrevealed elevated levels of chemical pollution at Sites 1 & 2

Measurements of PAHs and Metal concentrations in mussel tissuesrevealed elevated levels of chemical pollution at Sites 1 & 2

Representative chart for PAHs Representative chart for PAHs concentrations in mussel tissuesconcentrations in mussel tissues

0 100 200 300 400 500 600

PAHs concentration (ng/g dt)

Napth

Acy

Flu

Phe

Fla

Chry

Bkf

Bap

Ind

Ace

Ant

Pyr

Baa

Bbf

Per

Dba

Bzp Site 1 Site 2 Site 3 Site 4

0 100 200 300 400 500

PAHs concentration (ng/g dt)

Napth

Acy

Flu

Phe

Fla

Chry

Bkf

Bap

Ind

Ace

Ant

Pyr

Baa

Bbf

Per

Dba

Bzp Site 1 Site 2 Site 3 Site 4

Results for SOD activityResults for SOD activity

0

500

1000

1500

2000

2500

3000

3500

Summer Autum Winter Spring

SO

D A

ctiv

ity

(Unit

s/g

wt)

Site 1 Site 2 Site 3 Site 4

** **

*

**

***

**

0

500

1000

1500

2000

2500

Summer Autum Winter Spring

SO

D a

ctiv

ity

(Unit

s/g

wt)

Site 1 Site 2 Site 3 Site 4

* * *

*** ***

Results for CAT activityResults for CAT activity

0

0.5

1

1.5

2

2.5

3

3.5

Summer Autum Winter Spring

CA

T a

ctiv

ity

(A1/A

2 /m

in/m

g pro

t)

Site 1 Site 2 Site 3 Site 4

**

*

*

0

0.5

1

1.5

2

2.5

3

3.5

4

4.5

Summer Autum Winter Spring

CA

T a

ctiv

ity

(A1/A

2 /m

in/m

g pro

t)

Site 1 Site 2 Site 3 Site 4

**

*

Results for Lipid Peroxidation levelsResults for Lipid Peroxidation levels

0

200

400

600

800

1000

1200

1400

1600

1800

2000

Summer Autum Winter Spring

LPO

lev

els

ΜD

A (nm

ol/g

wt)

Site 1 Site 2 Site 3 Site 4***

** ** **

0

200

400

600

800

1000

1200

1400

1600

1800

2000

Summer Autum Winter Spring

LP

O le

vels

ΜD

A (

nm

ol/g

wt)

Site 1 Site 2 Site 3 Site 4

**

***

**

***

**

**

*

Conclusions Conclusions

Pollution in coastal waters of Saronikos gulfPollution in coastal waters of Saronikos gulf causes moderate levels of oxidative stresscauses moderate levels of oxidative stress

in tissues of musselsin tissues of mussels

The biomarkers of SOD, CAT & LPO are suitableThe 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 systemand can be used as cost effective test systemfor Environmental Risk Assessmentfor Environmental Risk Assessment

Factors that affect biomarker responsesFactors that affect biomarker responsesof oxidative stress in aquatic organismsof oxidative stress in aquatic organisms

BiomarkerResponsesBiomarkerResponses

Health conditionSexAge

Nutritional statusMetabolic activitiesMigratory behavior

Reproductive& Developmental status

Population density

SeasonAmbient temperatureHeterogeneity of the

Environmental Pollution