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