EULIMNOS: Successful International Cooperation in WBCSome History 2001 11stst Conference,...
Transcript of EULIMNOS: Successful International Cooperation in WBCSome History 2001 11stst Conference,...
EULIMNOS: EULIMNOS: Successful International Successful International
Cooperation in WBCCooperation in WBC
LotharLothar ErdingerErdingerUniversity of HeidelbergUniversity of Heidelberg
Prof. Lefter Kashta, Shkodra
Common topic of Interest
The LakeThe Lake
ConsortiumConsortium
University ofGraz
University of HeidelbergGermany
University of ShkodraAlbania
University of Montenegro
CG
HRK/Stability Pact
CETIPodgorica
CG
University ofLjubljanaSlovenia
Örebro University Sweden
University ofSalzburgAustria
University ofPristina
CG
University ofSouthhampton
UK
Some HistorySome History2001 2001 11stst Conference, ShkodraConference, Shkodra
Initiation of Scientific CooperationInitiation of Scientific CooperationJan. 2002Jan. 2002 22ndnd Conference, HeidelbergConference, HeidelbergMar. 2002Mar. 2002 Workshop, PodgoricaWorkshop, PodgoricaSept. 2002Sept. 2002 33rdrd Conference, PogradecConference, PogradecSept. 2003Sept. 2003 Participation in SETAC Conference, HeidelbergParticipation in SETAC Conference, HeidelbergFeb. 2004Feb. 2004 44thth Conference, ShkodraConference, Shkodra20042004--20082008 Common Participations in SETAC ConferencesCommon Participations in SETAC Conferences20072007 Successfull application within Successfull application within 20092009 Opening of Ecological Educational Center and Opening of Ecological Educational Center and
Biological Station, VirpazarBiological Station, VirpazarOngoingOngoing Exchange of Scientists and Students for shortExchange of Scientists and Students for short--
term integrated projectsterm integrated projects
Our WebsiteOur Website
Basic AimsBasic Aims
Basis forWater Management
Protection ofthe Lake
Implementation ofLaboratories
Common Scientific Projects
ImprovementOf Skills
CooperationCooperation
Tools: Integrated AssessmentTools: Integrated Assessment
Assessment ofFauna
Assessment ofFlora
Biotests of Water and Sediments
Analysis ofBioavailableCompounds
MicrobiologicalData for
Water Quality
ChemicalData for
Water Quality
Physico-chemical Parameters
PROJECTSPROJECTSUmbrella ProjectUmbrella Project
Monitoring of several sampling sites on both sides of the lake Monitoring of several sampling sites on both sides of the lake for basic chenical and microbiological Parameters.for basic chenical and microbiological Parameters.
Biological monitoring of Fauna and FloraBiological monitoring of Fauna and FloraTaxonomic studies on biodiversity of algae, waterplants, fish, Taxonomic studies on biodiversity of algae, waterplants, fish, and other species.and other species.
Triade ProjectTriade ProjectIntegrative assessment of water and sediment quality at Integrative assessment of water and sediment quality at selected sites on the lake.selected sites on the lake.
SPMD ProjectSPMD ProjectDevelopment and application of innovative sampling Development and application of innovative sampling methods for bioavailable compounds.methods for bioavailable compounds.
Lake monitoringLake monitoring
Industry
Houshold
Aerial
Oxygen Watercolour
pH
Turbidity
Alkalinity
Algae growth
Microbial growth
Nutrients
Temperature, Irradiation
Umbrella ProjectUmbrella Project
BenefitBenefitDescription of actual lake situation by scientific data.Description of actual lake situation by scientific data.Starting point and basis for furthergoing investigations.Starting point and basis for furthergoing investigations.
DevelopmentDevelopmentPossibly: Installation of permanent monitoring.Possibly: Installation of permanent monitoring.
Outcome so farOutcome so farBasic information on some routine parameters Basic information on some routine parameters –– basic basic evaluation of ecological conditions.evaluation of ecological conditions.
Future activitiesFuture activitiesIntegration into a wider monitoring concept which must be Integration into a wider monitoring concept which must be supported by national authorities.supported by national authorities.
PROJECTSPROJECTSUmbrella ProjectUmbrella Project
Monitoring of several sampling sites on both sides of the lake Monitoring of several sampling sites on both sides of the lake for basic chenical and microbiological Parameters.for basic chenical and microbiological Parameters.
Biological monitoring of Fauna and FloraBiological monitoring of Fauna and FloraTaxonomic studies on biodiversity of algae, waterplants, fish, Taxonomic studies on biodiversity of algae, waterplants, fish, and other species.and other species.
Triade ProjectTriade ProjectIntegrative assessment of water and sediment quality at Integrative assessment of water and sediment quality at selected sites on the lake.selected sites on the lake.
SPMD ProjectSPMD ProjectDevelopment and application of innovative sampling Development and application of innovative sampling methods for bioavailable compounds.methods for bioavailable compounds.
Biological monitoring of Flora and FaunaBiological monitoring of Flora and Fauna
BenefitBenefitScientific interest in both universities, long tradition, strongScientific interest in both universities, long tradition, strongbasis.basis.
DevelopmentDevelopmentStrengthening of cooperation between scientists on both Strengthening of cooperation between scientists on both sides of the lake.sides of the lake.
Outcome so farOutcome so farOngoing broadening of scientifically interesting specific Ongoing broadening of scientifically interesting specific knowledge.knowledge.
Future activitiesFuture activitiesFurther integration in ecotoxicological research activities.Further integration in ecotoxicological research activities.
PROJECTSPROJECTSUmbrella ProjectUmbrella Project
Monitoring of several sampling sites on both sides of the lake Monitoring of several sampling sites on both sides of the lake for basic chenical and microbiological Parameters.for basic chenical and microbiological Parameters.
Biological monitoring of Fauna and FloraBiological monitoring of Fauna and FloraTaxonomic studies on biodiversity of algae, waterplants, fish, Taxonomic studies on biodiversity of algae, waterplants, fish, and other species.and other species.
Triade ProjectTriade ProjectIntegrative assessment of water and sediment quality at Integrative assessment of water and sediment quality at selected sites on the lake.selected sites on the lake.
SPMD ProjectSPMD ProjectDevelopment and application of innovative sampling Development and application of innovative sampling methods for bioavailable compounds.methods for bioavailable compounds.
Triade ProjectTriade Project
BenefitBenefitApplication of modern ecotoxicological tools on a new Application of modern ecotoxicological tools on a new environment.environment.
DevelopmentDevelopmentInstallation of basic tools for ecotoxicological assessment in Installation of basic tools for ecotoxicological assessment in both university laboratories.both university laboratories.
Outcome so farOutcome so farProof of toxic compounds in sediment extracts by biotest Proof of toxic compounds in sediment extracts by biotest methods.methods.
Future activitiesFuture activitiesDevelopment of further skills, development of individual Development of further skills, development of individual research activities.research activities.
SEESEE--ERA NETERA NET
PROJECTSPROJECTSUmbrella ProjectUmbrella Project
Monitoring of several sampling sites on both sides of the lake Monitoring of several sampling sites on both sides of the lake for basic chenical and microbiological Parameters.for basic chenical and microbiological Parameters.
Biological monitoring of Fauna and FloraBiological monitoring of Fauna and FloraTaxonomic studies on biodiversity of algae, waterplants, fish, Taxonomic studies on biodiversity of algae, waterplants, fish, and other species.and other species.
Triade ProjectTriade ProjectIntegrative assessment of water and sediment quality at Integrative assessment of water and sediment quality at selected sites on the lake.selected sites on the lake.
SPMD ProjectSPMD ProjectDevelopment and application of innovative sampling Development and application of innovative sampling methods for bioavailable compounds.methods for bioavailable compounds.
SPMD ProjectSPMD Project
BenefitBenefitDevelopment and application of modern, scientifically Development and application of modern, scientifically challenging sampling technique .challenging sampling technique .
DevelopmentDevelopmentInstallation of basic tools for SPMD application in both Installation of basic tools for SPMD application in both university laboratories.university laboratories.
Outcome so farOutcome so farProof of toxic compounds in water phase, relevance not Proof of toxic compounds in water phase, relevance not known so far.known so far.
Future activitiesFuture activitiesDevelopment of further skills, development of individual Development of further skills, development of individual research activities.research activities.
EstablishmentEstablishment
•• Of a sustainable research and education Of a sustainable research and education facility localted directly on the lakeside...facility localted directly on the lakeside...
Ecological Station VirpazarEcological Station Virpazarfounded by A. and S. Perovicfounded by A. and S. Perovic
SummarySummary
With the help of HRK a cooperation network between With the help of HRK a cooperation network between Universities from Shkodra, Podgorica, Heidelberg, and Universities from Shkodra, Podgorica, Heidelberg, and Graz has been established.Graz has been established.A routine monitoring program has been initiated.A routine monitoring program has been initiated.New and modern tools of ecotoxicological risk New and modern tools of ecotoxicological risk assessment have been implemented and used to assess assessment have been implemented and used to assess Lake Shkodra.Lake Shkodra.Biotests show some interesting results mainly for Biotests show some interesting results mainly for sediments.sediments.Reasons for some highly toxic effects are yet unclear.Reasons for some highly toxic effects are yet unclear.
AcknowledgementAcknowledgement
We gratefully appriciate the help of We gratefully appriciate the help of HRK and Stability Pact for HRK and Stability Pact for
$upport$upport
Some DetailsSome Details
M10
M7
M1
M2M3
M4
M5
M6M8M9
A1A2
A3
A5
A4
Routine Routine Monitoring Monitoring --MicrobiologyMicrobiology
Sampling Sites - Albania
A1 A2 A3 A4 A5
Col
ony
Cou
nt
0
200
400
600
800
1000
1200
MPNTotal coliformsPseudomonas
Sampling Sites - Montenegro
M1 M2 M3 M4 M5 M6 M7 M8 M9 M10
Col
ony
Cou
nt
0
1000
2000
3000
4000
MPNTotal Coliforms
Some Chemical Monitoring Some Chemical Monitoring Data for M4Data for M4
(Radus)(Radus)
Month
Tota
l Col
iform
Cou
nt
0
500
1000
1500
2000
2500
3000
3500
4000
Oxy
gen
[mg/
l]
5
6
7
8
9
10
11
pH
7,6
7,7
7,8
7,9
8,0
8,1
8,2
Alk
alin
ity [m
mol
/l]
2,4
2,6
2,8
3,0
3,2
3,4
3,6
3,8
4,0
4,2
4,4
M10 M10 -- KarucKaruc
Month
May June July August September October
TC
0
1000
2000
3000
4000
NTU
0,6
0,8
1,0
1,2
1,4
1,6
1,8
2,0 Month
May June July August September October
TC
0
1000
2000
3000
4000
mg/
l
0,3
0,4
0,5
0,6
0,7
0,8
0,9
1,0
Month
May June July August September October
TC
0
1000
2000
3000
4000
pH
7,5
7,6
7,7
7,8
7,9
8,0
8,1
Alkalinity
Turbidity
COD
pH
EcotoxicologyEcotoxicology
The SedimentThe Sediment
Fate of Toxic CompoundsFate of Toxic Compounds
Toxic compounds
Outtake
Mineralisation
Metabolisation
Adsorption
Bioaccumulation
Significance of SedimentsSignificance of Sediments
Toxic Compounds
Sediments = Memory of the Lake
Food Chain
Sediment Triad
Bioassays
Fieldstudies
Chemicalanalysis
Bioassay-directedfractionation (BdF)
Extract
Bioassays
Fractionation
Active fractions
Hexa
n
Hexa
n - D
CM 2
:1
EtO
H
Ace
ton
DCM
Hexa
n - D
CM 1
:2
Hexa
n - D
CM 1
:1
DCM
- A
ceto
n 1:
1
0
200
400
600
800
1000
NR50
in m
g /m
L
Ch. analysis
IdentificationAbundance
0
10000
20000
30000
40000
0 20 40 60
time
Bioassays
Fieldstudies
Chemicalanalyses
Integrative
assessment
What are Bioassays good for?What are Bioassays good for?
Standardized Bioassays show the presence Standardized Bioassays show the presence of compounds with a certain toxic effect.of compounds with a certain toxic effect.Bioassays are sum parameters for the Bioassays are sum parameters for the measurement of toxic chemicals.measurement of toxic chemicals.Results of Bioassays are the starting point Results of Bioassays are the starting point for the identification of toxic compounds.for the identification of toxic compounds.
Biotesting of Sediment SamplesBiotesting of Sediment SamplesAmes Ames –– Test (several tester strains)Test (several tester strains)Comet Comet –– Assay (several eucaryontic cell lines)Assay (several eucaryontic cell lines)CytotoxicityCytotoxicity
Microtox assay using bacteriaMicrotox assay using bacteriaNeutral Red Assay using eucaryontic cellsNeutral Red Assay using eucaryontic cells
DHA Contact AssayDHA Contact AssayDehydrogenase ActivityDehydrogenase Activity
EROD AssayEROD AssayEthoxyresorufinEthoxyresorufin--OO--deethylasedeethylase(Indicator for Dioxin(Indicator for Dioxin--like compounds, AHH inducing like compounds, AHH inducing potency)potency)
YES AssayYES AssayHormonHormon--like activity using yeast cells with human receptorslike activity using yeast cells with human receptors
More Sampling......More Sampling......
Sites for
Sediment Sampling
Sediments – Memory of a lake
EROD Induction in Liver Cell Culture as a Biomarker of Dioxin-like Activity
Liver Cell Culture
Addition of medium and sample
Exposure: 48 hHomogenisation
EROD and protein
EC25TCDD determination
Reader 10-1 100 101 102 1030
10
20
30
40
50
60
70
Concentration extracted sample(mg/ml medium)
% o
f max
imum
ER
OD
indu
ctio
n
DioxinDioxin--like Organic Compounds like Organic Compounds (Dioxins, (Dioxins, DibenzofuransDibenzofurans, PCBs), PCBs)
Cl
Cl
Cl
Cl
O
Cl
Cl
Cl
Cl
O
O
Cl
ClCl
Cl
Polychlorinated Biphenyls TCDD
DibenzofuranPolycyclic aromatic hydrocarbon (PAH)
EROD Activity of Sediment ExtractEROD Activity of Sediment Extract(Radus (Radus -- Montenegro)Montenegro)
0.1 1 10 100
1
2
3
4
5
6
7
8
EC25: 2 mg/ml
T1 48h
2,3,7,8 TCDD
control
mg/ml
ERO
D a
ctiv
ity
Sediment contact assay
Arthrobacterglobiformis
Aquatox HD
Resazurin595 nm
Resorufin620 nm
Dehydrogenases
Controlls 4-NP Extract
w/o w w/o w w/o w
DHA contact testC
0,1250,2
50,5
12PC
T1T2T3
0
25
50
75
100
Inhi
bitio
n [%
]
Sediment [g]
Inhibition of microbial dehydrogenasesynthesis vs. concentration of sediment solid phase (Arthrobacterglobiformis)
YES YES ––Assay: SedimentsAssay: Sediments
T3
0
0,5
1
1,5
2
2,5
3
0,001 0,01 0,1 1 10
Extract Equivalent (g)
adju
sted
abs
orba
nce
at 5
40 n
m
T3upper 99% confidence intervallow er 99% confidence intervalE2 response
0
0,5
1
1,5
2
2,5
3
0,001 0,01 0,1 1 10
Extract Equivalent (g)
adju
sted
abs
orba
nce
at 5
40 n
m
T1upper 99% confidence intervallower 99% confidence intervalE2 response
0
0,5
1
1,5
2
2,5
3
0,001 0,01 0,1 1 10
Extract Equivalent (g)
adju
sted
abs
orba
nce
at 5
40 n
m
T2upper 99% confidence intervallower 99% confidence intervalE2 response
SPMDSPMD--ApproachApproach
Sampling of Bioavailable Organic Sampling of Bioavailable Organic CompoundsCompoundsAnalysis of Individual componentsAnalysis of Individual componentsBiotestingBiotesting
EROD EROD -- AssayAssayYES YES -- AssayAssay
Sampling Sites for SPMDSampling Sites for SPMD
AdriaticSea
Albania
MontenegroPodgorica
LakeSkadar/Shkodra
Shkodra
Bar
Virpazar
Golubovci
Koplik
R. Moraca
R.Buna
4
N
10 Km
ShirokaZogaj
Plavnica
Dajlon
5
6
12
3
0
2000
4000
6000
8000
10000
12000
Site 1 Site 2 Site 3 Site 4 Site 5 Site 6
Sum
PP
-PA
Hs
(ng/
SP
MD
)
Results of EROD Assay from SPMD Results of EROD Assay from SPMD DialysatesDialysates
0.01 0.1 1 10 1000
2
4
6
8
10
12
14
EC25: 36 %
Site 1
TCDD
DMSO control
SPMD dialysate concentration
ERO
D a
ctiv
ity
0.01 0.1 1 10 1000
2
4
6
8
10
12
14
EC25: 100 %
Site 2
TCDD
DMSO control
SPMD dialysate concentration
ERO
D a
ctiv
ity
0.01 0.1 1 10 1000
2
4
6
8
10
12
14
EC25: 22 %
Site 3
TCDD
DMSO control
SPMD dialysate concentration
ERO
D a
ctiv
ity
0.01 0.1 1 10 1000
2
4
6
8
10
12
14
EC25: n. d.
Site 4
TCDD
DMSO control
SPMD dialysate concentration
ERO
D a
ctiv
ity
0.01 0.1 1 10 1000
2
4
6
8
10
12
14
EC25: 42 %
Site 5
TCDD
DMSO control
SPMD dialysate concentration
ERO
D a
ctiv
ity
0.01 0.1 1 10 1000
2
4
6
8
10
12
14
EC25: 32 %
Site 6
TCDD
DMSO control
SPMD dialysate concentrationER
OD
act
ivity
TEQTEQ‘‘s for SPMD Sampless for SPMD Samples
0.0
0.1
1.0
10.0
100.0
1000.0
10000.0
1 2 3 4 5 6Site
TEQ
s (p
g/m
l)
Bio-TEQs PAH-TEQs
R2 = 0.74
0200400600800
10001200140016001800
3000 5000 7000 9000 11000
Sum PP-PAHs (ng/SPMD)TC
DD
-EQ
(pg/
ml)
YES YES -- ActivityActivity
0
0.5
1
1.5
2
2.5
3
0.01 0.1 1 10 100SPMD dialysate concentration
abs
orba
nce
at 5
40 n
m
Site 1E2 maximum response
0
0.5
1
1.5
2
2.5
3
0.01 0.1 1 10 100SPMD dialysate concentration
abs
orba
nce
at 5
40 n
m Site 2E2 maximum response
0
0.5
1
1.5
2
2.5
3
0.01 0.1 1 10 100SPMD dialysate concentration
abs
orba
nce
at 5
40 n
m
Site 3E2 maximum
response
0
0.5
1
1.5
2
2.5
3
0.01 0.1 1 10 100SPMD dialysate concentration
abs
orba
nce
at 5
40 n
m Site 4
E2 maximum response
0
0.5
1
1.5
2
2.5
3
0.01 0.1 1 10 100SPMD dialysate concentration
abs
orba
nce
at 5
40 n
m
Site 5E2 maximum response
0
0.5
1
1.5
2
2.5
3
0.01 0.1 1 10 100
abs
orba
nce
at 5
40 n
m
SPMD dialysate concentration
Site 6E2 maximum response
SummarySummary
With the help of HRK a cooperation network between With the help of HRK a cooperation network between Universities from Shkodra, Podgorica, Heidelberg, and Universities from Shkodra, Podgorica, Heidelberg, and Graz has been established.Graz has been established.A routine monitoring program has been initiated.A routine monitoring program has been initiated.New and modern tools of ecotoxicological risk New and modern tools of ecotoxicological risk assessment have been implemented and used to assess assessment have been implemented and used to assess Lake Shkodra.Lake Shkodra.Biotests show some interesting results mainly for Biotests show some interesting results mainly for sediments.sediments.Reasons for some highly toxic effects are yet unclear.Reasons for some highly toxic effects are yet unclear.
AcknowledgementAcknowledgement
We gratefully appriciate the help of We gratefully appriciate the help of HRK and Stability Pact for HRK and Stability Pact for
$upport$upport
Microbiological Monitoring Microbiological Monitoring ––Shkodra UniversityShkodra University