Welcome to the CLU-IN Internet Seminar Biological-based Assays - Indicators of Ecological Stress Sponsored by: National Institute of Environmental Health Sciences, Superfund Research Program Delivered: September 23, 2010, 2:00 PM - 4:00 PM, EDT (18:00-20:00 GMT) Instructors: Bruce Duncan, Senior Ecologist with EPA Region 10's Office of Environmental Assessment ([email protected]) Jim Shine, Associate Professor of Aquatic Chemistry, SRP Grantee ([email protected]) Moderator: Beth Anderson, Program Analyst, Superfund Research Program ([email protected]) Visit the Clean Up Information Network online at www.cluin.org 1
Moderator:Beth Anderson, Program Analyst, Superfund Research Program ([email protected])
Visit the Clean Up Information Network online at www.cluin.org1
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Bioavailability of Sediment Contaminants
1. NIEHS-sponsored Bioassay Network
2. Relationships between sediment, water, mussels, SPMEs, & fish
Bruce Duncan, EPA Region 10, Seattle, Office of Environmental AsBruce Duncan, EPA Region 10, Seattle, Office of Environmental AssessmentsessmentRisk Evaluation Unit Risk Evaluation Unit Session IV: BiologicalSession IV: Biological--based Assays based Assays –– Indicators of Ecological StressIndicators of Ecological StressEcological Risk: New Tools and Approaches Ecological Risk: New Tools and Approaches –– September 23, 2010September 23, 2010
Sept 2009 – Lisa Jackson
Water, OSWER and EJHelping communities – particularly underserved communities –reconnect with and revitalize their local urban waters
Cleaning Up Our CommunitiesProtecting America’s WatersExpanding the Conversation on Environmentalism and Working for Environmental Justice
EPA Collaboration: Water, Land-Based (OSWER) and EJ programs
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KC Donnelly
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National Network to Investigate the Utility of Short-term Bioassays for
Evaluating Sediment Quality
Investigate the utility of using SRP-developed assays to characterize the toxicity of complex mixtures in sediment
Hypothesis: SRP-developed assays will detect degraded sediment quality effectively and serve as an additional line of evidence if integrated into risk assessment
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Background
Superfund Research Program
◦ Created in 1986 under the Superfund Amendments and Reauthorization Act (SARA)◦ University-based grants program◦ Basic research◦ Complement EPA and ATSDR◦ Under National Institute of Environmental Health
Sciences
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Background
Collaboration between 5 University based Superfund Research Programs◦ Texas A&M University◦ Duke University◦ Michigan State University◦ University of California – Davis◦ University of California – San Diego
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Summary Table – “calibration” testing single contaminants and mixtures
BIOASSAY
Chemical
In vivo EROD (EC50)
Fish embryo teratogenicity
(EC10)GJIC
(EC50)CALUX(EC50)
BaP 1 ppb 200 ppb NA405 ppm(EC40)
Flu NA NA 4.4 ppm NA
BaP+Flu 1 ppb 100 ppb 4.8 ppm422 ppm(EC40)
Coal-tar .06 ppm 5 ppb 2.87 ppm 341 ppb
PCB 126 .03 ppb 0.1 ppb NA 49 ppt
PCB 153 NA NA 4.34 ppm NA
PCB mix -- -- -- --
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ConclusionsCalibration step was completed◦ Assays were not always more sensitive but can serve as an
additional line of evidence◦ Improved specificity
2nd Phase of project anticipated◦ Aliquots of homogenized sediment will be sent to Superfund
Research Program investigators for analysis
Study will attempt to “crosswalk” with biological effects data from sediment toxicity bioassays
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Bioavailability of Sediment Contaminants
Relationships between sediment, water, mussels, SPMEs, & fish
Bruce Duncan, EPA Region 10, Office of Environmental Assessment,Bruce Duncan, EPA Region 10, Office of Environmental Assessment, Risk Evaluation Unit Risk Evaluation Unit & Adjunct Professor, Texas A&M University, Health Science Center& Adjunct Professor, Texas A&M University, Health Science Center, School of Rural Public , School of Rural Public Health, Dept Environmental and Occupational Health Health, Dept Environmental and Occupational Health In collaboration with Matt Kelley, Postdoctoral Fellow In collaboration with Matt Kelley, Postdoctoral Fellow -- DugasDugas Lab, LSU Health Sciences Lab, LSU Health Sciences CenterCenter--ShreveportShreveport
Sept 2009 – Lisa Jackson
Water, OSWER and EJHelping communities – particularly underserved communities –reconnect with and revitalize their local urban waters
Cleaning Up Our CommunitiesProtecting America’s WatersExpanding the Conversation on Environmentalism and Working for Environmental Justice
EPA Collaboration: Water, Land-Based (OSWER) and EJ programs
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Partners
◦ EPA R10 – deployment, retrieval, designDive Team, Manchester Lab, Field support, Program volunteers
◦ Texas A&M University – design, tissue, water, sediment analysis
KC Donnelly (dec); Matt Kelley; Thomas McDonald◦ Southern California Coastal Water Research Project –
SPME design, analysisKeith Maruya, David Tsukada, Wayne Lao
- Accounts for metal:metal competitive interactions 56
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Gellyfish: Laboratory Development and TestingStep 4: Challenge Gellyfish/Model
A) Effect of varying salinity on Cu Uptake
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1e-10 1e-09 1e-08 1e-07 1e-06
[Zn2+] (Mol/L)
0.1
1
10
100C
uID
(µm
ol/L
)
Modeled
Measured
Step 4: Challenge Gellyfish/Model, cont’d
Gellyfish: Laboratory Development and Testing
B) Effect of high Zn2+ on Cu uptake by Gellyfish
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0.1 1 10 100 1000
Modeled Concentration (µmol/L)
0.1
1
10
100
1000
Mea
sure
d C
onc.
(µm
ol/L
)
Pb
Cu
Zn
Gellyfish: Laboratory Development and Testing
C) Mixed metal:metal competition experiments
Step 4: Challenge Gellyfish/Model, cont’d
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Field Experiment I: Spatial/Temporal Dynamics of Metal Speciation in Boston Harbor
- What is the concentration of free metal ions in Boston Harbor?
- Is this a problem?
- How does speciation vary during a year?
- Is there spatial variability in metal speciation?
- What factors most influence changes in metal speciation?
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Sample Locations:
Mystic River
Inner Harbor
Fort Point Channel
Savin Hill Cove
Marina Bay
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Results: Total Dissolved Copper
Date
0
50
100
150
Tota
l Dis
solv
ed C
u (n
Mol
/Kg)
Fort Pt Channel
Inner Harbor
Marina Bay
Mystic River
Savin Hill
J F M A M J J A S O N D
Acute WQC
Chronic WQC
- Water Quality Criteria (WQC) exceedances:
- 1 location (Marina Bay)62
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Results: Free Cu2+:
Date
0
10
20
30
40
50
Free
Cu2+
(pm
ol/k
g) Fort Pt Channel
Inner Harbor
Marina Bay
Mystic River
Savin Hill
J F M A M J J A S O N D
- More accurate assessment of effects
- Potential effects level: 10 pMol/kg (acute)
- exceeded for many months
- exceeded at multiple locations 63
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Date
1e-12
1e-11
1e-10
1e-09
1e-08
1e-07
Free
Me2+
(pm
ol/k
g)Cu
Zn
Pb
Ni
Cd
J F M A M J J A S O N D
Data For Other Metals: Free Metal Ion
Location: Fort Point Channel
- Correlations between metals?
- Independence of metal behavior? 64
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Date
0.01
0.1
1
10
100
1000
Me2+
/ M
e tota
l (%
)Cu
Zn
Pb
Ni
Cd
J F M A M J J A S O N D
Data For Other Metals: % Free Metal Ion
Location: Fort Point Channel
- Similar effects of environmental factors on speciation?65
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What We Can Do (And Have Done) With the Data…
-Correlation Structure of total metals and free metal ions
- Spatial and Temporal Autocorrelation Structure
- temporal component of variance larger- informs monitoring strategies
- Factors Influencing Metal Speciation (Regression Analyses)
- highlight: importance of antecedent rain/DOC interaction- rain associated DOC has less affinity for metals- more nuanced modeling needed?
- Ligand Specificity Experiments
- Are natural ligands metal specific?- to what extent can one metal alter the speciation of other metals?
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(a) Cu
5.0E-11
1.5E-10
2.5E-10
3.5E-10
4.5E-10
5.5E-10
1.E-09 1.E-08 1.E-07 1.E-06
Total Zn Addition (M)
Cu2+ in Surrounding
Solution (M)
(c) Pb
5.0E-11
2.5E-10
4.5E-10
6.5E-10
8.5E-10
1.E-09 1.E-08 1.E-07 1.E-06
Total Zn Addition (M)
Pb2+ in Surrounding
Solution (M)
(d) Ni
2.0E-10
8.0E-10
1.4E-09
2.0E-09
2.6E-09
3.2E-09
1.E-09 1.E-08 1.E-07 1.E-06
Total Zn Addition (M)
Ni2+ in Surrounding
Solution (M)
(e) Cd
3.0E-10
6.0E-10
9.0E-10
1.2E-09
1.5E-09
1.8E-09
1.E-09 1.E-08 1.E-07 1.E-06
Total Zn Addition (M)
Cd2+ in Surrounding
Solution (M)
Results of a Ligand Specificity Study:-Boston Harbor (Winter)- Water spiked with increasing Zn- Free metal ions of Cu, Pb, Ni, and Cd followed- Results: Free metal ions of Cu, Pb, Ni, Cd increase
- Implication: Ligands not metal specific- Summer Results Different!
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Me2+
Phytoplankton
Gellyfish
Concordance?Direct Uptake
TrophicTransfer
Shellfish
Experiment II: Does the Gellyfish Sampler Mimic the Uptake of Metals into Biological Organisms?
Megel (mol)M
e orga
nism
(mol
/g)
m = ??
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Co-Deployment of Gellyfish, Mussels in Boston Harbor and
Massachusetts Bay
Gellyfish Mounted in Baskets
Gellyfish Baskets and Mussel Cages on Deployment Line 69
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Locations:
Buoy B
Cape Cod Bay
Deer Island
Inner Harbor (Aquarium)
Outfall (OSM-1, OSM-4, OSM-6)
Quincy Bay
Savin Hill Cove
Sampling Locations:
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0.00 0.10 0.20 0.30 0.40 0.50(E-1)
Gellyfish Pb (mean µg)
0
2
4
6
8
10
Mus
sel P
b (m
ean
µg/g
)
Mussel Mean Pb = 161.76 Gellyfish Mean Pb + 1.29R2 = 0.95; p < 0.0001
0.00 0.16 0.32 0.48 0.64 0.80
Mean Gellyfish Cu (µg)
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6
9
12
15
Mea
n M
usse
l Cu
(µg/
g)
Mussel Mean Cu = 4.71 Gellyfish Mean Cu + 7.64R2 = 0.52; p = 0.02
Results: Gellyfish x Mussel Regressions
Pb: R2 = 0.95; p<0.001
Cu: R2 = 0.52; p=0.02
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Concluding Remarks – Gellyfish- Simple, effective tool for measurement of metal speciation
- Multiple Uses:
- Biogeochemistry studies
- Surrogate measures of biological uptake
- User groups:
- Geochemists
- speciation studies, competition studies
- Environmental Managers
- monitoring programs
- TMDL assessments
- New Sampler Designs:
- Shorter equilibration times (<10 hrs
- Metal specific ligands (biologically relevant ligands?) 72
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SRP would also like to thank the presenters and moderators of the Ecological Risk: New Tools and Approaches webinar series:Presenters:Gary Ankley, Toxicologist, USEPA/ORD Mid-Continent Ecology DivisionDavid Barber,* Associate Professor, Toxicology, University of FloridaNancy Denslow,* Professor Toxicology, University of FloridaKim Anderson,* Professor, Oregon State UniversityCelia Chen,* Research Associate Professor, Dartmouth CollegeMark Hahn,* Woods Hole Oceanographic InstitutionRichard Di Giulio,* Director, Duke University’s Integrated Toxicology Program, Duke UniversityBruce Duncan, Senior Ecologist, EPA Region 10Jim Shine,* Associate Professor of Aquatic Chemistry, Harvard University
Moderators:Heather Henry, Program Administrator, Superfund Research ProgramCharles Maurice, Superfund and Technology Liaison, EPA Office or Research and DevelopmentDiane Nacci, Senior Research Biologist, EPA Office or Research and DevelopmentBeth Anderson, Program Analyst, Superfund Research Program
*SRP Grantee (current or former)
Acknowledgements
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SRP would like to thank the Ecological Risk Planning Committee:
•Marc Greenberg (EPA, ERT Region 2)•Heather Henry (NIEHS, SRP)
•Sharon Thoms (EPA, Region 2)
•Jean Zodrow (EPA, Region 10)
And Members of the:•EPA Ecological Risk Assessment Forum (ERAF)
•DOD Tri-Services Ecological Risk Assessment Working Group (TSERAWG)
Acknowledgements
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