ENVIRONMENTAL CONTAMIBIOAVAILABILITY & EXPOSUR
NANT E
INNOINNOVVAATIVE TIVE TECHNOLTECHNOLOGIESOGIES T TO O QUQUANTIFYANTIFY
Kim A. Anderson, PhDProfessor, Environmental & Molecular ToxicologyDirector, Food Safety & Environmental Stewardship ProgramOregon State University
Whyy Bioavailable ?2
Environmental exposure and fate
Understanding environmental factors on diseases…
Must develop new bio-analytical tools to measure exposure
L.S. Birnbaum, EHP, 2010
Thinking outside the sampling jar
Few advancesIntelligent samplingEnvirEnviroonmental nmental exposureBiBioavaililabilitbility
BRIDGES
Environmental Toxicology
Environmental
Environmental Chemistry
Toxicology
Toxico-kineticsConcentrations
For example:source, route,
quantity
For example:Uptake, metabolism,
elimination
Bioavailabilityq y
Environmental Fate
For example:
Toxico-dynamics
For example: effects
Bioavailability
For example: transport,
distribution, degradation
For example: effects across levels of
biological organization
Exposure Effect
Adapted from: Anderson & Hillwalker, Ecotoxicology Bioavailability, Elsevier 20083
Multivariate patternrecognition
methods
ContaminantContaminantexposure fromenvironment
EnvironmentalEnvironmental
Exposure = ToxicResponse ?
Reduce uncertainty~bridge exposure
to biological response
Technolog De elopment and ValidationTechnology Development and Validation
PSD-BRIDGESenvironmental
tools
Zebrafishdevelopmental
model
GESAdapted from: Schwartz, D.A., et al BRID, 2005
BRIDGES B e y o n d Chemical Analysis
5
PSD Membrane Cell Membrane
Biological Response Indicator Devices for Gauging BRIDGES Environmental Stressors
Passive sampling devices q u a n t i t a t i ve technology for deployment at Superfund sitestechnology for deployment at Superfund sites
nts.
i ion
h
PSD theory: PSD represent an organic lipid membrane. Like a membrane, PSDs
discriminate against particulate bound material. As in situ time integrative passive
samplers, PSDs may be deployed for extended periods of time to sequester contamina
This overcomes potential issues such as detecti li it biThi t ti bioavaililabll i h d t tion limits, ble f ti ll t fraction collect
and fluctuating contaminant concentrations. In-situ, site specific calibration is throug
isotope labeled infusion into the PSD with performance reference compounds.
7
Air/Water Passive Sampling Device Air/water
k keC
ku ea/w Cm
uptake eliminateuptake eliminate
Rate to change of the concentration:
dCm/dt = kuCw – keCs
Conc at any t is determined by competing rates of uptake and releaseAnderson, et al; ES&T, 2008
Chemical Reaction Kinetics Modeluptake and release of contaminantuptake and release of contaminant
Holistic, Common Metricsame PSD PSD used d in water, air, seddiment, personall
Anderson, et al; ES&T, 2008
Bioavailable Passive Sampplers
Extraction and bioassays In situ sediment deployments
Developpment of Bio-analyytical Tools
Integrated Sampling Broad range of contaminants
CSuitable to LC and GPAHs, PCBs, Pesticides
1,200+ analytes screen
Oxygenated PAHsLayshock et al ETC, 2010
12
111111111111
float b
top bu
float b
top bu
float b
top bu
float b
top bu
float b
top bu
float b
top bu
float b
top bu
float b
top bu
float b
top bu
float b
top bu
float b
top bu
float b
top bu
PSD weig
float b
0 ft PSD weig
float b
0 ft PSD weig
float b
0 ft PSD weig
float b
0 ft PSD weig
float b
0 ft PSD weig
float b
0 ft PSD weig
float b
0 ft PSD weig
float b
0 ft PSD weig
float b
0 ft PSD weig
float b
0 ft PSD weig
float b
0 ft PSD weig
float b
0 ft
field
Toxicological ResponsesToxicological ResponsesEnvironmental ExposureEnvironmental Exposure Toxicological ResponsesToxicological ResponsesEnvironmental ExposureEnvironmental Exposure
prep
mix
prep
mix
prep
mix
l f l ield
ext
ract
prep
mix
prep
mix
prep
mix
prep
mix
prep
mix
prep
mix
l f l ield
ext
ract
ield
ext
ract
uoy
oy
uoy
oy
d ex
trac
t
uoy
oy
uoy
oy
d ex
trac
t
prep
mix
prep
mix
prep
mix
l f l ield
ext
ract
prep
mix
prep
mix
prep
mix
prep
mix
prep
mix
prep
mix
l f l ield
ext
ract
ield
ext
ract
uoy
oy
uoy
oy
uoy
oy
uoy
oy
d ex
trac
t
uoy
oy
uoy
oy
uoy
oy
uoy
oy
d ex
trac
t
p p p
test @ day 1
•mortality•morphology
1 embryo per
male female
VV
f ili d
test @ day 5
•mortality •morphology
fi pp pp pp
test @ day 1
•mortality•morphology
1 embryo per
male female
VV
f ili d
test @ day 5
•mortality •morphology
fifi
cageht
uoy
cageht
uoy
quantifyextraction
field
cageht
uoy
cageht
uoy
quantifyextraction
field p p p
test @ day 1
•mortality•morphology
1 embryo per
male female
VV
f ili d
test @ day 5
•mortality •morphology
fi pp pp pp
test @ day 1
•mortality•morphology
1 embryo per
male female
VV
f ili d
test @ day 5
•mortality •morphology
fifi
cageht
uoy
cageht
uoy
cageht
uoy
cageht
uoy
quantifyextractioncage
ht
uoy
cageht
uoy
cageht
uoy
cageht
uoy
quantifyextraction
field
•morphology•movement
fertilized eggs
•morphology •hatch rate•swimming
96 well plate
•morphology•movement
fertilized eggs
•morphology •hatch rate•swimming
96 well plate
PCBPAHPAH-metabolites
dialysisfield deployment
PCBPAHPAH-metabolites
dialysis
solvent exchange
field deployment
•morphology•movement
fertilized eggs
•morphology •hatch rate•swimming
96 well plate
•morphology•movement
fertilized eggs
•morphology •hatch rate•swimming
96 well plate
PCBPAHPAH-metabolites
dialysisfield deployment
PCBPAHPAH-metabolites
dialysis
solvent exchange
field deployment
BRIDGES: Reduce exposure uncertainty by analyzing biological responsesBRIDGES extracts with bioassay model (Zebrafish, Ames, etc) systems
Bioavailable, Bioassay Compatible
Complimentary PSD multi‐contaminantmulti‐contaminant,
LC and GChigh throughput
Comparable MetricComparable Metricquantified with in‐device
surrogates
Common Metricsediment, air, water,
personalpersonal
High Spatial and Temporal ResolutionTemporal Resol tion
time‐integrated
Nexu sIntegratedHolistic
Diagnostic
Zebrafishteratogenic and neurobehavioralneurobehavioral
AmesAmesgenotoxicity
Cometgenotoxicity DNA damage
Human Cellssurvival and growth
Human CellsHuman Cellsspecific responses
= Sampling site= SuperfundRM 1E
RM 3.5E
RM 7W RM 7E
WASHINGTON
OREGON
Portland
NORTHRM 17E
OREGON
CALIFORNIA
fSuperf
eund D
eploymmt
ent Sites
BRIDGES
15
Extraction and Analyysis of PSD
xtrE lE act clean
LC or GC compatible
HPLC, GCMS, GCMS-QQQ, LCMS
PAHs (EPA) and b e y o n d 33 PAHs
302 mw, dibenzopyrene isomers PAHsLayshock et al JEM 2010Layshock et al JEM, 2010
Oxygenated PAHs (ketones, quinones)Layshock and Anderson, ETC, 2010
Pesticides PCBsPesticides, PCBs,
1,200+ analyte screen
Effect of dredging at RM 6.3 on bioavailable PAHs at RM 7n=3 each a composite of 5 p<0 05 in situ calibration with labeled PRCn=3, each a composite of 5, p<0.05, in situ calibration with labeled PRC
4
3
3.5
4
g/L) **
**
2.5
3
ratio
n (µ
g
WetDry
1.5
2
H c
once
nt
y
0.5
1
ΣPA
H
0pre-tar tar removal post-tar
Spatial and Temporal at Supp p erfund sitesp
W li d ata f h Water quality d for the carcinogenic EPA PP PAHs.
/ = wet season
= dry season
The dashed lines represent the EPA Water Quality Guidelines for human health for for human health for consumption of water and organism.
N = 110, P<0.001, 3 week deployments = low flow (summer) = high river flow (>10,000 ft3/s, >1 in rain)
Sower & Anderson, EST, 2008, Sethajintanin and Anderson, EST, 2007
Site-specific Biological ResponsesAbnormal developmental morphological endpoints observed in embryonic zebrafish exposed to contaminant m ixtures f rom e xtracts o f LFTs d eployed at S f id i i f f LFT d l d Superfund sites.
Control 1% LFT Extract
hpf
Not
30 h
pf T
126
h T
PEYSEYSE
Not= notochord waviness; PE= pericardial edema; YSE= yolk sac edema; T= bent tail
PSD successfully bridged to in vivo full organism bio-assaynn=32 eac32 each h dosedose, PSD , PSD dose responsedose response, 0.8x , 0.8x 8x riv8x riveer wr waaterter
Positive control trimethyltinPositive control trimethyltin
Negative control 1% DMSO
PH Superfund site river mile=3.5WPH Superfund site river mile 3.5W
July 2010
10JUL29-01-017
100x
5uM TMT
0.8x
4x
20x
Percent of Total (%)0 20 40 60 80 100 120
1% DMSO
( )Mortality Adversely Affected Unaffected
10010JUL29-01-017
Res
pons
e (%
)
60
80
ence
of A
dver
se R
20
40
Concentration
0.8x 4x 20x 100x
Inci
d
0
PSD bridged to whole organism bio-assayCompC parison of the integratg ivve EZM (M (mean ±± 95%95% CCI)) of embryyonic zebrafish expposed to different concentrations of extract solution obtained from LFTs deployed M&B and Portland Superfund sites.
n = 941, Asterisks (*) indicate significant differences relative to control embryos (1% DMSO). Each dose total n=72, three different days 24 fish each day.
Hillwalker, Allan, Tanguay, and Anderson Chemosphere, 2010, 79, 1-7
Anticippated IMPACT
Common metric:
water, air, sediment, ppersonal
Bioavailable mixtures in
high throughput
screens
Calibrated, time-
integrintegrated ated estimate of
environmental eexposurexposure
22
Integrated with bioassays
linking linking exposure to biological endpointsendpoints
•
•
•
•
•
•
•
•
•
•
•
•
•
•
•
•
•
•
•
•
•
•
InnovationSequesters dissolved-bioavailable fraction
S t roab d f t tSequesters a ib d range of contaminants
Bioavailability changes of new remedial
technologies can be pre-tested in site-specific
sediment created microcosmsediment created microcosm
Has the ability to be use directly in bioassays
Linkable with integrated bioassay from in vitro
(mammalian cell) to in vivo vertebrate assays(mammalian cell) to in vivo vertebrate assays
Has the ability to be used to explore mechanism-
based understanding of bioavailability processes
Same technologySame technology as used for water and airas used for water and air
High spatial and temporal resolution possible
In situ sediment depth profiles possible
Captures episodic events
C ti d evet d lComparative data d loping rapi idlidly
Composites without mechanical power
No on-site power needed
Very low detection limits with analytical easeVery low detection limits with analytical ease
Less expensive
Greener technology
In situ assessment of contaminantsIn-situ assessment of contaminants mostmost
biologically relevant
Discriminates environment from other exposures
(e g diet)(e.g. diet)
Easier to replicate spatially than organism
Organisms in the environment are not sacrificed
Acknowledggements
Collaborators:Swinomish Indian Tribal CommunitySwinomish Indian Tribal CommunityConfederated Tribes of the Umatilla Indian Reservation
Oregon Dept of Environmental QualityKevin Parrot and Sott Manzano
Grand Isle State Park, LA, T. AugustineMS Gulf Port Harbor Master, DJ ZigglerMobile AL Ben Secour National Refugre, J. IssacsMobile AL Ben Secour National Refugre, J. IssacsPensacola FL Gulf Islands National Seashore, R. Hoggard
Funding:P42 ES016465 (PI Williams Project LeaderP42 ES016465 (PI Williams, Project Leader Anderson, Analytical Core leader Anderson)P30 ES000210 (PI Beckman)R21 ES020120 (PI Anderson)UN FAO GEF, (Co-PIs: Jepson, Anderson, Jenkins)
Collaborators:Oregon State UniversityProfessor Robert Tanguay, SRP Co-IProfessor Anna Harding, Co-IProfessor Anna Harding, Co IProfessor Dashwood, Linus Pauling Institute, CCP Core, David Yu, PhD. (Ames)
Pacific Northwest National LaboratoryKatrina Waters PhDKatrina Waters, PhD
Acknowledggements
http://fses.oregonstate.edu
GULF Outreach
http://oregonstate.edu/superfund/oilspill
Kevin Hobbie Nick Hamilton Greg Sower, PhDTed Haigh, PhD Jorge Padilla Angie Perez, PhDMelissa McCartney Kristin Pierre Lucas Quarles, MSMelissa McCartne Kristin Pierre L cas Q arles MSGlenn Wilson Nathan Rooney Solysa VisalliJennifer Przybyla Kyle Tidwell Margarett CorviSarah Allan Brian Smith, PhD O. Krissanakriang, PhDNorm Forsbergg D. SethajSteven O’Connell Not pictured:
jintanin, PhD
Lane Tidwell Jeremy Riggle, PhDPhil Janney Julie Layshock, PhD. Ricky Scott Hillwalker, W., PhD