Innovative Technologies to Quantify Environmental ......full organism bio-assay nn= 32 eac32 each h dosedose, PSD , PSD dose responsedose response, 0.8x , 0.8x 8x riv8x riveer wr waaterter

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

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Toxicological ResponsesToxicological ResponsesEnvironmental ExposureEnvironmental Exposure Toxicological ResponsesToxicological ResponsesEnvironmental ExposureEnvironmental Exposure

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p p p

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•mortality•morphology

1 embryo per

male female

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test @ day 5

•mortality •morphology

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•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) **

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2.5

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ratio

n (µ

g

WetDry

1.5

2

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once

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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

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Integrated with bioassays

linking linking exposure to biological endpointsendpoints

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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