Environmental Endocrine Disruption Environmental Endocrine Disruption In San Francisco Bay Fish In San Francisco Bay Fish

Effects AssessmentEffects Assessment

Kevin KelleyKevin Kelley1-31-3, Jesus Reyes, Jesus Reyes1-31-3, Rusty , Rusty FaireyFairey44, Zed Mason, Zed Mason2,32,3, Cathy , Cathy PropperPropper55, Julianne Kalman, Julianne Kalman1,2,1,2,33

11EEL --Environmental Endocrinology Laboratory , Marine Biology EEL --Environmental Endocrinology Laboratory , Marine Biology ProgramProgram

22IIRMES --Institute for Integrated Research on Materials, Environment & IIRMES --Institute for Integrated Research on Materials, Environment & SocietySociety

33CEPA --Center for Education in Proteomics AnalysisCEPA --Center for Education in Proteomics Analysis

California State University, Long BeachCalifornia State University, Long Beach

44Marine Pollution Laboratory, Moss Landing Marine LaboratoriesMarine Pollution Laboratory, Moss Landing Marine Laboratories

55Northern Arizona UniversityNorthern Arizona University

66Cabrillo Marine Aquarium, San Pedro, CACabrillo Marine Aquarium, San Pedro, CA

ENDOCRINE SYSTEMSENDOCRINE SYSTEMS

regulateregulate

essential physiological functionsessential physiological functions

Reproduction Reproduction - - Somatic GrowthSomatic Growth - - MetabolismMetabolism Defense/ImmuneDefense/Immune - - ThyroidThyroid

- - Developmental Developmental Stress Stress ResponsesResponses - - NeuroendocrineNeuroendocrine - - othersothers

……Homeostasis, Adaptation, Homeostasis, Adaptation, Survival…Survival…

Some points about endocrine Some points about endocrine systemssystems::

. . . highly sensitive & responsive. . . highly sensitive & responsive

. . . respond to presence of environmental . . . respond to presence of environmental factors (stressors) to a degree factors (stressors) to a degree commensurate with the need to maintain commensurate with the need to maintain homeostasishomeostasis

. . . endocrine disturbances observed are . . . endocrine disturbances observed are typically indicative of the type of typically indicative of the type of environmental factor(s) actingenvironmental factor(s) acting

Monitoring programs can take advantage of endocrine disruptionendocrine disruption as a sensitive sensitive

screening toolscreening tool to assess effects assess effects (impacts)(impacts)

of water/environmental quality & specific contaminant effects

Endocrine-disrupting Endocrine-disrupting chemicals (EDCs)chemicals (EDCs) are present in are present in

marine and marine and estuarine estuarine

environments, environments, particularly along highly particularly along highly

populated coastal populated coastal regionsregions: :

SF Bay EstuarySF Bay Estuary http://toxics.usgs.gov/photo_gallery/sanfran.html

1-1- determine incidence and magnitude of determine incidence and magnitude of endocrine disruption in wild fishesendocrine disruption in wild fishes

thyroid, stress responses, growth & repair, reproduction

2- 2- determine whether spatial differences can be established between different SF

Bay locations

3- 3- determine liver contaminant determine liver contaminant concentrations in individualsconcentrations in individuals

Correlate specific contaminant concentrations with types/magnitude of endocrine disruption

STUDY STUDY OBJECTIVESOBJECTIVES

Shiner Surfperch

Cymatogaster aggregata

Pacific Staghorn Sculpin Leptocottus armatus

pelagicpelagic

benthicbenthic

San Pablo BaySan Pablo Bay

Berkeley Berkeley WaterfrontWaterfront

Oakland Inner Oakland Inner HarborHarbor

Redwood CityRedwood City

far-field far-field referencereference:: Tomales BayTomales Bay

2006 Field 2006 Field SitesSites[pilot year][pilot year]

THYROID THYROID ENDOCRINENDOCRINE SYSTEME SYSTEM

Thyroid Hormones Thyroid Hormones

• broad physiological actionsbroad physiological actions• essential for essential for brain/neural brain/neural

developmentdevelopment• permissive to permissive to somatic growthsomatic growth• important regulators of important regulators of metabolismmetabolism

3,5,3’,5’-tetraiodothyronine (thyroxine)3,5,3’,5’-tetraiodothyronine (thyroxine) 3,5,3’-triiodothyronine 3,5,3’-triiodothyronine

T4T4 T3T3

0

20

40

60

80

100

San PabloBay

Berkeley Waterfront

RedwoodCity

Oakland Inner Harbor

Tomales Bay

(11) (19) (19) (9) (14)

Pla

sma T

hyro

xin

e

Pla

sma T

hyro

xin

e

(T4

) (T

4)

Conce

ntr

ati

on,

Conce

ntr

ati

on,

ng/m

lng/m

l

ShinerShiner

SurfpeSurfperchrch

Thryoxine (T4) Levels in Shiner Thryoxine (T4) Levels in Shiner SurfperchSurfperch

0

20

40

60

80

100

San PabloBay

Berkeley Waterfront

RedwoodCity

Oakland Inner Harbor

Tomales Bay

(8) (13) (17) (12) (5)

Pla

sma T

hyro

xin

e

Pla

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hyro

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e

(T4

) (T

4)

Conce

ntr

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Conce

ntr

ati

on,

ng/m

lng/m

l

Pacific Pacific StaghoStaghorn rn SculpiSculpinn

Thryoxine (T4) Levels in Pacific Thryoxine (T4) Levels in Pacific Staghorn SculpinStaghorn Sculpin

0

20

40

60

80

100

San PabloBay

Berkeley Waterfront

RedwoodCity

Oakland Inner Harbor

Tomales Bay

(8) (11) (13) (19) (17) (19) (12) (9) (5) (14)

Pla

sma T

hyro

xin

e

Pla

sma T

hyro

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e

(T4

) (T

4)

Conce

ntr

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on,

Conce

ntr

ati

on,

ng/m

lng/m

l

ShinerShiner

SurfpeSurfperchrch

Pacific Pacific StaghoStaghorn rn SculpiSculpinn

Thryoxine (T4) Levels in SF Bay Thryoxine (T4) Levels in SF Bay FishFish

Environmental Contaminants are Environmental Contaminants are Implicated in Thyroid Endocrine Implicated in Thyroid Endocrine

Disruption:Disruption:

……e.g., Phenols, Hg, Cd impair thyroxogenesis in thyroid e.g., Phenols, Hg, Cd impair thyroxogenesis in thyroid follicle cells in fish follicle cells in fish (Bhattacharya et al., ‘89)(Bhattacharya et al., ‘89)

……e.g., endosulfan, Aroclor 1254 and selected PCBs e.g., endosulfan, Aroclor 1254 and selected PCBs decrease T4 levels in fish, via effects on decrease T4 levels in fish, via effects on iodotyrosine iodotyrosine diodinasesdiodinases (Coimbra et al., ‘05; LeRoy et al., ‘06)(Coimbra et al., ‘05; LeRoy et al., ‘06)

……e.g., ioxynil, certain PCBs and PBDEs bind e.g., ioxynil, certain PCBs and PBDEs bind transthyretintransthyretin and TRs with high affinities, altering T4 and T3 levels/ and TRs with high affinities, altering T4 and T3 levels/ actions in fish actions in fish (Tomy et al., ‘04; Lema et al., ‘06; Morgado et al., (Tomy et al., ‘04; Lema et al., ‘06; Morgado et al., ‘07)‘07)

STRESS STRESS RESPONSE RESPONSE ENDOCRINENDOCRINE SYSTEME SYSTEM

0

400

800

1200

1600

Redwood City

Oakland Inner

Harbor

Berkeley Waterfro

nt

San Pablo Bay

21 13 21 10

Shiner SurfperchPla

sma C

ort

isol

Conce

ntr

ati

on (

ng/m

l)

aa aa

bb

bb

Stress ResponseStress Response

0

400

800

1200

Redwood City

Oakland Inner

Harbor

Berkeley Waterfro

nt

San Pablo Bay

17 16 13 19

Pacific Staghorn SculpinPacific Staghorn Sculpin

Pla

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Conce

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Stress ResponseStress Response

Other wild fish species living in Other wild fish species living in affected marine environments exhibit affected marine environments exhibit

impairment of their impairment of their neuroendocrine stress response neuroendocrine stress response

systemsystem

...experimental studies have ...experimental studies have shown that contaminants, shown that contaminants,

--such as PCBs-- --such as PCBs-- impair stress responses in fishimpair stress responses in fish

((Quabius et al., 2005; Vijayan et al., 2006Quabius et al., 2005; Vijayan et al., 2006))

Thyroxine/Thyroxine/thyroidthyroid

IGF-I/IGF-I/growthgrowth

San PabloBay

Berkeley Waterfront

RedwoodCity

Oakland Inner Harbor

Tomales Bay

Shiner PerchShiner PerchPacific Staghorn Pacific Staghorn SculpinSculpin

Shiner PerchShiner PerchPacific Staghorn Pacific Staghorn SculpinSculpin

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Endocrine Disruption—SF Bay Endocrine Disruption—SF Bay FishFish

S U M M A R Y –S U M M A R Y –Year 1Year 1

Cortisol/Cortisol/stress responsestress response Shiner PerchShiner PerchPacific Staghorn Pacific Staghorn SculpinSculpin

Testosterone/Testosterone/ReprodReprod

1717αα-Ethinylestradiol/-Ethinylestradiol/Pharm.Pharm.

Endocrine Disruption—SF Bay Endocrine Disruption—SF Bay FishFish

San PabloBay

Berkeley Waterfront

RedwoodCity

Oakland Inner Harbor

Tomales Bay

Pacific Staghorn Pacific Staghorn SculpinSculpin

Pacific Staghorn Pacific Staghorn SculpinSculpin

1717ββ-Estradiol/-Estradiol/Reprod.Reprod.

Pacific Staghorn Pacific Staghorn SculpinSculpin

S U M M A R Y –S U M M A R Y –Year 1Year 1

immatureimmature

Shiner Shiner Perch ?Perch ?

August August 2006 2006

Oakland Oakland Inner Inner

HarborHarbor

Endocrine Disruption—SF Bay Endocrine Disruption—SF Bay FishFish

San PabloBay

Berkeley Waterfront

RedwoodCity

Oakland Inner Harbor

Tomales Bay

Shiner PerchShiner PerchPacific Staghorn Pacific Staghorn SculpinSculpin

Shiner PerchShiner PerchPacific Staghorn Pacific Staghorn SculpinSculpin

Shiner PerchShiner PerchPacific Staghorn Pacific Staghorn SculpinSculpin

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S U M M A R Y –S U M M A R Y –Year 1Year 1

1717ββ-Estradiol/-Estradiol/Reprod.Reprod.

1717αα-Ethinylestradiol/-Ethinylestradiol/Pharm.Pharm.

Testosterone/Testosterone/ReprodReprod

Contaminant Analyses:Contaminant Analyses:• Feasible to measure individuals liver Feasible to measure individuals liver

contaminant levels (PCBs, PAHs, contaminant levels (PCBs, PAHs, pesticides)pesticides)

• Several contaminants measured at high Several contaminants measured at high concentrations in shiner perch and Pacific concentrations in shiner perch and Pacific staghorn sculpinstaghorn sculpin

• Relatively higher levels of contaminants in Relatively higher levels of contaminants in livers of Pacific staghorn sculpinlivers of Pacific staghorn sculpin

• Feasible to correlate individual liver Feasible to correlate individual liver contaminant concentrations with contaminant concentrations with endocrine and physiological indices endocrine and physiological indices

Year – 2 Year – 2 StudiesStudies

under under wayway

San Pablo Bay San Pablo Bay (Loch Lohmand Marina) (Loch Lohmand Marina)

Richmond/Berkeley WaterfrontRichmond/Berkeley Waterfront

Oakland Inner HarborOakland Inner Harbor

Redwood CityRedwood City

Tomales BayTomales Bay

Bodega BayBodega Bay

Study SitesStudy Sites [August 2007][August 2007]

RichmondRichmond (Lauritzen Channel, (Lauritzen Channel,

DDTsDDTs))

San Leandro BaySan Leandro Bay (PCBs)(PCBs)San Francisco San Francisco WaterfrontWaterfront Hunter’s PointHunter’s Point (POTW)(POTW)

Two Harbors –Santa Catalina IslandTwo Harbors –Santa Catalina Island (Pacific (Pacific

side)side) L.A. Harbor –Inner Cabrillo BeachL.A. Harbor –Inner Cabrillo Beach (collab. w/CMA)(collab. w/CMA)

THYROID THYROID Thyroxine & TriidothyronineThyroxine & Triidothyronine

REPRODUCTION REPRODUCTION Testosterone Testosterone & & 11-ketotestosterone11-ketotestosterone

REPRODUCTION REPRODUCTION 1717ββ-Estradiol, -Estradiol, 1717αα-Ethinylestradiol, Vitellogenin-Ethinylestradiol, Vitellogenin

STRESS STRESS Cortisol & CortisoneCortisol & Cortisone

GROWTH GROWTH IGF-I & IGFBPsIGF-I & IGFBPs

Endocrine Endocrine Parameters in Parameters in

2007-82007-8

• Altered thyroxine levels.Altered thyroxine levels. -- -- T4 measurementT4 measurement

• Altered T3 levels, T4:T3 ratios?Altered T3 levels, T4:T3 ratios? -- -- T3 measurement, calculate T3 measurement, calculate T4:T3T4:T3

• Altered thyroid gland morphology?Altered thyroid gland morphology? -- -- Thyroid histologyThyroid histology

• Impaired thyroid responsiveness?Impaired thyroid responsiveness? -- -- TSH-challenge TSH-challenge • Altered T4 metabolism or TH receptors?Altered T4 metabolism or TH receptors? -- -- peripheral peripheral

iodotyrosine deiodinasesiodotyrosine deiodinases (ID1-3),(ID1-3), TRTRαα, TR, TRββ

Emphasis:Emphasis: Thyroid Endocrine (HPT) Thyroid Endocrine (HPT) AxisAxis

• Impaired growth?Impaired growth? -- -- Growth & IGF system indicesGrowth & IGF system indices [[IGF-I, IGFBPs, cartilage growth activity]IGF-I, IGFBPs, cartilage growth activity]

• Relationships with specific bioaccumulated chemicals?Relationships with specific bioaccumulated chemicals? -- -- Hepatic contaminants measurement & correlationHepatic contaminants measurement & correlation

Underlying MechanismsUnderlying Mechanisms::

Environmental ContaminantsEnvironmental Contaminants::

Physiological ImpactsPhysiological Impacts::

Emphasis:Emphasis: Stress Endocrine (HPI) Stress Endocrine (HPI) AxisAxis

• Altered cortisol levels.Altered cortisol levels. -- -- Cortisol measurementCortisol measurement• Altered interrenal gland morphology?Altered interrenal gland morphology? -- -- Interrenal histologyInterrenal histology • Altered metabolism of cortisol?Altered metabolism of cortisol? -- -- cortisone measurementcortisone measurement• Impaired interrenal responsiveness?Impaired interrenal responsiveness? -- -- ACTH-challengeACTH-challenge• Impaired production of cortisol?Impaired production of cortisol? -- -- steroidogenic enzyme steroidogenic enzyme

expressionexpression (StAR, 11(StAR, 11ββ-hydroxylase, 11-hydroxylase, 11ββ-HSD-2)-HSD-2)

• Relationships with specific bioaccumulated chemicals?Relationships with specific bioaccumulated chemicals? -- -- Hepatic contaminants measurement & correlationHepatic contaminants measurement & correlation

• Impaired defense?Impaired defense? -- -- Parasitic infestation assessmentParasitic infestation assessment

• Impaired growth?Impaired growth? -- -- Growth & IGF system indicesGrowth & IGF system indices [[IGF-I, IGFBPs, cartilage growth activityIGF-I, IGFBPs, cartilage growth activity]]

Underlying MechanismsUnderlying Mechanisms::

Environmental Environmental ContaminantsContaminants::

Physiological ImpactsPhysiological Impacts::

I.I. Endocrine responses Endocrine responses and/orand/or endocrine disruption endocrine disruption serve as serve as sensitive bio-markerssensitive bio-markers of environmental of environmental perturbations perturbations

II.II. Endocrine systems areEndocrine systems are::

--highly --highly sensitivesensitive to environmental to environmental stressorsstressors

----respond proportionallyrespond proportionally to the need to to the need to maintain maintain homeostasishomeostasis

--show alterations --show alterations reflective of the kind of reflective of the kind of stressor(s) stressor(s) presentpresent (different EDCs affect (different EDCs affect different endocrine different endocrine axes via distinct axes via distinct underlying mechanisms)underlying mechanisms)

SUMMARY POINTSSUMMARY POINTS

III. III. Endocrine-disrupted conditions are observed in Endocrine-disrupted conditions are observed in SF Bay fishSF Bay fish (urban, industrial, boat marinas)(urban, industrial, boat marinas)

IV. Endocrine disruption is IV. Endocrine disruption is evident in different fish evident in different fish speciesspecies

V. Endocrine disruption is V. Endocrine disruption is occurring in different occurring in different endocrine systemsendocrine systems (stress, thyroid, reproduction, growth (stress, thyroid, reproduction, growth

& repair, & repair, defense, metabolism)defense, metabolism)

VI. There may be VI. There may be interactions among endocrine interactions among endocrine systemssystems (means wider impacts on physiological (means wider impacts on physiological performance)performance)

SUMMARY SUMMARY POINTSPOINTScontinuedcontinued

A.A. Identification of specific chemicals associated Identification of specific chemicals associated with endocrine disruptionwith endocrine disruption

B. Understand mechanisms and test candidate B. Understand mechanisms and test candidate EDCs, to establish cause and effectEDCs, to establish cause and effect

C. Build models to estimate ecologically relevant C. Build models to estimate ecologically relevant performance alterations from measures of performance alterations from measures of endocrine disruption endocrine disruption (growth, reproduction, (growth, reproduction, disease/parasitism)disease/parasitism)

D. Determine contaminant thresholds of effectsD. Determine contaminant thresholds of effects

E. Use new technologies (proteomics) to build E. Use new technologies (proteomics) to build integrated phenotypic biomarker fingerprintsintegrated phenotypic biomarker fingerprints (multifactorial screen) (multifactorial screen)

DIRECTIONDIRECTION

Long Beach Env’l. Long Beach Env’l. Endo. Lab.Endo. Lab.

Jesus Reyes, Kathy Sak, Andrew Hamilton, Becki Hagstrom, Maelanie Galima, Dawn Petschauer, Navdeep Brar, Tiffany Price, Tuan Truong, Dana Cianfrani, Shari Smolko, Tami Gonzalez, Claire Wingard, Kenna Koons, Julianne Kalman, Scott Adams, Wendy Hall

?

http://www.csulb.edu/depts/endo

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