Environmental Endocrine Environmental Endocrine Disruption Effects AssessmentDisruption Effects Assessment
In San Francisco Bay FishesIn San Francisco Bay FishesShiner Surfperch & Pacific Staghorn SculpinShiner Surfperch & Pacific Staghorn Sculpin
Kevin Kelley, Jesus Reyes, Kevin Kelley, Jesus Reyes, Rusty Fairey, Zed Mason, Cathy Rusty Fairey, Zed Mason, Cathy
Propper, Juli KalmanPropper, Juli KalmanEnvironmental Endocrinology LaboratoryEnvironmental Endocrinology Laboratory
Institute for Integrated Research on Materials, Environment & Institute for Integrated Research on Materials, Environment & Society (IIRMES)Society (IIRMES)
Center for Education in Proteomics Analysis (CEPA)Center for Education in Proteomics Analysis (CEPA)
Marine Biology Program, Dept of Biological Sciences, Calif. St. Marine Biology Program, Dept of Biological Sciences, Calif. St. Univ. Long BeachUniv. Long Beach
Moss Landing Marine Laboratories & Northern Arizona UniversityMoss Landing Marine Laboratories & Northern Arizona University
Endocrine-disrupting Endocrine-disrupting chemicals (EDCs)chemicals (EDCs) are present in are present in
marine and marine and estuarine estuarine
environments, environments, particularly along particularly along highly populated highly populated coastal regions: coastal regions:
SF Bay EstuarySF Bay Estuary
. . . measures of ‘endocrine disruption’ . . . measures of ‘endocrine disruption’ are increasingly used as sensitiveare increasingly used as sensitive
bioindicators for water/environmental bioindicators for water/environmental quality and specific contaminant effectsquality and specific contaminant effects
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, , emphasizing assessment of endocrine systems
regulating stress responses, growth & repair, thyroid & reproduction
2- 2- determine whether spatial differences can be established between different SF Bay locations, with respect to endocrine disrupted
condition(s)
3- 3- determine liver contaminant determine liver contaminant concentrations in individualsconcentrations in individuals, , to identify
correlations between specific contaminants and type(s) of endocrine disruption in fish
sampled from different SF Bay locations
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]
Tomales Tomales BayBay
Bodega Bodega BayBay
Far Field Far Field SitesSites
Thyroxine/HPT Thyroxine/HPT AxisAxis
IGF-I/Growth IGF-I/Growth AxisAxis
Cortisol/HPI Cortisol/HPI Axis Axis
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
++ ++++++
++++
++++++
++++
++++
?? ???? ??
Endocrine Disruption—SF Bay Endocrine Disruption—SF Bay FishFish
S U M M A R S U M M A R YY
Testosterone/HPG AxisTestosterone/HPG Axis
1717αα-Ethinylestradiol-Ethinylestradiol
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
1717ββ-Estradiol/HPG -Estradiol/HPG AxisAxis
Shiner PerchShiner PerchPacific Staghorn Pacific Staghorn SculpinSculpin
?? ?? ?? ?? ??
?? ?? ?? ?? ??
S U M M A R S U M M A R YY
Shiner Surfperch ?Shiner Surfperch ?A
ug
ust
2006
Oakla
nd
In
ner
Au
gu
st
2006
Oakla
nd
In
ner
Harb
or
Harb
or
0
50
100
WWTP-affected site (OCSD, turbot)
Shiner Surfperch
Pla
sma E
E2
Conce
ntr
ati
on (
pg/m
l)1717αα-Ethinylestradiol -Ethinylestradiol ((EE2) in Fish PlasmaEE2) in Fish Plasma
150
200
Pacific Staghorn Sculpin
(Tomales Bay, Redwood City, Oakland Inner Harbor, Berkeley, San Pablo Bay (Loch Lohmand)
N.N.D.D.
N.N.D.D.
Liver Contaminant Analysis, Liver Contaminant Analysis, GC/MSGC/MS
Selected Contaminants* Shiner Surfperch
Pacific Staghorn Sculpin
Chlordane-gamma 1 – 4 2 – 5
Chlordane-alpha 3.5 – 15 6 – 10
trans-Nonachlor 3.5 – 12 5 – 20
cis-Nonachlor 1.5 – 5 4 – 13
4,4’-DDE 20 – 47 48 – 54 *range of ng/g wet weight values shown
So-Calif would have 4, 4’-DDTSo-Calif would have 4, 4’-DDT
Selected Contaminants* Shiner Surfperch
Pacific Staghorn Sculpin
Naphthalene 13 – 19.5 15 – 20
2-methylnaphthalene 9 – 15 6 – 17
1-methylnaphthalene 5 – 13 6 – 8
Biphenyl 1 – 5.5 2 – 5
Fluorine 2.5 – 11 3.5 – 5
Phenanthrene 6.5 – 11 6.5 – 8
*range of ng/g wet weight values shown
Liver Contaminant Analysis, Liver Contaminant Analysis, GC/MSGC/MS
Selected Contaminants* Shiner Surfperch
Pacific Staghorn Sculpin
PCB 095 6 – 12 8 – 14
PCB 149 11 – 35 25 – 35
PCB 138 27 – 70 56 – 63
PCB 180 14 – 47 28 – 58
PCB 194 nd – 8 6 – 21
PCB 153 27 – 66 56 – 68
Other congeners measured:28, 31, 44, 49, 52, 66, 70, 74, 87, 99,101,105, 110, 118, 128+167, 141, 151, 156, 158, 168+132, 170, 177, 183, 187, 200
1 – 16 2 – 35
*range of ng/g wet weight values shown
Liver Contaminant Analysis, Liver Contaminant Analysis, GC/MSGC/MS
ThyroxineThyroxine & & TriidothyronineTriidothyronine
TestosteroneTestosterone & 11- & 11-ketotestosterone ketotestosterone
1717ββ-Estradiol-Estradiol
IGF-I & IGF-I & IGFBPs IGFBPs
Endocrine Disruption—SF Endocrine Disruption—SF Bay FishBay Fish
Cortisol Cortisol & Cortisone & Cortisone
** **
POTW, POTW, seasonaseasona
l l effects?effects?
????
[[YellowYellow=Year-1 fish =Year-1 fish analyses]analyses]
**=spatial differences detected=spatial differences detected
Hepatic contaminants Hepatic contaminants (chlorinated pesticides, PCB (chlorinated pesticides, PCB congeners, PAHs)congeners, PAHs)
1717αα-Ethinylestradiol-Ethinylestradiol
Year – 2 Year – 2 StudiesStudies
under under wayway
San Pablo Bay San Pablo Bay (Loch Lohmand (Loch Lohmand Marina)Marina)
Richmond/Berkeley Richmond/Berkeley WaterfrontWaterfrontOakland Inner HarborOakland Inner Harbor
Redwood CityRedwood City
far-field far-field referencereference:: Tomales BayTomales Bay
Bodega BayBodega Bay
Study Sites Study Sites 20072007 July 29-July 29-
August 4August 4
RichmondRichmond (Lauritzen Channel, (Lauritzen Channel, DDTs)DDTs)
San Leandro BaySan Leandro Bay (PCBs)(PCBs)San Francisco San Francisco WaterfrontWaterfront Hunter’s Point Hunter’s Point (POTW)(POTW)
Shiner Shiner PerchPerch
Pacific Staghorn Pacific Staghorn SculpinSculpin
Redwood CityRedwood City
San Francisco San Francisco WaterfrontWaterfrontS.F. Hunter’s PointS.F. Hunter’s Point (POTW (POTW outfall)outfall)Oakland Inner Oakland Inner HarborHarbor
Loch Lomand MarinaLoch Lomand Marina (San Pablo (San Pablo Bay)Bay)Richmond WaterfrontRichmond Waterfront
Richmond –Lauritzen ChannelRichmond –Lauritzen Channel (DDT-(DDT-contaminated)contaminated)
Bodega BayBodega Bay (far-field (far-field reference)reference)
San Leandro BaySan Leandro Bay (PCB-(PCB-contaminated)contaminated)
Tomales BayTomales Bay (far-field (far-field reference)reference)
3131 20202121 2020
---- 2323
2525 99
2929 99
1313 1717
2020 1111
2727 1212
22 11111212 66
180180 138138
FIELD DATA - AUGUST 2007FIELD DATA - AUGUST 2007
Two Harbors –Santa Catalina IslandTwo Harbors –Santa Catalina Island (Pacific side, far- (Pacific side, far-field reference)field reference)L.A. Harbor –Inner Cabrillo BeachL.A. Harbor –Inner Cabrillo Beach (collab. with Cabrillo Marine (collab. with Cabrillo Marine Aquarium)Aquarium)
Added sitesAdded sites::
Shiner Shiner PerchPerch
Pacific Staghorn Pacific Staghorn SculpinSculpin
Redwood CityRedwood City
San Francisco San Francisco WaterfrontWaterfrontS.F. Hunter’s PointS.F. Hunter’s Point (POTW (POTW outfall)outfall)Oakland Inner Oakland Inner HarborHarbor
Loch Lomand MarinaLoch Lomand Marina (San Pablo (San Pablo Bay)Bay)Richmond WaterfrontRichmond Waterfront
Richmond –Lauritzen ChannelRichmond –Lauritzen Channel (DDT-(DDT-contaminated)contaminated)
Bodega BayBodega Bay (far-field (far-field reference)reference)
San Leandro BaySan Leandro Bay (PCB-(PCB-contaminated)contaminated)
Tomales BayTomales Bay (far-field (far-field reference)reference)
3131 20202121 2020
---- 2323
2525 99
2929 99
1313 1717
2020 1111
2727 1212
22 11111212 66
180180 138138
LowLow infestation infestation rates by rates by EthusaEthusa
californicacalifornica (Isopoda)(Isopoda)
HighHigh infestation infestation rates by rates by ElthusaElthusa
californicacalifornica (variable across (variable across
sites & sites & individuals)individuals)
55.255.2%%
60.060.0%%
60.060.0%%
PARASITE PARASITE PREVALENCEPREVALENCE::
[#infected hosts/#examined [#infected hosts/#examined hosts x100%]hosts x100%]
20.220.2%%
<10%<10%
strongly strongly correlatecorrelate
d with d with gill gill
damagedamage
FIELD DATA - AUGUST 2007FIELD DATA - AUGUST 2007
Cortisol & Interrenal histology Cortisol & Interrenal histology --under --under wayway
Other parasites Other parasites -to be -to be evaluatedevaluated
Shiner Shiner PerchPerch
Pacific Staghorn Pacific Staghorn SculpinSculpin
Redwood CityRedwood City
San Francisco San Francisco WaterfrontWaterfrontS.F. Hunter’s PointS.F. Hunter’s Point (POTW (POTW outfall)outfall)Oakland Inner Oakland Inner HarborHarbor
Loch Lomand MarinaLoch Lomand Marina (San Pablo (San Pablo Bay)Bay)Richmond WaterfrontRichmond Waterfront
Richmond –Lauritzen ChannelRichmond –Lauritzen Channel (DDT-(DDT-contaminated)contaminated)
Bodega BayBodega Bay (far-field (far-field reference)reference)
San Leandro BaySan Leandro Bay (PCB-(PCB-contaminated)contaminated)
Tomales BayTomales Bay (far-field (far-field reference)reference)
3131 20202121 2020
---- 2323
2525 99
2929 99
1313 1717
2020 1111
2727 1212
22 11111212 66
180180 138138
Immature, Immature,
or early or early developing developing
gonadsgonads
Variety of Variety of reproductive reproductive
statesstates
[immature, [immature, mature, mature,
gravid/pregnantgravid/pregnant]]
FIELD DATA - AUGUST 2007FIELD DATA - AUGUST 2007
3/180 shiners were pregnant3/180 shiners were pregnant
Sex ratiosSex ratios:: TBA TBA
Gonadal somatic indexGonadal somatic index:: TBA TBA
HistologyHistology:: under way under way
Contaminant Analyses:Contaminant Analyses:• Feasible to measure individuals liver Feasible to measure individuals liver
contaminant levelscontaminant levels• Key contaminants measured at high Key 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
• Liver contaminant concentrations will be Liver contaminant concentrations will be correlated with indices of endocrine correlated with indices of endocrine disruption within individualsdisruption within individuals
Allows analysis of whether pollutants, individually or in Allows analysis of whether pollutants, individually or in combination, are associated with endocrine disruption & associated combination, are associated with endocrine disruption & associated reductions in physiological viabilityreductions in physiological viability
Different endocrine systems will likely exhibit different sensitivities Different endocrine systems will likely exhibit different sensitivities to contaminantsto contaminants
Will point to potential contaminant thresholds of effects in fish Will point to potential contaminant thresholds of effects in fish (further testing needed)(further testing needed)
Emphasis:Emphasis: Stress (HPI) Axis Stress (HPI) Axis Continuing Analyses & Emphasis 2007Continuing Analyses & Emphasis 2007
• a) field work -Aug 2007, a) field work -Aug 2007, completed completed
• b) cortisol measurement, b) cortisol measurement, completed, data analysis phase completed, data analysis phase underwayunderway
• c) interrenal histology, c) interrenal histology, underwayunderway
• d) hepatic contaminants measurement (d) hepatic contaminants measurement (underwayunderway) & correlation ) & correlation
• d) d) cortisonecortisone measurement measurement (11(11ββ-HSD-2)-HSD-2)
• e) ACTH-challenge (is interrenal responsiveness impaired?)e) ACTH-challenge (is interrenal responsiveness impaired?)
• f) ectoparasitic infestation assessmentf) ectoparasitic infestation assessment
• g) growth endocrine system impacts (g) growth endocrine system impacts (indicesindices: IGF-I, IGFBPs, : IGF-I, IGFBPs, cartilage growth activity) impactscartilage growth activity) impacts
• h) steroidogenic enzyme expression h) steroidogenic enzyme expression (StAR, 11(StAR, 11ββ-hydroxylase, 11-hydroxylase, 11ββ-HSD-2)-HSD-2)
b-d = potential bioassays for disruption b-d = potential bioassays for disruption (screening)(screening)
Thyroid (HPT) Axis Thyroid (HPT) Axis Disruption Disruption
––Studies proposed for Studies proposed for 20072007