1. Environmental Endocrine Disruption Effects Assessment
In San Francisco Bay Fishes
Shiner Surfperch & Pacific Staghorn Sculpin
Kevin Kelley, Jesus Reyes, Rusty Fairey, Zed Mason, Cathy Propper, Juli Kalman
Environmental Endocrinology Laboratory
Institute for Integrated Research on Materials, Environment & Society (IIRMES)
Center for Education in Proteomics Analysis (CEPA)
Marine Biology Program, Dept of Biological Sciences, Calif. St. Univ. Long Beach
Moss Landing Marine Laboratories & Northern Arizona University

2. . . . measures of ‘endocrine disruption’ are increasingly used as sensitive bioindicators for water/environmental quality and specific contaminant effects
Endocrine-disrupting chemicals (EDCs) are present in marine and estuarine environments, particularly along highly populated coastal regions:
SF Bay Estuary

3. STUDY OBJECTIVES
1- determine incidence and magnitude of endocrine disruption in wild fishes, emphasizing assessment of endocrine systems regulating stress responses, growth & repair, thyroid & reproduction
2- determine whether spatial differences can be established between different SF Bay locations, with respect to endocrine disrupted condition(s)
3- determine liver contaminant concentrations in individuals, to identify correlations between specific contaminants and type(s) of endocrine disruption in fish sampled from different SF Bay locations

4. Pacific Staghorn Sculpin Leptocottus armatus
Shiner Surfperch
Cymatogaster aggregata
pelagic
Pacific Staghorn Sculpin Leptocottus armatus
benthic

5. 2006 Field Sites [pilot year] San Pablo Bay Berkeley Waterfront
Oakland Inner Harbor
Redwood City
far-field reference: Tomales Bay

6. Far Field Sites
Bodega Bay
Tomales Bay

7. Endocrine Disruption—SF Bay Fish
S U M M A R Y
Endocrine Disruption—SF Bay Fish
Tomales
Bay
San Pablo
Bay
Berkeley
Waterfront
Redwood
City
Oakland
Inner Harbor
Cortisol/HPI Axis
Shiner Perch + +
Pacific Staghorn Sculpin ++ ++
Thyroxine/HPT Axis
Shiner Perch +
Pacific Staghorn Sculpin ++ ++ ++
IGF-I/Growth Axis
Shiner Perch ? ?
Pacific Staghorn Sculpin ? ?

8. Endocrine Disruption—SF Bay Fish
S U M M A R Y
Endocrine Disruption—SF Bay Fish
Tomales
Bay
San Pablo
Bay
Berkeley
Waterfront
Redwood
City
Oakland
Inner Harbor
17β-Estradiol/HPG Axis
Shiner Perch ? ? ? ? ?
Pacific Staghorn Sculpin
17α-Ethinylestradiol
Shiner Perch
Pacific Staghorn Sculpin
Testosterone/HPG Axis
Shiner Perch ? ? ? ? ?
Pacific Staghorn Sculpin

9. Shiner Surfperch ?
August Oakland Inner Harbor

10. Pacific Staghorn Sculpin
17α-Ethinylestradiol (EE2) in Fish Plasma
200
150
Plasma EE2 Concentration (pg/ml)
100 50
N.D.
N.D.
Shiner Surfperch
Pacific Staghorn Sculpin
WWTP-affected site (OCSD, turbot)
(Tomales Bay, Redwood City, Oakland Inner Harbor, Berkeley, San Pablo Bay (Loch Lohmand)

11. Liver Contaminant Analysis, GC/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’-DDT

12. Liver Contaminant Analysis, GC/MS
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

13. Liver Contaminant Analysis, GC/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, , 141, 151, 156,
158, , 170, 177, 183, 187, 200
1 – 16
2 – 35
*range of ng/g wet weight values shown

14. * * * ? Endocrine Disruption—SF Bay Fish POTW, seasonal effects??
[Yellow=Year-1 fish analyses] *
Cortisol & Cortisone *
Thyroxine & Triidothyronine
POTW, seasonal effects??
17β-Estradiol
17α-Ethinylestradiol
Testosterone & 11-ketotestosterone ?
IGF-I & IGFBPs
Hepatic contaminants (chlorinated pesticides, PCB congeners, PAHs) *
=spatial differences detected

15. Year – 2 Studies
under way

16. Study Sites 2007 July 29-August 4
San Pablo Bay (Loch Lohmand Marina)
Richmond (Lauritzen Channel, DDTs)
Richmond/Berkeley Waterfront
Oakland Inner Harbor
San Leandro Bay (PCBs)
San Francisco Waterfront
Hunter’s Point (POTW)
Redwood City
far-field reference: Tomales Bay
Bodega Bay

17. Pacific Staghorn Sculpin
FIELD DATA - AUGUST 2007
Shiner Perch
Pacific Staghorn Sculpin
Redwood City 31 20
San Francisco Waterfront 21 20
S.F. Hunter’s Point (POTW outfall) -- 23
Oakland Inner Harbor 25 9
San Leandro Bay (PCB-contaminated) 29 9
Loch Lomand Marina (San Pablo Bay) 13 17
Richmond Waterfront 20 11
Richmond –Lauritzen Channel (DDT-contaminated) 27 12
Bodega Bay (far-field reference) 2 11
Tomales Bay (far-field reference) 12 6
180
138
Added sites:
Two Harbors –Santa Catalina Island (Pacific side, far-field reference)
L.A. Harbor –Inner Cabrillo Beach (collab. with Cabrillo Marine Aquarium)

18. FIELD DATA - AUGUST 2007 Shiner Perch Pacific Staghorn Sculpin 31 20
Redwood City 31
60.0% 20
San Francisco Waterfront 21 20
S.F. Hunter’s Point (POTW outfall) --
strongly correlated with gill damage 23
Oakland Inner Harbor 25 9
<10%
San Leandro Bay (PCB-contaminated) 29
55.2% 9
Loch Lomand Marina (San Pablo Bay) 13 17
Richmond Waterfront 20
60.0% 11
Richmond –Lauritzen Channel (DDT-contaminated) 27
20.2% 12
Bodega Bay (far-field reference) 2 11
Tomales Bay (far-field reference) 12 6
PARASITE PREVALENCE:
[#infected hosts/#examined hosts x100%]
180
138
High infestation rates by Elthusa californica (variable across sites & individuals)
Low infestation rates by Ethusa californica (Isopoda)
Cortisol & Interrenal histology --under way
Other parasites -to be evaluated

19. FIELD DATA - AUGUST 2007 Shiner Perch Pacific Staghorn Sculpin 31 20
Redwood City 31 20
San Francisco Waterfront 21 20
S.F. Hunter’s Point (POTW outfall) -- 23
Oakland Inner Harbor 25 9
San Leandro Bay (PCB-contaminated) 29 9
Loch Lomand Marina (San Pablo Bay) 13 17
Richmond Waterfront 20 11
Richmond –Lauritzen Channel (DDT-contaminated) 27 12
Bodega Bay (far-field reference) 2 11
Tomales Bay (far-field reference) 12 6
180
138
3/180 shiners were pregnant
Sex ratios: TBA
Gonadal somatic index: TBA
Histology: under way
Variety of reproductive states
[immature, mature, gravid/pregnant]
Immature,
or early developing gonads

20. Contaminant Analyses:
Feasible to measure individuals liver contaminant levels
Key contaminants measured at high concentrations in shiner perch and Pacific staghorn sculpin
Relatively higher levels of contaminants in livers of Pacific staghorn sculpin
Liver contaminant concentrations will be correlated with indices of endocrine disruption within individuals
Allows analysis of whether pollutants, individually or in combination, are associated with endocrine disruption & associated reductions in physiological viability
Different endocrine systems will likely exhibit different sensitivities to contaminants
Will point to potential contaminant thresholds of effects in fish (further testing needed)

21. Emphasis: Stress (HPI) Axis Continuing Analyses & Emphasis 2007
a) field work -Aug 2007, completed
b) cortisol measurement, completed, data analysis phase underway
c) interrenal histology, underway
d) hepatic contaminants measurement (underway) & correlation
d) cortisone measurement (11β-HSD-2)
e) ACTH-challenge (is interrenal responsiveness impaired?)
f) ectoparasitic infestation assessment
g) growth endocrine system impacts (indices: IGF-I, IGFBPs, cartilage growth activity) impacts
h) steroidogenic enzyme expression (StAR, 11β-hydroxylase, 11β-HSD-2)
b-d = potential bioassays for disruption (screening)

22. Thyroid (HPT) Axis Disruption –Studies proposed for 2007