Download Item_3_-_Endocrine_Disruptors_in_Fish

Environmental Endocrine Disruption
Effects Assessment
In San Francisco Bay Fishes
Shiner Surfperch & Pacific Staghorn Sculpin
Kevin M. Kelley
Environmental Endocrinology Laboratory
Inst. 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
ENDOCRINE SYSTEMS
regulate
essential physiological functions
Reproduction - Growth & Repair - Metabolism
Defense/Immune - Thyroid - Developmental
Stress Responses - Neuroendocrine - others
Homeostasis, Adaptation, Survival
Because endocrine systems are . . .
. . . highly sensitive, responsive
. . . respond to presence of environmental
stressors to a degree commensurate with the
need to maintain homeostasis
. . . reflective in their response to the
specific type(s) of stressor present
. . . measures of ‘endocrine disruption’ are
increasingly being pursued 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
http://toxics.usgs.gov/photo_gallery/sanfran.html
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
Shiner Surfperch
Cymatogaster aggregata
Pacific Staghorn Sculpin
Leptocottus armatus
2006 Field Sites
San Pablo Bay
Berkeley Waterfront
Oakland Inner Harbor
Redwood City
far-field reference:
Tomales Bay
Bodega Bay
Far Field Sites
Bodega Bay
Tomales Bay
STRESS
RESPONSE
AXIS
Stress!
BRAIN
HYPOTHALAMUS
CRH
PITUITARY
anterior
posterior
circulation
HypothalamoPituitaryInterrenal
Axis
(HPI Axis)
ACTH
Interrenal
tissues
↑↑CORTISOL
adaptational responses
Stress Response
Shiner Surfperch
a
Plasma Cortisol
Concentration (ng/ml)
a
1200
b
b
800
400
21
13
21
10
Berkeley
Waterfront
San Pablo
Bay
0
Redwood
City
Oakland
Inner
Harbor
Plasma Cortisol
Concentration (ng/ml)
Stress Response
a
a
1200
Pacific Staghorn Sculpin
800
b
400
c
17
16
13
Redwood
City
Oakland
Inner
Harbor
Berkeley
Waterfront
19
0
San Pablo
Bay
STRESSOR
impaired
BRAIN
HPI Axis
HYPOTHALAMUS
CRH
PITUITARY
anterior
posterior
circulation
ACTH
Interrenal
tissues
EDCs ?
REDUCED
cortisol
response
Other wild fish species living in affected
marine environments exhibit
impairment of their neuroendocrine
stress response system
...experimental studies have
shown that contaminants,
such as PCBs,
impair stress responses in fish
(Quabius et al., 2005; Vijayan et al., 2006)
THYROID
ENDOCRINE
SYSTEM
BRAIN
HYPOTHALAMUS
TRH
PITUITARY
anterior
posterior
circulation
HypothalamoPituitaryThyroid
Axis
(HPT Axis)
TSH
Thyroid
gland
T4 , T3
variety of physiological actions
Thyroid Hormones
T4
3,5,3’,5’-tetraiodothyronine (thyroxine)
T3
3,5,3’-triiodothyronine
• broad physiological actions
• essential for brain/neural development
• permissive to somatic growth
• important regulators of metabolism
Concentration, ng/ml
Plasma Thyroxine (T4)
Thryoxine (T4) Levels in Shiner Surfperch
80
Shiner
Surfperch
60
40
20
0
(11)
(19)
(19)
San Pablo
Bay
Berkeley
Waterfront
Redwood
City
(9)
Oakland
Inner Harbor
(14)
Bodega
Bay
Concentration, ng/ml
Plasma Thyroxine (T4)
Thryoxine (T4) Levels in Pacific Staghorn Sculpin
Pacific
Staghorn
Sculpin
80
60
40
20
0
(8)
San Pablo
Bay
(13)
(17)
Berkeley
Waterfront
Redwood
City
(12)
Oakland
Inner Harbor
(5)
Tomales
Bay
Thryoxine (T4) Levels in SF Bay Fish
Concentration, ng/ml
Plasma Thyroxine (T4)
100
80
Pacific
Staghorn
Sculpin
60
40
Shiner
20
0
(8)
(11)
San Pablo
Bay
(13)
(19)
Berkeley
Waterfront
(17)
(19)
Redwood
City
(12)
(9)
(5)
(14)
Oakland Tomales/ Bodega
Inner Harbor
Bay
Surfperch
BRAIN
impaired
HYPOTHALAMUS
HPT Axis
TRH
PITUITARY
anterior
posterior
circulation
TSH
Thyroid
Gland
EDCs ?
Altered
T4 (T3) production
altered physiology
PBDEs and PCBs are implicated in
thyroid endocrine disruption:
…in mammalian studies, PBDE exposure causes
reduction of thyroid hormone level and production
(e.g., Skarman et al., 2005; Boas et al., 2006)
…in fish, PBDE exposure causes reduction in T4, but
not T3 (Tomy et al., 2004 –lake trout; Lema et al., 2006 –
fathead minnow)
…in fish, PCB exposure alters T4 and T3 levels (LeRoy
et al., 2006 –Atlantic croaker)
Reproductive
Steroids
Plasma 17β-Estradiol Concentrations
E2 (pg/ml)
400
300
Pacific Staghorn Sculpin
200
N.D.
for
Testosterone
(selected samples)
100
0
Female
[winter]
Female
[summer]
Male
[winter]
English Sole
[Orange County]
[n=13-29/bar]
Male
Female
[immature]
all SF Bay
locations
tested
August 2006 Oakland Inner Harbor
Shiner Surfperch ?
17α-Ethinylestradiol (EE2)
in Fish Plasma
Plasma EE2 Concentration
(pg/ml)
200
150
100
50
N.D. N.D.
0
Turbot
WWTP-affected site
Shiner
Surfperch
Pacific Staghorn
Sculpin
(Tomales Bay, Redwood City,
Oakland Inner Harbor, Berkeley, San
Pablo Bay (Loch Lohmand)
The
Growth
Endocrine
Axis
PITUITARY
Growth Hormone
LIVER
IGFBP-3
IGFBP-1
IGF-I
(IGFBPs 2,4-6)
IGFBP
“IGF Axis”
IGF
IGF
IGF
REC
target
cell
GROWTH
Jack Mackerel
Trachurus symmetricus
Stress Effects on Plasma
IGFBP Levels
Stressed
Rat Plasma
Control
45 kDa
30 kDa
IGFBP-1
30 kDa
28 kDa
increased IGFBP-1 >> growth inhibition
cell culture work supports this
Endocrine Disruption—SF Bay Fish
[Yellow=Year-1 fish analyses]
**
Cortisol & Cortisone
Thyroxine & Triidothyronine
17β-Estradiol & 17α-Ethinylestradiol
Testosterone & 11-ketotestosterone
?!
POTW,
seasonal
effects??
IGF-I & IGFBPs (June 2007)
Hepatic contaminants (chlorinated pesticides, PCB congeners, PAHs)
=spatial differences detected
Study Sites 2007
July-August
San Pablo Bay
(Loch Lohmand Marina)
Richmond (Laurentian Channel, DDTs)
Berkeley Waterfront
Oakland Inner Harbor
San Leandro Bay (PCBs)
Redwood City
South of Dunbarton (POTW)
far-field reference:
Tomales Bay
Bodega Bay
Stress (HPI) Axis –Continuing Analyses 2007
Field season July-Aug 2007
•
•
•
•
•
•
•
a) interrenal histology
b) hepatic contaminants correlation
c) ACTH-challenge*
d) cortisone measurement* (11β-HSD-2)
e) ectoparasitic infestation assessment 
f) growth endocrine system (IGF-I, IGFBPs)
g) steroidogenic enzyme expression*
(StAR, 11β-hydroxylase, 11β-HSD-2)
With an emphasis on Oakland Inner Harbor (San Pablo Bay, San Leandro Bay??)
vs. control sites
*b-d = potential bioassays for disruption (EDC screening?)
Thyroid (HPT) Axis –Continuing Analyses 2007
Field season July-Aug 2007
•
•
•
•
•
•
•
a) T3 measurement*
b) hepatic contaminants correlation
c) thyroid histology [?]
d) TSH-challenge* [?]
e) peripheral deiodinases (D1-3)* [?]
f) thyroid hormone receptors (TRα & TRβ)* [?]
g) growth endocrine system (IGF-I, IGFBPs)
*b-d = potential bioassays for disruption (EDC screening?)
?? QUESTIONS
Liver Contaminant Analysis, GC/MS
Selected Contaminants* Shiner
Surfperch
Pacific
Staghorn
Sculpin
Chlordane-gamma
1–4
Chlordane-alpha
3.5 – 15
2–5
6 – 10
trans-Nonachlor
cis-Nonachlor
4,4’-DDE
3.5 – 12
1.5 – 5
20 – 47
5 – 20
4 – 13
48 – 54
*range of ng/g wet
weight values shown
So-Calif would have 4, 4’-DDT
Nov. 2006
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:
1 – 16
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
2 – 35
*range of ng/g wet weight values
shown
Nov. 2006
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
Nov. 2006
Russell Fairey et al.
Moss Landing Marine
Laboratories
Southern California Sea Grant Program
San Francisco Estuary Institute
steroidogenic targets
cholesterol
mitochondria
StAR, PBR
P450scc
pregnenolone
progesterone
17a-hydroxypregnenolone
dehydroepiandrosterone
17a-hydroxyprogesterone
11-deoxycorticosterone
11-deoxycortisol
P45011b
corticosterone
aldosterone
cortisol
11b-HSD-2
Cortisone
(inactive)
androstenedione
testosterone
P450arom
estradiol-17b
Thyroxogenesis
Stress!!
Stress!!
handling
water quality
crowding / captivity
Stimulate the HPI Axis
elevated CORTISOL levels
Catabolic Pathways
↑ Glucose, other fuels
↑ Hepatic glucose production
↑ AA release from muscle
↑ Lipid breakdown
increased fuel
mobilization & availability
Overall:
Anabolic Pathways
↓Growth & Repair
↓Reproduction
↓Immune function
Overall: reduced
energyexpensive processes
Elthusa californica
. . . was commonly found in opercular chambers of shiner
surfperch (staghorn sculpin, to lesser degree)