ecological effects of endocrine disruption: quantifying generational effects david walker, phd...
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![Page 1: Ecological Effects of Endocrine Disruption: Quantifying Generational Effects David Walker, PhD University of Arizona David Walker 1, Nick Paretti 2, Gail](https://reader034.vdocuments.site/reader034/viewer/2022051316/56649d395503460f94a12bc5/html5/thumbnails/1.jpg)
Ecological Effects of Endocrine Disruption: Quantifying Generational Effects
David Walker, PhDUniversity of Arizona
David Walker1, Nick Paretti2, Gail Cordy2, Timothy S. Gross3, Edward T. Furlong4, Steven D. Zaugg4
Dana W. Kolpin5, and Dennis McIntosh6
1 University of Arizona, Environmental Research Laboratory, 2601 E. Airport Dr., Tucson, AZ 85706 [email protected]
2 USGS., WRD, 520 N. Park Ave, Suite 221, Tucson, AZ 85719 [email protected]
3 USGS-Florida Caribbean Science Center, 7920 NW 71st St., Gainesville Florida, 32653 [email protected]
4 USGS, National Water Quality Laboratory, Denver Federal Center, P.O. Box 25046, MS 407, Lakewood, CO 80225-0046 [email protected]
5 USGS, WRD, P.O. Box 1230, Iowa City, IA 52240 [email protected]
6 Delaware State University, 1200 N. DuPont Highway Dover, DE 19901 [email protected]
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Aquatic Ecology and Endocrine Disruption
• An organisms ability to better-exploit a resource (or group of resources) in the face of environmental stress and inter-specific competition, coupled with conservation of the genetic material enabling this exploitation, is what drives speciation.
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• Genetic conservation of traits is initiated, and sustained by, subtle behavioral cues for mating, spawning, aggression, territoriality, avoidance, etc.
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Quantification and Research Design Issues; One Size Does Not Fit All
Mech
anistic U
nd
erstand
ing
Ecological Significance
Genetic
Biochemical
Physiological
Behavioral
Reproductive
Assemblages
Histopathological
Immunological
Bioenergetic
Populations
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• Unlike semi-arid or north-temperate regions, effluent-dependent water’s (EDW’s) in arid regions usually contain 100% effluent year-round.
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Roger Road WWTP
Tucson
Santa Cruz River
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This Study
• Laboratory study with control, replication, and randomization.
• Use fish native to the region (largely pollution-tolerant).
• Concentrate on long-term, persistent compounds.
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•Morphology epitomizes adaptations to living in turbid, flashy rivers with extremes in environmental conditions.
•Pollution-tolerant
•Omnivorous, adults grow to 2 feet in length.
•Large enough to survive multiple venipuncture events.
•Readily available supply from USFWS Deter National Fish Hatchery and Technology Center
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Treatment Treatment
Control
Control
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Water temperature maintained between 25-29o C.
Photoperiod was maintained at 12 hours of light and dark
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Treatment/Dosages
• Fish in raceways exposed for 3 months per treatment
• 1st treatment = 1/3 by volume treated ww and 2/3 water treated by RO
• 2nd treatment = 2/3 by volume treated ww and 1/3 water treated by RO
• 3rd treatment = full strength treated ww
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0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
Dose 1 Dose 2 Dose 3
Detergent Metabolites
Fire/Flame Retardants
Fragrances/flavors
Fuels/PAHs
Plasticizers/antioxidants
Herbicides/insecticides
Non-prescription drugs
Prescription Drugs SteroidsHousehold Wastewater Compounds
Results in µg/L
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Males
17β-estradiol (pg/mL)
Control (n = 6): 217.3
Treatment (n = 13): = 547.4
11-ketotestosterone (pg/mL)
Control (n = 6): = 820.8
Treatment (n = 13): = 473.5
Vitellogenin (mg/mL
Control (n = 6): = 0.09
Treatment (n = 13): = 0.32
17-B
ETA
(pg/
ml)
100
200
300
400
500
600
700
800
900
1000
1100
Control Treatment
T/C
11-K
ETO
(pg/
ml)
0
200
400
600
800
1000
1200
1400
Control Treatment
T/C
VIT
ELLO
(pg/
ml)
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
Control Treatment
T/C
P = 0.0016 P = 0.021 P = 0.036
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Females
17β-estradiol (pg/mL)
Control (n = 54): 568.2
Treatment (n = 47): 403.7
11-ketotestosterone (pg/mL)
Control (n = 54): 591.3
Treatment (n = 47): 530.4
Vitellogenin (mg/mL)
Control (n = 54): 0.18
Treatment (n = 47): 0.18
17-B
ETA
(pg/
ml)
0
200
400
600
800
1000
1200
Control Treatment
T/C
Missing Rows 25
Oneway Anova
Oneway Analysis of 17-BETA (pg/ml) By T/C
11-K
ETO
(pg/
ml)
100200300400500600700800900
1000110012001300
Control Treatment
T/C
Missing Rows 25
Oneway Anova
Oneway Analysis of 11-KETO (pg/ml) By T/C
VIT
ELLO
(pg/
ml)
-0.1
0.1
0.3
0.5
0.7
0.9
1.1
Control Treatment
T/C
Missing Rows 25
Oneway Anova
Oneway Analysis of VITELLO (pg/ml) By T/C
P = 0.001 P = 0.286 P = 0.986
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5
5.5
6
6.5
7
6
6.25
6.5
6.75
7
7.25
0.1
0.3
0.5
0.7
0.9
ln 17-beta
5 5.5 6 6.5 7
ln 11-keto
6 6.25 6.5 6.75 7 7.25
sqrtvitello
.1 .2 .3 .4 .5 .6 .7 .8 .9 1
5.5
6
6.5
7
5.5
6
6.5
7
0.1
0.2
0.3
0.4
0.5
0.6
0.7
ln 17ß
5.5 6 6.5 7
ln 11Kt
5.5 6 6.5 7
ln vtg
.1 .2 .3 .4 .5 .6 .7
ln 17β ln 11-keto
ln Vtg
ln 17β 1.00 -0.89 0.74
ln 11-keto
-0.89 1.00 -0.73
ln Vtg 0.74 -0.73 1.00
Control Males Treatment Malesln 17β ln 11-
ketoln Vtg
ln 17β 1.00 -0.50 0.69
ln 11-keto
-0.50 1.00 -0.28
ln Vtg 0.69 -0.28 1.00
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5
5.5
6
6.5
7
5
5.5
6
6.5
7
0.1
0.3
0.5
0.7
0.9
ln 17ß
5 5.5 6 6.5 7
ln 11Kt
5 5.5 6 6.5 7
ln vtg
.1 .2 .3 .4 .5 .6 .7 .8 .9 1
5
5.5
6
6.5
7
5
5.5
6
6.5
7
0.1
0.3
0.5
0.7
0.9
ln 17ß
5 5.5 6 6.5 7
ln 11Kt
5 5.5 6 6.5 7
ln vtg
.1 .2 .3 .4 .5 .6 .7 .8 .9 1
ln 17β ln 11-keto
ln Vtg
ln 17β 1.00 -0.67 0.52
ln 11-keto
-0.67 1.00 -0.70
ln Vtg 0.52 -0.70 1.00
Control Females Treatment Females
ln 17β ln 11-keto
ln Vtg
ln 17β 1.00 0.11 0.28
ln 11-keto
0.11 1.00 -0.36
ln Vtg 0.28 -0.36 1.00
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We believe this study is highly representative of the biological effect of
endocrine-disrupting compounds on aquatic organisms at the landscape scale.
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• This study quantified hormonal impairment; but not fertility or fecundity
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New (Ongoing) Research
• Repeat design. • Quantify impairment following a 4 month
exposure .• Aromatase (?),GnRh, GtH I and II,
steroidal hormones, protein development, GSI, HSI, and histology.
• Provide environmental stimuli for spawning.
• Collect eggs and quantify fertility and fecundity of parent generation.
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• F1 generation grown in effluent for one year for the determination of sex ratio and hormonal/morphological change.
• Compounds identified using POCIS and SPMD (David Alvarez/USGS-CERC).
• Sperm quality (Jill Jenkins/USGS)
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Summary
• Biomarkers infer reproductive impairment, they don’t quantify it.
• Physiological impairment can occur without morphological change.
• Bio-indicator species selection should be ecosystem and objective-specific.
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Acknowledgements
USFWS Dexter National Fish Hatchery and Technology Center.
USGS Toxics and Hydrology Program
This work was supported by State of Arizona funding,
through the Technology and Research Initiative..