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Quantifying Endocrine Disruption in a Threatened and Endangered Fish Species Dr. David Walker University of Arizona David Walker 1, Nick Paretti 2, Gail.

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Presentation on theme: "Quantifying Endocrine Disruption in a Threatened and Endangered Fish Species Dr. David Walker University of Arizona David Walker 1, Nick Paretti 2, Gail."— Presentation transcript:

1 Quantifying Endocrine Disruption in a Threatened and Endangered Fish Species Dr. David Walker University of Arizona David Walker 1, Nick Paretti 2, Gail Cordy 2, Timothy S. Gross 3, Dana W. Kolpin 5, and Dennis McIntosh 6 1 University of Arizona, Environmental Research Laboratory, 2601 E. Airport Dr., Tucson, AZ 85706 dwalker@ag.arizona.edu 2 USGS., WRD, 520 N. Park Ave, Suite 221, Tucson, AZ 85719 nvp@email.arizona.edu 3 USGS-Florida Caribbean Science Center, 7920 NW 71 st St., Gainesville Florida, 32653 tim_s_gross@usgs.gov 5 USGS, WRD, P.O. Box 1230, Iowa City, IA 52240 dwkolpin@usgs.gov 6 Delaware State University, 1200 N. DuPont Highway Dover, DE 19901 dmcintosh@desu.edu

2 Quantification and Research Design Issues; One Size Does Not Fit All Mechanistic Understanding Ecological Significance Genetic Biochemical Physiological Behavioral Reproductive Assemblages Histopathological Immunological Bioenergetic Populations

3 Unlike semi-arid or north-temperate regions, effluent-dependent water’s (EDW’s) in arid regions usually contain 100% effluent year- round.

4

5 Roger Road WWTP Built in 1951. Treated effluent is discharged into the Santa Cruz River or diverted into the city’s reclaimed water system. Treats the wastewater generated by a population of about 419,000. A capacity of 41 mgd and treated an average of 38 mgd from 2004 to 2005. Produces secondarily-treated wastewater

6 Roger Road WWTP Tucson Santa Cruz River

7 This Study Laboratory study with controls, replicates, and randomization. Use fish native to the region (largely pollution-tolerant). Framework or foundation for refinement of future studies. Varying doses of effluent (“treatments”). Concentrate on long-term, persistent compounds.

8 Bonytail Chub (Gila elegans)

9 Treatment Control

10 Water temperature maintained between 25-29 o C. Photoperiod was maintained at 12 hours of light and dark

11 Treatment/Dosages Fish in raceways exposed for 3 months per treatment 1 st treatment = 1/3 by volume treated ww and 2/3 water treated by RO 2 nd treatment = 2/3 by volume treated ww and 1/3 water treated by RO 3 rd treatment = full strength treated ww

12 Experimental Design Fish are PIT tagged Fish randomly assigned to tanks Blood collected from 10 randomly selected individuals within each raceway Blood drawn from the same individual fish contiguously over the extent of experiment 17β-estradiol, 11-ketotestosterone, and vitellogenin

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14

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16 Males - Overall 17β-Estradiol Control (n = 6): 217.3 Treatment (n = 13): = 547.4 11-ketotestosterone Control (n = 6): = 820.8 Treatment (n = 13): = 473.5 Vitellogenin Control (n = 6): = 0.09 Treatment (n = 13): = 0.32

17 Females - Overall 17β-Estradiol Control (n = 54): 568.2 Treatment (n = 47): 403.7 11-ketotestosterone Control (n = 54): 591.3 Treatment (n = 47): 530.4 Vitellogenin Control (n = 54): 0.18 Treatment (n = 47): 0.18

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20 Synergistic Effects Ratios of primary male and female sex hormones, in “undisturbed” populations would be expected to have an inverse relationship i.e. as one increased, the other would decrease. We could therefore assume that major deviations from this inverse relationship between male and female primary sex hormones, could be attributed to impairment.

21 ln 17βln 11- keto ln Vtg ln 17β 1.00-0.890.74 ln 11- keto -0.891.00-0.73 ln Vtg 0.74-0.731.00 Control MalesTreatment Males ln 17βln 11- keto ln Vtg ln 17β 1.00-0.500.69 ln 11- keto -0.501.00-0.28 ln Vtg 0.69-0.281.00

22 ln 17βln 11- keto ln Vtg ln 17β 1.00-0.670.52 ln 11- keto -0.671.00-0.70 ln Vtg 0.52-0.701.00 Control Females Treatment Females ln 17βln 11- keto ln Vtg ln 17β 1.000.110.28 ln 11- keto 0.111.00-0.36 ln Vtg 0.28-0.361.00

23 Eigenvalue2.49941.59831.35200.78430.43640.23060.0990 Percent35.705922.832819.314511.20386.23413.29501.4140 Eigenvectors Impairment0.032070.47871-0.599710.031980.52405-0.28832-0.22668 Det. Met-0.02386-0.69182-0.127190.137210.651810.205030.13755 Fire Ret.0.272060.146640.488350.742880.19262-0.10884-0.25528 Fragrances-0.453150.458510.068260.203180.170530.609720.37099 Herbicides-0.243540.130710.61232-0.518830.45492-0.26042-0.06864 HH Waste0.592040.168670.05748-0.082590.11987-0.159140.75579 Plasticizers0.556070.122270.05280-0.333150.114090.63094-0.38854 Males

24 Eigenvalue2.54241.37851.11610.94300.65820.27600.0858 Percent36.319919.695215.944013.47109.40303.94331.2262 Eigenvectors Impairment0.292450.18743-0.641530.257360.51884-0.35362-0.08546 Det. Met0.05636-0.564190.380590.434440.557230.180170.04430 Fire Ret.0.041970.511510.472400.59978-0.16230-0.31014-0.17655 Fragrances-0.434530.42877-0.210060.275960.176820.664760.18408 Herbicides-0.268750.341610.37217-0.519330.59522-0.22040-0.00263 HH Waste0.578080.218460.15670-0.069810.017240.100290.76047 Plasticizers0.560640.191470.11504-0.185900.090080.49575-0.58936 Females

25 Summary Significant hormonal impairment of both sexes, as compared to controls, at very low concentration of compounds. This impairment could have never been determined in a field study. Commonly-used parametric analyses are often inadequate in determining impairment.

26 Summary (cont) Determination of impairment using phased biomarkers. –Phase 1: Aromatase/GnRH –Phase 2: GtH I, GtH II –Phase 3: Sex hormones –Phase 4: Protein development (vtg, oocyte, spermiation) –Phase 5: Intersex/Morphological change

27 This study is highly representative of the biological effect of endocrine-disrupting compounds at the landscape scale.

28 Current and Future Research Fertility/fecundity and sex ratio/development of F2 generation (funded and initiated). Behavior. Treatments using streambed sediment from affected EDW’s. Assemblage Effects

29 Questions?

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