1. Filename.ppt East Canyon Creek Health: Impacts from Reduced Flows and Endocrine Disrupting Compounds Swaner Preserve and EcoCenter Utah State University August 29, 2011 Michael Luers mluers@sbwrd.org

2. Filename.ppt Presentation Summary  Background Information  Issues facing East Canyon Creek  Introduction to Endocrine Disrupting Compounds (EDCs)  SBWRD’s research efforts

3. Filename.ppt

4. Brown Trout (Salmo trutta) Bonneville Cutthroat (Oncorhynchus clarki) East Canyon Creek Provides Habitat for Sensitive Trout Species

5. Filename.ppt East Canyon Creek is Drying Up Year7Q10 198816.0 cfs 19933.5 cfs 20031.8 cfs

6. Filename.ppt What Is The Endocrine System? The Endocrine System: System of glands each of which secrete a type of hormone Compounds that block, mimic, stimulate or inhibit the endocrine system – Endocrine Disrupting Compounds (EDCs)

7. Filename.ppt Potential Sources of Endocrine Disrupting Compounds (EDCs) Natural Hormones Pharmaceuticals and Personal Care Products Pesticides Detergents Industrial Compounds (Natural and synthetic estrogen based hormones are of greatest concern in our situation)

8. Filename.ppt insect repellent chemical DEET antibacterial hand soap triclosan birth control pill ethinyl estradiol polycarbonate plastic bisphenol-a Some of these products contain Endocrine Disrupting Compounds (EDCs) coffee caffeine fire extinguisher TDCPP, TCEP, TCPP

9. Filename.ppt Detection of EDCs Possible by Advances in Analytical Methods Liquid Chromatography/Mass Spectrometer

10. Filename.ppt Public Perception of EDCs in Water is a Growing Concern

11. Filename.ppt Natural and Synthetic Hormones Thought to be the EDCs of Greatest Concern MicroconstituentType/Purpose AcetaminophenPain Relief CaffeineStimulant CarbamazepineAnti-Epileptic CotinineStimulant DiazepamAnti-Anxiety Estrone Natural Hormone Estradiol Ethinyl Estradiol - 17  Synthetic Hormone FluoxetineAnti-Depressant Progesterone Natural Hormone SulfamethoxazoleAntibiotic Testosterone Natural Hormone TrimethoprimAntibiotic TriclosanAnti-Microbial

12. Filename.ppt Sources of Estrogens  Mature woman 4.8 μg estriol 3.5 μg estradiol 8.0 μg estrone  Post Menopause Woman 7 μg/day (all 3)  Men 7 μg/day (all 3)  Pregnant Woman 6,000 μg estriol 600 μg estrone 259 μg estradiol  Synthetic Hormones 26% of ethyl estradiol in birth control pills is excreted  Hormone & estrogen replacement therapy 65% estradiol excreted 15% estrone excreted

13. Filename.ppt Current Literature Describes Potential Effects of EDC Exposure for Male Fish Estrogenicity (feminization) Vitellogenin induction in male fish Intersex fish Skewed sex ratios Population collapse (Nash et al, 2004) GREATERIMPACTGREATERIMPACT

14. Filename.ppt Concentrations of Estrogens that begin to affect Male Fish Inducement of vitellogenin production ≈ 5 ng/L 17β-estradiol estriol is 30 times less potent than above ≈ 3.2 ng/L for estrone ≥ 1 ng/L for 17α-ethinyl estradiol Inducement of intersex ≈ 10 ng/L for estrone, or 17β-estradial estriol is 100 times less potent than above ≈ 4 ng/L for 17α-ethinyl estradiol estrogenic substances are additive Boulder Creek White sucker

15. Filename.ppt Estrogenicity and the Environment “The occurrence of feminized fish is associated with effluent discharges … the incidence and severity is positively correlated with the proportion of treated sewage effluent in receiving waters.”

16. Filename.ppt Vitellogenin (Vtg) is a Biomarker of Estrogen Exposure in Male Fish Egg yolk protein Vtg not normally found in male fish Vtg detected after exposures less than 1.0 ng/L (ppt)

17. Filename.ppt Estrogen Activity Measured by E-Screen Bioassay Breast cancer cell line with growth response to estrogen Negative Control Positive Control Reported as estradiol equivalents (MRL = 0.030 ppt)

18. Filename.ppt Timeline of Research Efforts

19. Filename.ppt SBWRD Collects Influent & Effluent Grab Samples Initial Sampling

20. Filename.ppt Despite Advanced Treatment, EDCs Detected in Treated Wastewater

21. Filename.ppt Summary of Effluent Test Results ConstituentSamples Detection Frequency AVG (ng/L) MRL (ng/L) E-Screen Bioassay18100%0.690.03 Cotinine15100%291.0 Sulfamethoxazole24100%8461.0 Gemfibrozil1995%851.0 Trimethoprim2295%731.0 Carbamazepine3494%815.0 Triclosan2090%365.0 Ibuprofen1989%291.0 Caffeine2488%233.0 Fluoxetine2488%501.0 Diazepam1747%871.0 Ethinyl Estradiol- 17  3429%5.51.0 Estrone3426%471.0 Estradiol3418%1.61.0 Acetaminophen2413%2.91.0 Iopromide20n/a12 1 5.0 Testosterone28n/a1.2 1 1.0 1 Result shown is a single detection (not an average)

22. Filename.ppt Ozone Oxidation Recommended as Treatment Technology of Choice GAC Filtration Ozone/Peroxide UV/Peroxide NF/RO not considered due to cost and concentrate disposal constraints

23. Filename.ppt What Should the Treatment Target Be? No Federal or State Guidance. Literature review of environmentally safe levels for the two most common constituents: Estrogen Activity Carbamazepine Measurable estrogen effects at 1 ppt (Purdom et al, 1994) Carbamazepine toxicity values at ppm (acute) and ppb (chronic)

24. Filename.ppt Sex Ratio Investigation Recent Efforts to Identify Impacts of EDCs on Downstream Trout Sentinel Study

25. Filename.ppt Sentinel Study conducted to determine if EDC concentrations in treated wastewater (effluent) are high enough to induce vitellogenesis in sentinel (caged) fish? Sex Ratio Study conducted to determine if there is a sex ratio problem (males vs. females) downstream? – worse case scenario! Are EDCs accumulating in the tissues of downstream fish? Study Objectives

26. Filename.ppt Sentinel Study Methods Holding pen in effluent aeration basin Holding pen at fish hatchery

27. Filename.ppt Sentinel Study Methods Blood Sample (0.5mL) Collected From Each Fish

28. Filename.ppt Sentinel Study Methods Vtg Analysis Requires Blood Plasma

29. Filename.ppt Sentinel Study Results (Rainbow Trout) ControlECWRF Baseline3 weeksBaseline3 weeks Vitellogenin (ng/ml) 0.343 ± 0.090.110 ± 0.030.136 ± 0.05 Total length (mm)255.2 ± 3.6265.6 ± 3.9250.0 ± 4.9263.9 ± 5.3 Total weight (g)188.3 ± 8.0207.9 ± 9.8181.4 ± 9.6216.6 ± 11.8 GSI 0.21 ± 0.07 0.19 ± 0.06

30. Filename.ppt Sentinel Study Results (Rainbow Trout) ControlECWRF Baseline3 weeksBaseline3 weeks Vitellogenin (ng/ml) 0.343 ± 0.090.110 ± 0.030.136 ± 0.0526.049 ± 9.02 Total length (mm)255.2 ± 3.6265.6 ± 3.9250.0 ± 4.9263.9 ± 5.3 Total weight (g)188.3 ± 8.0207.9 ± 9.8181.4 ± 9.6216.6 ± 11.8 GSI 0.21 ± 0.07 0.19 ± 0.06

31. Filename.ppt Sex Ratio Investigation Methods ElectrofishingTransport Holding

32. Filename.ppt Sex Ratio Investigation Methods Gender Determination: Palpitation or Necropsy

33. Filename.ppt Summary of Brown Trout Collected For the Sex Ratio Investigation 71 Fish Captured 43 Released (Sexually Mature) 28 Harvested (Gender by Necropsy) 5 Additional Fish Sampled Upstream (Negative Control)

34. Filename.ppt Excess Number of Female Fish Not Seen in the Downstream Population

35. Filename.ppt Sex Ratio Investigation Results Altered Sex Ratio Not Seen in Brown Trout 38 Female (54%) to 33 Male (46%) Vtg Not Detected in Male Brown Trout ( 4 to 1 ratio stream flow/effluent)

36. Filename.ppt 4:1 Dilution of Effluent in East Canyon During Sex Ratio Study Future Research: Sample Vitellogenin During Periods of Low Stream Flow Upstream Flow 75% Effluent Flow 25% Downstream Flow 100%

37. Filename.ppt Pharmaceuticals Found in Sentinel and Stream Fish Tissues * Common Name Pharmaceutical Name Fillet Downstream Liver Downstream Sentinel Fillet Sentinel Liver Caffeine 1.22.4 BenadrylDiphenhydramine0.2952.850.1876.77 HypertensionPropranolol0.901.2 Blood pressure control Diltiazem0.2050.030.46 TegretolCarbamazepine0.491.0 AntidepressantParoxetine0.668.8 Prozac (metabolite) Norfluoxetine20.081.7 ProzacFluoxetine18.561.0 Zoloft (metabolite)Desmethylsertraline140.010.2533.3 ZoloftSertraline92.0 ValiumDiazepam2.69.0 Cholesterol control Gemfibrozil22.3 *Results are presented as an average of all positive test results, results in units of nanograms/gram

38. Filename.ppt EDCs Can Be Regulated at Several Points in Their Lifecycle

39. Filename.ppt What is SBWRD Doing Now to Control EDCs? In the general provisions of SBWRD’s pretreatment program it states: “To prevent the introduction … of pollutants which may pass through the system, inadequately treated, into receiving waters…”

40. Filename.ppt Given the Potential Impact to Fish SBWRD has Implemented Steps for Source Control of EDCs Established unused medication take back programs with local law enforcement and Recycle Utah Working to establish a minimum base stream flow Identified all health care facilities and met with hospital officials to review disposal practices Continue with research during low stream flow periods of the year

41. Filename.ppt Summary and Conclusions:  EDCs detected at low concentrations in tertiary treated wastewater  Effluent (100%) EDCs concentrations are high enough to induce vitellogenesis in Rainbow Trout  Concentration of EDCs don’t appear to have altered the sex ratio of downstream Brown Trout (4:1, stream flow/effluent ratio)  Stream flow can attenuate the EDC problem

42. Filename.ppt Recent Federal Activities: Endocrine-Disrupting Chemicals Exposure Elimination Act of 2011 (S. 1361) (H.R. 2521) on July 13, 2011 to facilitate cooperation between the National Institute of Environmental Health Sciences and EPA, and other regulatory agencies to reduce exposure to chemicals identified as endocrine disruptors.

43. Filename.ppt For more info visit: http://www.sbwrd.com Questions?

44. Filename.ppt Pharmaceuticals Found in Both Sentinel and Downstream Fish Tissues Results in units of nanograms/gram AnalyteBrand Name DownstreamSentinel FilletLiverFilletLiver DiphenhydramineBenadryl2.8 0.1 DiltiazemCardizem, Dilacor 0.3 CarbamazepineTegretol 0.61.1 ParoxetinePaxil 6.4 NorfluoxetineProzac (metabolite) 25 69 FluoxetineProzac 19 48 DesmethylsertralineZoloft (metabolite) 14012540 SertralineZoloft 75 DiazepamValium 9.8 GemfibrozilLopid 10.6