Impact of EDCs on Trout in an pEffluent Dominated Stream

2009 GWPC Water/Energy Sustainability SymposiumMi h l LMichael Luers

mluers@sbwrd.org

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Presentation Summary

SBWRD Background information Introduction to endocrine disrupting

compounds (EDCs), pharmaceuticals and personal care products (PPCPs)

Implications for aquatic life Implications for aquatic life SBWRD’s research efforts

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Morgan

Salt Lake

r Cre

ek

To Evanston, WY&Morgan

Summit

District

Boundary

ÉW RFEast Canyon

Silv

er

East Canyon Creek

§¦80

§¦80

&

To Salt Lake City

ÉW RF Water ReclamationFacility

ÉW R FSilver CreekWater ReclamationFacility

E C C kEas t Canyon Trunk

Silver Creek Trunk

McLeod Cr

Kimball Creek

"º2 42£¤40

Old Ranch Rd

Summit

Salt Lake

kUtah Olympic Park

The Canyons Resort

East Canyon Creek

Silver CreekMcLeod C

Silver Creek"º2 84

ToKamas

&

&

ky

Park CityMountain Resort k

Park

City

Silver Creek

Thayn

es C

anyo

n

To Heber

&

Mountain Resort k

Deer Valley Resortk

City

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SummitWasatch0 1 2

Miles

What are Endocrine Disrupting C d (EDC )?Compounds (EDCs)?

Natural and Synthetic yHormones

Pharmaceuticals and Personal Care Products

PesticidesPesticides

Detergents

Industrial Compounds

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SBWRD is Concerned About EDCs Discharged to East Canyon CreekDischarged to East Canyon Creek

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SBWRD’s Reason for Concern

Brown TroutBrown Trout(Salmo trutta)

Upstream reach of East

Bonneville Cutthroat(Oncorhynchus clarki)

pCanyon Creek in late summer

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(Oncorhynchus clarki)

Detection of Trace Level EDCs Possible by Ad i A l ti l M th dAdvances in Analytical Methods

Liquid Chromatography/Mass Spectrometer

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Public Perception of EDCs in Water is a Ch ll f W t P f i lChallenge for Water Professionals

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Natural and Synthetic Hormones Thought to be the EDCs of Greatest Concernto be the EDCs of Greatest Concern

Microconstituent MRL1 (ng/L) Type/PurposeAcetaminophen 1.0 Pain Relief

Caffeine 3.0 StimulantCarbamazepine 5.0 Anti-Epileptic

Cotinine 1.0 StimulantDiazepam 1.0 Anti-AnxietyEstroneEstrone 1.01.0 Natural HormoneNatural Hormone

EstradiolEstradiol 1.01.0 Natural HormoneNatural HormoneEthinyl EstradiolEthinyl Estradiol-- 1717 1.01.0 Synthetic HormoneSynthetic Hormone

Fluoxetine 1.0 Anti-DepressantProgesteroneProgesterone 1.01.0 Natural HormoneNatural Hormone

Sulfamethoxazole 1.0 AntibioticTestosteroneTestosterone 1.01.0 Natural HormoneNatural HormoneTrimethoprim 1.0 Antibiotic

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Triclosan 5.0 Anti-Microbial1Method Reporting Limit

Sources of EstrogensSources of Estrogens Mature woman

4 8 t i l S th ti H4.8 μg estriol3.5 μg estradiol8.0 μg estrone

Synthetic Hormones26% of ethyl estradiol in birth control pills is

Post Menopause Woman7 μg/day (all 3)

Men

birth control pills is excreted

7 μg/day (all 3) Pregnant Woman

6,000 μg estriol

Hormone & estrogenreplacement therapy65% estradiol excreted

600 μg estrone259 μg estradiol

65% estradiol excreted15% estrone excreted

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Timeline of Research Efforts

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EDCs Detected at Low Concentrations in ECWRF Effluentin ECWRF Effluent

JUNE 2007 SBWRD Collects Influent & Effluent Grab Samples

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SBWRD Collects Influent & Effluent Grab Samples

Historical EDC/PPCP DetectionsConstituent Samples Detection

FrequencyAVG(ng/L)

MRL(ng/L)Frequency (ng/L) (ng/L)

E-Screen Bioassay 18 100% 0.69 0.03Cotinine 15 100% 29 1.0

Sulfamethoxazole 24 100% 846 1.0G fib il 19 95% 85 1 0Gemfibrozil 19 95% 85 1.0

Trimethoprim 22 95% 73 1.0

Carbamazepine 34 94% 81 5.0

Triclosan 20 90% 36 5.0Ibuprofen 19 89% 29 1.0Caffeine 24 88% 23 3.0

Fluoxetine 24 88% 50 1.0

Diazepam 17 47% 87 1.0

Ethinyl EstradiolEthinyl Estradiol-- 1717 34 29% 5.5 1.0EstroneEstrone 34 26% 47 1.0

EstradiolEstradiol 34 18% 1.6 1.0Acetaminophen 24 13% 2.9 1.0p

Iopromide 20 n/a 121 5.0TestosteroneTestosterone 28 n/a 1.21 1.0

DEET 1 n/a 4371 25TDCPP 1 n/a 2221 25

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TDCPP 1 n/a 222 25Tris (2-chloroethyl) phosphate 1 n/a 1661 25

1Result shown is a single detection (not an average)

Current Literature Describes Potential Eff t f EDC E f Fi hEffects of EDC Exposure for Fish

Estrogenicity (feminization)Estrogenicity (feminization)Vitellogenin induction in male fishIntersex fishIntersex fishSkewed sex ratiosP l ti llPopulation collapse

BioaccumulationChronic Toxicity

(Nash et al 2004)

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(Nash et al, 2004)

Vitellogenin (Vtg) is a Biomarker of E t E i M l Fi hEstrogen Exposure in Male Fish

Egg yolk protein

Vtg not normally found inVtg not normally found in male fish

Vtg detected after exposures l 1 0 /L ( t)

g pas low as 1.0 ng/L (ppt)

No known short-term health consequencesconsequences

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Connection Between Vtg Induction and M S Eff t N t Y t U d t dMore Severe Effects Not Yet Understood

FEMALES:MALES

?

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Estrogen Activity Measured by E Screen BioassayE-Screen Bioassay

Breast cancer cell line with growth response to estrogen

Negative Control

Positive Control

Reported as estradiol equivalents (MRL = 0.030 ppt)

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Valuable Lessons Learned Along the Way

OCT 200 NOV 2007OCT 2007GAC Benchscale Test

NOV 2007Brown Trout Investigation

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Ozone Oxidation Recommended as Treatment Technology of ChoiceTreatment Technology of Choice

GAC FiltrationOzone/Peroxide UV/PeroxideGAC Filtration

MAR 2008

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Treatment Technology TestingNF/RO not considered due to cost and concentrate disposal constraints

EDC Removal Using Existing Infrastructure: GMF to GACInfrastructure: GMF to GAC

Existing Granular Media Filter

GAC Contactor

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(Parkson Dynasand)

Efforts to Identify Impacts of Effluent on D t T tDownstream Trout

NOV 2008Sex Ratio Investigation

DEC 2008Sentinel Study

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Study Objectives

Determine if EDC concentrations are high enough to

induce vitellogenesis by holding sentinel fish in the

effluent

Determine if downstream fish populations were being

feminized by conducting a field investigation of sex ratios

Evaluate fish tissue for bioaccumulation of EDCs Evaluate fish tissue for bioaccumulation of EDCs

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Sentinel Study Methods

Holding pen in effluent

i

Holding pen at fish

h h aeration basin

hatchery

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Sentinel Study Methods

Blood Sample (0.5mL) Collected From Each Fish

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Sentinel Study MethodsVtg Analysis Requires Blood Plasma

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Vtg Levels Increased in Fish Exposed to ECWRF EffluentECWRF Effluent

Control ECWRFControl ECWRFBaseline 3 weeks Baseline 3 weeks

Vitellogenin 0 343 ± 0 09 0 110 ± 0 03 0 136 ± 0 05 26 049 ± 9 02Vitellogenin (ug/ml)

0.343 ± 0.09 0.110 ± 0.03 0.136 ± 0.05 26.049 ± 9.02

Total length (mm) 255.2 ± 3.6 265.6 ± 3.9 250.0 ± 4.9 263.9 ± 5.3

Total weight (g) 188.3 ± 8.0 207.9 ± 9.8 181.4 ± 9.6 216.6 ± 11.8

GSI 0.21 ± 0.07 0.19 ± 0.06

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Objectives of Brown Trout Sex Ratio InvestigationInvestigation

Compare Females vs MalesCompare Females vs Males

Sample Size Goal (n=70)

Expected 50:50 Ratio

Blood Samples (Vtg)

Tissue Samples (Bioaccumulation)( )

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Sex Ratio Investigation Methods

Electrofishing Transport Holding Pen

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Sex Ratio Investigation Methods

Gender Determination: Palpitation or Necropsy

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Sex Ratio Investigation Methods

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Summary of Fish Collected For the Sex R ti I ti tiRatio Investigation

71 Fish Captured71 Fish Captured

43 Released (Sexually Mature)

28 Harvested (Gender by Necropsy)

5 Additional Fish Sampled ( C )Upstream (Negative Control)

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Sex Ratio Investigation Results

Altered Sex Ratio Not Seen

38 Female (54%) to 33 Male (46%)

Vtg Not Detected in Male Fish

4 t 1 Dil ti f Effl t i St4 to 1 Dilution of Effluent in Stream

Fish Tissue Results – Still Pending

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Conclusions

EDCs detected at low concentrations (ppt) in ECWRF(ppt) in ECWRF

Effluent EDCs concentrations high enough i d i ll i i lto induce vitellogenesis in male trout

ECWRF effluent does not appear to have altered the sex ratio of the brown trout

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Observations

Additional research needed to understand which of the compounds are responsible forwhich of the compounds are responsible for vitellogenin induction in male fish

Bioassays and biomarkers are cost effective Bioassays and biomarkers are cost effective indicators of estrogenicity potential

EDC’s can be removed using advanced treatment technologies

Water professionals need to be prepared to address EDC concerns with customers

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Questions?

For more info visit: http://www sbwrd com

Questions?

For more info visit: http://www.sbwrd.com

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Concentrations of Estrogens that begin t ff t M l Fi hto affect Male Fish

Inducement of vitellogenin production• ≈ 5 ng/L 17β-estradiol• estriol is 30 times less potent than above

3 2 /L f t• ≈ 3.2 ng/L for estrone• ≥ 1 ng/L for 17α-ethinylestradiol

Inducement of intersex• ≈ 10 ng/L for estrone, or 17β-estradial

t i l i 100 ti l t t th b• estriol is 100 times less potent than above• ≈ 4 ng/L for 17α-ethinylestradiol• estrogenic substances are additive

White

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White sucker

Fish Tracking Results for Negative Control Test Group

H t h G (N ti C t l)Hatchery Group (Negative Control):

50 Fish

7 Mortalities

4 Lost Tags4 Lost Tags

39 Pre & Post Exposure

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Fish Tracking Results for Effluent Exposure Group

ECWRF Effl t GECWRF Effluent Group:

50 Fish

10 Mortalities

9 Lost Tags9 Lost Tags

31 Pre & Post Exposure

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