1. From Treatment Plants to Turbots: Data Suggesting Effects From Endocrine Disrupting Chemicals in Wastewater Discharged into the Pacific Ocean Jeffrey L. Armstrong, Ph.D. Environmental Assessment Division Orange County Sanitation District Jeffrey L. Armstrong, Ph.D. Environmental Assessment Division Orange County Sanitation District

2. Love those Acronyms!  CEC = Contaminants of Emerging Concern  CPC = Contaminants of Potential Concern  PPCP = Pharmaceuticals and Personal Care Products  EDC = Endocrine Disrupting Chemical  PPCP ≠ EDC, CEC/CPC ≠ EDC  CEC = Contaminants of Emerging Concern  CPC = Contaminants of Potential Concern  PPCP = Pharmaceuticals and Personal Care Products  EDC = Endocrine Disrupting Chemical  PPCP ≠ EDC, CEC/CPC ≠ EDC

3. What are EDC’s???  EDCs directly affect the endocrine system  e,.g., birth control, thyroid medication  Also called hormonally active agents, endocrine active chemicals, estrogen mimics,  Xenoestrogens = man-made compounds (e.g., birth control)  Phytoestrogens = naturally occurring estrogen compounds (e.g., lilac)  EDCs directly affect the endocrine system  e,.g., birth control, thyroid medication  Also called hormonally active agents, endocrine active chemicals, estrogen mimics,  Xenoestrogens = man-made compounds (e.g., birth control)  Phytoestrogens = naturally occurring estrogen compounds (e.g., lilac)

4. EDCs are not just hormones!  Of course, they include hormones, but they’re also:  Pharmaceuticals (  -blockers, birth control)  Personal care products (sun screen)  Industrial chemicals (plasticizers)  Pesticides (organochlorine compounds)  Legacy contaminants (DDT, PCB)  Of course, they include hormones, but they’re also:  Pharmaceuticals (  -blockers, birth control)  Personal care products (sun screen)  Industrial chemicals (plasticizers)  Pesticides (organochlorine compounds)  Legacy contaminants (DDT, PCB)

5. Endocrine disruption is not just “girly fish”  Gender Effects  Feminization or masculinization  Stress response (Cortisol inhibition)  Growth and repair, immune function, reproduction  Behavioral changes  Decreased territoriality, color changes, predator response  Reproductive impairment  Sperm DNA Damage  Gender Effects  Feminization or masculinization  Stress response (Cortisol inhibition)  Growth and repair, immune function, reproduction  Behavioral changes  Decreased territoriality, color changes, predator response  Reproductive impairment  Sperm DNA Damage

6. What are the issues?  Where are EDCs coming from?  Are endocrine disrupting chemicals adversely affecting wildlife? Humans?  What can be done about them?  Does anything need to be done about them?  What about water reclamation efforts?  Where are EDCs coming from?  Are endocrine disrupting chemicals adversely affecting wildlife? Humans?  What can be done about them?  Does anything need to be done about them?  What about water reclamation efforts?

7. Sources of EDCs  Agricultural Runoff  Industrial Waste  Prescription / Non-prescription pharmaceuticals  Wastewater Effluent  Many references in the scientific literature  Ole’ Miss Study  Agricultural Runoff  Industrial Waste  Prescription / Non-prescription pharmaceuticals  Wastewater Effluent  Many references in the scientific literature  Ole’ Miss Study

8. Ocean Discharge of Effluent Newport Pier Newport Pier Balboa Pier Balboa Pier Huntington Pier Huntington Pier 120-inch Dia. Pipeline 4 ½ Miles Out 200 Feet Deep 120-inch Dia. Pipeline 4 ½ Miles Out 200 Feet Deep 1 Mile Long Diffuser with 500 ports 1 Mile Long Diffuser with 500 ports Huntington Beach Newport Beach Corona Del Mar PLANT NO.1 PLANT NO. 2 S a n t a A n a R i v e r Newport Bay

9. What is OCSD doing about EDCs?  Conducting collaborative studies with universities and other researchers.  Universities provide expertise OCSD lacks  OCSD provides:  Ecological expertise  In-kind services (vessel, crew and equipment)  Funding of graduate students  Conducting collaborative studies with universities and other researchers.  Universities provide expertise OCSD lacks  OCSD provides:  Ecological expertise  In-kind services (vessel, crew and equipment)  Funding of graduate students

10. EDC Exposure to Flatfish  Cortisol Inhibition  Dr. Kevin Kelley CSU Long Beach, OCSD  Estrogenicity  Dr. Dan Schlenk UC Riverside, OCSD  Sperm DNA Damage  CSC Biomarker Lab (San Diego, CA), OCSD  Cortisol Inhibition  Dr. Kevin Kelley CSU Long Beach, OCSD  Estrogenicity  Dr. Dan Schlenk UC Riverside, OCSD  Sperm DNA Damage  CSC Biomarker Lab (San Diego, CA), OCSD

11. OCSD Monitoring Area O O

13. Cortisol Production Effects  Produced via the HPI Axis in response to a stressor  Stressors = chemical, physical, environmental  Cortisol production is:  metabolically expensive  Inhibited by chronic stress  Produced via the HPI Axis in response to a stressor  Stressors = chemical, physical, environmental  Cortisol production is:  metabolically expensive  Inhibited by chronic stress

14. HORNYHEAD TURBOT ENGLISH SOLE Peak Cortisol Concentrations in Post-trawl Flatfish Ref.Outfall Outfall Ref.

15. Orange C0. Peak Cortisol Hornyhead Turbot from Different Locales T11 T1 cortisol (ng/ml) 4006 Santa Monica Bay 40364320 San Diego Co. 160 - 140 - 120 - 100 - 80 - 80 - 60 - 60 - 40 - 40 - 20 - 20 - 4037

16. Reproductive/Gender Effects  Reproductive Effects  DNA Damage  Behavioral Effects  Gender Effects  Estrogenicity  Androgenicity  Reproductive Effects  DNA Damage  Behavioral Effects  Gender Effects  Estrogenicity  Androgenicity

17. Sperm DNA Damage  Studies conducted 2000–2004  Comet Assay  DNA damage consistently greater at Outfall Station over Reference Station  Studies conducted 2000–2004  Comet Assay  DNA damage consistently greater at Outfall Station over Reference Station

18. Sperm DNA Damage English Sole Gonad Reference Station Outfall Station Tail Moment

19. Proportion of plasma vitellogenin levels at given concentrations in male hornyhead turbot

20. Proportion of plasma vitellogenin levels at given concentrations in male English sole

21. Proportion of hornyhead turbot and English sole males collected at the outfall and reference site: 1988 - 2004

22. Conclusions of EDC Studies to Date  Indications of Feminization  Increased [vitellogenin] in males – both species  Increased sperm DNA damage in HT  Indications of Masculinization  Increased male GSI – both species  Higher proportion of male HT  ES Stress and Growth Factor Hormone Production Inhibited at the Outfall  HT expression inhibited at all sites  Indications of Feminization  Increased [vitellogenin] in males – both species  Increased sperm DNA damage in HT  Indications of Masculinization  Increased male GSI – both species  Higher proportion of male HT  ES Stress and Growth Factor Hormone Production Inhibited at the Outfall  HT expression inhibited at all sites

23. Conclusions in Context to OCSD OCSD effluent is a source of EDCs Not the only source Flatfish in OCSD monitoring area show evidence of EDC exposure Does exposure equal effect? No population-level effects observed Yet??? Other effects??? Endpoints??? Definitive cause-effect studies needed linking specific chemicals to receiving water impacts OCSD effluent is a source of EDCs Not the only source Flatfish in OCSD monitoring area show evidence of EDC exposure Does exposure equal effect? No population-level effects observed Yet??? Other effects??? Endpoints??? Definitive cause-effect studies needed linking specific chemicals to receiving water impacts

24. Is mitigation necessary?  Pharmaceutical Take-back Programs  Public Education  Proper disposal of pharmaceuticals, personal care products, and pesticides/herbicides  Treatment Options  Reverse Osmosis  Microfiltration  Ozonation  Pharmaceutical Take-back Programs  Public Education  Proper disposal of pharmaceuticals, personal care products, and pesticides/herbicides  Treatment Options  Reverse Osmosis  Microfiltration  Ozonation

25. Studies in Progress  Estrogenicity in Flatfish UC Riverside, Doctoral Student Research  Correlation of EDCs in Fish Tissues to POTW Effluent, Sediments, and Infauna (Fish food) CSU Long Beach, OCSD, and City of LA  Cortisol Inhibition and Fish Parasitization CSU Long Beach  Endocrine Disruption in Coastal Flatfish SCCWRP, OCSD, LACSD, CLAEMD, CSD, UC Riverside, CSU Long Beach, UC San Diego  Estrogenicity in Flatfish UC Riverside, Doctoral Student Research  Correlation of EDCs in Fish Tissues to POTW Effluent, Sediments, and Infauna (Fish food) CSU Long Beach, OCSD, and City of LA  Cortisol Inhibition and Fish Parasitization CSU Long Beach  Endocrine Disruption in Coastal Flatfish SCCWRP, OCSD, LACSD, CLAEMD, CSD, UC Riverside, CSU Long Beach, UC San Diego

26. Acknowledgements  CSU Long Beach, CA Dr. Kevin Kelley Professor of Endocrinology Dr. Julianne Kalman Post Doctoral Researcher, Lecturer Jesus Reyes L aboratory Researcher Kathy Sak Laboratory Researcher  U.C. Riverside, CA Dr. Dan Schlenk Professor of Aquatic Toxicology Dr. Luke Roy Masters Graduate (UCR) Mary Ann Rempel Doctoral Candidate  Computer Sciences Corp., San Diego, CA Scott Steinert  CSU Long Beach, CA Dr. Kevin Kelley Professor of Endocrinology Dr. Julianne Kalman Post Doctoral Researcher, Lecturer Jesus Reyes L aboratory Researcher Kathy Sak Laboratory Researcher  U.C. Riverside, CA Dr. Dan Schlenk Professor of Aquatic Toxicology Dr. Luke Roy Masters Graduate (UCR) Mary Ann Rempel Doctoral Candidate  Computer Sciences Corp., San Diego, CA Scott Steinert

27. Jeff Armstrong (714) 593-7455 jarmstrong@ocsd.com Jeff Armstrong (714) 593-7455 jarmstrong@ocsd.com Visit Our Website: www.OCSD.com