1. Detection of Estrogenic Hormones in BC Sewage Treatment Plant Effluents Heather Osachoff PhD candidate, Simon Fraser University Toxicogenomics Analyst, Environment Canada hosachof@sfu.ca Vesna Furtula (Environment Canada, N. Vancouver, BC) Joanne Harkness (Urban Systems Ltd., Kamloops, BC) Graham van Aggelen (Environment Canada, N. Vancouver, BC) Chris Kennedy (Simon Fraser University, Burnaby, BC)

2. Research Objective Research Objective  Overall objective: to evaluate the effects of xenoestrogens on fish.  Limited information available on the estrogenicity of British Columbia (BC) sewage treatment plant (STP) effluents.  The objective of this study was to profile estrogen hormones in BC STP effluents:  STPs chosen outside of Vancouver area  Site locations kept confidential  3 of 6 STPs evaluated discharge to the Fraser River system

3. What is Sewage? What is Sewage?  Domestic, industrial and agricultural wastes are degraded/digested/cleaned up at sewage treatment plants (STPs) and ‘cleaner’ effluents discharged.  STP effluents contribute contaminants (TSS, BOD, metals, nutrients, pharmaceuticals) to the environment at the discharge site.  A key research focus is on pharmaceuticals and personal care products (PPCPs)  Much interest in endocrine disrupting chemicals (EDCs) as well

4. The Issue The Issue  Estrogen hormones (EDCs) are found in STP effluents  Estrogen hormones:  Coordinate reproduction, behaviour, female processes  Well-documented that male or juvenile fish exposed to estrogen hormones can be feminized, or even exhibit partial or complete intersex gonads amongst other effects Estrogen

5. Contributors to Estrogenicity Contributors to Estrogenicity Three natural estrogen hormones:  Estradiol (E2): main vertebrate estrogen hormone  Estrone (E1): a stable form of E2 and less potent  Estriol (E3): pregnancy estrogen hormone One synthetic estrogen hormone:  Ethinylestradiol (EE2): found in birth control pill and hormone replacement therapy pharmaceuticals Additional less potent compounds:  Other minor xenoestrogens: surfactants, pesticides, industrial compounds, pharmaceuticals.

6. Paradox of Sewage Treatment Paradox of Sewage Treatment Sewage treatment works use multiple tiers of sequential processes:  Primary = remove large solids by settling or screening  Secondary = use microbes to digest nutrients  Tertiary = processes to kill microbes and remove nutrients The relationship between sewage treatment and estrogen levels: The higher the level of sewage treatment, the more estrogen hormones can (sometimes) be found.

7. General Information – 6 BC STPs General Information – 6 BC STPs Table of STP info a Biological nutrient removal plant; b Activated sludge plant; c Rotating biological contactor STPs that discharge to the Fraser River system PlantTypeInfluent Flow (m 3 d -1 ) Treatment Type Treatment Level BODTSSNH 3 NO 3 Advanced P Removal STP 1Tertiary30,300LagoonsSecondary No Yes STP 2Tertiary9,700BNR a Secondary Yes STP 3Tertiary1,000Modified ASP b Secondary YesLimitedNo STP 4Secondary4,800LagoonsSecondary No STP 5Secondary5,500RBC c Secondary No STP 6Secondary900ASP b Secondary YesNo

8. Materials and Methods Materials and Methods  Collected duplicate 1 L influent and/or effluent grab samples in November 2009, March/April 2010 and June 2010  Analyzed via GC-MS by Dr. Vesna Furtula (EC-N. Van)  Profiled each sample for 23 sterol compounds (this includes hormones as well as fecal or plant sterols)  Data is average of duplicates and of multiple dilutions  Influent is a difficult matrix to work with since it is so “dirty”  Published in Furtula et al. (2011) Journal of Water, Air and Soil Pollution (accepted 5Aug2011, online epub)

9. Results Results  No estrogen hormones were detected in influent  In the effluent, only Estrone (E1) was detected (BC may be different from other parts of the world where EE2 is detected)  An evaluation of the efficiency of each STP process in removing estrogen hormones was not possible  STP efficiencies evaluated using other sterol compounds profiled in the chemical analysis (data in Furtula et al. 2011)  Not surprisingly, the tertiary STPs removed more sterol compounds than the secondary STPs

10. Results: Estrogenic Chemical Concentrations Results: Estrogenic Chemical Concentrations

11. Implications Implications  The frequency of Estrone (E1) detection was 50% (in effluent)  We have a BC Water Quality Guideline for Ethinylestradiol (EE2) but not for E1  Suggest development of an E1 guideline  EE2 is a common occurrence in effluents from other nations; where is it in BC? Is there a critical population (prescription) necessary, bound to solids, or degraded with BC STP processes?

12. Implications to the Salish Sea Implications to the Salish Sea  Calculation of the daily loadings of Estrone (E1) to the environment for: STP #3: 0.04 grams/day (0.0014 oz/day) STP #2: 0.57 grams/day (0.02 oz/day)  STPs on the Fraser River system ultimately discharge into the Salish Sea. Additionally, there are STPs directly adjacent to the Salish Sea that discharge into it.  Sewage effluent is chronically discharged (winter exceptions).  Numerous reports of E1 effects on aquatic life: Metcalfe et al. (2001) showed intersex fish at 10 ng/L (comparable to levels found in this study).

13. Final Conclusions Final Conclusions  In 50% of the effluents tested, the estrogen hormone Estrone (E1) is present.  This is of concern since this stable, active hormone has effects on numerous physiological systems and has been shown to cause “feminization” of some aquatic organisms.

14. Future Directions Future Directions  We propose that more BC STP effluents need to be evaluated for estrogen hormones content.  Discharge locations should be evaluated for estrogen hormone profiles as well to characterize the level in the environment.

15.  Sewage treatment plant operators for collection and provision of the samples  Funding from the BC Ministry of Environment, Water Protection and Sustainability Branch Gabriele Matscha and Cindy Meays  Scholarships from NSERC and Pacific Leaders Fellowship (to Heather Osachoff)  Environment Canada co-workers + students: Johnny Liu and the Chemistry Section of PESC Thank you to … Thank you to … hosachof@sfu.ca