1. Endocrine Disruptors in the aquatic environment from non- wastewater sources Poul Bjerregaard Ecotoxicology Group Institute of Biology University of Southern Denmark EEA ’Chemicals and Water’ Workshop, December 6-7, 2010

2. UK and many other countries Discharges from waste water treatments plants cause feminisation in male fish

3. Feminisation expressed as: Intersex –Ovotestes

4. Intersex in roach Rutilus rutilus

5. ♀♂

6. Normal testis ♂ ♂ - intersex Bjerregaard et al. 2006. Ecotox. Environ Safety

7. Intersex-frequency among British roach Jobling et al. 1998. Env. Sci. Technol. 32, 2498

8. Intersex in roach 0 5 10 15 20 25 30 Intersex frequency (%) Lake AlmindLake Ravn Aarhus EgaaKristrup landkanal Control sites *** Streams receiving sewage effluents Bjerregaard et al. 2006

9. Recent French investigation 474 roach examined in uncontaminated area Frequency of intersex: 0 –Geraudie et al. 2010. Fish Physiol. Biochem. 36, 767-777

10. Feminisation expressed as: Intersex –Ovotestes Elevated vitellogenin levels Ovary Liver Gonadotropins Estrogen Vitellogenin

11. Estrogenic effect of UK discharges Harries et al. 1996. Environ. Toxicol. Chem. 15, 1993-2002

13. Danish WWTP effluents No nitrifi- cation Nitrifi- cation E1: 60-70% E2: 30-40% EE2: Less importance Stuer-Lauridsen et al. 2005. Danish EPA-report PNEC

14. Little estrogenic effect from Danish WWTPs Still endocrine disrupted fish

15. Brown trout vitellogenin levels Bjerregaard et al. Ecotoxicol. 15, 315-327, 2006 & Bjerregaard et al. Environ.Toxicol.Chem. 27, 2387-2396, 2008

16. Other possibilities Leaching of estrogens from agriculture

17. Leaching from pig manure Kjær et al. 2007. Env. Sci. Technol. 41, 3911-3917

18. Leaching from pig manure Kjær et al. 2007. Env. Sci. Technol. 41, 3911-3917

19. EC 50 values for brown trout Induction of vitellogenin synthesis after 8- 10 days’ exposure –E2: 15 ng/l –E1:88 ng/l

20. Phytoestrogens Clover AlfalfaPeasLupine

21. Naturally produced estrogens Biochanin A 17β-estradiol

22. Biochanin A Effects in brown trout

23. Phytoestrogens from silage Suggested as a potential source of estrogenic activity in UK streams in farmland Matthiessen et al. 2006. Sci. Tot. Environ. 367, 616-630

24. Phytoestrogen concentrations in water Most determinations: –Nanogrammes per litre range Some determinations: –Microgrammes per litre.

25. Other possibilities? Discharges from scattered houses in the open land with simple waste water treatment

26. Danish survey Up to 414 ng/l E2-equivalents in drains from septic tanks Stuer-Lauridsen et al. 2005. Danish EPA report

27. Pesticides Numerous pesticides have endocrine disrupting potential Many pesticides detected in freshwater systems

28. Other natural sources? Extracts from oak leves have anti- androgenic activity Hermelink et al. 2010 Saponins from horse chestnut?

29. Endocrine disruption in the aquatic environment Potentially a mix various sources: –Natural estrogens From humans or livestock –Synthetic estrogens and androgens –Pesticides –Phytoestrogens –Other natural products

30. Endocrine disrupting effects – also extending into coastal areas UK flounders Matthiessen et al. UK mussels Langston & Chesman (2006, 2007) DK flounders Unpublished results Baltic eelpout? Unpublished results