Endocrine Disruptors in the aquatic environment from non-

wastewater sources

Poul Bjerregaard

Ecotoxicology GroupInstitute of Biology

University of Southern Denmark

EEA ’Chemicals and Water’ Workshop, December 6-7, 2010

UK and many other countries

• Discharges from waste water treatments plants cause feminisation in male fish

Feminisation expressed as:

• Intersex– Ovotestes

Intersex in roach

Rutilus rutilus

♀ ♂

Normal testis ♂ - intersex

Bjerregaard et al. 2006. Ecotox. Environ Safety

Intersex-frequency among British roach

Jobling et al. 1998. Env. Sci. Technol. 32, 2498

Intersex in roach

0

5

10

15

20

25

30

Inte

rsex

fre

qu

ency

(%

)

Lake Almind Lake Ravn Aarhus Egaa Kristrup landkanal

Control sites

***

Streams receiving sewage effluents

Bjerregaard et al. 2006

Recent French investigation

• 474 roach examined in uncontaminated area• Frequency of intersex:• 0

– Geraudie et al. 2010. Fish Physiol. Biochem. 36, 767-777

Feminisation expressed as:

• Intersex– Ovotestes

• Elevated vitellogenin levels

OvaryLiver

Gonadotropins

Estrogen

Vitellogenin

Estrogenic effect of UK discharges

Harries et al. 1996. Environ. Toxicol. Chem. 15, 1993-2002

Danish WWTP effluents

Nonitrifi-cationNitrifi-

cation

E1: 60-70% E2: 30-40% EE2: Less importance

Stuer-Lauridsen et al. 2005. Danish EPA-report

PNEC

Little estrogenic effect from Danish WWTPs

• Still endocrine disrupted fish

Brown trout vitellogenin levels

10

102

103

104

105

106

Vit

ello

ge

nin

(n

g /

ml)

*

*

*

*

EE2

*

*

***

***

*

Kongs

høj Å

BMale fish only

Fig. 3

Voel

Bæ

kU

sser

ød Å

Gib

er Å

Knubb

ro B

æk

Gra

nslev

ÅØ

rum

ÅHo

ed Å

Høj

en Å

Mar

en M

ølleb

æk

Tang

e Å

Hund

stru

p Å

Brend

e Å

Sand

er B

æk

Stokk

ebæ

kken

Vind

inge

ÅBar

brek

ær G

røft

Hat

tebæ

kken

Hald

rup

Møl

leå

Bjerg

skov

Bæ

kSt

avis

Å

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

Other possibilities

• Leaching of estrogens from agriculture

Leaching from pig manure

Kjær et al. 2007. Env. Sci. Technol. 41, 3911-3917

Leaching from pig manure

Kjær et al. 2007. Env. Sci. Technol. 41, 3911-3917

EC50 values for brown trout

• Induction of vitellogenin synthesis after 8-10 days’ exposure– E2: 15 ng/l– E1: 88 ng/l

Phytoestrogens

Clover Alfalfa PeasLupine

Naturally produced estrogens

O

OH

OH

OCH3O

OHCH3

OHBiochanin A

17β-estradiol

Biochanin AEffects in brown trout

Con

trol

Sol

vent

cont

rol

31±2

2

51±3

9

120±

8623

5±14

850

ng

E2

l-1

100

101

102

103

104

105

106

107

108

Vite

llog

enin

leve

l (n

g m

l-1 p

lasm

a)

Con

trol

Sol

vent

cont

rol

0.8±

0.08

1.2±

0.2

1.8±

0.5

3.8±

1.0

9.4±

2.5

100

101

102

103

104

105 B

Fig. 5

***

11 13 15 1515 15

**

*

* *

A

16 14 14 14 151316

15

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

Phytoestrogen concentrations in water

• Most determinations:– Nanogrammes per litre range

• Some determinations: – Microgrammes per litre.

Other possibilities?

• Discharges from scattered houses in the open land with simple waste water treatment

Danish survey

• Up to 414 ng/l E2-equivalents in drains from septic tanks

• Stuer-Lauridsen et al. 2005. Danish EPA report

Pesticides

• Numerous pesticides have endocrine disrupting potential

• Many pesticides detected in freshwater systems

Other natural sources?

• Extracts from oak leves have anti-androgenic activity

• Hermelink et al. 2010

• Saponins from horse chestnut?

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

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