02, Monday 05OCT2009, J Parrott, Environment Canada 1

Effects of Pharmaceuticals in the Canadian Environment

Joanne Parrott, Science and Technology Branch, Environment Canada, 867 Lakeshore Road, Burlington, Ontario L7R 4A6

Joanne Parrott, Science and Technology Branch, Environment Canada, 867 Lakeshore Road, Burlington, Ontario L7R 4A6

• Pharmaceuticals are designed to have a

therapeutic (=biological) effect

– Effects on non-target organisms are mostly

unknown

• Aquatic organisms are chronically exposed

• Potential for multigenerational exposure

• Little known about environmental persistence, fate

Why Worry about Pharmaceuticals?

02, Monday 05OCT2009, J Parrott, Environment Canada 2

Pharmaceuticals and Personal Care Products (PPCPs) in Municipal Wastewater Effluents - Thames River, ON

0

500

1,000

1,500

2,000

2,500

3,000

Napro

xen

Ibupro

fen

Dicolfe

nac

Indom

ethac

in

Ketopro

fen

Salic

ylic

Aci

d

Gemfib

rozil

Estra

diol

Estro

ne

Triclo

san

Galax

olide

Tonalid

e

Compound

ng

/L

MAX

Mean

Painkillers NSAIDS

Chol ↓Hormones

Antibacterial

Musk fragrances

Lishman et al 2006 Sci Tot Environ

Perso

nal C

are

Produ

cts

Effects in lab organisms at low concentrations (low ug/L

or below)… compare to levels in municipal wastewater

effluents (MWWEs) and Great Lakes rivers and waters

• anti-inflammatory - Diclofenac

• antidepressants - Fluoxetine, Fluvoxamine

• synthetic hormone - Ethinylestradiol

• complex effluent mixtures - MWWEs

Pharmaceuticals to Examine

02, Monday 05OCT2009, J Parrott, Environment Canada 3

Non-steroidal, Anti-inflammatory drug (NSAID)

Diclofenac

Expose trout for 28 d to 1 ug/L - examine tissues for changes

Liver - Hepatocytes - solvent control rainbow trout

• round nuclei (n) • regularly arranged ER (er) • glycogen storage areas (g) (magnification: 4200×).

Liver of a fish exposed to 1 ug/L diclofenac for 28 days.

• compartmentation changed• glycogen areas depleted

Triebskorn R, et al. 2004. Aquatic Toxicology 68:151–166.

control

1 ug/L diclofenac

02, Monday 05OCT2009, J Parrott, Environment Canada 4

ng/L ug/L

500 ng/L gill changes, 1 ug/L liver changes, rainbow trout

Diclofenac

Non-steroidal, Anti-inflammatory drug (NSAID)

∑ mean NSAIDs = 1.45 ug/L (Thames R, Lishman et al 2006)

MAX at one of 12 plants Thames R, 940 ng/L Diclofenac

430 ng/L, MWWE, fall(Lishman et al, 2005)

215 ng/L, MWWE, summer(Lishman et al, 2005)

50-150 ng/L, rivers(Metcalfe et al, 2003)

360 ng/L, max MWWE(Metcalfe et al, 2003)

Fluoxetine (Prozac™)Antidepressant, anti-anxiety drug

Selective serotonin reuptake inhibitors, SSRIs

02, Monday 05OCT2009, J Parrott, Environment Canada 5

Antidepressants in MWWE on Grand R., ON (Metcalfe et al., ET&C in Press 2009)

0.01

0.1

1

10

Venla

faxi

ne

Effexo

r™

Citalo

pram

Celex

a™Bupro

pion

Wel

lbutri

n™

Fluoxe

tine

Proza

c™Ser

tralin

e

Zoloft™

Sum o

f all

MW

WE

antid

epre

ssan

ts

Antidepressants

µg

/LDifferent antidepressants inhibit reuptake of different neurotransmitters: serotonin, dopamine and norepinephrine

S SSN SDNS

Humans – serotonin regulates appetite, sleep, arousal, and mood.

Bivalve mollusks - serotonin regulates reproductive processes such as spawning, oocyte maturation and parturition.

02, Monday 05OCT2009, J Parrott, Environment Canada 6

0

20

40

60

80

100

0 0.01 0.1 1 10 100 1,000 10,000 100,000

Fluvoxamine (ug/L)

% P

artu

riti

on

*

* *

* All significant, p<0.01

3 ug/L

control clams

% Parturition (release of young) in fingernail clams exposed to

fluvoxamine for 4 hours (Fong et al 1998)

Fish age (days)

Fis

h w

ith

bla

ck s

po

t (%

)

Fluoxetine exposure delayed maturation in female medaka (from Henry et al, 2004)

0

5

10

15

20

25

30

35

30 40 50 60 70 80 90

0

6 ug/L

60 ug/L

Fluoxetine exposure started

02, Monday 05OCT2009, J Parrott, Environment Canada 7

ng/L ug/L

Fluoxetine

3 ug/L Fluvoxamine,

clam spawning

Antidepressant, anti-anxiety drug

6-60 ug/L, medaka

maturation

100 ng/L, MWWE(Metcalfe et al 03)

45 ng/L, rivers d/s(Metcalfe et al 03)

3.2 ug/L, sum SSRI antidepressants in MWWE (Metcalfe et al 07)

EthinylestradiolSynthetic estrogen in birth control pills

EE2

02, Monday 05OCT2009, J Parrott, Environment Canada 8

fryfry survival, length, weight

survival, % hatch Fathead minnow lifecycle exposure –

sampling times and endpoints

Fathead minnow lifecycle exposure –

sampling times and endpoints

30 and 60 days post-hatch

survival, length, weight, abnormalitiesjuvenilesjuveniles

160 days-post-hatch

2o sex characteristicslength, weight, liver weight, gonad weight

adult fathead minnowsadult fathead minnows

egg prod, fertilization, % hatch

eggseggs

% m

ale

adu

lts

EE2 (ng/L)

water eth 0.32 1.0 3.5 9.6 23

Percent male fathead minnows at maturity

No male fish

Controls

Male

Female

0

10

20

30

40

50

60

70Fewer

male fish

* *

02, Monday 05OCT2009, J Parrott, Environment Canada 9

0

20

40

80

100

p<0.001

*

p=0.025

*

water eth 0.32 1.0

EE2 (ng/L)

Eg

gs

Fer

tili

zed

(%

)

60

no eggs, no males

Reduced egg fertilization success

Controls

EE2 – effects on egg fertilization at 0.32 to 1 ng/L (Parrott & Blunt, 2005)

EE2 – effects on fish populations 5-6 ng/L(Kidd et al 07)

ng/L ug/L

EE2 – 12 Kg/yr Canada = MWWE ~ 0.2 ng/L

avg 9 ng/L(Ternes et al 99)

< 1 ng/L (Lee et al 04)

4 of 70 US

streams,

> 5 ng/L(Kolpin et al 02)

EthinylestradiolSynthetic estrogen in birth control pills

02, Monday 05OCT2009, J Parrott, Environment Canada 10

What’s in a Municipal Wastewater Effluent

(MWWE)?

Nutrients (N, P), metals, oils,Trace levels of SOME….

Natural/synthetic hormones, household chemicals, pharmaceutical drugs, personal care products…

City of Hamilton (0.5 million people)

Municipal Wastewater Effluent (MWWE)

Lake Ontario

2 km0

Western tip of Lake Ontario

02, Monday 05OCT2009, J Parrott, Environment Canada 11

0

2,000

4,000

6,000

8,000

10,000

12,000

14,000

16,000

90 100 110 120 130 140 150 160

Days post-hatch

Eg

g P

rod

uct

ion Lab control water

10% H MWWE

100% H MWWE

Egg Production in Fathead Minnows Exposed to MWWE for a Lifecycle

Per 12-13 females

Vajda et al 2008, ES&T

White sucker collected downstream of MWWE, Boulder, Colorado

MWWE is 10 % to 75 % of stream flow

High EE2, natural estrogens, bisphenol A, nonylphenols

Fewer males downstream of MWWE

Intersex Males downstream of MWWE

02, Monday 05OCT2009, J Parrott, Environment Canada 12

Conclusions

Some PPCPs appear to cause effects in lab

organisms at concns of low ng/L to low ug/L.

Some MWWEs in Great Lakes discharge PPCP

concentrations in these ranges.

Thresholds for effects of synthetic estrogens very

low (ng/L)…. Other PPCPs – higher thresholds

(ug/L and above).

Some evidence of effects with lab fish and wild

fish exposure to MWWE.

What we don’t know…Which compounds should we monitor in MWWEs and rivers?

Are there widespread effects in wild fish, frogs, turtles, etc. in Canadian Rivers?

Are we looking at the right biological endpoints?

02, Monday 05OCT2009, J Parrott, Environment Canada 13

EthinylestradiolSynthetic estrogen in birth control pills

Ministry of the Environment, British Columbia, Canada

Proposed draft water quality guideline 0.5 ng/L

Most PPCPs – not enough data to set limits or do environmental risk assessment

“For the protection of freshwater aquatic life, it is recommended that the 30-day average concentration of 17α-ethinylestradiol (EE2) in water, based on 5 weekly samples, should not exceed 0.5 ng L-1 with no single value to exceed 0.75 ng L-1 (no more than 50% above the guideline value).”

Ambient Water Quality Guidelines for the Protection of Pharmaceutically-Active Compounds (PhACs) – 17α-Ethinlyestradiol

Environmental Assessment Work Group (EAWG) – Health Canada, Environment Canada, industry stakeholders, non-government organizations

Effects Assessment based on longer testsrelevant to exposure route and mechanism of action (not short standard tests of fish survival etc.)

Propose new Environmental Risk

Assessment Regulations under Food & Drug Act

…PPCPs and other compounds

Exposure Assessment based on DOSES of pharmaceutical (not AMOUNT produced/imported)

02, Monday 05OCT2009, J Parrott, Environment Canada 14

Thank You!!