Alan Kolok

Department of Biology Department of Environmental, Agricultural

& Occupational Health

Fate and transport of steroids in the environment:

What goes in, must come out?

Institute of Agriculture and Natural Resources

October 14, 2010 University of Nebraska – Lincoln

East Campus Union.

Steroids and the U.S. beef cattle industry

More than 90% of US beef cattle are given growth promoting implants.

The most common implant contains trenbolone acetate and

estradiol.

28 mg 200 mg Synovex Plus

Estradiol Trenbolone

Acetate

Brand

Name

Endogenous steroid metabolites

(but not Testosterone)

Unimplanted

17 -Trenbolone, 17 -Trenbolone

17 -estradiol, estrone

Trenbolone acetate

Estradiol benzoate

Implanted

Na

no

gra

ms r

eco

vere

d

Estrone

Androstenedione

0.05

0.10

0.15

0.2 17 Trenbolone

Sellin et al. 2009. Aq. Toxicol.

.1

.2

.3

.4

.5

.6

.7

0

.04

.08

.12

.16

I

I

U

Exposures to exogenous steroids and steroid

mimics disrupt the endocrine function of fish.

Normal Female

17-estradiol

Vitellogenin

17-estradiol

Vitellogenin

Trenbolone

Ankley et al. 2003. Environ Toxicol Chem 22:1350-1360.

17-TB reduces female plasma vitellogenin levels

Exposures to steroids and steroid mimics disrupt

the endocrine function of fish.

Normal Female

17-estradiol

Vitellogenin

Normal Male

17-estradiol

Vitellogenin

Seki et al. 2006. Environ Toxicol Chem. 25:2742-2752.

Vitellogenin

Estrogens

Vit

ello

gen

in (

mg

/mL

)

Estradiol concentration (ng/L)

17-E2 increases male plasma vitellogenin levels

Rela

tive

Gen

e E

xp

ressio

n

Steer Feces (mg/L)

0

.5

1

1.5

2

2.5

3

3.5

Control 0.012 1.25 125 6250

Vtg/L8

Sellin et al. 2009. Aq.Tox. 92:221-227.

Steer Feces (mg/L)

Vtg/L8

Rela

tive

Gen

e E

xp

ressio

n

0

.2

.4

.6

.8

1

1.2

1.4

0 1.25 125 6250 1250

*

Implanted

Unimplanted

Sellin et al. 2009. Aq.Tox. 92:221-227.

Laboratory

Field

No

TBA,

Estrogenic

effect

* detection limit = 1 ng

Compound

17-estradiol

Estrone

Estriol

17-trenbolone

17-trenbolone

Pearl Cr.

ND

ND

ND

ND

ND

Bow Cr.

ND

ND

ND

ND

ND

E. Bow Cr.

ND

ND

ND

ND

ND

Compound

17-estradiol

Estrone

Estriol

17-trenbolone

17-trenbolone

Niobrara

ND

ND

ND

ND

ND

Elkhorn

ND

ND

ND

ND

ND

Platte

ND

ND

ND

ND

ND

Elkhorn River Bow Creek

Niobrara

River

Re

lative

gene

exp

ressio

n

Elkhorn

River

Platte

River

0.4

0.8

1.2

Field

Laboratory R

ela

tive

ge

ne

exp

ressio

n

0

0.8

1.6

Elkhorn

River

Lab H2O

Rela

tive

ge

ne

exp

ressio

n

0.2

0.6

1

East Bow

Creek

Pearl

Creek

Re

lative

ge

ne

exp

ressio

n

0.1

0.4

0.7

East Bow

Creek Lab water

controls

Sellin et al. 2009. ET&C. 28:2443-2448. Sellin et al. 2010. Aq Tox 96:103-108.

Laboratory

Field

No

TBA,

Estrogenic

effect

No

TBA,

Anti-

estrogenic

effect

Water

Sediment

Elkhorn River -Vtg R

ela

tive g

ene

expre

ssio

n

0.4

0.8

1.2

1.6

2

Bow Creek - ER

Rela

tive g

ene

expre

ssio

n

0

0.2

0.4

0.6

0.8

Sellin et al. 2010. Aq Tox 96:103-108.

Defeminization of female fathead minnows and sediment exposure.

Lab

water Field

water Field

exposure Sediment,

field water

Sediment,

lab water

Is the trenbolone in the sediment ?

Tb in

Water

(+ control)

A

Tb in

sediment

B

Lab

Water

(neg. control)

Vitellogenin

0

Rela

tive

gene

exp

resso

in

0.4

0.8

1.2

1.6

C

Kolok et al. In prep. Env Biol Fish