A LINK BETWEEN REPRODUCTIVE ABNORMALITIES AND

CONTAMINANTS IN FISH FROM THE ROCKY MOUNTAINS

Julie DenhamProf. Staci Simonich,

Department of Chemistry and

Environmental Molecular Toxicology

Oregon State University

HHMI 2009

History of Persistent Organic Pollutants

Industrially made in the early 1930’s

Short term affects eliminated agricultural problem

Persistent contaminants toxic to ecosystems

Linked to diseases, abnormalities, and endocrine disruption

Environmental Fate Accumulate in the ecosystem

Undergo atmospheric transport

Volatilize at high temperatures and condense at lower temperatures

Settle remote high altitude ecosystems

POPs in the environment

Contamination source

Transport

Partitions into ecosystem

Soil

Release into environment

Water Tissue Atmosphere

http://www.epa.gov/glnpo/atlas/glat-ch4.html

Bioaccumulation

Fractionate into lipids

Concentrations increase through food web

Fish are a useful tool of which POPs are present in the environment and the impacts they have on specific ecosystems

http://io.uwinnipeg.ca/~simmons/16cm05/1116/54-25-DDTInFoodChain-L.jpg.

Impacts on fish A hormone imbalance of intersex

characteristics and increased vitellogenin levels in males

Intersex- an abnormality that can be found in animals containing both sexual reproductive organs

Vitellogenin (VG)- a protein synthesized in the liver in reaction to excess estrogen levels

- precursor for egg-yolk synthesis

Spirit Lake, Rocky Mountain National Park

Reproductive Abnormalities in Trout from Western U.S. National Parks. Schwindt, A. R., et al. Spermatocysts with spermatogonia

Developing vitellogenic eggs

Why Rocky Mountain NP?

• Signs of intersex fish in Rocky Mountain Nat. Park

• Pesticide concentrations highest among the Western National parks

• Black Circles Containing white numbers represent 2003-2006 sampling• White circles are historic samples collected from 1800’s- 1934

Reproductive Abnormalities in Trout from Western U.S. National Parks. Schwindt, A. R., et al.

Previous research showed:

POPs Measured in Rocky Mountain Fish

• Highest Concentration

• One currently in use

• Four compounds are xenoestrogens

p,p-DDE Dieldrin trans-Nonachlor

Dacthal PCB’s

Hexachlorobenzene (HCB) Endosulfan Sulfate

Xenoestrogens Xenoestrogens -

organic compounds that have estrogen like affects

p,p-DDE Dieldrin

PCB’s Endosulfan Sulfate

• Not produced as a natural hormone

• Cause excess vitellogenin concentrations

Scope

Measure POPs in fish samples collected previously

To interpret the POP concentrations in the fish with respect to intersex.

Hypothesis

The accumulation of persistent organic pollutants in male trout of remote high altitude lakes caused intersex characteristics.

Method

Fish Homogenate

X g Fish + X g Na2SO4

Extractionof analytes

(ASE)

Gel permeation chromatography

Fraction B: target analytes

Fraction 2 (archived for recovery)

Fraction 1(target analytes)

Removal of polar

compounds (SPE)

Fraction A: lipid archive

Quantification of target analytes

GCMS

Method Validation

Use of certified Standard Reference Material (SRM)

SRM is certified by NIST to have a known value of pesticides

Used in analytical methods for precision and accuracy

SRM 1946 - Lake Superior Trout tissue- Certified concentrations for chlorinated

pesticides and PCBs

SRM Data

Performed in Triplicate

Average concentrations of triplicate data were compared to the known quantities in SRM samples

- standard deviation of Triplicate vs. SRM known values

POPs Measured

trans-NonachlorDieldrinp,p-DDE

Hexachlorobenzene (HCB)Dacthal PCB’s

-In SRM and previous studies from Rocky Mountain National Park

SRM Data

SRM Data

Future Work

Finish extracting pesticides from intersex and control male fish

Analysis on GC/MS to quantify the concentrations of POPs

Use of statistical correlations between the POPs and intersex to determine if a relationship exists

Acknowledgements Dr. Staci Simonich

Howard Hughes Medical Institute

Simonich lab-

Jill, Jessica, Wentao, Carlos, Leah, On, &

Dr. Steve Sylvester

Dr. Kevin Ahern

National Park Service

Oregon State University Department of EMT

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