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 UniversityHHMI 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 SimonichHoward Hughes Medical InstituteSimonich lab-
Jill, Jessica, Wentao, Carlos, Leah, On, & Dr. Steve SylvesterDr. Kevin AhernNational Park ServiceOregon State University Department of EMTFamily and Friends
