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A Link Between Reproductive Abnormalities and Contaminants in Fish from the Rocky Mountains

Julie Denham Prof. Staci Simonich, Department of Chemistry and Environmental Molecular Toxicology Oregon State University HHMI 2009 . A Link Between Reproductive Abnormalities and Contaminants in Fish from the Rocky Mountains. History of Persistent Organic Pollutants.

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A Link Between Reproductive Abnormalities and Contaminants in Fish from the Rocky Mountains

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  1. Julie Denham Prof. Staci Simonich, Department of Chemistry and Environmental Molecular Toxicology Oregon State University HHMI 2009 A Link Between Reproductive Abnormalities and Contaminants in Fish from the Rocky Mountains

  2. 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

  3. Environmental Fate • Accumulate in the ecosystem • Undergo atmospheric transport • Volatilize at high temperatures and condense at lower temperatures • Settle remote high altitude ecosystems

  4. POPs in the environment Contamination source Release into environment Transport Partitions into ecosystem Water Soil Tissue Atmosphere http://www.epa.gov/glnpo/atlas/glat-ch4.html

  5. 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.

  6. 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

  7. Developing vitellogenic eggs Spirit Lake, Rocky Mountain National Park Reproductive Abnormalities in Trout from Western U.S. National Parks. Schwindt, A. R., et al. Spermatocysts with spermatogonia

  8. Why Rocky Mountain NP? Previous research showed: • 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.

  9. POPs Measured in Rocky Mountain Fish p,p-DDE Dieldrin trans-Nonachlor Highest Concentration One currently in use Four compounds are xenoestrogens PCB’s Dacthal Hexachlorobenzene (HCB) Endosulfan Sulfate

  10. Xenoestrogens • Xenoestrogens - organic compounds that have estrogen like affects p,p-DDE • Not produced as a natural hormone Dieldrin • Cause excess vitellogenin concentrations PCB’s Endosulfan Sulfate

  11. Scope • Measure POPs in fish samples collected previously • To interpret the POP concentrations in the fish with respect to intersex.

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

  13. Fish Homogenate Extraction of analytes (ASE) Method X g Fish + X g Na2SO4 Removal of polar compounds (SPE) Fraction 1 (target analytes) Fraction 2 (archived for recovery) Gel permeation chromatography Fraction A: lipid archive Fraction B: target analytes Quantification of target analytes GCMS

  14. 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

  15. 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

  16. POPs Measured -In SRM and previous studies from Rocky Mountain National Park p,p-DDE Dieldrin trans-Nonachlor PCB’s Dacthal Hexachlorobenzene (HCB)

  17. SRM Data

  18. SRM Data

  19. 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

  20. 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 Family and Friends

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