Fish Bioaccumulation Studies Associated with the Kingston Fly Ash Release

1. Fish Bioaccumulation Studies Associated with the Kingston Fly Ash Release Marshall Adams - Oak Ridge National Lab Tyler Baker - TVA Allison Fortner - Arcadis Terry Matthews and Mary McCracken - ORNL

2. Presentation Outline - Research objectives - Study Design - Results and major findings - causal relationships - Relevance to overall fly ash project - integration with other studies

3. Objectives of Bioaccumulation Studies Assess spatial and temporal patterns of metal bioaccumulation in sentinel fish species Evaluate potential causal relationships between exposure to fly ash-associated metals and bioaccumulation in representative fish species Determine if fly ash exposure is causing short and/or long- term health effects on representative (sentinel) fish populations (tomorrows presentation)

4. Study Design Sample Frequency spring and fall each year since spring 2009 Sample Sites - 6-7 sample sites along downstream gradient of Emory and Clinch Rivers - includes 2 instream reference sites Fish Species Sampled Spring - bluegill, largemouth bass, redear sunfish, and while crappie Fall - bluegill, largemouth bass, and channel catfish Metals Analyzed - ICP suite of 25 metals including selenium, arsenic, etc. - metals measured in fillets (muscle) from all fish, liver and ovaries (spring), whole body (shad) Special Studies - whole body composite gizzard shad - reconstruction analysis on bluegill and LMB - trophic analysis on redear using stable isotopes

5. Fly Ash Spill Exposure of Fish to Metals Focus on sentinel species (different trophic levels and home ranges) - sunfish - bass - catfish - crappie Bioaccumulation Analysis of Fish Health Assess Effects and Causality 25 metals + Hg Physio- logical Repro- ductive Histo- path Bio- energetic Whole fish Muscle Liver Ovary

6. ERM 8 (Ref) Little Emory (Ref) ERM 3 CRM 25.0 (ref) Ash spill CRM 8 ERM 0.9 CRM 7 CRM 1.5

7. Sentinel species for bioaccumulation and fish health studies Types of Studies Bioaccumulation, fish health, and reconstruction analysis Bluegill - mid trophic level/omnivore - restricted home range Largemouth bass - upper trophic level/predator - intermediate home range Bioaccumulation, fish health, and reconstruction analysis Channel catfish - bottom feeder/omnivore - large home range Bioaccumulation and fish health Redear Sunfish Bioaccumulation, fish health, & trophic analysis (stable isotopes) - exclusive molluscavore - high site fidelity Gizzard Shad Bioaccumulation and role in food chain transfer of metals - feeds on detritus & periphyton - large home range

8. Spatial Pattern Ash spill Ash spill Refs Refs Refs Refs Downstream

9. Spatial Pattern Ash spill Ash spill Refs Refs References below detection

10. Temporal Pattern Selenium in Bluegill Muscle EPA tier 1 action level for assessing toxicity of selenium on fish and birds EPA tier 1 action level for assessing toxicity of selenium on fish and birds Reference Sites Ash spill References

11. Selenium in Ovarian Tissue EPA tier 2 action level for assessing toxicity of selenium on fish and birds References

12. Temporal Pattern Ash Spill References

13. Temporal Pattern

14. Temporal Pattern Selenium in whole body gizzard shad References

15. References

16. Selenium in whole body Gizzard Shad Selenium in Largemouth Bass (muscle) Range of Se in shad

17. Composite Bioaccumulation Index (Fillets—integrates 4 species and 4 seasons) DOE Legacy Positive control Composite Bioaccumulation Index Selenium Arsenic Ash spill References Sample Site

18. Composite Bioaccumulation Index (Liver– integrates 3 species and 2 seasons) DOE Legacy Positive control Selenium Composite Bioaccumulation Index Arsenic Ash spill Sample Site References

19. Relationship between selenium in fillet and whole body of bluegill based on reconstruction analysis

20. Relevance and Application

21. Integrated Aquatic Food Chain Studies Tree swallow Racoon Piscivorous birds Frogs Adult mayflies GB heron Osprey Mussel emergence Mayfly nymph LM bass Shad Sunfish Periphyton Metals, detritus, periphyton To periphyton Se-2 Se+4 Ash

22. Establishing causal relationships among environmental variables Bird bioaccumulation Bird Health Metals in other biota - turtles - amphibians - raccoons Fish bioaccumulation Fish Health Macroinvert bioaccumulation Metals in water and sediment Possible direct cause & effect relationship Possible spatial correlations (indirect causality)

23. Summary and Conclusions - For all fish species studied, there is spatial gradient in bioaccumulation of selenium and arsenic with highest levels downstream of the ash spill - In most cases, levels of selenium in fish collected below the ash spill are not high enough to trigger the EPA tiered monitoring scheme for assessing toxicity of selenium in fish - Spatial and temporal patterns in bioaccumulation suggest there are causal relationships between exposure to fly ash-associated metals and levels of metals in fish - Food chain studies provide a weight-of-evidence approach for evaluating possible causal relationships between exposure to fly ash metals and effects on ecological resources