Influence of Selenium in Upper Mud River Watershed on Bluegill ( Lepomis macrochirus )

1. Influence of Selenium in Upper Mud River Watershed on Bluegill (Lepomismacrochirus) Alison M. Anderson Patricia M. Mazik Paul Ziemkiewicz

2. Introduction • Selenium • Naturally occurring metalloid • Associated with: • Coal seams • Phosphate deposits • Other mineral formations

3. Introduction • Increased mobilization due to: • Irrigation • Sewage • Fly ash/coal combustion • Mining

4. Selenium Cycling • 4 Oxidation states • Selenite (SeO3-2) • Selenate (SeO4-2) • Selenides (Se -2) • Elemental selenium (Se) • Insoluble • Poorly assimilated by aquatic plants

5. Selenium Cycling • Immobilized • Selenate is oxidized to Selenite • Selenite binds to: • Clay • Organic particles • Forms precipitate with iron • Precipitate settles out • Sediment can be buried • Remobilized by: • Plant root absorption • Consumption by inverts and fish

6. Effects on Fish Benefits: • Protects DNA, lipids, and proteins If fish are deficient: • Loss coordination • Reduced growth • Reduced immune response

7. Effects on Fish: Adults • Impairs gas exchange • Swollen lamellae • Constricts blood vessels • Binds to hemoglobin • Unable to carry oxygen • Induce cataracts • Edema • Seleno-proteins

8. Seleno-proteins • Similar to sulfides • Cells cannot discriminate • Selenium substituted for sulfur • Results in dysfunctional amino acids/proteins • Accumulates in gonads, kidney, and liver

9. Effects on Fish: Parent to Progeny • Seleno-proteins stored in ovarian tissues • Proteins transferred to eggs • Building blocks for tissue synthesis • Metabolic fuel • External feeding • Critical period • High mortality

10. Effects on Fish: Parent to Progeny • Faulty proteins used for development • Results in teratogenesis: • Craniofacial deformities • Spinal deformities • Missing fins • Edema • Inability to feed leading to death

11. Spinal Deformities Kyphosis Lordosis Scoliosis

12. Objectives • Determine the effects of selenium on the aquatic ecosystem of the Upper Mud River Watershed, WV • Determine the effects of selenium on the survival of bluegill larvae

13. Study Site: Upper Mud River • Lower Guyandotte River Watershed • Lincoln and Boone Counties, WV • 124 ha reservoir • 4.5-6 m depth • Inhabitants: • Bluegill • Largemouth bass • Crappie • Channel catfish

14. Study Site: Plum Orchard Lake(Reference Site) • Paint Creek Watershed • Fayette County, WV • 81.75 ha reservoir • 12.2 m depth • Inhabitants: • Bluegill • Largemouth bass • Channel catfish • White crappie

15. Methods

16. Water Quality • October 2008-November 2010 • Mud River (main stem and tributaries) • 20 total sites • Mined (13) • Unmined (7) • Reference Site • 2 sites

17. Water quality

18. Plum Orchard Lake (Reference Site)

19. Parameters Measured • YSI: DO, pH, temperature, conductivity • Total and dissolved Fe, total Se, TDS, sulfates, alkalinity, hardness, TSS and TOC Picture credit: Melissa O’Neal

20. Sediments • July 2009 • Stantec, Inc. • Stream (13) • Reservoir (5) • Transects • Top 5 cm Picture credit: Stantec, Inc., Lexington, KY

23. Plankton • July, August, September 2010 • Mud River Reservoir (2) • Reference Site (1) • 10-20 min/ tow • Surface and mid depth • 95% ethanol

24. Benthic Macroinvertebrates • November 2009 and 2010 • Mud River (11) • Reference Site (2) • 2009: 4 kick samples • 2010: 6-8 kick samples • Sorted into functional feeding group (FFG) Photo credit: Melissa O’Neal

27. Bluegill Tissue • June 2009 and 2010 • Spawning • Mud River Reservoir • Whole Body (n=118) • Gonad (n=31) • Reference Site • Whole Body (n=86) • Gonad (n=10) • Egg selenium calculated Photo credit: John Howell

28. Bluegill Larval Lab Study • June 2009 and 2010 • 2009: Entire redd collected • 2010: Only shells, rocks, and branches • Individual redds collected • Transported to WVU wet lab

30. Larval Laboratory Study • Nest split into tanks • A Tanks: 28 day early life stage test for chronic toxicity • B Tanks: 9 day swim-up success • Tanks contained water from original location • 3-days after hatching fed fine fish food and brine shrimp twice daily

31. Larval Laboratory Study • Multiple full water changes done daily • Ammonia (daily) • Nitrite, alkalinity, hardness (weekly) • Temperature, DO, and pH (subset daily)

32. Results

33. WaterQuality

34. Water Quality

35. 1.4 1.2 1.0 0.8 Selenium (µg/g DW) 0.6 0.4 0.2 0.0 Unmined Mined Site Type Stream Sediments a a a a

36. Mud Reservoir Sediments a a a a a

37. Plankton a a b

38. Benthic Macroinvertebrates

39. 35 30 25 20 Selenium (µg/g DW) 15 10 5 0 Mud Reservoir Reference Site Bluegill Whole Body Levels * a b

40. 30 25 20 Selenium (µg/g DW) 15 10 5 0 Mud Reservoir Reference Site Bluegill Gonad Levels * a b

41. Larval Lab Study

42. Discussion

43. Water Quality • The mine boundary is a good delineation for site types • Sites in the mine and downstream were of high (> 5 µg/L) or moderate hazard (3-5 µg/L) based on the hazard profile by Lemly (1995) • Levels as low as 2 µg/L can be bioaccumulated in the food chain and cause reproductive failure

44. Sediments • All sediment samples considered no to low hazard (≤ 2 µg/g DW) • Reservoir transects • All samples also considered low to no hazard at < 2 µg/g DW • Selenium seems to remain in the water column

45. Plankton • Mud River Reservoir (mined and unmined) sites differed from the reference site • Not sorted to species (zooplankton, etc.)

46. Benthic Macroinvertebrates • Mined areas greater than dietary threshold for fish (3 µg/g) and birds (7 µg/g) • Did not see a significant difference between the reference site and unmined sites

47. Bluegill Tissues • No hazard category for whole body/gonad selenium concentrations • Egg selenium (68.76 ± 1.95 µg/g ) • High hazard (>20 µg/g)

48. Bluegill Tissues Toxic Effect Threshold: Reference Mud Reservoir 5 ug/g is considered the threshold for the health and reproductive success of freshwater fish (Lemly 1995)

49. Larval Lab Study • Did not observe deformities in larval fish • Low survival possibly due to temperature and ammonia levels

50. Summary • No observations of deformaties • Mined areas had higher selenium levels in the following categories: • Water selenium concentrations • Benthic macroinvertebrates • Whole body/gonad/egg concentrations