1. Toxicity of Daphnia to Manganese Compared to Manganese Concentrations in Local Waters Kristel Fijolek Senior Capstone, Spring 2005

2. Overview Introduction and Background Introduction and Background Hypothesis Hypothesis Materials and Methods Materials and Methods Results Results Conclusion Conclusion

3. Introduction and Background Manganese Manganese –Found in the natural environment –Range found in potable water: 0.0019 – 0.0091 mg/L –Range found in environmental fresh water: 0.00097 – 1.835 mg/L –Levels increase with acid mine drainage –Eramet ~ 4,000,000 pounds/yr into the air

4. Introduction and Background continued Eramet, Rt. 7.

5. Introduction and Background continued Manganese Manganese –Toxicity causes Parkinson’s like symptoms –Normal blood level < 10 µg/L –Can move up the food chain by concentrating in the tissues – passed on to the organism which eats it

6. Introduction and Background continued Daphnia magna as bioindicators Daphnia magna as bioindicators –Require little space and care –Easy to acquire and inexpensive –Short life cycle –Important link in the food chain –Sensitive to toxins –Commonly used in acute toxicity studies

7. Introduction and Background continued http://www.cisba.it/images/DaphniaMagna.gif

8. Hypothesis My questions: My questions: –How much of the manganese in the air is getting into the water? –What is the LC 50 for Daphnia, and how does this compare with local manganese levels? –Null hypothesis

9. Materials and Methods 48 hour acute toxicity testing 48 hour acute toxicity testing 20 Daphnia per culture dish – number dead counted per dish and averaged per concentration 20 Daphnia per culture dish – number dead counted per dish and averaged per concentration 3 dishes per MnSO 4 concentration 3 dishes per MnSO 4 concentration Testing carried out in culture medium and filtered Ohio River water Testing carried out in culture medium and filtered Ohio River water

10. Materials and Methods continued Daphnia cultures

11. Materials and Methods continued Daphnia culture medium Daphnia culture medium –CaSO 4 * H 2 O: 192 mg/L –NaHCO 3 : 192 mg/L –MgSO 4 : 120 mg/L –KCl: 8 mg/L

12. Materials and Methods continued Dosing set-up

13. Materials and Methods continued Analysis of water samples Analysis of water samples Perkin-Elmer 1100B Flame Atomic Absorption Spectrophotometer - Limit of detection: 0.052 mg/L www.york.ac.uk/depts/eeem/pics/photos/Dept/perkinelmeraa.jpg

14. Materials and Methods continued

15. All samples filtered through Whatman # 1 filter paper All samples filtered through Whatman # 1 filter paper –Ohio River –Veto Lake –Marietta College tap water

16. Results – acute toxicity RW and CM: Y =.0233 X + 11.442

17. Results – LC 50 From the equations of the trendlines, 50 (% dead) used for the Y value to calculate LC 50 From the equations of the trendlines, 50 (% dead) used for the Y value to calculate LC 50 –LC 50 in river water: 165 mg/L –LC 50 in culture medium: 165 mg/L

18. Results – water analysis

19. Veto Lake – below limit of detection Veto Lake – below limit of detection Ohio River – below limit of detection Ohio River – below limit of detection Tap water – below limit of detection Tap water – below limit of detection –Limit of detection: 0.052 mg/L

20. Conclusion LC 50 for Daphnia was much higher than the Mn levels found in the water samples. LC 50 for Daphnia was much higher than the Mn levels found in the water samples. Mn levels in the water samples were not high enough to kill Daphnia Mn levels in the water samples were not high enough to kill Daphnia

21. Acknowledgements Dr. Brown, Dr. McShaffrey, and the Marietta College biology department. Dr. Brown, Dr. McShaffrey, and the Marietta College biology department. Photographs (unless otherwise indicated) by Bill Fijolek Photographs (unless otherwise indicated) by Bill Fijolek

22. References Carroll, KC, Lopez, DL, and Stoertz MW. 2003. Solute Transport at Low Flow in an Acid Stream in Appalachian Ohio. Water, Air, and Soil Pollution. 144:195-222. Carroll, KC, Lopez, DL, and Stoertz MW. 2003. Solute Transport at Low Flow in an Acid Stream in Appalachian Ohio. Water, Air, and Soil Pollution. 144:195-222. Centers for Disease Control. Health Consultation. Washington County Air Quality (a/k/a Marietta Air Emissions) Marietta, Washington County, Ohio. 7 Sept. 2004. Centers for Disease Control. Health Consultation. Washington County Air Quality (a/k/a Marietta Air Emissions) Marietta, Washington County, Ohio. 7 Sept. 2004. Crompton TR. 1997. Toxicants in the Aqueous Ecosystem. West Sussex, England: Wiley & Sons. Crompton TR. 1997. Toxicants in the Aqueous Ecosystem. West Sussex, England: Wiley & Sons. Environmental Defense. Scorecard. 13 Sept. 2004. Environmental Defense. Scorecard. 13 Sept. 2004.

23. References continued Environmental Inquiry. Bioassays Using Daphnia. 28 Aug. 2004. http://ei.cornell.edu/toxicology/bioassays/daphnia/ analyze.asp> Environmental Inquiry. Bioassays Using Daphnia. 28 Aug. 2004. http://ei.cornell.edu/toxicology/bioassays/daphnia/ analyze.asp> Guilhermino L, Diamantino M, Silva C, and Soares AMVM. 2000. Acute Toxicity Test with Daphnia magna: An Alternative to Mammals in the Prescreening of Chemical Toxicity? Ecotoxicology and Environmental Safety. 46: 357-362. Guilhermino L, Diamantino M, Silva C, and Soares AMVM. 2000. Acute Toxicity Test with Daphnia magna: An Alternative to Mammals in the Prescreening of Chemical Toxicity? Ecotoxicology and Environmental Safety. 46: 357-362. Miller WE, Greene, JR, and Shiroyama T. 1978, Environmental Protection Agency, Office of Research and Development, Corvallis Environmental Research Laboratory. Miller WE, Greene, JR, and Shiroyama T. 1978, Environmental Protection Agency, Office of Research and Development, Corvallis Environmental Research Laboratory. Rand, G. ed. 1995. Fundamentals of Aquatic Toxicology: Effects, Environmental Fate, and Risk Assessment. Washington DC: Taylor and Francis. Rand, G. ed. 1995. Fundamentals of Aquatic Toxicology: Effects, Environmental Fate, and Risk Assessment. Washington DC: Taylor and Francis.

24. Questions?