1. Growth and Morphological Variation in Microorganisms Exposed to Lead JOE TONEY*, DANIELLE DAY, JENNIFER SEABAUGH, ANDREW CORCORAN, JENNIFER ARNOLD, and JAMES E. CHAMPINE Southeast Missouri State University

2. Lead Mining in Missouri Lead-belt region (Mineral Area College) Chat: mine tailings covering acres of land Hypothesis: Natural enrichment for lead- resistant microorganisms

3. Lead Content, ppm, of Soil and Chat FractionSample 12 Soil Sample 14 Chat Sample 15 Soil Total Bioavailable 2,72516,7331,289 Total4,02316,7332,294 IsolatesPb4, Pb5CPA 1,2,6 CPC 2,3,4,5 Pb1, Pb3

4. Approach Bacteria were eluted from chat and applied to the surface of agar plates. Defined chemical media containing lead called “Roane Media” This media is formulated to minimize precipitation of lead. Equation –[Pb]sol = 0.045[Pb]tot-0.12 mM

5. My Isolates CPC2 CPC3 CPC5 CPC4 TSA RM RM 1mM RM 2.5mM

6. CPC5=CPA6=Pb5 Rhodococcus fascians (luteus)

7. CPC3=CPA1 Gram positive species not in the MIDI database Microbacterium (Corynebacterium) laevinformans Lead No Lead

8. Serratia marcescens No Lead Lead

9. Enterobacter Pb 3 O 4 PbO PbO 2 No Lead Lead

10. Gradient Plates Safranin Crystal Violet Gradient Discontinuous Gradient

11. CPC5 0 mM Pb 5 mM Pb No Growth

12. Reasons for Color Variation Secondary Metabolism –Pigments produced in response to stress –Serratia marcescens –Test by pigment extraction procedures Precipitation of Pb –Found in Fungal Isolate Pb3 –Coloration resembles Lead Oxides and/or Phosphate –Test by Electron Microscopy and Electron Diffraction Studies

13. Acknowledgements Coauthors –Jennifer Seabaugh –Danielle Day –James Collins Jennifer Arnold, Andrew Corcoran Maija Bluma Funding from SEMO Undergraduate Research Program