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Growth and Morphological Growth and Morphological Variation in Microorganisms Variation in Microorganisms
Exposed to Lead Exposed to Lead
JOE TONEY*, DANIELLE DAY, JENNIFER JOE TONEY*, DANIELLE DAY, JENNIFER SEABAUGH, ANDREW CORCORAN, SEABAUGH, ANDREW CORCORAN, JENNIFER ARNOLD, and JAMES E. JENNIFER ARNOLD, and JAMES E.
CHAMPINECHAMPINESoutheastSoutheast Missouri State University Missouri State University
Lead Mining in MissouriLead Mining in Missouri
• Lead-belt region (Mineral Area College)
• Chat: mine tailings covering acres of land
• Hypothesis: Natural enrichment for lead-resistant microorganisms
Lead Content, ppm, of Soil and ChatLead Content, ppm, of Soil and Chat
Fraction Sample 12 Soil
Sample 14 Chat
Sample 15 Soil
Total
Bioavailable
2,725 16,733 1,289
Total 4,023 16,733 2,294
Isolates Pb4, Pb5 CPA 1,2,6
CPC 2,3,4,5
Pb1, Pb3
ApproachApproach
• 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
My IsolatesMy IsolatesCPC2 CPC3
CPC5CPC4
TSA RM
RM 1mM
RM 2.5mM
CPC5=CPA6=Pb5CPC5=CPA6=Pb5Rhodococcus fascians (luteus)Rhodococcus fascians (luteus)
CPC3=CPA1CPC3=CPA1
• Gram positive species not in the MIDI database
• Microbacterium (Corynebacterium) laevinformans
Lead
No Lead
Serratia marcescensSerratia marcescens
No Lead Lead
EnterobacterEnterobacter
Pb3O4PbOPbO2
No Lead
Lead
Gradient PlatesGradient Plates
Safranin Crystal Violet
Gradient Discontinuous Gradient
CPC5
0 mM Pb
5 mM Pb
NoGrowth
Reasons for Color VariationReasons 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
AcknowledgementsAcknowledgements
• Coauthors– Jennifer Seabaugh– Danielle Day– James Collins
• Jennifer Arnold, Andrew Corcoran
• Maija Bluma• Funding from SEMO
Undergraduate Research Program