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This research project at Davis Mine investigates microbial communities below tailings piles and in mine groundwater. Utilizing sterile anaerobic techniques, the study identified sulfur-reducing bacteria and Desulfobulbus in different media and pH conditions. Challenges included time constraints, equipment limitations, and fungal contamination. Future research involves dilution transfers, DNA extraction, and exploring iron-reducing bacteria. The findings contribute to a better understanding of long-term environmental impacts and provide valuable lab and research experience. This project aims to integrate findings into educational curriculums. Have questions? Reach out for more information.
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Davis Mine Project By: Jason Jean and Janice Wing
Presentation Outline • Reasons for our research project • Methods • Findings • Obstacles • Research continuation • Impact on us
Reasons for our research project • Personal • Professional • Enrichment media
Methods • Sterile, anaerobic technique • pH adjustment • Inoculation • Incubation • Observations and Microscopy
Sterilize Postgate and Pfennig media before inoculation in the autoclave. • Davis Mine Well media was not sterilized in the autoclave before inoculation.
Opening sediment jars and inoculating sediment into media bottles under anoxic conditions
Findings • Sulfur-reducing bacteria grew in Postgate pH 7.0 • Sulfur-reducing bacteria grew in Davis Mine Well 1 Groundwater media at pH 7.0 • Thread-like filaments were visible in sample jars that developed a black precipitate, but in no others. • Desulfobulbus was successfully cultivated in Postgate media but was not successful in Pfennig. • Orange film developed in Pfennig enrichment bottles.
Enrichment bottles with Postgate’s medium at pH 3.0 4.0 5.0 7.0
Enrichment bottles with Pfennig’s medium at pH 3.0 4.0 5.0 7.0
Davis Mine Well 1 media at pH 3.0 4.0 5.0 7.0
Davis Mine Well 3 media at pH 3.0 4.0 5.0 7.0
Davis Mine Well 4 media at pH 3.0 4.0 5.0 7.0
Black precipitate formed in Postgate media of pH 7.0 inoculated with Desulfobulbus
Black precipitate formed in transfers of Postgate media at pH 7.0
Black precipitate formed in transfers of Davis Mine Well 1 media at pH 7.0
Microscopy using a live/dead bio-stain with this fluorescent microscope. Pictures were taken.
Microscopy of Postgate pH 3.0 sample inoculated with Well 1 sediment
Microscopy of Postgate pH 4.0 inoculated with Well 4 sediment
Postgate pH 7.0 sediment from Well 4 Thread-like filament
Postgate pH 7.0 sediment from Well 4 Dark spots of pyrite forming
What is the orange film and sediment? (Seen in all Pfennig media, here the sample is pH 3.0)
Obstacles • Time • Equipment • Anaerobic technique is rigorous
Research continuation • Dilution transfers and DNA extraction • Continued incubation • Postgate media with a variety of donors • DM Well media with a variety of donors • Mercy • Iron-reducers
Impact on us • Better understanding of long-range research • Lab and project research experience • Integrate into classroom curriculum aligned with state frameworks • Biology • Environmental Science