AN INTEGRATED STUDY OF A HOT SPRINGS MICROBIAL COMMUNITY IN THERMOPOLIS, WY

The biogeochemical characterization of the Big Spring in Thermopolis, Wyoming has been an ongoing study since 2005. The spring is characterized as a stable pH neutral, low nutrient, high alkalinity, high hydrogen sulfide and no dissolved oxygen environment. Minerals are deposited rapidly within the microbial mats, trimming the filaments and forming thick carbonate and sulfate terraces that drip over the cliffs into the Big Horn River. This year’s studies emphasized the genomics of the 52^oC hot springs microbial thermophile population. Thermophiles are thought to be related to the first life on Earth, and understanding the extreme environments in which they live may help us understand the evolution of early life on, and perhaps give us clues in searching for Earth life beyond our own planet. As an integrated field and lab project, the students in a sophomore-level Earth Systems Science course at Central Wyoming College, were able to extract DNA from the microbes 12 samples, of geographic variability, to determine the types of bacteria present in this environment.

In preparation for amplification (PCR) and genetic sequencing the DNA was extracted using the PowerSoil DNA Isolation Kit from Mo Bio Laboratories, Inc. Once the DNA extraction process was completed the samples were placed into a -70^oC freezer for storage until sent for off-site processing. The 16S rRNA genomes were sequenced by Research Testing Laboratory (RTL) of Lubbock, Texas. One-step PCR and Reads 454 MSC sequencing was performed utilizing Roche 454 FLX instruments and Titanium reagents to produce a tag-encoded FLX amplicon pyrosequencing analysis, and identified by comparisons with the NCBI (v.1-1-11) archives.

Results of sequencing show a microbial community populated by over 200 possible species, with moderate to high variability between sites. Dominant species include Chloroflexus sp and Spirulina sp with others identified as Pedobacter saltans, Thiofaba tepidiphila, Sulfurovum sp, Venenivibrio stagnispumantis, Enterobacter asburiae::Enterobacter sp, Flavobacterium sp, Sporobacterium sp. In addition to the microbial analysis, the distribution of microbe mats were mapped, water geochemistry was analyzed (and was consistent with earlier work) and underwater photography of the source spring was collected.