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Item Bacterial community dynamics and variability in shallow aquifers(Montana State University - Bozeman, College of Letters & Science, 2019) Zelaya, Anna Jesus; Chairperson, Graduate Committee: Matthew Fields; Albert E. Parker, Kathryn L. Bailey, Ping Zhang, Joy Van Nostrand, Ning Daliang, Dwayne A. Elias, Jizhong Zhou, Terry C. Hazen, Adam P. Arkin and Matthew W. Fields were co-authors of the article, 'High spatiotemporal variability of bacterial diversity over short time scales with unique geohydrochemistry within a shallow, pristine aquifer' which is contained within this thesis.; Albert E. Parker, Kathryn L. Bailey, Ping Zhang, Joy Van Nostrand, Ning Daliang, Dwayne A. Elias, Jizhong Zhou, Terry C. Hazen, Adam P. Arkin and Matthew W. Fields were co-authors of the article, 'Population filtering in sediment biofilms from dynamic, source planktonic communities' which is contained within this thesis.; Sara Altenburg, Kathryn L. Bailey, Heidi J. Smith, Dwayne A. Elias and Matthew W. Fields were co-authors of the article, 'Community structure across particle size in-vitro and in-situ' which is contained within this thesis.; Heidi J. Smith, Frederick von Netzer and Matthew W. Fields were co-authors of the article, 'Contribution of total and viable DNA pools on diversity estimates of contaminated and non-contaminated subsurface sediments and groundwater' which is contained within this thesis.The shallow, terrestrial subsurface plays an important role in sustaining life above ground. Globally, subsurface environments are becoming increasingly threatened by anthropogenic sources of contamination and disturbance. The Oak Ridge Reservation (ORR) played an important role in the development of nuclear weapons during World War II. In the process, the underlying subsurface became contaminated with radioactive and hazardous wastes. Strategies to remediate polluted subsurface environments such as Oak Ridge have included natural attenuation by resident microbes such as bacteria. Such studies would be aided by a thorough understanding of the natural variability of microbial diversity over space and time in uncontaminated environments. We examined microbial community diversity and variability in both contaminated and non-contaminated sites of the ORR via ss-rRNA paired-end sequencing. An initial in-situ spatiotemporal survey of non-contaminated groundwater was conducted in order to understand how diversity changes over time in an undisturbed aquifer. Moreover, different models were used to estimate possible causal relationships between geochemical parameters and population distribution. Additionally, surrogate sediment samplers were filled with native sediment to assess the diversity of the attached bacterial fraction. Communities observed in the attached fraction were a subset of groundwater communities, although the dominant fractions of each were distinct. After initial assessment, in-vitro groundwater was used as both a nutrient source and microbial inoculum for bioreactors filled with glass beads of various sizes in order to understand particle size effects on community dynamics. Potential viability of resident microbes in both contaminated and non-contaminated groundwater and cored sediments was assessed using PMA-Seq and other complementary methods. Collectively, results show that 1) microbial communities in groundwater are highly dynamic over short timescales, 2) corresponding changes in geochemistry are mostly weakly related to changes in community structure (except perhaps after a disturbance or stress period), 3) community assembly may be affected by pore space volume, 4) PMA-viable populations differ between solid and aqueous fractions, 5) most subsurface cells are intact, and 6) traditional estimates of largely abundant populations may be influenced by the presence of DNA from non-viable members, resulting in less abundant populations being unmeasured or underestimated. The results of this study have implications for sampling and appropriate estimations of microbial populations in situ as well as the inherent variability in an uncontaminated shallow aquifer.Item Investigation of microbial population biology during the degradation of organic contaminant mixtures(Montana State University - Bozeman, College of Agriculture, 2001) Kern, Eric AnthonyItem Community analysis of groundwater and surrogate sediment samples during electron donor and electron acceptor injections into a chromium-contaminated site in Hanford, Washington, USA(Montana State University - Bozeman, College of Letters & Science, 2013) Bowen De Leon, Kara Beth; Chairperson, Graduate Committee: Matthew Fields; Bradley D. Ramsay was a co-author and Matthew W. Fields was the corresponding author of the article, 'Quality-score refinement of SSU rRNA gene pyrosequencing differs across gene region for environmental samples' in the journal 'Microbial ecology' which is contained within this thesis.; Boris Faybishenko, Bradley D. Ramsay, Darrell R. Newcomer and Terry C. Hazen were co-authors and Matthew W. Fields was the corresponding author of the article, 'Stimulation for in situ Cr(VI) bioreduction causes convergence of groundwater and sediment-adhered bacterial populations with differing population networks' submitted to the journal 'ISME journal' which is contained within this thesis.; Boris Faybishenko, Bradley D. Ramsay, Darrell R. Newcomer and Terry C. Hazen were co-authors and Matthew W. Fields was the corresponding author of the article, 'Microbial community changes with episodic nitrate during in situ stimulation for Cr(VI) reduction at the Hanford 100-H site' submitted to the journal 'Applied and environment microbiology' which is contained within this thesis.; Robin Gerlach and Brent M. Peyton were co-authors and Matthew W. Fields was the corresponding author of the article, 'Archaeal and bacterial communities in alkaline hot springs in Heart Lake Geyser Basin, Yellowstone National Park' in the journal 'Extremophiles' which is contained within this thesis.; Mary Lynn Young, Laura B. Camilleri, Steven D. Brown, Jeffrey M. Skerker and Adam M. Deutschbauer were co-authors and Matthew W. Fields was the corresponding author of the article, 'Draft genome sequence of Pelosinus fermentans JBW45 isolated during in situ stimulation for Cr(VI) reduction' in the journal 'Journal of bacteriology' which is contained within this thesis.Heavy-metal contamination is a common problem in the industrialized world today. Bioremediation, the use of microorganisms or microbial processes to degrade or reduce contaminants, has been found to be successful for many different contaminants and environments. As part of the World War II Manhattan Project, eight single-pass reactors in the Hanford Nuclear Reservation in Washington were used for plutonium production for ~43 years. Columbia River water, used to cool these reactors, was contaminated with heavy metals and radionuclides and stored in retention basins before discharge or leakage into the ground or river. Hexavalent chromium (Cr(VI)), a carcinogen and mutagen, was used as a corrosion inhibitor in the cooling systems and is a common contaminant at the Hanford Site. In 2008, a polylactate compound was injected into Cr(VI)-contaminated groundwater in the Hanford 100-H area to stimulate the resident microbial community and resulted in Cr(VI) levels below background levels for ~3 months. Temporal and spatial community analyses of the groundwater and surrogate sediment samples via small-subunit rRNA gene pyrosequencing indicated an enrichment of Pseudomonas and fermentative organisms upon injection. The sediment and groundwater communities, while different pre-injection, converged on a similar, fermentative community after injection. Correlation analyses suggested that nitrogen-metabolism and fermentation might be driving community changes. Nitrate is another common contaminant at Hanford and an influx of nitrate, a competing terminal electron acceptor, into the Hanford 100-H site may be detrimental to the bioremediation process. To test this, nitrate was injected into the system following a lactate injection to stimulate the resident community. The nitrate injection resulted in a drastic community shift from metal-reducing organisms to denitrifiers and an increase in Cr(VI) concentrations to above background levels. Long after the nitrate injection, denitrifiers continued to be predominant in the groundwater community; however, the sediment community was comprised of denitrifying and metal-reducing populations. This suggests that the effects of an episodic nitrate event are long lasting, but the sediment community is more resilient than the groundwater community. These community analyses have revealed organisms and metabolic properties of interest during polylactate or nitrate perturbation and can be used to improve future bioremediation strategies.