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- Creators: Beethoven, Ludwig van, 1770-1827
Description
The taxonomic and metabolic profile of the microbial community inhabiting a natural system is largely determined by the physical and geochemical properties of the system. However, the influences of parameters beyond temperature, pH and salinity have been poorly analyzed with few studies incorporating the comprehensive suite of physical and geochemical measurements required to fully investigate the complex interactions known to exist between biology and the environment. Further, the techniques used to classify the taxonomic and functional composition of a microbial community are fragmented and unwieldy, resulting in unnecessarily complex and often non-consilient results.
This dissertation integrates environmental metagenomes with extensive geochemical metadata for the development and application of multidimensional biogeochemical metrics. Analysis techniques including a Markov cluster-based evolutionary distance between whole communities, oligonucleotide signature-based taxonomic binning and principal component analysis of geochemical parameters allow for the determination of correlations between microbial community dynamics and environmental parameters. Together, these techniques allow for the taxonomic classification and functional analysis of the evolution of hot spring communities. Further, these techniques provide insight into specific geochemistry-biology interactions which enable targeted analyses of community taxonomic and functional diversity. Finally, analysis of synonymous substitution rates among physically separated microbial communities provides insights into microbial dispersion patterns and the roles of environmental geochemistry and community metabolism on DNA transfer among hot spring communities.
The data presented here confirms temperature and pH as the primary factors shaping the evolutionary trajectories of microbial communities. However, the integration of extensive geochemical metadata reveals new links between geochemical parameters and the distribution and functional diversification of communities. Further, an overall geochemical gradient (from multivariate analyses) between natural systems provides one of the most complete predictions of microbial community functional composition and inter-community DNA transfer rates. Finally, the taxonomic classification and clustering techniques developed within this dissertation will facilitate future genomic and metagenomic studies through enhanced community profiling obtainable via Markov clustering, longer oligonucleotide signatures and insight into PCR primer biases.
This dissertation integrates environmental metagenomes with extensive geochemical metadata for the development and application of multidimensional biogeochemical metrics. Analysis techniques including a Markov cluster-based evolutionary distance between whole communities, oligonucleotide signature-based taxonomic binning and principal component analysis of geochemical parameters allow for the determination of correlations between microbial community dynamics and environmental parameters. Together, these techniques allow for the taxonomic classification and functional analysis of the evolution of hot spring communities. Further, these techniques provide insight into specific geochemistry-biology interactions which enable targeted analyses of community taxonomic and functional diversity. Finally, analysis of synonymous substitution rates among physically separated microbial communities provides insights into microbial dispersion patterns and the roles of environmental geochemistry and community metabolism on DNA transfer among hot spring communities.
The data presented here confirms temperature and pH as the primary factors shaping the evolutionary trajectories of microbial communities. However, the integration of extensive geochemical metadata reveals new links between geochemical parameters and the distribution and functional diversification of communities. Further, an overall geochemical gradient (from multivariate analyses) between natural systems provides one of the most complete predictions of microbial community functional composition and inter-community DNA transfer rates. Finally, the taxonomic classification and clustering techniques developed within this dissertation will facilitate future genomic and metagenomic studies through enhanced community profiling obtainable via Markov clustering, longer oligonucleotide signatures and insight into PCR primer biases.
ContributorsAlsop, Eric Bennie (Author) / Raymond, Jason (Thesis advisor) / Anbar, Ariel (Committee member) / Farmer, Jack (Committee member) / Shock, Everett (Committee member) / Walker, Sarah (Committee member) / Arizona State University (Publisher)
Created2014
Description
The greatest barrier to understanding how life interacts with its environment is the complexity in which biology operates. In this work, I present experimental designs, analysis methods, and visualization techniques to overcome the challenges of deciphering complex biological datasets. First, I examine an iron limitation transcriptome of Synechocystis sp. PCC 6803 using a new methodology. Until now, iron limitation in experiments of Synechocystis sp. PCC 6803 gene expression has been achieved through media chelation. Notably, chelation also reduces the bioavailability of other metals, whereas naturally occurring low iron settings likely result from a lack of iron influx and not as a result of chelation. The overall metabolic trends of previous studies are well-characterized but within those trends is significant variability in single gene expression responses. I compare previous transcriptomics analyses with our protocol that limits the addition of bioavailable iron to growth media to identify consistent gene expression signals resulting from iron limitation. Second, I describe a novel method of improving the reliability of centroid-linkage clustering results. The size and complexity of modern sequencing datasets often prohibit constructing distance matrices, which prevents the use of many common clustering algorithms. Centroid-linkage circumvents the need for a distance matrix, but has the adverse effect of producing input-order dependent results. In this chapter, I describe a method of cluster edge counting across iterated centroid-linkage results and reconstructing aggregate clusters from a ranked edge list without a distance matrix and input-order dependence. Finally, I introduce dendritic heat maps, a new figure type that visualizes heat map responses through expanding and contracting sequence clustering specificities. Heat maps are useful for comparing data across a range of possible states. However, data binning is sensitive to clustering cutoffs which are often arbitrarily introduced by researchers and can substantially change the heat map response of any single data point. With an understanding of how the architectural elements of dendrograms and heat maps affect data visualization, I have integrated their salient features to create a figure type aimed at viewing multiple levels of clustering cutoffs, allowing researchers to better understand the effects of environment on metabolism or phylogenetic lineages.
ContributorsKellom, Matthew (Author) / Raymond, Jason (Thesis advisor) / Anbar, Ariel (Committee member) / Elser, James (Committee member) / Shock, Everett (Committee member) / Walker, Sarah (Committee member) / Arizona State University (Publisher)
Created2017
ContributorsBeethoven, Ludwig van, 1770-1827 (Composer)
ContributorsBeethoven, Ludwig van, 1770-1827 (Composer)
Description
Amazonia, inhabited and investigated for millennia, continues to astonish scientists with its cultural and natural diversity. Although Amazonia is rapidly changing, its vast and varied landscape still contains a complex natural pharmacopeia. The Amazonian tribes have accrued valuable environmental and geological knowledge that can be studied. This dissertation demonstrates that Indigenous Knowledge considered alongside Western Science can enhance our understanding of the relationship of people to geological materials and hydrological resources, and reveal mineral medicines with practical applications.
I used methods from anthropology and geology to explore the geological knowledge of the Uitoto, a tribe of the Colombian Amazon. The Uitoto use two metaphors to describe Earth systems: 1. the earth is a body, and 2. the Amazon is a tree. I found that they classify surface-water systems according to observable characteristics and use mineral clays to treat various maladies. I argue that Uitoto knowledge about Amazonian mineral resources and surface water is practical, empirically–based and, in many cases, more nuanced than mainstream scientific knowledge.
I studied the mode of action of a natural antibacterial clay from the Colombian Amazon (AMZ) to discover whether the Uitoto’s claims about the clay’s medicinal values was verifiable using the methods of Western Science. Natural antibacterial clays can inhibit the growth of human pathogens. Methods from microbiology and geochemistry were combined to evaluate the mineral-microbe interactions that inhibit growth of model Gram-negative (Escherichia coli) and Gram-positive (Bacillus subtilis) bacteria. The AMZ antibacterial clay contains 45 % kaolinites and 30 % smectites. Its high surface area maintains an acidic environment (pH 4.5) and releases high concentrations of aluminum. Aluminum accumulates in the outer membrane of E. coli by binding to phospholipids. Furthermore, the membrane’s permeability increases due to synergistic effects between aluminum and transition metals released from the AMZ (i.e. Fe, Cu). The changes in the membrane may compromise its function as a barrier. Understanding the antibacterial mechanism of AMZ is key for its safe use as a natural product. These findings can help us harness the capabilities of antibacterial clays more efficiently.
Lastly, I integrated the results of this work in place-based, cross-cultural educational materials tailored for the tribal schools in the Colombian Amazon. The design of the units was informed by principles of curriculum design and successful pedagogic approaches for Native American students. The purpose of these educational materials is to return the results of research, enhance learning and participation of indigenous peoples in geosciences, and respond to the multicultural and plurilingual educational needs in countries such as Colombia.
I used methods from anthropology and geology to explore the geological knowledge of the Uitoto, a tribe of the Colombian Amazon. The Uitoto use two metaphors to describe Earth systems: 1. the earth is a body, and 2. the Amazon is a tree. I found that they classify surface-water systems according to observable characteristics and use mineral clays to treat various maladies. I argue that Uitoto knowledge about Amazonian mineral resources and surface water is practical, empirically–based and, in many cases, more nuanced than mainstream scientific knowledge.
I studied the mode of action of a natural antibacterial clay from the Colombian Amazon (AMZ) to discover whether the Uitoto’s claims about the clay’s medicinal values was verifiable using the methods of Western Science. Natural antibacterial clays can inhibit the growth of human pathogens. Methods from microbiology and geochemistry were combined to evaluate the mineral-microbe interactions that inhibit growth of model Gram-negative (Escherichia coli) and Gram-positive (Bacillus subtilis) bacteria. The AMZ antibacterial clay contains 45 % kaolinites and 30 % smectites. Its high surface area maintains an acidic environment (pH 4.5) and releases high concentrations of aluminum. Aluminum accumulates in the outer membrane of E. coli by binding to phospholipids. Furthermore, the membrane’s permeability increases due to synergistic effects between aluminum and transition metals released from the AMZ (i.e. Fe, Cu). The changes in the membrane may compromise its function as a barrier. Understanding the antibacterial mechanism of AMZ is key for its safe use as a natural product. These findings can help us harness the capabilities of antibacterial clays more efficiently.
Lastly, I integrated the results of this work in place-based, cross-cultural educational materials tailored for the tribal schools in the Colombian Amazon. The design of the units was informed by principles of curriculum design and successful pedagogic approaches for Native American students. The purpose of these educational materials is to return the results of research, enhance learning and participation of indigenous peoples in geosciences, and respond to the multicultural and plurilingual educational needs in countries such as Colombia.
ContributorsLondoño Arias, Sandra Carolina (Author) / Williams, Lynda B (Thesis advisor) / Semken, Steven (Thesis advisor) / Brandt, Elizabeth A. (Committee member) / Hartnett, Hilairy H (Committee member) / Raymond, Jason (Committee member) / Arizona State University (Publisher)
Created2016
ContributorsBeethoven, Ludwig van, 1770-1827 (Composer)
ContributorsBeethoven, Ludwig van, 1770-1827 (Composer)
ContributorsBeethoven, Ludwig van, 1770-1827 (Composer)
ContributorsBeethoven, Ludwig van, 1770-1827 (Composer)