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Microorganisms can produce metabolites in the gut including short chain fatty acids, vitamins, and amino acids. Certain metabolites produced in the gut can affect the brain through changes in neurotransmitter concentrations. Serotonin, a neurotransmitter, is associated with mood, appetite, and sleep. Up to 90% of serotonin synthesis is located in the gut, by human enterochromaffin cells. Bacteria known to biosynthesize tryptophan, precursor to serotonin, include Escherichia coli, Enterococcus and Streptococcus. Tryptophan is synthesized by bacteria with the enzyme tryptophan synthase and requires Vitamin B6 (Pyridoxal). We hypothesize that gut isolates from surgical weight loss patients can enhance tryptophan production, which relies on vitamin B6 availability. Our goal was to isolate bacteria in order to test for tryptophan production and to determine how Vitamin B6 concentrations could affect tryptophan production. We isolated gut bacteria was from successful surgical weight loss patient with selective pressures for Enterobacter isolates and Enterococcus isolates. We tested the isolates were tested to determine if they could biosynthesize tryptophan in-vitro. Bacterial cultures were enriched with yeast and enriched with serine and indole, substrates necessary for tryptophan biosynthesis. We analyzed the supernatant samples for tryptophan production using GC-FID. Bacterial isolates most closely related to E. coli and Klebsiella based on 16S rRNA gene sequences, produced tryptophan in vitro. While under serine & indole media conditions, R1, the isolate most similar to Klebsiella produced more tryptophan than R14, the isolate most similar to E. coli. We tested the R1 isolate with a gradient of vitamin B6 concentrations from 0.02 µg/mL to 0.2 µg/mL to determine its effect on tryptophan production. When less than 0.05 µg/mL of Vitamin B6 was added, tryptophan production at 6 hours was higher than tryptophan production with Vitamin B6 concentrations at 0.05 µg/mL and above. The production and consumption of tryptophan by Klebsiella under 0 µg/mL and 0.02 µg/mL concentrations of Vitamin B6 occurred at a faster rate when compared to concentrations 0.05 µg/mL or higher of Vitamin B6.
ContributorsYee, Emily L. (Author) / Krajmalnik-Brown, Rosa (Thesis director) / Ilhan, Zehra (Committee member) / W. P. Carey School of Business (Contributor) / School of Life Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2016-05
Description
Under current climate conditions northern peatlands mostly act as C sinks; however, changes in climate and environmental conditions, can change the soil carbon decomposition cascade, thus altering the sink status. Here I studied one of the most abundant northern peatland types, poor fen, situated along a climate gradient from tundra (Daring Lake, Canada) to boreal forest (Lutose, Canada) to temperate broadleaf and mixed forest (Bog Lake, MN and Chicago Bog, NY) biomes to assess patterns of microbial abundance across the climate gradient. Principal component regression analysis of the microbial community and environmental variables determined that mean annual temperature (MAT) (r2=0.85), mean annual precipitation (MAP) (r2=0.88), and soil temperature (r2=0.77), were the top significant drivers of microbial community composition (p < 0.001). Niche breadth analysis revealed the relative abundance of Intrasporangiaceae, Methanobacteriaceae and Candidatus Methanoflorentaceae fam. nov. to increase when MAT and MAP decrease. The same analysis showed Spirochaetaceae, Methanosaetaceae and Methanoregulaceae to increase in relative abundance when MAP, soil temperature and MAT increased, respectively. These findings indicated that climate variables were the strongest predictors of microbial community composition and that certain taxa, especially methanogenic families demonstrate distinct patterns across the climate gradient. To evaluate microbial production of methanogenic substrates, I carried out High Resolution-DNA-Stable Isotope Probing (HR-DNA-SIP) to evaluate the active portion of the community’s intermediary ecosystem metabolic processes. HR-DNA-SIP revealed several challenges in efficiency of labelling and statistical identification of responders, however families like Veillonellaceae, Magnetospirillaceae, Acidobacteriaceae 1, were found ubiquitously active in glucose amended incubations. Differences in metabolic byproducts from glucose amendments show distinct patterns in acetate and propionate accumulation across sites. Families like Spirochaetaceae and Sphingomonadaceae were only found to be active in select sites of propionate amended incubations. By-product analysis from propionate incubations indicate that the northernmost sites were acetate-accumulating communities.
These results indicate that microbial communities found in poor fen northern peatlands are strongly influenced by climate variables predicted to change under current climate scenarios. I have identified patterns of relative abundance and activity of select microbial taxa, indicating the potential for climate variables to influence the metabolic pathway in which carbon moves through peatland systems.
ContributorsSarno, Analissa Flores (Author) / Cadillo-Quiroz, Hinsby (Thesis advisor) / Garcia-Pichel, Ferran (Committee member) / Krajmalnik-Brown, Rosa (Committee member) / Childers, Daniel (Committee member) / Arizona State University (Publisher)
Created2022
Description
Widespread use of halogenated organic compounds for commercial and industrial purposes makes halogenated organic pollutants (HOPs) a global challenge for environmental quality. Current wastewater treatment plants (WWTPs) are successful at reducing chemical oxygen demand (COD), but the removal of HOPs often is poor. Since HOPs are xenobiotics, the biodegradation of HOPs is usually limited in the WWTPs. The current methods for HOPs treatments (e.g., chemical, photochemical, electrochemical, and biological methods) do have their limitations for practical applications. Therefore, a combination of catalytic and biological treatment methods may overcome the challenges of HOPs removal.This dissertation investigated a novel catalytic and biological synergistic platform to treat HOPs. 4-chlorophenol (4-CP) and halogenated herbicides were used as model pollutants for the HOPs removal tests. The biological part of experiments documented successful co-oxidation of HOPs and analog non-halogenated organic pollutants (OPs) (as the primary substrates) in the continuous operation of O2-based membrane biofilm reactor (O2-MBfR). In the first stage of the synergistic platform, HOPs were reductively dehalogenated to less toxic and more biodegradable OPs during continuous operation of a H2-based membrane catalytic-film reactor (H2-MCfR). The synergistic platform experiments demonstrated that OPs generated in the H2-MCfR were used as the primary substrates to support the co-oxidation of HOPs in the subsequent O2-MBfR. Once at least 90% conversation of HOPs to OPs was achieved in the H2-MCfR, the products (OPs to HOPs mole ratio >9) in the effluent could be completely mineralized through co-oxidation in O2-MBfR. By using H2 gas as the primary substrate, instead adding the analog OP, the synergistic platform greatly reduced chemical costs and carbon-dioxide emissions during HOPs co-oxidation.
ContributorsLuo, Yihao (Author) / Rittmann, Bruce (Thesis advisor) / Krajmalnik-Brown, Rosa (Committee member) / Torres, Cesar (Committee member) / Arizona State University (Publisher)
Created2022
Description
Understanding why animals form social groups is a fundamental aim of sociobiology. To date, the field has been dominated by studies of kin groups, which have emphasized indirect fitness benefits as key drivers of grouping among relatives. Nevertheless, many animal groups are comprised of unrelated individuals. These cases provide unique opportunities to illuminate drivers of social evolution beyond indirect fitness, especially ecological factors. This dissertation combines behavioral, physiological, and ecological approaches to explore the conditions that favor group formation among non-kin, using as a model the facultatively social carpenter bee, Xylocopa sonorina. Using behavioral and genetic techniques, I found that nestmates in this species are often unrelated, and that non-kin groups form following extensive inter-nest migration.Group living may arise as a strategy to mitigate constraints on available breeding space. To test the hypothesis that nest construction is prohibitively costly for carpenter bees, I measured metabolic rates of excavating bees and used imaging techniques to quantify nest volumes. From these measurements, I found that nest construction is highly energetically costly, and that bees who inherit nests through social queuing experience substantial energetic savings. These costs are exacerbated by limitations on the reuse of existing nests. Using repeated CT scans of nesting logs, I examined changes in nest architecture over time and found that repeatedly inherited tunnels become indefensible to intruders, and are subsequently abandoned. Together, these factors underlie intense competition over available breeding space. The imaging analysis of nesting logs additionally revealed strong seasonal effects on social strategy, with social nesting dominating during winter. To test the hypothesis that winter social nesting arises from intrinsic physiological advantages of grouping, I experimentally manipulated social strategy in overwintering bees. I found that social bees conserve heat and body mass better than solitary bees, suggesting fitness benefits to grouping in cold, resource-scarce conditions. Together, these results suggest that grouping in X. sonorina arises from dynamic strategies to maximize direct fitness in response to harsh and/or competitive conditions. These studies provide empirical insights into the ecological conditions that favor non-kin grouping, and emphasize the importance of ecology in shaping sociality at its evolutionary origins.
ContributorsOstwald, Madeleine (Author) / Fewell, Jennifer H (Thesis advisor) / Amdam, Gro (Committee member) / Harrison, Jon (Committee member) / Pratt, Stephen (Committee member) / Kapheim, Karen (Committee member) / Arizona State University (Publisher)
Created2022
Description
Electroactive bacteria connect biology to electricity, acting as livingelectrochemical catalysts. In nature, these bacteria can respire insoluble compounds like
iron oxides, and in the laboratory, they are able to respire an electrode and produce an
electrical current. This document investigates two of these electroactive bacteria:
Geobacter sulfurreducens and Thermincola ferriacetica. G. sulfurreducens is a Gramnegative iron-reducing soil bacterium, and T. ferriacetica is a thermophilic, Grampositive bacterium that can reduce iron minerals and several other electron acceptors.
Respiring insoluble electron acceptors like metal oxides presents challenges to a
bacterium. The organism must extend its electron transport chain from the inner
membrane outside the cell and across a significant distance to the surface of the electron
acceptor. G. sulfurreducens is one of the most-studied electroactive bacteria, and despite
this there are many gaps in knowledge about its mechanisms for transporting electrons
extracellularly. Research in this area is complicated by the presence of multiple pathways
that may be concurrently expressed. I used cyclic voltammetry to determine which
pathways are present in electroactive biofilms of G. sulfurreducens grown under different
conditions and correlated this information with gene expression data from the same
conditions. This correlation presented several genes that may be components of specific
pathways not just at the inner membrane but along the entire respiratory pathway, and I
propose an updated model of the pathways in this organism. I also characterized the
composition of G. sulfurreducens and found that it has high iron and lipid content
independent of growth condition, and the high iron content is explained by the large
abundance of multiheme cytochrome expression that I observed. I used multiple
microscopy techniques to examine extracellular respiration in G. sulfurreducens, and in
the process discovered a novel organelle: the intracytoplasmic membrane. I show 3D
reconstructions of the organelle in G. sulfurreducens and discuss its implications for the
cell’s metabolism. Finally, I discuss gene expression in T. ferriacetica in RNA samples
collected from an anode-respiring culture and highlight the most abundantly expressed
genes related to anode-respiring metabolism.
ContributorsHowley, Ethan Thomas (Author) / Torres, César I (Thesis advisor) / Krajmalnik-Brown, Rosa (Thesis advisor) / Nannenga, Brent (Committee member) / Arizona State University (Publisher)
Created2022
Description
The microorganisms that colonize the gastrointestinal tract have been recognized over the last several decades to have a significant bearing on the health trajectories of the hosts that harbor them. The collection of these gut microbes display links with acute and chronic disease, garnering substantial interest in leveraging the microbiome for improved health states. How these microbes assemble as a complex community and interact with each other, and the host depends on a multitude of factors. In adulthood, diet is one of the main moderators, having a significant influence on community composition and the functional output captured in the metabolites produced and/or modified by the gut microbiome. Thus, the assembly of microbes in the gut are tightly intertwined with health. In this dissertation, I examine the impact of diet and feeding behaviors on the gut microbiome and what features may be grounding or responsive under such pressures. Specifically, I first explore the avian gut microbiome as a barometer of nutritional and environmental influence on host health. Birds have continually displayed robust physiology under dietary pressures, placing them in an important, though underutilized, position within the translational science framework. Second, I describe the association of food insecurity on gut microbiome and metabolome profiles in a diverse college-based sample. Food insecurity provides its own set of unique pressures, such as unintentional calorie restriction, and inconsistent dietary intake and access to healthy food options. Third, I examine the effect of a one vs. two-consecutive days of intermittent fasting on the gut microbiome, the plasma metabolome, and associated clinical outcomes in overweight and obese adults. Growing in scientific and lay popularity, dietary fasting has been noted to induce changes in the diversity of gut microflora and gut motility, though different fasting lengths have not been assessed in the context of the human microbiome. Overall, this collection of work underscores that the community of microbes in the gut are individualized, resilient, and baseline composition and functioning are germane to how an individual may react to a particular dietary intervention.
ContributorsMohr, Alex (Author) / Sweazea, Karen L. (Thesis advisor) / Johnston, Carol S. (Committee member) / Sears, Dorothy D. (Committee member) / Whisner, Corrie M. (Committee member) / Krajmalnik-Brown, Rosa (Committee member) / Arizona State University (Publisher)
Created2022
Description
Trichloroethene (TCE) and hexavalent chromium (Cr (VI)) are ubiquitous subsurface contaminants affecting the water quality and threatening human health. Microorganisms capable of TCE and Cr (VI) reductions can be explored for bioremediation at contaminated sites. The goal of my dissertation research was to address challenges that decrease the efficiency of bioremediation in the subsurface. Specifically, I investigated strategies to (i) promote improve microbial reductive dechlorination extent through the addition of Fe0 and (ii) Cr (VI) bio-reduction through enrichment of specialized microbial consortia. Fe0 can enhance microbial TCE reduction by inducing anoxic conditions and generating H2 (electron donor). I first evaluated the effect of Fe0 on microbial reduction of TCE (with ClO4– as co-contaminant) using semi-batch soil microcosms. Results showed that high concentration of Fe0 expected during in situ remediation inhibited microbial TCE and ClO4– reduction when added together with Dehalococcoides mccartyi-containing cultures. A low concentration of aged Fe0 enhanced microbial TCE dechlorination to ethene and supported complete microbial ClO4– reduction. I then evaluated a decoupled Fe0 and biostimulation/bioaugmentation treatment approach using soil packed columns with continuous flow of groundwater. I demonstrated that microbial TCE reductive dechlorination to ethene can be benefitted by Fe0 abiotic reactions, when biostimulation and bioaugmentation are performed downstream of Fe0 addition. Furthermore, I showed that ethene production can be sustained in the presence of aerobic groundwater (after Fe0 exhaustion) by the addition of organic substrates. I hypothesized that some lessons learned from TCE Bioremediation can be applied also for other pollutants that can benefit from anaerobic reductions, like Cr (VI). Bioremediation of Cr (VI) has historically relied on biostimulation of native microbial communities, partially due to the lack of knowledge of the benefits of adding enriched consortia of specialized microorganisms (bioaugmentation). To determine the merits of a specialized consortium on bio-reduction of Cr (VI), I first enriched a culture on lactate and Cr (VI). The culture had high abundance of putative Morganella species and showed rapid and sustained Cr (VI) bio-reduction compared to a subculture grown with lactate only (without Morganella). Overall, this dissertation work documents possible strategies for synergistic abiotic and biotic chlorinated ethenes reduction, and highlights that specialized consortia may benefit Cr (VI) bio-reduction.
ContributorsMohana Rangan, Srivatsan (Author) / Krajmalnik-Brown, Rosa (Thesis advisor) / Delgado, Anca G (Thesis advisor) / Torres, César I (Committee member) / van Paassen, Leon (Committee member) / Arizona State University (Publisher)
Created2022
Description
This dissertation encompasses the interaction of antimicrobial chemicals and emerging contaminants with multi-drug resistant (MDR) bacteria and their implications in engineered systems. The aim is to investigate the effect of combination antimicrobials on MDR bacteria E. coli, evaluate the extent of synergism and antagonism of utilizing two distinct biocidal chemicals, and evaluate the influence of endocrine-disrupting chemicals (EDCs) on protein production in response to stressors. Resistance mechanisms of bacteria such as E. coli include the use of protein systems that efflux excess nutrients or toxic compounds. These efflux proteins activate in response to environmental stressors such as contaminants and antimicrobials to varying degrees and are major contributors to antibiotic resistance in pathogenic bacteria. As is the case with engineered microbial environments, large quantities of emerging contaminants interact with bacteria, influencing antibiotic resistance and attenuation of these chemicals to an unknown degree. Interactions of antimicrobials on MDR bacteria such as E. coli have been extensively studied for pathogens, including synergistic combinations. Despite these studies in this field, a fundamental understanding of how chemicals influence antibiotic resistance in biological processes typical of engineered microbial environments is still ongoing. The impacts of EDCs on antibiotic resistance in E. coli were investigated by the characterization of synergism for antimicrobial therapies and the extrapolation of these metrics to the cycling of EDCs in engineered systems to observe the extent of antibiotic resistance proteins to the EDCs. The impact of this work provides insight into the delicate biochemistry and ongoing resistance phenomena regarding engineered systems.
ContributorsNovoa, Diego Erick (Author) / Conroy-Ben, Otakuye (Thesis advisor) / Abbazadegan, Morteza (Committee member) / Krajmalnik-Brown, Rosa (Committee member) / Arizona State University (Publisher)
Created2022
Description
Speciation, or the process by which one population diverges into multiple populations that can no longer interbreed with each other, has brought about the incredible diversity of life. Mechanisms underlying this process can be more visible in the early stages of the speciation process. The mechanisms that restrict gene flow in highly mobile species with no absolute barriers to dispersal, especially marine species, are understudied. Similarly, human impacts are reshaping ecosystems globally, and we are only just beginning to understand the implications of these rapid changes on evolutionary processes. In this dissertation, I investigate patterns of speciation and evolution in two avian clades: a genus of widespread tropical seabirds (boobies, genus Sula), and two congeneric passerine species in an urban environment (cardinals, genus Cardinalis). First, I explore the prevalence of gene flow across land barriers within species and between sympatric species in boobies. I found widespread evidence of gene flow over all land barriers and between 3 species pairs. Next, I compared the effects of urbanization on the spatial distributions of two cardinal species, pyrrhuloxia (Cardinalis sinuatus) and northern cardinals (Cardinalis cardinalis), in Tucson, Arizona. I found that urbanization has different effects on the spatial distributions of two closely related species that share a similar environmental niche, and I identified environmental variables that might be driving this difference. Then I tested for effects of urbanization on color and size traits of these two cardinal species. In both of these species, urbanization has altered traits involved in signaling, heat tolerance, foraging, and maneuverability. Finally, I tested for evidence of selection on the urban populations of both cardinal species and found evidence of both parallel selection and introgression between the species, as well as selection on different genes in each species. The functions of the genes that experienced positive selection suggest that light at night, energetics, and air pollution may have acted as strong selective pressures on these species in the past. Overall, my dissertation emphasizes the role of introgression in the speciation process, identifies environmental stressors faced by wildlife in urban environments, and characterizes their evolutionary responses to those stressors.
ContributorsJackson, Daniel Nelson (Author) / McGraw, Kevin J (Thesis advisor) / Amdam, Gro (Committee member) / Sweazea, Karen (Committee member) / Taylor, Scott (Committee member) / Arizona State University (Publisher)
Created2023
Description
The objective of this meta-analysis is to holistically evaluate the existing body of literature on the anti-neoplastic potential of snake and bee venom. In recent years, venom-based therapeutics have emerged as a promising solution for combating cancer, generating a notable rise in publications on the topic. Consequently, this comprehensive study aims to assess the current state of research and identify trends that may guide future investigations. Following the guidelines established by PRISMA, a total sample of 26 research papers were extracted from the electronic databases, PubMed and Scopus. These papers were categorized based on their publication dates, and research questions were formulated regarding three main topics: venom type, cancer-targeting mechanism, and cancer type. Statistical analysis of the research questions was performed using 2x2 contingency tables for a chi-square test. The results of the analysis reveal a statistically significant increase in publications focused on cell death mechanisms and breast cancer in recent years. This increase in publications reflects a growing interest in the potential for venom to induce apoptosis in cancer cells and target the unique biological properties of breast cancer. Overall, this meta-analysis offers insight into the evolving sphere of venom-based cancer research, providing a glimpse into the potential trajectory of this field.
ContributorsHolder, Marina (Author) / Amdam, Gro (Thesis director) / Mana, Miyeko (Committee member) / Barrett, The Honors College (Contributor) / School of Life Sciences (Contributor) / Economics Program in CLAS (Contributor)
Created2023-12