Microbiology Commons^™

Developing Field And Spaceflight Microscopy, Nikki Johnston

Dissertations and Theses

Designing for spaceflight favors the reduction of unnecessary features and cost-cutting, whether thisbe computation, hardware, or procedure. Resources are spent toward fortification and mission success over convenience. Such design goals paired with the relatively smaller budgets of academia lead us to do as much as we can in-house and use open-source as much as we can without compromising mission success. We also believe that thoroughly testing technology in extreme environments on earth is necessary for the success of a flight instrument. We focus on microscopy for life detection. I have spent my time here learning methods of hand-building microscopes to …

Exploring The Durability Of Marine Picocyanobacterial Microcolonies, Erik Swanson

University Honors Theses

Picocyanobacteria of the genus Synechococcus are widespread aquatic photoautotrophs that play essential roles in primary production and carbon cycling. Some Synechococcus strains form multicellular microcolonies, but the mechanisms of cell-to-cell attachments and structural durability of these microcolonies remain poorly understood. This research investigated the physical durability of microcolonies in two marine Synechococcus strains (CC9605 and CC9311) using physical disruption treatments. Cultures were subjected to vortexing and sonication treatments, and populations of single cells and microcolonies were quantified using flow cytometry based on differences in measures of size. Results indicated that strain CC9605 showed no significant treatment effect, whereas CC9311 displayed …

Developing A High-Resolution Off-Axis Common-Mode Digital Holographic Microscope, Lucy Cook

University Honors Theses

Off-axis digital holographic microscopy (DHM) is a powerful tool for 3D, non-invasive live-cell tracking without moving parts. However, traditional setups face an inherent dilemma: split-path interferometers offer high spatial resolution but poor temporal stability, while more stable common-mode configurations are historically limited to lower numerical aperture (NA) regimes. This thesis bridges that gap by scaling a common-mode DHM architecture into a high-resolution benchtop instrument featuring NA = 0.65 objectives, paired with a high-power 520 nm laser source to combat transmission losses and sustain imaging frame rates across an expanded optical footprint. We map the multi-variable design space required to satisfy …

Investigating A Link Between Dna Repair And Dna Replication In Escherichia Coli Using A Bacterial Two-Hybrid System, Lo Cooper

University Honors Theses

Cells rely on nucleotide excision repair (NER) to remove bulky DNA lesions, such as those caused by UV light. In Escherichia coli, NER involves the UvrABC complex, and includes two main pathways, global genomic repair and transcription-coupled repair (TCR). While TCR, which preferentially removes lesions that block RNA polymerase on the transcribed strand of active genes, is well studied, less is known about whether a similar coupling between repair proteins and replisomes blocked by damaged DNA exists. Such a process has been suggested by a high throughput interactome study, using the replisome subunit HolC as bait and the damage …

Digesting The Details: How Cooking Shapes The Microbiome And Influences Obesity, Tracy Lieu

University Honors Theses

Obesity is a growing global health concern linked to chronic diseases and changes in the gut microbiome. While diet is known to affect gut microbes, less attention has been given to how food processing methods, such as heating, fermentation, and cold processing, change food in ways that influence the gut microbiome and health. This thesis reviews research on how different cooking and preparation techniques affect the availability of nutrients for microbial fermentation and the resulting effects on microbial diversity and metabolism. The findings show that fermentation and cold processing are the most supportive of gut health. These methods help preserve …

Culturable Microbial Communities In Root Nodules Of Red Alder (Alnus Rubra Bong.) And Scotch Broom (Cytisus Scoparius L.): Patterns Of Diversity And Composition, Felisha (Fay) Rene Pooler

Dissertations and Theses

Endophytes are a hyper diverse group of bacterial or fungal organisms residing within plant tissues without causing visible symptoms of disease. While their overall diversity and ecological functions are not well-understood, some endophytes can provide a range of important benefits to their host such as enhanced growth and increased tolerance to environmental stress. Nitrogen-fixing bacteria residing within nodules--specialized structures on the roots of legumes and actinorhizal plants--play an integral role in the plant microbiome by facilitating nitrogen availability in nutrient-poor environments. Remarkably, the patterns of diversity including both bacterial and fungal endophytes in root nodules are virtually known, except in …

The Experience Of Homelessness And Its Effects On Gut Eubiosis, Kamerin Villagomez

University Honors Theses

Microorganisms which populate the human gastrointestinal tract (GIT) have a profound effect on overall human health. The relationship between host health and eubiosis, or balance, of the GIT microbiome is symbiotic and can have reciprocal relationships with quality of sleep, quality of diet, stress and mental health, and the experience of pain. These specific aspects of health are unique in populations experiencing homelessness due to the multi-dimensional and complex nature of their lived experience. Very few studies explore populations experiencing homelessness, with even fewer investigating the intersection between this population and the human GIT microbiome. It is important to do …

Predation Of Appendicularians On Prochlorococcus, Carey Sweeney

Dissertations and Theses

Prochlorococcus is the most abundant photosynthetic cell on Earth. This picocyanobacterium is critical to primary productivity and biogeochemical cycles of the open ocean. While abiotic environmental factors that influence Prochlorococcus are relatively well-understood, biotic controls are less well known. Appendicularians are ubiquitous gelatinous zooplankton that feed on marine microorganisms including Prochlorococcus. However, the details of this feeding interaction are extremely understudied relative to its potential importance. In this study, I characterized the relationship between appendicularians and Prochlorococcus with laboratory experiments. I showed that feeding on Prochlorococcus changes with the life stage of the appendicularians and concentration of prey. I …

Dietary Saturated Fatty Acids Modulate Host Innate Immunity, Amy Lorraine Seufert

Dissertations and Theses

Nutrition, cellular metabolism, and inflammatory regulation of the immune system are interconnected, and advancing our understanding of this will greatly improve treatment of metabolic and inflammatory disease. Specifically, dietary saturated fats are becoming increasingly appreciated for their ability to modify innate immune cell function in mammals following absorption into the blood and lymphatic circulation systems. The work herein focuses on the capacity for lipids in the diet to modulate innate immune memory. My thesis first broadly outlines fundamental background, including: the evolution of human nutrition, the discovery of innate immune memory, and key topics that reoccur throughout the text surrounding …

Prochlorococcus Death By A Ubiquitous Zooplankton Filter Feeder, Avery Harman

University Honors Theses

Prochlorococcus is a globally abundant cyanobacterium that can help reveal the ecology of microorganisms in the ocean. Specifically, mortality by zooplankton such as appendicularians is globally important and Prochlorococcus can help reveal some of the choices made around feeding for this ubiquitous filter feeder. Two ecotypes of Prochlorococcus were introduced to appendicularians to determine if appendicularian feed selectively between these ecotypes. Further, a single ecotype of Prochlorococcus was fed to appendicularians over a variety of life stages to determine if retention rate changed with appendicularian life stage. My results reveal that there is no significant selection between the two Prochlorococcus …

Hell's Tails: Localizing The Tail Proteins Of The Hyperthermophile-Infecting Virus Ssv1, Jane Arterberry

University Honors Theses

The tail of Saccharolobus spindle-shaped virus 1 (SSV1) has long been thought to be composed solely of viral protein 4 (VP4), but recent SSV1 structural studies have implicated the presence of additional structural proteins C166 and B78 in SSV1's unique tail. The exact locations of these proteins have not been confirmed. Structural proteins VP4 (C-terminus) and B78 are hypothesized to contribute to an adapter region bridging the viral capsid to the tail, whereas C166 is thought to comprise the majority of an internal extended structure, respectively. SSV1 structural proteins lack cysteine residues providing targets to determine their locations in the …

The Influence Of A Ubiquitous Filter Feeder On Coastal Microbial Communities, Melissa Steinman

Dissertations and Theses

Doliolids are a unique study system for investigating the intricate relationship between pelagic tunicates and the marine microbial community. This study identified the eukaryotic and prokaryotic microbial taxa in wild-caught doliolids in the Northern California Current system. Doliolids were collected during bloom events identified at three different shelf locations with variable upwelling intensity. This study found that doliolids host a microbiome unique from that of the seawater surrounding them and that feeding by doliolids includes a range of prokaryotic microbial functional groups. This is the first study to identify that doliolids are a source of mortality for pelagic archaea, and …

The Influence Of A Ubiquitous Filter Feeder On Coastal Microbial Communities., Melissa Steinman, Moritz S. Schmid, Robert K. Cowen, Su Sponaugle, Kelly R. Sutherland, Anne W. Thompson

Student Research Symposium

Doliolids have a unique ability to impact the marine microbial community through bloom events and high filtration rates. Their predation on large eukaryotic microorganisms is established and evidence of predation on smaller prokaryotic microorganisms is beginning to emerge. We studied the retention of both eukaryotic and prokaryotic microbial taxa by wild-caught doliolids in the northern California Current system. We use qPCR to quantify the impact of doliolids on three important and globally abundant taxa: Synechococcus, SAR11 and diatoms. Doliolids were collected during bloom events identified at three different shelf locations with variable upwelling intensities. We discovered that in addition …

Diving Into Aquatic Microbial Ecology And Evolution With Anne Thompson, Anne Thompson

PDXPLORES Podcast

On this episode of PDXPLORES, Assistant Professor of Biology, Anne Thompson, discusses her award-winning research work studying the diverse ecology of microorganisms found in Earth’s oceans. Thompson’s research examines microbial mortality impacts on the structure of oceanic food webs, carbon flow, and cell interactions that create dynamic patterns of nutrient and energy abundance within diverse aquatic ecosystems.

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Investigating The Mammillary Bodies As An Early Target Of Alzheimer's Disease, Cole Martinson

University Honors Theses

Alzheimer's disease (AD) is the most common form of dementia. With minimal treatment options and no cure, developing a deeper understanding of the pathology of the disease is crucial. For nearly four decades, the accumulation of beta-amyloid (Aβ) plaques has been correlated with the disease and its progression. Previous studies mapping this accumulation show that the mammillary bodies (MB) are an early target of the disease. Glutamatergic neurons are of key interest due to their prevalence in the central nervous system, specifically the MB. To understand the effects of AD on the glutamatergic system, we used vesicular glutamate transporter 2 …

Understanding Ubiquitin-Like Signaling At The Host-Pathogen Interface, Danna Eloisa Guzman Gonzalez

University Honors Theses

Research on pathogen intervention and ubiquitin modification has increased over the last few years but a lot is still unknown about ubiquitin-like modifiers (Ubl) and how or if they are modified by pathogens in the same way as Ubiquitin. In this work, two different studies conducted over the course of 2 years are highlighted. The first study focused on purifying and synthesizing four activity based probes in order to study enzymatic activity in the Ubl-modification systems. These probes were reacted with virulence factors isolated from different bacteria and it was concluded that there were unexpected bands formed between some of …

The Influence Of Microorganisms On Human Health, Tiffany Bui

University Honors Theses

The purpose of this thesis is to explore the interplay between the environment, lifestyle, and composition of human microbiomes across different communities, with a specific focus on immune and digestive health. This literature review investigates three distinct types of communities: isolated/traditional, transitional, and urban/industrialized communities, each characterized by unique environments, societal structures, and interactions. These factors contribute to the distinct microbial populations and compositions within each community. It is impossible to avoid contact with microorganisms as they are present in every aspect of our lives, from the air we breathe, the water we drink, the soil we walk on, the …

Exploring Extreme Viruses With Ken Stedman, Kenneth Stedman

PDXPLORES Podcast

In this episode of PDXPLORES, Ken Stedman, Professor of Biology at Portland State University, discusses his distinguished virology career. From discovering his love of extreme viruses at Max Planck Institute for Biochemistry in Munich, Germany, to heading the Advanced Molecular and Cell Biology Research Laboratory and the eXtreme Virus Lab, Steadman has investigated extreme virus structures, function, genetics, and evolution for more than 25 years. He is the co-founder of the research lab Center for Life in Extreme Environments as well as the Chief Scientific Officer of StoneStable, Inc. Adding to his scientific accomplishments and accolades, Professor Stedman …

Tools For Quantifying Bacterial Motility Using Digital Holographic Microscopy As Applied To Studying The Simulated Microgravity Environment, Jacqueline Marie Acres

Dissertations and Theses

Digital holographic microcopy (DHM) is a label-free technique that has gained attention in recent years as a tool for volumetric imaging. One application of DHM is for the study of microbial motility with the advantage being that organisms may freely move within their environment. Images created from DHM are in the form of holograms. Holograms are time recordings showing XY information with the Z information contained within. Z information can be retrieved from the holograms directly through a variety of numerical techniques or through reconstruction. Datasets generated from DHM are large and processing remains a challenging task. Here, we show …

Viruses In Astrobiology, Ignacio De La Higuera, Ester Lázaro

Biology Faculty Publications and Presentations

Viruses are the most abundant biological entities on Earth, and yet, they have not received enough consideration in astrobiology. Viruses are also extraordinarily diverse, which is evident in the types of relationships they establish with their host, their strategies to store and replicate their genetic information and the enormous diversity of genes they contain. A viral population, especially if it corresponds to a virus with an RNA genome, can contain an array of sequence variants that greatly exceeds what is present in most cell populations. The fact that viruses always need cellular resources to multiply means that they establish very …

System Design For The Quantification Of Microbial Motility In Extreme Environments, Megan Marie Dubay

Dissertations and Theses

Motility of microorganisms is understudied but provides useful insights into their behavior. Organisms' ability to move autonomously changes how they interact with their environment--finding nutrients, interacting with other organisms, and avoiding unfavorable conditions. Understanding motility features can also be used to identify specific species, such as the identification of Vibrio cholerae in human samples. Motility might also be used as evidence of life existing in even the most extreme environments on Earth, and possibly beyond. Specialized microscopy systems can be required to examine the motility of microorganisms due to the nature of the environments to which the instruments are exposed. …

The Role Of The Gut-Brain Axis In Alzheimer's Disease: A Narrative Review, Leila Karout

University Honors Theses

The importance of the gut microbiome in health has become increasingly recognized as more research points to the strong association between an imbalanced gut microbial composition and a diseased state. Alzheimer's Disease burdens millions, and the rate of growth is expected to increase. However, current treatment methods are less than ideal and do not offer a substantial benefit to patients' quality of life. Additionally, there are gaps in the current understanding of Alzheimer's Disease pathogenesis. Research on the relationship between gut microbiota and neurodegeneration provides a different perspective on the pathogenesis and treatment methods for dementia, such as Alzheimer's Disease. …

Virology: Mutagenizing The Proteolytic Cleavage Site In The Major Capsid Protein In Ssv1, Thejanee Liyanaarachchi

University Honors Theses

SSV1, spindle-shaped virus 1, is an archaeal virus with a unique shape that belongs to the family of fuselloviruses and infects hyperthermophilic archaea that thrive at 80°C and pH 3. SSV1 has two capsid proteins, VP1 and VP3, that presumably produce the lemon-shaped virion structure characteristic of fuselloviruses. The VP1 amino acid sequence is highly conserved among SSVs. Here, we investigate the importance of the proteolytic cleavage site in position E66 in VP1 by changing its glutamic acid to the other 19 amino acids. We have developed a new protocol for creating SSV1 mutants through HiFi Assembly (NEB) to increase …

A Bioinformatic And Biochemical Analysis Of Cruciviruses, George William Kasun

Dissertations and Theses

Cruciviruses are novel ssDNA viruses discovered through metagenomics and direct environmental DNA amplification and cloning. The genomes of cruciviruses suggest that gene transfer between RNA and DNA viruses occurred due to the presence of putative protein-encoding genes that are homologous to both ssRNA and ssDNA viruses. In order to gain a better understanding of this group of viruses both bioinformatic analyses and in vitro biochemical experiments were employed. The results of the bioinformatic analyses show that cruciviruses are a highly diverse group of ssDNA viruses. Their placement within established ssDNA phylogenies is difficult due to heterogeneity in their putative replication-associated …

A Small Rna Is Functional In Escherichia Fergusonii Despite Containing A Large Insertion., Austin P. Wright, H. Auguste Dutcher, Brianna Butler, Timothy J. Nice, Rahul Raghavan

Biology Faculty Publications and Presentations

Bacterial small RNAs (sRNAs) are important regulators of gene expression; however, the impact of natural mutations on sRNA functions has not been studied extensively. Here we show that the sRNA MgrR contains a unique 53 bp insertion in Escherichia fergusonii, a close relative of Escherichia coli and Salmonella enterica. The insertion is a repetitive extragenic palindromic (REP) sequence that could block transcription, but full-length MgrR is produced in E. fergusonii, showing that the insertion has not affected sRNA production. Additionally, despite containing the large insertion, the sRNA appears to be functional because deletion of mgrR made E. fergusonii …

Microbes On The Mountain: Plant-Microbe Associations And Interactions On Mount St. Helens, Emily Rose Wolfe

Dissertations and Theses

Plant-microbe associations and interactions provide critical context to studies in both community and ecosystem ecology, especially in systems that are relatively new and still undergoing early successional processes. Microbes can colonize the surfaces and interiors of all plant tissues, and these assemblages vary in composition both spatially and temporally, even within the same plant. Endophytes are bacteria or fungi that spend most of their lifecycles living within plant tissues asymptomatically--typically, "endophyte" refers specifically to aboveground tissues such as leaves and stems, and therefore may have direct influences on defenses against herbivory, pathogen or pest tolerance, and even afterlife effects on …

Novel Small Rnas Expressed By Bartonella Bacilliformis Under Multiple Conditions Reveal Potential Mechanisms For Persistence In The Sand Fly Vector And Human Host, Shaun Wachter, Linda D. Hicks, Rahul Raghavan, Michael F. Minnick

Biology Faculty Publications and Presentations

Bartonella bacilliformis, the etiological agent of Carrión’s disease, is a Gram-negative, facultative intracellular alphaproteobacterium. Carrión’s disease is an emerging but neglected tropical illness endemic to Peru, Colombia, and Ecuador. B. bacilliformis is spread between humans through the bite of female phlebotomine sand flies. As a result, the pathogen encounters significant and repeated environmental shifts during its life cycle, including changes in pH and temperature. In most bacteria, small non-coding RNAs (sRNAs) serve as effectors that may post-transcriptionally regulate the stress response to such changes. However, sRNAs have not been characterized in B. bacilliformis, to date. We …

Direct Binding Of Tfeα Opens Dna Binding Cleft Of Rna Polymeras, Sung-Hoon Jun, Jaekyung Hyun, Hoyoung Kim, Michael S. Bartlett, Hyun-Soo Cho, Katsuhiko S. Murakami

Biology Faculty Publications and Presentations

Opening of the DNA binding cleft of cellular RNA polymerase (RNAP) is necessary for transcription initiation but the underlying molecular mechanism is not known. Here, we report on the cryo-electron microscopy structures of the RNAP, RNAP-TFEα binary, and RNAPTFEα-promoter DNA ternary complexes from archaea, Thermococcus kodakarensis (Tko). The structures reveal that TFEα bridges the RNAP clamp and stalk domains to open the DNA binding cleft. Positioning of promoter DNA into the cleft closes it while maintaining the TFEα interactions with the RNAP mobile modules. The structures and photo-crosslinking results also suggest that the conserved aromatic residue in the extended winged-helix …

Host-Dependent Differences In Replication Strategy Of The Sulfolobus Spindle-Shaped Virus Strain Ssv9 (A.K.A., Ssvk1): Infection Profiles In Hosts Of The Family Sulfolobaceae, Ruben M. Ceballos, Coyne Gareth Drummond, Carson Len Stacy, Elizabeth Padillo-Crespo, Kenneth M. Stedman

Biology Faculty Publications and Presentations

The Sulfolobus Spindle-shaped Virus (SSV) system has become a model for studying thermophilic virus biology, including archaeal host-virus interactions and biogeography. Several factors make the SSV system amenable to studying archaeal genetic mechanisms (e.g., CRISPRs) as well as virus-host interactions in high temperature acidic environments. Previously, we reported that SSVs exhibited differential infectivity on allopatric vs. sympatric hosts. We also noticed a wide host range for virus strain SSV9 (a.k.a., SSVK1). For decades, SSVs have been described as “non-lytic” double-stranded DNA viruses that infect species of the genus Sulfolobus and release virions via budding rather than host lysis. In this …

Unveiling Crucivirus Diversity By Mining Metagenomic Data, Ignacio De La Higuera, George Kasun, Ellis L. Torrance, Alyssa A. Pratt, Amberlee Maluenda, Jonathan Colombet, Maxime Bisseux, Viviane Ravet, Anisha Dayaram, Daisy Stainton, Simona Kraberger, Peyman Zawar-Reza, Sharyn Goldstien, James V. Briskie, Robyn White, Helen Taylor, Christopher Gomez, David G. Ainley, Jon S. Harding, Rafaela S. Fontenele, Joshua Schreck, Simone Ribeiro, Stephen A. Oswald, Jennifer M. Arnold, François Enault, Arvind Varsani, Kenneth M. Stedman

Biology Faculty Publications and Presentations

The discovery of cruciviruses revealed the most explicit example of a common protein homologue between DNA and RNA viruses to date. Cruciviruses are a novel group of circular Rep-encoding single-stranded DNA (ssDNA) (CRESSDNA) viruses that encode capsid proteins that are most closely related to those encoded by RNA viruses in the family Tombusviridae. The apparent chimeric nature of the two core proteins encoded by crucivirus genomes suggests horizontal gene transfer of capsid genes between DNA and RNA viruses. Here, we identified and characterized 451 new crucivirus genomes and 10 capsid-encoding circular genetic elements through de novo assembly and mining of …