Microbiology Commons^™

Poop! There It Is! Anti-Germinants And Biological Variables As Modulators Of Clostridioides Difficile Infection (Cdi), Jacqueline Renee Phan

UNLV Theses, Dissertations, Professional Papers, and Capstones

Clostridioides difficile infection (CDI) is a major cause of antibiotic-associated diarrhea. Due to its insidious nature, the Centers for Disease Control and Prevention (CDC) has declared CDI an urgent threat. A key characteristic of C. difficile is its ability to form dormant spores that act as the infectious vehicles for disease. In the gut, spores recognize bile salts to germinate into toxin-producing cells.Dysbiosis of the gut microbiome is a key factor in allowing the C. difficile spores to germinate. Normal gut microbiota naturally protects from CDI. However, biological variables such as diet and sex have been found to modulate to …

Effects Of Cations And Dipicolinic Acid On B. Anthracis Spore Physiology And Cytotoxicity, Chandler P. Hassan

UNLV Theses, Dissertations, Professional Papers, and Capstones

Inhalation of Bacillus anthracis spores leads to the most severe form of anthrax. Following phagocytosis by alveolar macrophages, spores germinate inside the phagolysosome. The mechanism used by the newly germinated B. anthracis cells to survive within macrophages is not completely understood.B. anthracis spores contain large concentrations of calcium in complex with dipicolinic acid (Ca-DPA). Upon germination, the cell excretes the large depot of Ca-DPA. DPA is an amphipathic molecule that could act as a buffer and modulate phagolysosome acidification. Intracellular calcium overload can disrupt signaling pathways required for normal macrophage function and trigger cell death. We hypothesized the release of …

Desert Spring Prokaryotes And Benthic Macroinvertebrates: Geological And Ecological Controls, Ariel D. Friel

UNLV Theses, Dissertations, Professional Papers, and Capstones

Desert springs are aquatic oases, being considered among the most diverse and imperiled ecosystems on Earth. Despite the ecological significance and threatened status of desert springs, few studies have surveyed the microorganisms in these systems or their interactions with other community members. The primary goal of this dissertation was to describe the major controls influencing the community ecology of prokaryotic and BMI communities in cold- and moderate-temperature desert springs of the southern hydrographic Great Basin. Chapter 1 provides a broad overview of spring ecosystems and of each chapter included in this dissertation. Chapter 2 documents the first-ever, regional-scale survey of …

Biochemical Characterization Of Fsa1572 From Fervidibacter Sacchari, The First Hyperthermophilic Gh50 With Β-1, 4-Glucanase Activity, Jonathan Covington

UNLV Theses, Dissertations, Professional Papers, and Capstones

The bacterium Fervidibacter sacchari is an aerobic hyperthermophile that catabolizes various polysaccharides and is the only cultivated member of the class Fervidibacteria within the phylum Armatimonadota. Among its glycoside hydrolase (GH) cache is an enzyme from GH family 50 (GH50), an understudied family with only 25 characterized representatives and two known activities from 1,518 predicted members in the Carbohydrate-Active EnZyme (CAZy) Database. Here, we expressed, purified, and functionally characterized an extracellular GH50 from F. sacchari called Fsa1572. Using colorimetric assays, we show it has novel β-1,4-glucanase activity and only weak agarose activity that is typical for GH50 enzymes. The purified …

Oral Prevalence Of Akkermancia Muciniphila Among Pediatric And Adult Orthodontic And Non-Orthodontic Patients, Ching Shen

UNLV Theses, Dissertations, Professional Papers, and Capstones

Akkermansia muciniphila (AM) is one of many highly abundant intestinal microbes that influences homeostasis and metabolic disorders and may also play a role in oral disorders. However, there is little evidence regarding the oral prevalence of this organism. Based upon this lack of evidence, the primary goal of this project is to survey an existing saliva repository to determine the overall prevalence of this organism and any associations with demographic or patient characteristics (age, sex, body mass index, race/ethnicity, orthodontic therapy). Using an approved protocol, a total n = 141 pediatric samples from an existing saliva repository were screened using …

Calcite Spherulite Formations And Their Controls In Halophilic Bacterial Cultures, Zakaria Jibrin

UNLV Theses, Dissertations, Professional Papers, and Capstones

This study arose from an accidental discovery of calcite spherulites forming on colonies of various halophilic bacteria grown on solid Luria-Bertani medium made containing water from a brine pool in Death Valley, their natural habitat. Further studies with a large spherulite forming bacteria, Pseudoalteromonas haloplanktis, showed that calcite precipitation conditions were the consequence of bacteria using amino acids as a source of carbon and energy, a metabolism that produces NH3 and CO2 waste products. NH3 raises the pH of the medium, turning CO2 into CO32-. Calcium in the natural brine combines with CO32-. The resulting spherulitic calcite precipitation is nano-acicular …

Comparing The Incidence And Prevalence Of Oral Microbial Pathogens Selenomonas Noxia And Streptococcus Mitis Within The Unlv-Sdm Clinical Patient Population, Namgu Kim

UNLV Theses, Dissertations, Professional Papers, and Capstones

Introduction: Selenomonas noxia (SN) is a gram-negative, anaerobic bacteria, which contributes to development and progression of periodontal disease. Some evidence now suggests Streptococcus mitis (SM), a gram-positive, facultative bacterium contributing to the etiology of dental caries and periodontal disease, may also influence the prevalence of SN within subgingival complexes. Based upon the overall lack of prevalence data, the objective of this study was to evaluate presence of SN and SM using qPCR among saliva samples taken from pediatric, adult, and orthodontic dental school clinics.

Methods: This study involved a retrospective analysis of previously collected saliva samples from an existing biologic …

Implications Of The Evolutionary Past On The Activities Of Virb: A Key Transcriptional Regulator Of Virulence Genes In Shigella Flexneri, Jillian Nicole Socea

UNLV Theses, Dissertations, Professional Papers, and Capstones

Shigella species are bacterial agents that are the leading cause of diarrheal diseases, the second leading cause of child mortality worldwide. One key protein in Shigella is VirB, a transcriptional regulator of virulence genes that is essential for pathogenicity. VirB functions to antagonize H-NS, a histone-like nucleoid structuring protein that silences many genes found on the large virulence plasmid (pINV). The interplay between H-NS, the silencer, and VirB, the anti-silencer, is fundamental for Shigella virulence. Furthermore, this mechanism of gene regulation appears widespread in many bacteria including other enteric pathogens.

The goal of my dissertation was to explore the activities …

Genomic Analysis Of Paenibacillus Larvae Bacteriophages, Casey Stamereilers

UNLV Theses, Dissertations, Professional Papers, and Capstones

American Foulbrood is the most destructive bacterial infection of the honeybee (Apis mellifera) and is caused by the Gram-positive, spore forming bacterium Paenibacillus larvae. Current treatment methods rely on antibiotics, but antibiotics treatments are experiencing a reduction in efficacy due to the recent rise in antibiotic resistant strains of P. larvae. This has been a major catalyst for exploration of alternative treatment methods. Phage therapy is an alternative treatment method that uses viruses that exclusively infect bacteria, known as bacteriophages (phages), to combat bacterial infections. Several experimental studies have shown that phages P. larvae phages are effective at lysing P. …

Transcriptional Control Of Virulence Genes In The Bacterial Pathogen Shigella Flexneri, Joy Asami Mckenna

UNLV Theses, Dissertations, Professional Papers, and Capstones

Shigella species, the causal agents of bacillary dysentery, use a type III secretion system (T3SS) to inject two waves of virulence proteins, known as effectors, into the human colonic epithelium to subvert host cell machinery. Transcriptional regulation of these virulence genes is controlled by the three-tiered VirF/VirB/MxiE signaling cascade. Of these, VirB has the largest regulon (~50 genes); however, VirB regulatory properties are poorly understood. To date, VirB is known to function to counter or 'anti-silence' transcriptional silencing mediated by the histone-like nucleoid structuring protein H-NS. To better understand VirB regulation, my colleagues and I chose to better define the …

Rapid Detection Of Toxin-Producing Cyanobacteria, Timothy Alba

UNLV Theses, Dissertations, Professional Papers, and Capstones

Lake Mead provides drinking water to millions of people in Nevada, California, and Arizona. In 2015, the Southern Nevada Water Authority detected the cyanobacteria-produced toxin microcystin in the lake for the very first time. This toxin is lethal in large doses, and in small doses it causes a myriad of serious health effects. Detecting microcystin directly is a time-consuming and expensive process that requires liquid chromatography tandem mass spectrometry or immunological analyses, which require a full day or more to process samples. In order to provide water managers with the methodology for a toxin monitoring plan, this work developed a …

Oral Localization Of Scardovia Wiggsiae, Graydon Ramos Carr

UNLV Theses, Dissertations, Professional Papers, and Capstones

Early childhood caries is one of the most prevalent diseases in the United States among children. The formation of caries is a complex, multifactorial process that is still being studied. Researchers have thought for years that Streptococcus mutans was the primary causative agent of early childhood caries. The recent discovery of a novel cariogenic pathogen, Scardovia wiggsiae and its significant contribution to the etiology of early childhood caries has led oral health researchers to re-evaluate this microorganism and its link to this disease. While there have been several projects undertaken within the University of Nevada, Las Vegas School of Dental …

The Influence Of G4 Dna Structures On Stationary Phase Mutagenesis In Bacillus Subtilis, Tatiana Ermi

UNLV Theses, Dissertations, Professional Papers, and Capstones

Until the late 1980’s it was widely understood that bacterial variance emerges randomly during growth. Research that followed has convincingly shown evidence for mutations arising in non-growing conditions, a phenomenon known as stationary-phase mutagenesis. In Bacillus subtilis, an elegant mutagenic mechanism has been elucidated in non-growing cells that biases mutations to transcribed regions of a subpopulation. One interesting possibility is that mutations can be further biased to hotspots within genes through alternate DNA structures known as non-B DNA. Non-B DNA have been linked to genomic instability and disease in humans, lesser is known about its role in bacteria. Here we …

The Significance Of Mfd And Oxidative Damage In Stationary-Phase Mutagenesis In Bacillus Subtilis, Holly Anne Martin

UNLV Theses, Dissertations, Professional Papers, and Capstones

The process of stationary-phase mutagenesis, also known as adaptative or stress-induced mutagenesis, is a phenomenon where bacterial cells accumulate mutations in non-replicative conditions. This process has mainly been studied in Escherichia coli and Bacillus subtilis; however, the underlying mechanisms found in each of these systems differ. Here, I use B.msubtilis to study previously understudied aspects of stationary-phase mutagenesis. In this dissertation, I describe work that has led to three major discoveries which are described below.

First in B. subtilis, Mfd is important for stationary-phase mutagenesis and its mutagenic function at regions of the genome that are transcriptionally upregulated has been …

The Perplexing Paradox Of Clostridioides (Clostridium) Difficile Infection (Cdi) - Analysis Of Anti-Germinants As Part Of Cdi Prophylaxis, Christopher Yip

UNLV Theses, Dissertations, Professional Papers, and Capstones

Clostridioides (Clostridium) difficile infections (CDI) have become the leading cause of nosocomial antibiotic-associated diarrhea worldwide. Under normal circumstances, bacteria found in the gastrointestinal tract provide a barrier against C. difficile colonization. Upon antibiotic therapy, the protective barrier is lost as the microbial community becomes depleted thus providing the opportunity for C. difficile to colonize the human gut. Exposure to taurocholate, a bile acid produced within the mammalian gastrointestinal tract, causes C. difficile spores to begin their transition, a process known as germination, from metabolically dormant structures to toxin-producing cells. As germination is required for the onset of CDI, anti-germination compounds …

Prevalence Of Aggregatibacter Actinomycetemcomitans And Fusobacterium Nucleatum Among Clinical Orthodontic Saliva Samples, Jason Robert Klingler

UNLV Theses, Dissertations, Professional Papers, and Capstones

Changes to the oral microflora occur when there are shifts in the levels of the numerous bacterial species. Changes in bacterial load occur in health, disease, and dental treatments such as orthodontics and can be detected through saliva. Many studies dealing with saliva have centered around detecting bacteria known for correlation with chronic periodontitis and caries. Fewer have focused on bacterial species that contribute to microbial shifts not strictly correlated with disease. Measuring the degree of disease progression or future susceptibility is not always possible with traditional clinical parameters alone.

Aggregatibacter actinomycetemcomitans (AA) is a bacterial strain that serves as …

Effect Of Common Oxidative Water Treatments On Acanthamoeba Internalized Legionella, James Park

UNLV Theses, Dissertations, Professional Papers, and Capstones

Legionella pneumophila can cause pneumonic and non-pneumonic disease in humans. Infections occur from aerosolized contaminated water. This bacterium is an opportunistic intracellular pathogen able to infect both protozoans, such as Acanthamoeba polyphaga, and human macrophages. Both L. pneumophila and A. polyphaga resist commonly used water treatments, such as chlorination, but L. pneumophila has displayed greater resistance in the presence of A. polyphaga. Therefore, there is concern that L. pneumophila could become established in plumbing systems after water treatment, leading to infections. The objective of this study was to show the effect of chlorine and chlorine dioxide exposures on the survival …

Beyond Genetics: Addressing Activity In Microbial Ecology, Scott Christopher Thomas

UNLV Theses, Dissertations, Professional Papers, and Capstones

The field of microbial ecology is on the cusp of another revolution, one that uses synergy to address the limitations of previous studies. My dissertation is representative of the new era of research to come, through combining the latest technological advances with established classical methods to provide a more complete understanding of microbial ecosystems. In my first study, I evaluate the conservation of central carbon metabolic pathways and activity along a temperature and diversity gradient (60-95 ℃) through the use of 16S rRNA gene amplicon data, metagenome sequencing, and position-specific 13C-labeled metabolite probing. In my second study, I provide the …

Prokaryotic Diversity And Aqueous Geochemistry Of Subsurface Environments Of The Death Valley Regional Flow System, Joshua David Sackett

UNLV Theses, Dissertations, Professional Papers, and Capstones

This dissertation summarizes over four years of effort towards the completion of a Ph.D. in Biological Sciences. The work presented in this document covers a broad range of topics, but a central unifying theme is prokaryotic life in the continental subsurface. The work presented in each chapter relied heavily on cultivation-independent methods for assessing prokaryotic communities, including prokaryotic community structure reconstruction from high-throughput sequencing of 16S rRNA gene libraries and single-cell genome analysis of novel uncultivated bacteria.

Chapter 2 examines the aqueous geochemistry and prokaryotic diversity of Devils Hole, a cavernous limnocrene and sole natural habitat for the critically endangered …

Transcriptional Silencing And Anti-Silencing Of Virulence Genes In The Bacterial Pathogen Shigella Flexneri: Virb, Dna Supercoiling, And The Histone-Like Nucleoid Structuring Protein, Michael Adam Picker

UNLV Theses, Dissertations, Professional Papers, and Capstones

Transcriptional silencing and anti-silencing affect many aspects of bacterial physiology, including virulence in bacterial pathogens. In Shigella species, a group of gram-negative pathogens that cause bacillary dysentery in humans, the histone-like nucleoid structuring protein (H-NS) transcriptionally silences virulence genes found on the large virulence plasmid while VirB anti-silences these genes. However, the mechanistic details of their interplay are not fully understood. To elucidate their regulatory mechanisms, I use the icsP virulence locus, which shares a long intergenic region with the divergently transcribed ospZ gene (1535 bp from TSS to TSS). Prior to this work, two discrete H-NS binding regions had …

Screening For The Novel Cariogenic Pathogen Scardovia Wiggsiae Among Orthodontic Patients, Adam Whiteley

UNLV Theses, Dissertations, Professional Papers, and Capstones

Screening for the Novel Cariogenic Pathogen Scardovia wiggsiae among Orthodontic Patients

by

Adam Whiteley

Dr. Karl Kingsley, Examination Committee Chair Professor of Biomedical Sciences University of Nevada, Las Vegas School of Dental Medicine

The recent discovery of a novel cariogenic pathogen Scardovia wiggsiae has led many scientists and oral health researchers to re-evaluate and re-examine existing saliva repositories to determine the prevalence among patient populations. Recent efforts at this institution have used existing saliva samples to determine the prevalence among both adults and pediatric patients. These studies have revealed this organism may be found in approximately one-quarter of all samples …

Complete Sequencing And Comparative Analysis Of The Genomes Of The First Magnetotactic Gammaproteobacteria Isolated In Pure Culture: Strains Bw-2 And Ss-5, Corey Geurink

UNLV Theses, Dissertations, Professional Papers, and Capstones

The genomes of the first two discovered magnetotactic bacteria (MTB) belonging to the ammaproteobacteria, strains BW-2 and SS-51, were sequenced, sealed, annotated and compared to MTB of other phylogenetic groups. Cells of both strains are rod-shaped and biomineralize cuboctahedral and elongated octahedral crystals of magnetite, respectively, that are enveloped in a protein-embedded, lipid-bilayer membrane referred to as the magnetosome membrane or vesicle. The crystals and their associated membranes are known as magnetosomes. Magnetosome crystals consist of either magnetite (Fe3O4) or greigite (Fe3S4) and, because of their specific mineral compositions, crystal morphologies and sizes, the biomineralization processes involved in magnetosome formation …

A Potential Solution To A Poopy Problem: Bile Salt Analogs As Prophylactics Against Clostridium Difficile Infection, Jacqueline Renee Phan

UNLV Theses, Dissertations, Professional Papers, and Capstones

Clostridium difficile infection (CDI) is a major cause of antibiotic-associated diarrhea. In 2011, over 500,000 patients were diagnosed with CDI in the United States and over 29,000 people died of CDI-related complications. With an average of $35,000 to treat a single case of inpatient CDI, cost burden to the healthcare system can reach up to $3.2 billion annually. As both hospital- and community-acquired CDI incidences rise due to the emergence of hypervirulent strains and CDI reoccurrences of up to 25%, standard treatments are rendered less effective and new methods of prevention are critical.

CDI is caused by bacteria called Clostridium …

Evaluation Of A Fluorescence Method For Quantifying Bioaerosol Concentrations On Air Quality Filter Samples, Rachel Kolberg

UNLV Theses, Dissertations, Professional Papers, and Capstones

Airborne particulate matter (PM) in outdoor environments contains many components that cause adverse human health effects. The size of the particulates determine in what manner the particles would bypass the body’s defense mechanisms to enter the respiratory system and is directly related to their health impacts. Currently the United States Environmental Protection Agency is enforcing the National Ambient Air Quality Standards (NAAQS) to regulate the annual and 24-hour average concentrations of PM2.5 and PM10 in the air. PM2.5 are fine particles with aerodynamic diameter <2.5μm, small enough to reach the deepest parts of the bronchi and lungs. PM10 include PM2.5 and larger particles with aerodynamic diameter of 2.5-10μm. Both PM2.5 and PM10 contain multiple components from multiple sources. Bioaerosols are an important component of PM, but there is limited knowledge about how bioaerosols contribute to PM2.5 and PM10 concentrations. There is also a lack of research about the incidence and prevalence of disease caused by bioaerosols and about the limits of exposure to bioaerosol particulates. The main barrier to assess bioaerosol concentrations and health-related effects is the absence of quick and inexpensive methodology for quantifying bioaerosols. This study explored the feasibility of using fluorescence microscopy to quickly quantify bioaerosols in PM2.5 and PM10 collected on polycarbonate filters. Bioaerosols were stained with a DNA marker directly on a filter, followed by fixation, microscopic imaging, and automatic counting. The method was first validated using reference samples prepared by depositing different known concentrations of E. coli onto blank polycarbonate filters. The results indicated a linear response over two orders of magnitude (R2 = 0.9) and an accuracy within ±25%. E. coli were also deposited onto selected ambient PM10 and PM2.5 filter samples to determine if pre-loaded particles would interfere with bioaerosol imaging and counting. It was found that despite an increase in uncertainty (variability), the calibration slope remained within ±10% of unity for both PM2.5 and PM10 samples. Bioaerosol concentrations in ambient samples, as quantified by this method, were on average 14% higher for PM10 than for PM2.5 acquired concurrently in a desert environment of Las Vegas, Nevada. The application of this method to other types of compliance filters, such as Teflon filters and tapes of a Beta Attenuation Monitor (BAM) were also explored in this study. By means of a high-yield approach this method is expected to facilitate bioaerosol research, support exposure and health assessments, and help refine NAAQS for PM2.5 and PM10.

Development Of The K-State (Competence) And Mutagenesis In Stressed Bacillus Subtillis Cells, Amanda Kidman

UNLV Theses, Dissertations, Professional Papers, and Capstones

Mutagenesis is central to the evolutionary process. We currently view evolution as a gradual process affecting all cells within a population. However, my project studied an underestimated part of the evolutionary process: mutations generated during stationary phase (caused by nutritional stress or growth arrest) within a subpopulation of cells. I used Bacillus subtilis, a bacterial model for cell growth and differentiation. Stationary-phase cultures of B. subtilis differentiate subpopulations that exhibit different survival strategies including competence, secondary metabolite production, biofilm formation, cannibalism, and endospore formation. The development of competence permits cells to uptake exogenous DNA and incorporate it into their genome. …

Oral Microbial Burden Of Periodontal Pathogens Among Orthodontic Patients, David Jolley

UNLV Theses, Dissertations, Professional Papers, and Capstones

Objectives

Many research studies involving orthodontic patients have a natural inclination to focus on changes in levels of cariogenic pathogens after bracket placement, and very few studies examine the role of changes of periodontal pathogens – particularly among adult patients. Interestingly, recent evidence suggests that increased levels of a specific periodontal pathogen, Fusobacterium nucleatum, may elevate risk for development of colon cancer in adults through direct pathways. Based upon this new evidence, the objective of the current study was to screen saliva samples taken from orthodontic patients to determine the prevalence of periodontal pathogens, including F. Nucleatum, T. denticola, and …

Incomplete Denitrification In Thermus Species, Chrisabelle Mefferd

UNLV Theses, Dissertations, Professional Papers, and Capstones

Members the bacterial genus Thermus have been shown to be incomplete denitrifiers, terminating with nitrite or nitrous oxide (N2O). However, the ability to carry out denitrification and the evolution of nitrogen oxide reductase genes in Thermus remains poorly understood. This study tests the hypothesis that incomplete denitrification is common in Thermus and seeks to uncover patterns in the evolution of denitrification pathways in Thermus. Denitrification capacity was determined in a collection of 25 strains representing ten species of Thermus and phylogenetic analysis was performed to determine whether denitrification genes evolved horizontally in Thermus. No strains in this study reduced nitrate …

The Role Of Mfd In Stationary-Phase Oxidative Damage Repair In Bacillus Subtilis, Katelyn E. Porter

UNLV Theses, Dissertations, Professional Papers, and Capstones

Since the 1950’s it has been shown that bacterial cells accumulate mutations even in non- dividing conditions, but how this type of mutation occurs is still highly debated. In Bacillus subtilis, Mfd, a precursor of the nucleotide excision repair (NER) system, mediates the formation of mutations in stationary-phase or non-replicating cells. In growing cells, Mfd recruits repair when RNA polymerase is stalled during transcription; it then recruits proteins from NER to repair damage. Here, we examine the hypothesis that Mfd mediates the formation of mutations by interacting with cellular components that repair reactive oxygen species (ROS), a natural byproduct of …

Biogeochemistry Of A Saline, Alkaline, Terminal Lake Ecosystem In Transition; Walker Lake, Nevada, Katherine Lynn Willever

UNLV Theses, Dissertations, Professional Papers, and Capstones

Walker Lake is a saline, alkaline, terminal lake ecosystem located in west-central Nevada. For over one hundred years, anthropogenic streamflow diversions within the Walker River Basin have ultimately led to little or no water reaching Walker Lake, the basin’s terminus for water flow. These diversions have resulted in a >46 meter decrease in the lake surface altitude and increases in salinity and dissolved salt constituents that have caused the elimination of native fish species. This study examines how the lack of freshwater inflow has altered the physical, chemical, and microbiological structure of Walker Lake during the lake’s ongoing desiccation.

Between …

Effectiveness Of Commercially-Available Cosmetic Cleaners On Cosmetics And Cosmetic Brushes, Vanessa Ortiz

UNLV Theses, Dissertations, Professional Papers, and Capstones

The complex nature of skin contributes to the microbial population present on its surface. While normal skin flora is either beneficial or has no effect on the body, there are instances where pathogenic microorganisms are present and can cause infections. Damaged skin is more susceptible to infections from these microbes. Behavioral characteristics, such as the use of cosmetics, can affect the microbial population present on the skin. Staphylococcus aureus is the organism most commonly isolated from cosmetics, and it can be responsible for conjunctivitis, impetigo, boils, and folliculitis.

There are many ways microbial contamination of cosmetics can occur, such as …