Microbiology Commons^™

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 …

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 …

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 …

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 …

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 …

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 …

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 …

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 …

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 …

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 …

Cultivation-Dependent Analysis Of Microorganisms Associated With Various Hydraulic Fracturing Fluids, Anthony Harrington

UNLV Theses, Dissertations, Professional Papers, and Capstones

The overarching goal of this study is to gain more insight to the microbial interactions associated with hydraulic fracturing by studying the culturable microorganisms present in various types of hydraulic fracturing fluids. Extraction of natural gas and other unconventional resources using hydraulic fracturing has increased in the last decade and very few studies have been conducted on the microorganisms associated with the various water types used in the process. From the very few published studies, the only cultivation-dependent method used involved determining the most probable number (MPN) of various metabolic groups, however the researchers failed to ask more in-depth questions …

Natural And Lignocellulose-Enriched Microbial Communities In Great Boiling Spring, Nv, Jessica Cole

UNLV Theses, Dissertations, Professional Papers, and Capstones

The natural microbial communities present in Great Boiling Spring were investigated and contrasted against those present after in situ enrichment with lignocellulose. High-throughput cultivation-independent DNA sequencing of the V8 region of the small subunit (SSU) rRNA gene generated a total of 274,119 quality-filtered pyrosequencing fragments. Twelve natural spring samples were analyzed, including four high-temperature water samples and eight sediment samples ranging from 87 - 62 °C. Eight lignocellulosic enrichments incubated in the spring sediment and water at two high-temperature sites were analyzed. The natural water communities were found to be extremely uneven but relatively constant throughout time. The natural sediment …