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 …

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 …

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 …

Rasagiline Effects On Glucose Metabolism, Cognition, And Tau In Alzheimer’S Dementia, Dawn C. Matthews, Aaron Ritter, Ronald G. Thomas, Randolph D. Andrews, Ana S. Lukic, Carolyn Revta, Jefferson W. Kinney, Babak Tousi, James B. Leverenz, Howard Fillit, Kate Zhong, Howard H. Feldman, Jeffrey Cummings

Integrated Health Sciences Faculty Publications

Background: A Phase II proof of concept (POC) randomized clinical trial was conducted to evaluate the effects of rasagiline, a monoamine oxidase B (MAO-B) inhibitor approved for Parkinson disease, in mild to moderate Alzheimer's disease (AD). The primary objective was to determine if 1 mg of rasagiline daily for 24 weeks is associated with improved regional brain metabolism (fluorodeoxyglucose–positron emission tomography [FDG-PET]) compared to placebo. Secondary objectives included measurement of effects on tau PET and evaluation of directional consistency of clinical end points. Methods: This was a double-blind, parallel group, placebo-controlled, community-based, three-site trial of 50 participants randomized 1:1 to …

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 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 …

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 …

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 …

Alternate Germinants Of C. Difficile, A Leading Hospital Pathogen, Sean Tanzey, Ernesto Abel-Santos

McNair Poster Presentations

Clostridium difficile infections (CDI) are the leading nosocomial infections worldwide. Humans are asymptomatic carriers of C. difficile spores in the intestinal tract. The process known as germination occurs when otherwise harmless C. difficile spores are converted to toxin-producing cells upon recognition of bile salts in humans. This distinctive transition ultimately leads to the onset of disease and recurrent CDI. Germination profiles will be characterized in response to peptidoglycan (PG) fragments isolated from various bacterial species. These specific peptidoglycan fragments contain different amino acid residues that may induce different germination responses. Purification and structural determination of the peptidoglycan fragments will be …

Determining The Presence Of Carbapenem Antibiotic Resistance In Clinical Isolates, Fidelis Uzoma Enyinnaya

UNLV Theses, Dissertations, Professional Papers, and Capstones

Carbapenems are β-lactam antibiotics reserved for the treatment of severe microbial infections, especially those targeting the Enterobacteriaceae. Introduced in the 1980s, carbapenems have been used successfully in hospitals, and in the 1990s resistance was discovered. Carbapenem resistance is conferred through the production of carbapenemases. In the U.S., the most common carbapenemase is Klebsiella pneumoniae carbapenemase (KPC). In 2012, the National Healthcare Safety Network reported a carbapenem resistance rate of 13.0% among Klebsiella pneumoniae infections, and indicated that the mortality rate associated with carbapenem resistant Enterobacteriaceae (CRE) infections ranged from 48.0-71.9%. According to the Food and Drug Administration (FDA), carbapenem resistance …

Anti-Germinants As A New Strategy To Prevent Clostridium Difficile Infections, Amber Janece Howerton

UNLV Theses, Dissertations, Professional Papers, and Capstones

Clostridium difficileinfections (CDI) have emerged as a leading cause of hospital-associated complications. CDI is the major cause of antibiotic-related cases of diarrhea and nearly all cases of pseudomembranous colitis. The infective form of C. difficileis the spore, a dormant and hardy structure that forms under stress. Germination of C. difficile spores into toxin producing bacteria in the GI tract of susceptible patients is the first step in CDI establishment. Patient susceptibility occurs with a disruption of the natural gut microbiota by broad-spectrum antibiotics. Antibiotic treatments usually resolve CDI but refractory cases are on the rise. Of great concern is the …

A Polymerase Chain Reaction Method For The Detection Of Selenomonas Noxia, Arthuro Mehretu

UNLV Theses, Dissertations, Professional Papers, and Capstones

In recent studies, periodontal health has been linked to being overweight and/or obese. Among common oral bacteria, Selenomonas noxia has been implicated in converting periodontal health to disease. Selenomonas spp. have also been found in gastric ulcers, and were misdiagnosed as Helicobacter -like organisms, but no further studies were conducted. The increasing clinical and epidemiological importance of S. noxia necessitates the development of a rapid detection method. In this study, a TaqMan 16S rRNA based real-time Polymerase Chain Reaction (PCR) method was developed, optimized and evaluated for the rapid and specific detection of S. noxia . The 16S PCR assay …

Dna Secondary Structures And Their Contribution To Mutagenesis In B. Subtilis Stationary Phase Cells, Carmen Vallin, Holly Martin, Christian Ross, Ronald Yasbin, Eduardo Robleto

Undergraduate Research Opportunities Program (UROP)

It is widely known and accepted that the cause of many mutations in cells are generated during the replication process of actively dividing cells, however more recent research has shown that mutations also arise in non growing conditions, a phenomenon known as stationary phase mutagenesis. Much of what is known come from studies in eukaryotic and bacterial models. It has been proposed that in non~growing cells, the process of transcription plays an important role in mutagenesis. We test the hypothesis that DNA secondary structures, formed during transcription, promote mutagenesis. The transcription-generated structures are speculated to be prone to mutations by …

Inactivation Of Spo0a Gene Increases Stationary Phase Mutagenesis In Bacillus Subtilis, Denisse Reyes, Amanda Prisbrey, Holly Martin, Eduardo Robleto

Undergraduate Research Opportunities Program (UROP)

Stationary phase mutagenesis occurs when a population of cells acquires mutations conferring escape from nongrowing or stress conditions. This type of mutations is observed in nutritionally starved cells. Because the mutations occur after the onset of stress and in cells that are in non-replicative conditions, elucidating the underlying mechanisms contributes novel views to the process of evolution and apply to the formation of cancer in human cells and antibiotic resistance in microbial pathogens. Studies have shown that in Bacillus subtilis, the Mfd protein which is a transcription repair coupling factor is necessary for this phenomenon to occur. Here, we investigate …

Synthesis Of Chimeric Receptors Essential For Spore Germination, Christopher Yip, Christian Ross, Eduardo Robleto, Ernesto Abel-Santos

Undergraduate Research Opportunities Program (UROP)

Various species of bacteria have been reported to form an endospore, a metabolically dormant cell, during times of nutrient deficiencies and extreme stress. These said structures are outstandingly resistant to harsh chemicals, extreme temperatures, and can revert back to a metabolically active cell, through a process known as germination, when the necessary conditions are met. The rigid membrane of the endospore contains various germination (Ger) receptors which sense the external environment for necessary metabolites and germinants. Ger receptors are encoded by tricistronic operons that produce three distinct membrane proteins, the A, B, and C subunits. Although the function of the …

Regulation Of The Shigella Flexneri Icsp Gene And H-Ns Dependent Repression, Rosa Ojeda, Amanda Wigley, Dustin Harrison, Helen Wing

Undergraduate Research Opportunities Program (UROP)

The gram negative bacterium Shigella flexneri is known to cause dysentery in humans and primates. In order to help prevent the spread of shigellosis, gene regulation must be understood. Studies show that the virulence genes in S .flexneri are thermo regulated. At 30°C histone-like nucleoid structuring protein (H-NS) represses transcription of virulence genes and at 37°C VirB derepresses virulence genes. One of the genes that contribute to the virulence of S. flexneri is icsP. My project focuses on the regulation of the S. flexneri icsP gene and has two main goals. The first is to identify the sequence of H-NS …

Identification Of Nitrifying Bacteria Contained In A Commercial Inoculant Using Molecular Biology Techniques, Anthony Harrington, John Perry, Penny S. Amy

Undergraduate Research Opportunities Program (UROP)

Nitrifying bacteria play an important role in aquatic and terrestrial environments through the nitrogen cycle. Nitrification, one of the processes of the nitrogen cycle, refers to the oxidation of ammonia to nitrate. This process requires two types of chemoautotrophic bacteria, ammonia-oxidizing bacteria (AOB), and nitrite-oxidizing bacteria (NOB). These bacteria are essential in maintaining an optimal environment for plants and aquatic organisms, such as fish. Current applications of nitrifiers include: inoculants for aquariums, biofertilizers, and nitrogen removal in wastewater treatment plants. This study wants to identify a consortium of nitrifers that can be used to produce sufficient nitrate for plants in …

Transcriptional Regulation Of Shigella Virulence Plasmid-Encoded Genes By Virb And Crp, Christopher Thomas Hensley

UNLV Theses, Dissertations, Professional Papers, and Capstones

Shigella flexneri is a species of Gram-negative intracellular pathogens that causes bacillary dysentery in humans. Shigella relies on the precise transcriptional regulation of virulence genes, encoded by a large virulence plasmid, for invasion and infection of human colonic epithelial cells. The transcription of most identified virulence genes are regulated through a cascade controlled by the primary regulator of virulence genes, VirF, and the global transcriptional regulator, VirB. Currently, few studies have addressed how individual Shigella virulence genes are precisely regulated for optimal expression during specific stages of pathogenesis and within the constraints of the regulatory cascade. This work addresses how …

Bacillus Cereus And Bacillus Anthracis Germination Kinetics: A Michaelis-Menten Approach, Helen Luu

UNLV Theses, Dissertations, Professional Papers, and Capstones

Bacillus species are rod-shaped, gram-positive bacteria that are capable of producing endospores. In this dormant stage, the endospores can persist in hostile physical and chemical environments. Once conditions become favorable, the spores germinate into actively dividing cells, vegetative cells. Germination is a crucial step for the pathogenicity of the Bacilli in affecting a host organism.

Our study applies mathematical approaches to spore germination to determine whether the binding of one germinant will affect the binding of another germinant. We pursued this approach with two different species, B. cereus and B. anthracis, both pathogenic organisms. B. cereus is a widely known …

Protection Of Macrophages J774a.1 By Purine Nucleoside Analogues From Bacillus Anthracis Mediated Necrosis, Zadkiel R. Alvarez

UNLV Theses, Dissertations, Professional Papers, and Capstones

Eight years after the lamentable anthrax attacks, major scientific effort continues to be done, in order to stop imminent acts of bioterrorism. Innovative ways of therapy against the anthrax disease are being investigated. B. anthracis, the etiological agent of the infection, has a dormant stage in its life cycle known as the endospore. When conditions become favorable spores germinate, transforming into vegetative bacteria. In inhalational anthrax, the most fatal manifestation of the disease, spores enter the organism through the respiratory track, and are phagocytosized by alveolar macrophages of the lungs. Spores are able to sense nutrient availability, activating their germination …

Microbial Nitrogen Cycling In Nevada Geothermal Springs, Mitchell G. Chaires, Jeremy A. Dodsworth, Brian P. Hedlund

Undergraduate Research Opportunities Program (UROP)

Hot spring habitats above maximum photosynthetic temperature (73 ºC) are not well understood with respect to nitrogen (N) cycling. Few predictions have been made, and even fewer measurements of in situ activities have been reported. Thermodynamic calculations based on in situ chemical and temperature measurements will be used to predict the occurrence of the specific N-cycling reactions. In addition, these measurements in two springs will aid in an attempt to cultivate ammonia oxidizing species.

Report Of Significant Findings--Las Vegas Bay/Boulder Basin Investigations, James F. Labounty, Michael Horn, Bureau Of Reclamation

Publications (WR)

Field sampling was carried out between 0830 and 1500 beginning at the confluence of Las Vegas Wash and the Inner Las Vegas Bay. Ten (10) locations were sampled, each in a similar manner. Locations of sampling stations are in line from the Wash-Bay confluence to a point midway between Saddle and Black Islands. In addition, sampling was done at a location midway between Sentinel Island and the base of Fortification Hill, and at the buoy line in front of Hoover Dam. Sampling stations are labeled from LV01, at Wash-Bay confluence, to LV17 at Hoover Dam. A significant data collection point …

Characterization Of The Aquatic Environment In Lake Mead Near The Proposed Spring Canyon Pumped-Storage Project, And Assessment Of Potential Aquatic Impacts, Charles R. Liston, Stephen J. Grabowski, Bureau Of Reclamation

Publications (WR)

A pumped storage system consists of an upper reservoir and lower reservoir separated by an elevation difference. During low demand energy periods such as nights and weekends water is pumped from the lower to the upper reservoir using available energy from conventional steam electric power plants. During high energy demand periods, such as mornings and afternoons of weekdays, upper reservoir water is allowed to drop back down through the same system of water conduits and turbines, generating electricity to conveniently meet abrupt electrical energy requirements. The same water turbines thus act both as pumps and as conventional hydroelectric turbines.

Because …

A Review Of The Limnology Of And Water Quality Standards For Lake Mead, Charles R. Goldman

Publications (WR)

1. The waters of Las Vegas Bay, a heavily utilized recreational resource, receive discharges from a variety of municipal and industrial waste sources. The U.S. Environmental Protection Agency has determined, on the basis of numerous studies, that the present water quality violates state and federal standards and constitutes a public nuisance. Consultants have advised the Sewage and Wastewater Advisory Committee that rapid abatement of the alleged pollution conditions can be achieved by an advanced wastewater treatment (AWT) plant.

2. The major problems in Las Vegas Bay are an objectionable water color, excessive turbidity, noxious odors, and oxygen depletion in certain …

Report On Wastewater Disposal To Board Of County Commissioners, Clark County Nevada, Clair N. Sawyer

Publications (WR)

This report is concerned with the management of waste-waters and subsurface drainage from the City of Las Vegas and its environs and the BMI complex, all in Clark County, Nevada. The prime objective is to recommend a plan of action which will utilize the natural resources of the area in the least costly manner and still protect Lake Mead for recreational purposes and use as a public water supply, At this writing, conditions in the upper Las Vegas arm of Boulder Basin are quite unsatisfactory for some recreational purposes due to the extensive blooms of algae which develop. Experience at …

Phytoplankton Distribution And Water Quality Indices For Lake Mead (Colorado River), Robert D. Staker, Robert W. Hoshaw, Lorne G. Everett

Publications (WR)

Phytoplankton samples were collected in Lake Mend 6 times from September 1910 to June 1971 for 8 stations at depths of 0. 3, 5, 10, 20, and 30 m. These samples were processed through a Millipore filter apparatus and 79 planktonic algae were identified. Algal divisions represented were Bacillariophyta, 42 species; Chlorophyta, 18 ; Cyanophyta, 9; Chrysophyta, 3; Cryptophyta, 3; Pyrrophyta, 2; and Euglenophyta, 2. Blue-green algae were dominant in late summer and fall; green algae, diatoms, and, cryptomonads in winter; and green algae in spring. The early summer flora was best represented by the Chlorophyta, Cryptophyta, and Chrysophyta. Palmer's …

A Diurnal Zooplankton Migration Study In Lake Mead, Robert D. Staker

Publications (WR)

The diurnal vertical movement of zooplankton was first recorded in freshwater lakes by Weismann (1877) in Lake Constance (Bodensee), although Cuvier was credited with observing the migration of Daphniae in 1817 (Gushing, 1955). The migration is best observed in deep oligotrophic lakes and migrations of 50 m per day are described (Worthington, 1931). In addition, two species of chaetognaths are reported to migrate 400 m a day in marine waters off of Lisbon (Waterman and Berry, 1967) while Birge (1895) found little evidence of the phenomenon at all in Lake Mendota, Wisconsin.

Most zooplankton that migrate rise at night and …

Phytoplankton Successions And Lake Dynamics In Las Vegas Bay, Lake Mead, Nevada, Ervon R. Koening, Richard W. Tew, James E. Deacon

Publications (WR)

Phytoplankton successions, applications of the general growth equation, and physical measurements have been employed to investigate events occurring at the interface between industrial and sewage effluent contained in Las Vegas Wash and the waters of Lake Mead, Nevada. The data indicate that the entering waters tend to form a density current interrupted at intervals by dynamic effects generated in the lake. The dynamic relationships described here for the spring months suggest that a much more thorough understanding of physical, chemical and biological interactions is necessary to permit solution of the numerous problems of Las Vegas Bay discussed or alluded to …

A Mathmatical Model Of Primary Productivity And Limnological Patterns In Lake Mead, Lorne G. Everett

Publications (WR)

The temporal and spatial changes in chemical and biological properties of Lake Mead have been investigated, thereby indicating the sources of water pollution and the time of highest pollution potential. Planktonic organisms have been shown to indicate the presence of water problems. Macro- and micro-nutrient analyses have shown that primary productivity is not inhibited by limiting concentrations. A mathematical model has been developed, tested with one set of independent data, and shown worthy of management utility. Although the model works very well for the Lake Mead area, the physical reality of the Multiple Linear Regression equation should be tested on …