Bacteria Commons^™

Developing Coexpression Systems To Introduce Hydroxyproline Into Protein Engineered Collagen Peptides Utilizing Hydroxylase From Acanthamoeba Polyphaga Mimivirus, Jennifer Soldatich

Theses and Dissertations

A major challenge of developing collagen mimetic peptides (CMPs) by bacterial expression is to include hydroxyproline for biomedical applications. Coexpression of Prolyl-4-hydroxylase from A.mimivirus with CMPs was investigated. Additionally, four expression designs were created for potential applications in mammalian cells to utilize their natural ability of proline-hydroxylation by post-translational modification.

Studying Gene Function In Streptococcus Sanguinis Using Rna-Sequencing Analysis, Ahmed Ismail

Theses and Dissertations

The oral cavity hosts hundreds of bacterial species, amongst them are the Streptococci family. More specifically, S. sanguinis is a benign bacteria in the oral cavity to protect the tooth surface from S. mutans. which is a pathogen causing dental caries. While S. sanguinis is harmless in the oral cavity, it becomes pathogenic once entering the bloodstream and causing Infective Endocarditis (IE). To further study S. sanguinis and eventually develop an effective drug to treat IE caused by S. sanguinis, a gene expression study was conducted. In order to study gene expression, RNA-sequencing was conducted, and appropriate methods to remove …

Dendrimer-Based Antibiotics For The Treatment Of Bacterial Biofilm In Cystic Fibrosis (Cf), Younan Ma

Theses and Dissertations

Pseudomonas aeruginosa (PA) is the predominant pathogen in chronic lung infections of cystic fibrosis (CF) patients. The most important mechanism of adaptation of PA to host defense and antibiotic treatment is the formation of biofilms within the mucus layer covering the lung bronchi. The effectiveness of antibiotics such as aminoglycosides is significantly attenuated by their limited penetration through thick mucus and embedded biofilm matrix in patients’ lung. Inhaled tobramycin (Tobra), which is the most commonly used antibiotics in the treatment of PA infections for CF patients, is usually found to be in very high concentration in patients’ lung, and yet …

Detection And Quantification Of Bacterial Species Important To Mental And Physical Health, David G. Leonard

Theses and Dissertations

The human gut microbiome contains an abundance of microorganisms which could influence mental health as well as physical health. These microorganisms produce chemicals which affect the brain and the body in various ways. Probiotic bacteria and yeasts have been studied to determine effects they have on mice, rats, and humans to illustrate the importance these microorganisms on health. Studies have shown that adding beneficial microorganisms to the human diet can have positive effects on mental and physical health, to include lessening symptoms of depression and anxiety, lessen gastrointestinal inflammation, displacing pathogens, and improving immunomodulatory response. A quantitative way to identify …

The Role Of Manganese In Streptococcus Sanguinis, Tanya M. Puccio

Theses and Dissertations

Streptococcus sanguinis is primarily associated with oral health as a commensal bacterium. As an opportunistic pathogen, S. sanguinis is capable of colonizing heart valve vegetations, leading to the disease infective endocarditis. Previous studies from our lab have identified the high-affinity manganese transporter SsaACB as important for endocarditis virulence. The impact that manganese depletion has on S. sanguinis had never been evaluated and a secondary manganese transporter has not been identified. Thus, we employed the use of a fermentor to control large-scale growth over time and depleted manganese in an ΔssaACB mutant using a metal chelator, EDTA. The changes in …

Prediction Of The Vaginal Microbiome Using Urine Samples, Christopher R. Diachok

Theses and Dissertations

The vaginal microbiome is associated with women’s health, including but not limited to the pathogenesis of bacterial vaginosis, adverse pregnancy outcomes including preterm birth and sexually transmitted infections. However, there has been very limited research on the urinary microbiome and its relationship to the vaginal microbiome. We sought to predict the vaginal microbiome profile using clean catch urine samples from the same person. Here, paired vaginal and urine samples were collected and sequenced by 16S ribosomal RNA sequencing. Alpha and beta diversity analysis showed that there was no significant difference between the paired vaginal and urine microbiomes. In fact, a …

Characterization Of The Gonococcal Calprotectin Zn-Acquisition System, Tdfh, And The Efficacy Of Metal Starvation For Treating Gonococcal Disease, Michael T. Kammerman

Theses and Dissertations

Abstract

Characterization of the Gonococcal Calprotectin Zn-Acquisition System, TdfH, and the Efficacy of Metal Starvation for Treating Gonococcal Disease.

By Michael T. Kammerman, B.S.

A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy at Virginia Commonwealth University

Virginia Commonwealth University, 2020

Major Director: Cynthia Nau Cornelissen, Ph.D.

Professor, Department of Microbiology and Immunology

Neisseria gonorrhoeae, the bacterial agent that is responsible for the human disease gonorrhea, has had a steady increase in the number of infections per year. In 2018 the WHO estimated over 87 million infections occurred world-wide, and the CDC …

The Relationship Between Biofilm Production And Human Respiratory Cell Attachment And Invasion By Acinetobacter Baumannii, Rachel A. Pearson

Theses and Dissertations

Acinetobacter baumannii is an opportunistic gram-negative bacterial pathogen that causes many nosocomial infections in immunocompromised individuals. Common infections include catheter-associated urinary tract infections, ventilator-associated pneumonia, skin and soft tissue infections, and bloodstream infections that often lead to septicemia. Increasing multidrug resistance (MDR) in A. baumannii warrants new approaches to understanding its virulence mechanisms and pathogenicity. As a first step in infection, A. baumannii can attach to host cells, providing a surface for the bacteria to grow and perhaps facilitating biofilm formation and subsequent tissue invasion. In this study, we evaluated antibiotic resistance and characterized biofilm formation, attachment, invasion, and surface …

Examination Of Strain-Dependent Differences In S. Sanguinis Virulence And Growth, Shannon Baker

Theses and Dissertations

Streptococcus sanguinis, an abundant and benign inhabitant of the oral cavity, is an important etiologic agent of infective endocarditis, particularly in people with pre-disposing cardiac valvular damage. Although commonly isolated from patients with IE, little is known about the factors that make any particular S. sanguinis isolate more virulent than another or, indeed, whether significant differences in virulence exist among isolates. To investigate the virulence of multiple isolates, a variation of the Bar-seq (barcode sequencing) method was employed. A conserved chromosomal site was identified for subsequent insertion of a barcode identifier, unique for each strain. Barcode insertion did not …

Usefulness Of The Captia Syphilis Igg Eia Test Method And Reverse Algorithm For Detection Of Syphilis Infection In A Public Health Setting, Patricia Armour

Theses and Dissertations

Syphilis, a systemic sexually transmitted disease, is on the rise in the US, with infection rates the highest recorded since 1994 according to the CDC. Useful laboratory testing is an important diagnostic tool for determining individual syphilis infection and preventing community-wide disease spread.

The purpose of this study was to determine the usefulness of a specific automated treponemal test method, the Captia^TM Syphilis IgG EIA, and the syphilis reverse algorithm interpretation for detecting syphilis infection among patients seeking care in a public health clinic. The study employed a retrospective, nonexperimental descriptive correlational design with data collected between 2012-2013 from …

A Systems Biology Approach For Predicting Essential Genes And Deciphering Their Dynamics Under Stress In Streptococcus Sanguinis, Fadi El-Rami

Theses and Dissertations

Infectious diseases are the top leading cause of death worldwide. Identifying essential genes, genes indispensable for survival, has been proven indispensable in defining new therapeutic targets against pathogens, major elements of the minimal set genome to be harnessed in synthetic biology, and determinants of evolutionary relationships of phylogenetically distant species. Thus, essentiality studies promise valuable revenues that can decipher much of biological complexities.

Taking advantage of the available microbial sequences and the essentiality studies conducted in various microbial models, we proposed a framework for the prediction of essential genes based on our experimentally verified knowledge of the pathways involved in …

The Effects Of Amixicile, A Pyruvate Ferredoxin Oxidoreductase Inhibitor, On Oral Treponemes, Lucas A. Reed

Theses and Dissertations

Periodontal disease (PD) is a polymicrobial infection characterized by inflammation of the gingiva, alveolar bone resorption, and tooth loss (edentulism). Treponema denticola along with Porphyromonas gingivalis and Tannerella forsythia are among the “Red Complex” and are main etiological agents in PD. Treponemes are a member of the Spirochaeta phylum and are obligate anaerobes, that express pyruvate ferredoxin oxidoreductase (PFOR). The enzyme catalyzes the oxidation of pyruvate to acetyl-CoA and reduced ferredoxin. Amixicile is a novel bacteriostatic derivative of nitazoxanide and an inhibitor of PFOR. In light of the fact that Treponemes express PFOR, this study was conducted to investigate the …

Investigation Of Anaplasma Phagocytophilum And Anaplasma Marginale Adhesin-Host Cell Interactions, Kathryn S. Hebert

Theses and Dissertations

Anaplasma phagocytophilum and A. marginale are the etiologic agents of bovine anaplasmosis and human granulocytic anaplasmosis, respectively. As obligate intracellular pathogens, binding and entry of host cells is a prerequisite for survival. The molecular events associated with these processes are poorly understood. Identifying the adhesins mediating binding, delineating their key functional domains, and determining the molecular determinants to which they bind not only benefits better understanding of Anaplasma spp. pathobiology, but could also benefit the development of novel approaches for protecting against infection. We previously demonstrated that A. phagocytophilum outer membrane protein A (ApOmpA) is critical for bacterial binding and …

Characterization Of A Novel Protease In Staphylococcus Aureus, Adam L. Johnson

Theses and Dissertations

A newly discovered cysteine protease, Prp, has been shown to perform an essential, site-specific cleavage of ribosomal protein L27 in Staphylococcus aureus. In Firmicutes and related bacteria, ribosomal protein L27 is encoded with a conserved N-terminal extension that must be removed to expose residues critical for ribosome function. Uncleavable and pre-cleaved variants were unable to complement an L27 deletion in S. aureus, indicating that this N-terminal processing event is essential and likely plays an important regulatory role. The gene encoding the responsible protease (prp) has been shown to be essential, and is found in all organisms …