Microbiology Commons^™

Fluorescence-Reported Allelic Exchange Mutagenesis Reveals A Role For Chlamydia Trachomatis Tmea In Invasion That Is Independent Of Host Ahnak, M. J. Mckuen, Konrad E. Mueller, Y. S. Bae, Kenneth A. Fields

Microbiology, Immunology, and Molecular Genetics Faculty Publications

Development of approaches to genetically manipulate Chlamydia is fostering important advances in understanding pathogenesis. Fluorescence-reported allelic exchange mutagenesis (FRAEM) now enables the complete deletion of specific genes in C. trachomatis L2. We have leveraged this technology to delete the coding sequences for a known type III effector. The evidence provided here indicates that CT694/CTL0063 is a virulence protein involved in chlamydial invasion. Based on our findings, we designate the gene product corresponding to ct694-ctl0063 translocated membrane-associated effector A (TmeA). Deletion of tmeA did not impact development of intracellular chlamydiae. However, the absence of TmeA manifested as a decrease in infectivity …

Mechanism Of Candida Albicans Biofilm And Virulence Inhibition By A Bacterial Secreted Factor, Carrie Graham

Dissertations and Theses (Open Access)

The human microbiome is a diverse polymicrobial population comprised of both fungi and bacteria. Perturbations of the normal microbiome can have a profound impact on health, including the development of infections. Exploitation of these polymicrobial interactions has the potential to provide novel treatment and prevention strategies for infectious diseases. Enterococcus faecalis, a Gram-positive bacterium, and Candida albicans, a polymorphic fungus, occupy overlapping niches as ubiquitous constituents of the gastrointestinal and oral microbiome. Both species are also amongst the most important and problematic, opportunistic nosocomial pathogens and are often co-isolated during infection. Surprisingly, these two species antagonize each other’s …

Type Ix Secretion System: Characterization Of An Effector Protein And An Insight Into The Role Of C-Terminal Domain Dimeration In Outer Membrane Translocation., Lahari Koneru

Electronic Theses and Dissertations

Porphyromonas gingivalis and Tannerella forsythia are two of the primary pathogens that are associated in the etiology and progression of chronic periodontitis. In T. forsythia, KLIKK proteases are the recently identified group of proteolytic enzymes that are secreted through Type IX secretion system (T9SS). Among, these KLIKK proteases a synergistic relationship was observed between karilysin and mirolysin in invading the host complement system for the survival of the bacteria. Since, karilysin has been already characterized, in this study we propose to study about mirolysin through structural, biochemical and biological characterization. The obtained results from the experiments has shown the …

Experimental Approaches To Understand And Control Salmonella Infection In Poultry, Yichao Yang

Graduate Theses and Dissertations

Salmonella is a major foodborne pathogen around the world and chickens are the major reservoir to transmit Salmonella into the human food chain. For decreasing the infection of Salmonella, we developed six attenuated live vaccines based on Salmonella Enteritidis (SE) and Typhimurium (ST) for testing the cross-serovar and cross-serogroup protection from the challenge of Salmonella Heidelberg and Campylobacter jejuni. One of the constructed vaccine strain showed ability to protect against challenge from Salmonella Heidelberg. Even though some preventive approaches are able to decrease Salmonella colonization in the gastrointestinal tract of chickens or other farm animals, Salmonella transmission mechanisms remain unclear. …

Influence Of Epstein-Barr Virus On Systemic Lupus Erythematosus Disease Development And The Role Of Depression On Disease Progression, Caleb Cornaby

Theses and Dissertations

Systemic Lupus Erythematosus (SLE) is an autoimmune disease affecting 20 to 250 individuals per 100,000 worldwide. Symptomology includes dermatological manifestations such as discoid lesions, acute cutaneous rashes, and oral and nasal ulcers, along with musculoskeletal, pulmonary, and renal complications. Abnormal T and B lymphocyte function and apoptosis, immune complex clearance, complement function, and nucleosome processing are typical of disease pathophysiology. SLE is the result of both environmental and genetic factors, which together create the conditions leading to disease onset and progression. Of these environmental factors, Epstein-Barr virus (EBV) infection is known to cause the genesis of cross-reactive antibodies in SLE …

Single Nucleotide Polymorphisms In Efflux Pumps Genes In Extensively Drug Resistant Mycobacterium Tuberculosis Isolates From Pakistan, Akbar Kanji, Rumina Hasan, Asho Ali, Ambreen Zaver, Ying Zhang, Kehkashan Imtiaz, Wanliang Shi, Taane G. Clark, Ruth Mcnerney, Jody Phelan

Department of Pathology and Laboratory Medicine

It is challenging to understand mechanisms of drug resistance in Mycobacterium tuberculosis (MTB) due to the large variability in resistance associated genes. Efflux pump genes contribute to drug resistance and thus add to this complexity. Efflux pump gene protein superfamilies have been characterized by genome analysis of drug resistant strains and through invitro transcriptional studies. However, there is limited information regarding efflux pump genes in extensively drug resistant (XDR) tuberculosis (TB) isolates. Whole genome sequencing (WGS) based analysis of 37 extensively drug resistant (XDR) and five drug sensitive (DS) MTB clinical isolates was performed. Single nucleotide polymorphisms (SNPs) in efflux …

Acetobacter Fabarum Genes Influencing Drosophila Melanogaster Phenotypes, Kylie Makay White

Theses and Dissertations

Research in our lab has predicted hundreds of bacterial genes that influence nine different traits in the fruit fly, Drosophila melanogaster. As a practical alternative to creating site-directed mutants for each of the predicted genes, we created an arrayed transposon insertion library using a strain of Acetobacter fabarum DsW_054 isolated from fruit flies. Creation of the Acetobacter fabarum DsW_054 gene knock-out library was done through random transposon insertion, combinatorial mapping and Illumina sequencing. Successful mapping of transposon insertion was achieved for 6418 mutants with hits within 63% of annotated genes within Acetobacter fabarum DsW_054. Insertion sites were verified in …

Embracing Bacterial Cellulose As A Catalyst For Sustainable Fashion, Luis Quijano

Senior Honors Theses

Bacterial cellulose is a leather-like material produced during the production of Kombucha as a pellicle of bacterial cellulose (SCOBY) using Kombucha SCOBY, water, sugar, and green tea. Through an examination of the bacteria that produces the cellulose pellicle of the interface of the media and the air, currently named Komagataeibacter xylinus, an investigation of the growing process of bacterial cellulose and its uses, an analysis of bacterial cellulose’s properties, and a discussion of its prospects, one can fully grasp bacterial cellulose’s potential in becoming a catalyst for sustainable fashion. By laying the groundwork for further research to be conducted …

The Algz/R Two-Component System Is Responsible For Attenuation Of Virulence In Pseudomonas Aeruginosa, Danielle A. Williams

Electronic Theses and Dissertations

Pseudomonas aeruginosa is an important opportunistic pathogen. Many P. aeruginosa virulence factors are regulated by the AlgZ/R two component system. AlgZ is the sensor histidine kinase which phosphorylates AlgR, the response regulator. AlgR activates transcription of different gene targets based upon its phosphorylation state. The genes that encode AlgZ and AlgR are transcribed in an operon. While regulation of algR expression has been well studied, regulation of algZ expression has not. Using a pilW mutant in concert with algZTF-lacZ transcriptional fusion, we conducted a transposon mutagenesis to identify algZ regulators. We identified an unknown autoregulatory loop. The type …

Phosphate Signaling Through Alternate Conformations Of The Pstscab Phosphate Transporter, Ramesh Krishna Vuppada

Theses and Dissertations

Phosphate is an essential compound for life. Escherichia coli employs a signal transduction pathway that controls the expression of genes that are required for the high-affinity acquisition of phosphate and the utilization of alternate sources of phosphorous. These genes are only expressed when environmental phosphate is limiting. The seven genes for this signaling pathway encode the two-component regulatory proteins PhoB and PhoR, as well as the high-affinity phosphate transporter PstSCAB and an auxiliary protein called PhoU. As the sensor kinase PhoR has no periplasmic sensory domain, the mechanism by which these cells sense environmental phosphate is not known. This paper …

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 …

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 …

Role Of Incompatibility Group 1 (Inci1) Plasmid-Encoded Factors On Salmonella Enterica Antimicrobial Resistance And Virulence, Pravin Raghunath Kaldhone

Graduate Theses and Dissertations

Foodborne illnesses are a leading cause of infectious diseases in the world. Among enteric organisms Salmonella is a key pathogen. It’s high prevalence in poultry and other food-animal sources make it imperative to study. Salmonella has the ability to modify its genetic content with help of mobile genetic elements such as plasmids. Incompatibiltiy group 1 (IncI1) plasmids are commonly reported in Salmonella. This study evaluates role on IncI1 plasmids in antimicrobial resistance and virulence in Salmonella. Genetic determinants of resistance and virulence are noted among our IncI1-containing Salmonella isolates. These genetic elements are also transferable and reported to carry respective …

Development Of A Cholera Toxin Cta2/B Based Staphylococcus Aureus Vaccine To Prevent Bovine Mastitis, Neha Misra

Boise State University Theses and Dissertations

Staphylococcus aureus is an important pathogen causing chronic and invasive disease worldwide. This bacterium is a leading cause of community and hospital acquired infections in humans, and is also known to infect wild and domestic animals. Bovine mastitis, or inflammation of the udder, is one of the most economically relevant diseases of the dairy industry, with a high incidence worldwide. S. aureus is a major etiological agent causing this disease. S. aureus mastitis is highly contagious and difficult or impossible to treat. Management practices at dairy farms, that include good sanitation and antibiotic use, have been partially successful in reducing …

Complete Genome Sequence Of A New Isolate Of Solenopsis Invictavirus 3 From Solenopsis Invicta × Richteri Hybrid Ants, Steven M. Valles, Jason B. Oliver, Karla M. Addesso

Agricultural and Environmental Sciences Faculty Research

Solenopsis invicta virus 3 (SINV-3) is a positive-sense, single-stranded RNA virus that infects the red imported fire ant, Solenopsis invicta Buren. We report here the full genome (10,383 nucleotides) of an isolate infecting Solenopsis invicta× richteri hybrid ants, which we have identified as SINV-3 hybrid.

Development Of A Broadly Protective Modified-Live Virus Vaccine Candidate Against Porcine Reproductive And Respiratory Syndrome Virus, Haiyan Sun, Aspen Workman, Fernando A. Osorio, David J. Steffen, Hiep L.X. Vu

Nebraska Center for Virology: Faculty Publications

Modified-live virus (MLV) vaccines are widely used to protect pigs against porcine reproductive and respiratory syndrome virus (PRRSV). However, current MLV vaccines do not confer adequate levels of heterologous protection, presumably due to the substantial genetic diversity of PRRSV isolates circulating in the field. To overcome this genetic variation challenge, we recently generated a synthetic PRRSV strain containing a consensus genomic sequence of PRRSV-2. We demonstrated that our synthetic PRRSV strain confers unprecedented levels of heterologous protection. However, the synthetic PRRSV strain at passage 1 (hereafter designated CON-P1) is highly virulent and therefore, is not suitable to be used as …

Evaluation Of A Microsphere-Based Immunoassay (Mia) In Measuring Diagnostic And Prognostic Markers Of Dengue Virus Infection, Jason H. Ambrose

USF Tampa Graduate Theses and Dissertations

Infections with dengue viruses (DENV) constitute both a global problem as well as locally in Florida. DENV comprise four distinct serotypes of single-stranded RNA viruses and belong to the family Flaviviridae. DENV are among the most medically important arboviruses in the world and cases may currently exceed 400 million per annum. Additionally, dengue established its first recorded endemic transmission cycle in the state of Florida in over a half century, first within the Florida Keys during 2009-10 followed by an unrelated outbreak in Martin County in 2013. The clinical profile of DENV infections ranges from a mild febrile illness …

About Logan Weihe And Beloved Microcosm, Logan M. Weihe

Steeplechase: An ORCA Student Journal

No abstract provided.

Both Adhe And A Separate Nadph-Dependent Alcohol Dehydrogenase Gene, Adha, Are Necessary For High Ethanol Production In Thermoanaerobacterium Saccharolyticum, Tianyong Zheng, Daniel G. Olson, Sean J. Murphy, Xiongjun Shao, Liang Tian, Lee Lynd

Dartmouth Scholarship

Thermoanaerobacterium saccharolyticum has been engineered to produce ethanol at ∼90% theoretical yield and titer of 70 g/L. Its ethanol-producing ability has drawn attention to its metabolic pathways, which could potentially be transferred to other organisms of interest. Here we report that the iron-containing AdhA is important for ethanol production in the high-ethanol strain of T. saccharolyticum (LL1049). A single-gene deletion of adhA in LL1049 reduced ethanol production by ∼50%, whereas multiple gene deletions of all annotated alcohol dehydrogenases except adhA and adhE did not affect ethanol production. Deletion of adhA in wild-type T. saccharolyticum reduced NADPH-linked ADH activity (acetaldehyde-reducing) by …

Sweetpotato Virus C And Its Contribution To The Potyvirus Complex In Sweetpotato (Ipomoea Batatas), Favio E. Herrera Eguez

LSU Doctoral Dissertations

In Louisiana, sweetpotato (Ipomoea batatas) is infected in Louisiana by the four ubiquitous potyviruses: Sweetpotato feathery mottle virus (SPFMV), Sweetpotato virus G (SPVG), Sweetpotato virus 2 (SPV2) and the strain of SPFMV previously known as the common strain, recently renamed as Sweetpotato virus C (SPVC). These four viruses belong to the Potyviridae family, with single stranded RNA of ~11kb. In this group of plant viruses, a single polyprotein is coded entirely but later cleaved into ten mature proteins: P1, HC-pro, P3, 6K1, CI, 6K2, NIa-VPg, NIa-Pro Nib and Coat Protein (CP). In sweetpotato potyviruses, two additional open reading …

Efficacy Of An Adenoviral Vectored Multivalent Centralized Influenza Vaccine, Amy Lingel, Brianna L. Bullard, Eric A. Weaver

Nebraska Center for Virology: Faculty Publications

Mice were immunized with Adenovirus expressing the H1-con, H2-con, H3-con and H5-con HA consensus genes in combination (multivalent) and compared to mice immunized with the traditional 2010–2011 FluZone and FluMist seasonal vaccines. Immunized mice were challenged with 10–100 MLD50 of H1N1, H3N1, H3N2 and H5N1 influenza viruses. The traditional vaccines induced robust levels of HA inhibition (HI) titers, but failed to protect against five different heterologous lethal influenza challenges. Conversely, the multivalent consensus vaccine (1 × 1010 virus particles (vp)/mouse) induced protective HI titers of ≥40 against 8 of 10 influenza viruses that represent a wide degree of divergence within …

The Ecology Of Antibiotic Resistance: Sources And Persistence Of Vancomycin-Resistant Enterococci And Antibiotic Resistant Genes In Aquatic Environments, Suzanne M. Young

USF Tampa Graduate Theses and Dissertations

The growing crisis of antibiotic resistance is a major threat to ecosystems and human health. Infections caused by known and emerging antibiotic resistant pathogens are on the rise globally, with approximately 700,000 deaths per year caused by antibiotic resistant bacteria (1). In the United States, infections from antibiotic resistant bacteria cause more than 2 million illnesses and 23,000 deaths (2). Antibiotic resistant bacteria and antibiotic resistance genes are released into aquatic ecosystems through hospital waste, residential sewer lines and animal agricultural waste streams. Animal agriculture accounts for approximately 70% of antibiotic use in the United States (3). In agricultural ecosystems, …

Emergence Of The L Phenotype In Group B Streptococci In The South Of Ireland, Katherine Hayes, Lesley Cotter, L. Barry, Fiona O'Halloran

Department of Biological Sciences Publications

Group B Streptococcal isolates (n = 235) from the South of Ireland were characterised by serotyping, antimicrobial susceptibility and determination of the phenotypic and genotypic mechanisms of resistance. Resistance to erythromycin and clindamycin was observed in 21·3% and 20·4% of the total population, respectively. The c-MLSB phenotype was the most common phenotype detected (62%), with ermB being the predominant genetic determinant, present in 84% of resistant isolates. The rare L phenotype was observed in 2·9% (n = 7) of isolates, four of which harboured the lsaC gene responsible for clindamycin resistance. Serotypes Ia, III and II were the most common …

Mechanism Elucidation And Inhibitor Discovery Against Serine And Metallo-Beta-Lactamases Involved In Bacterial Antibiotic Resistance, Orville A. Pemberton

USF Tampa Graduate Theses and Dissertations

The emergence and proliferation of Gram-negative bacteria expressing β-lactamases is a significant threat to human health. β-Lactamases are enzymes that degrade the β-lactam antibiotics (e.g., penicillins, cephalosporins, monobactams, and carbapenems) that we use to treat a diverse range of bacterial infections. Specifically, β-lactamases catalyze a hydrolysis reaction where the β-lactam ring common to all β-lactam antibiotics and responsible for their antibacterial activity, is opened, leaving an inactive drug. There are two groups of β-lactamases: serine enzymes that use an active site serine residue for β-lactam hydrolysis and metalloenzymes that use either one or two zinc ions for catalysis. Serine enzymes …

Packaging Of Genetic Material By Gene Transfer Agents (Gtas) Produced By Marine Roseobacter Species And Their Effect On Stimulating Bacterial Growth, Shahd Bader Aljandal

USF Tampa Graduate Theses and Dissertations

Horizontal gene transfer is one of the most important mechanisms for prokaryotic genome innovation and evolution. Gene Transfer Agents (GTAs) are phage-like particles that package small fragments of the genome of a GTA-producing bacterial cell. GTA chromosomal gene clusters usually contain 15-conserved open reading frames (ORFs) and are present in most of the sequenced marine alpha-proteobacteria genomes. Some marine strains have been shown to produce GTA particles that were biologically active in marine environment.

GTA particles range in size, morphology and the amount of host DNA they package. To date, the characteristics of GTAs are largely based on observations of …

Defining Electron Bifurcation In The Electron-Transferring Flavoprotein Family, Amaya M. Garcia Costas, Saroj Poudel, Anne-Frances Miller, Gerrit J. Schut, Rhesa N. Ledbetter, Kathryn R. Fixen, Lance C. Seefeldt, Michael W. W. Adams, Caroline S. Harwood, Eric S. Boyd, John W. Peters

Chemistry Faculty Publications

Electron bifurcation is the coupling of exergonic and endergonic redox reactions to simultaneously generate (or utilize) low- and high-potential electrons. It is the third recognized form of energy conservation in biology and was recently described for select electron-transferring flavoproteins (Etfs). Etfs are flavin-containing heterodimers best known for donating electrons derived from fatty acid and amino acid oxidation to an electron transfer respiratory chain via Etf-quinone oxidoreductase. Canonical examples contain a flavin adenine dinucleotide (FAD) that is involved in electron transfer, as well as a non-redox-active AMP. However, Etfs demonstrated to bifurcate electrons contain a second FAD in place of the …

Susceptibility Pattern Of Methicillin Resistant Staphylococcus Aureus To Vancomycin And Other Alternate Agents: Report From A Private Sector Hospital Laboratory, Faryal Saleem, Naima Fasih, Afia Zafar

Department of Pathology and Laboratory Medicine

With increasing prevalence of methicillin resistant Staphylococcus aureus in clinical settings and injudicious use of antibiotics, resistance among MRSA against commonly used antibiotics is increasing. To assess the susceptibility pattern of MRSA against vancomycin, linezolid, tigecycline, rifampicin, fosfomycin fusidic acid, clindamycin, trimethoprim-sulfamethoxazole and teicoplanin, minimum inhibitory concentrations (MICs) for given antimicrobials were performed on 234 MRSA clinical isolates using automated VITEK 2 system. Vancomycin, linezolid, rifampicin, clindamycin, co-trimoxazole and teicoplanin susceptibilities were interpreted according to CLSI breakpoints, while tigecycline, fosfomycin and fusidic acid were interpreted according to BSAC breakpoints. All isolates were found susceptible to vancomycin, tigecycline, teicoplanin and linezolid. …

Susceptibility Pattern Of Methicillin Resistant Staphylococcus Aureus To Vancomycin And Other Alternate Agents: Report From A Private Sector Hospital Laboratory, Faryal Saleem, Naima Fasih, Afia Zafar

Department of Pathology and Laboratory Medicine

With increasing prevalence of methicillin resistant Staphylococcus aureus in clinical settings and injudicious use of antibiotics, resistance among MRSA against commonly used antibiotics is increasing. To assess the susceptibility pattern of MRSA against vancomycin, linezolid, tigecycline, rifampicin, fosfomycin fusidic acid, clindamycin, trimethoprim-sulfamethoxazole and teicoplanin, minimum inhibitory concentrations (MICs) for given antimicrobials were performed on 234 MRSA clinical isolates using automated VITEK 2 system. Vancomycin, linezolid, rifampicin, clindamycin, cotrimoxazole and teicoplanin susceptibilities were interpreted according to CLSI breakpoints, while tigecycline, fosfomycin and fusidic acid were interpreted according to BSAC breakpoints. All isolates were found susceptible to ancomycin, tigecycline, teicoplanin and linezolid. …

High-Throughput Single-Molecule Telomere Characterization, Jennifer Mccaffrey, Eleanor Young, Katy Lassahn, Justin Sibert, Steven Pastor, Harold Riethman, Ming Xiao

School of Medical Diagnostics & Translational Sciences Publications

We have developed a novel method that enables global subtelomere and haplotype-resolved analysis of telomere lengths at the single-molecule level. An in vitro CRISPR/Cas9 RNA-directed nickase system directs the specific labeling of human (TTAGGG) n DNA tracts in genomes that have also been barcoded using a separate nickase enzyme that recognizes a 7bp motif genome-wide. High-throughput imaging and analysis of large DNA single molecules from genomes labeled in this fashion using a nanochannel array system permits mapping through subtelomere repeat element (SRE) regions to unique chromosomal DNA while simultaneously measuring the (TTAGGG) n tract length at the end of each …

Blood Meal Acquisition Enhances Arbovirus Replication In Mosquitoes Through Activation Of The Gabaergic System, Y Zhu, R Zhang, B Zhang, T Zhao, Penghua Wang, G Liang, G Cheng

NYMC Faculty Publications

Mosquitoes are hematophagous insects that carry-on and transmit many human viruses. However, little information is available regarding the common mechanisms underlying the infection of mosquitoes by these viruses. In this study, we reveal that the hematophagous nature of mosquitoes contributes to arboviral infection after a blood meal, which suppresses antiviral innate immunity by activating the GABAergic pathway. dsRNA-mediated interruption of the GABA signaling and blockage of the GABAA receptor by the specific inhibitors both significantly impaired arbovirus replication. Consistently, inoculation of GABA enhanced arboviral infection, indicating that GABA signaling facilitates the arboviral infection of mosquitoes. The ingestion of blood by …