Microbiology Commons^™

Pathogenicity Of Acinetobacter Calcoaceticus, Kaitlan A. Sullivan

MUSC Theses and Dissertations

Acinetobacter is a genus of gram-negative bacteria that have been appearing frequently in hospitals contributing to infections in the blood, lungs, urinary tract, and other parts of the body. It infects patients with weakened immune systems that are placed on ventilators, after the use of catheters, or have any other open wounds produced by prolonged hospital stays. This genus of bacteria is problematic due to its high probability of becoming resistant to multiple classes of antibiotics. Thus, we are determining the pathogenicity of clinical isolates of Acinetobacter calcoaceticus using the organism Caenorhabditis elegans as a model.

We are testing …

Acinetobacter Baumannii - The Perfect Pathogen, Jesse Guzik, Myrna Rezcallah, Alexcia Zeller, Kaite Mattson

Research and Scholarship Symposium Posters

Acinetobacter was discovered in 1911 by Martinus Beijerinck. Acinetobacter baumannii didn't receive its scientific name until 1986. A. baumannii is now commonly referred to as "Iraqibacter" due to a rise in infections among US military personnel returning from Iraq and Afghanistan. After the Iraq War began in 2003, the frequency of cases began to rise, especially among patients in intensive care units. Patients using ventilators, catheters, have postoperative wounds, stay in the hospital for an extended period of time, or are immunocompromised are at a considerably higher risk of getting A. baumannii. Because of its large number of virulence …

Distribution Patterns Of Allorhizobium Vitis In Missouri Vineyards And Non-Vineyard Soils, Jacquelyn M. Wray

MSU Graduate Theses

Crown gall disease causes significant economic loss to the grape and wine industry. Preventive strategies are most effective for mitigating the loss of grapevines in vineyards, as there is no known cure for this disease. The bacterium Allorhizobium vitis carrying a tumor-inducing (Ti) plasmid is the causative agent. A. vitis bacteria live systemically in the grapevine before causing visible symptoms and can survive in residual plant tissues and soil for more than two years. Diagnostic methods have been developed to detect A. vitis bacteria in grapevines and soil. However, more reliable, specific, and high-throughput diagnostics are still needed for screening …

Resolving The Repression Pathway Of Virulence Gene Hila In Salmonella, Alexandra King, Lon Chubiz Phd, Brenda Pratte, Lauren Daugherty

Undergraduate Research Symposium

Salmonella is a relatively abundant, virulent species of bacteria that is most known for spreading gastrointestinal diseases through food. These illnesses result in approximately 1.35 million infections, including over 25,000 hospitalizations each year, in the U.S. alone (CDC.gov). As antibiotic resistance becomes an increasingly urgent public health problem, the importance of developing alternative treatment methods is only becoming more crucial. One of the genes responsible for this virulence is known as hilA. HilA is the main transcriptional regulator of Salmonella Pathogenicity Island-1 gene (UniProt). SPI-1 plays an important role in the invasion of Salmonella into epithelial cells. The proteins encoded …

Mara Repression Of Virulence Gene Hila In Salmonella, Alexandra King, Lauren Daugherty, Lon Chubiz Phd

Undergraduate Research Symposium

Salmonella is a bacteria most commonly known for causing the eponymous food-related illness. Due to their rapid reproduction rate and their ability to be propogated and maintained in a lab setting, they are commonly used in lab studies so that we can better understand how Salmonella causes disease in organisms that are more difficult to study. One area of interest is analyzing how Salmonella controls expression of the mechanisms that actually cause disease, called virulence traits, in response to the environment. In this study, antibiotic stress was used to analyze virulence gene expression. MarA is a gene that regulates ampicillin …

Increasing The Frequency Of Periodic Spatial Disturbance Decreases Surface Attachment Protein Expression In Staphylococcus Aureus, Ivana M. Barraza

All HCAS Student Capstones, Theses, and Dissertations

Staphylococcus aureus is an opportunistic pathogen that can cause a variety of acute and chronic illnesses. The severity of these illnesses such as sepsis, necrotizing pneumonia, and toxic shock syndrome is measured through the virulence that S. aureus inflicts on its host. Methicillin-resistant S. aureus (MRSA) is commonly associated with secondary infections and is challenging to treat given the limited selection of antibiotics that are effective against it. Accordingly, novel approaches to reduce S. aureus pathogenicity are required. S. aureus regulates pathogenesis through a cell-to-cell communication system referred to as quorum sensing. Effective communication determines the production of two broad …

Molecular Mechanisms Of Antimicrobial Resistance In Multi-Drug Resistant Enterococci, Ayesha Khan

Dissertations & Theses (Open Access)

Antibiotic resistance is a major global public health threat. Enterococci are recalcitrant, nosocomial pathogens that can be intrinsically resistant to valuable antibiotics, like beta-lactams, or evolve resistance to all existing antimicrobials. The LiaFSR system regulates resistance to cell membrane (CM) stressors like daptomycin (DAP), a front-line drug for multi-drug resistant infections. DAP resistance (DAP-R) in E. faecalis is mediated by CM phospholipid alterations. Emergence of DAP-R often leads to b-lactam resensitization, a phenomenon called the seesaw effect. The molecular mechanism of DAP-R and the seesaw effect are unknown. Here we show that LiaX is a surface exposed protein whose C-terminal …

Genome Sequencing Analysis Of Laboratory Isolate Of Francisella Noatunensis Subs. Orientalis, Joseph Paquette

Senior Honors Projects

Francisella noatunensis subs. orientalis is a known fish pathogen that has been most notably isolated from tilapia (Oreochromis niloticus) in Costa Rica. The genome of this Francisella species pathogen has been sequenced using Next-Generation Sequencing and been made available for the scientific community. Dr. Kathryn Ramsey’s research laboratory in the Department of Cell and Molecular Biology at the University of Rhode Island works with several Francisella species pathogens and is interested in identifying the differences, if any, between the known genome sequence of Francisella noatunensis and that of a laboratory isolate of the same species. With the use …

Transposon Mutagenesis Facilitates Discovery Of Genotype-Phenotype Associations And Functional Interrogation Of The Mycobacterium Kansasii Genome, William C. Budell

Dissertations, Theses, and Capstone Projects

Mycobacterium kansasii (Mk) is a nontuberculous mycobacterium (NTM) and medically relevant opportunistic human pathogen. Mk causes dangerous disease pathologies ranging from tuberculosis-like chronic pulmonary disease (CPD) to non-pulmonary focal or disseminated infections that are exacerbated by comorbidities such as chronic obstructive pulmonary disease (COPD), co-infection with HIV, or cancer. Among the most frequently identified cause of NTM-linked CPD, Mk infections contribute to a globally increasing NTM disease burden and are difficult to treat, requiring a long-term, multi-drug regimen. Although a less virulent pathogen than Mycobacterium tuberculosis (Mtb), Mk elicits similar disease features and shares in vitro …

Characterization Of The Role Of Transcriptional Regulator Of Arac/Xyls Family In Tularemia Pathogenesis, Dina Marghani

NYMC Student Theses and Dissertations

The Tier 1 Select Agent, Francisella tularensis causes an acute and fatal disease known as tularemia. Many studies have devoted enormous efforts to understand how F. tularensis avoids host defense mechanisms, replicates within an extremely secure immune system, and eventually causes the deadly disease tularemia. The extremely high virulence of Francisella depends on its ability to manipulate gene expression according to the surrounding environment. This process requires the involvement of unique transcriptional regulators. Francisella possesses very few transcriptional regulators, and a majority of them characterized to-date have been shown to regulate genes involved in virulence and cellular functions. The role …

Oxygen Deprivation Influences The Survival Of Listeria Monocytogenes In Gerbils, Jillian Harris, Oindrila Paul, Si Hong Park, Sally White, Khemraj Budachetri, Daniel M. Mcclung, Jessica G. Wilson, Alicia K. Olivier, Justin A. Thornton, Paul R. Broadway, Steven C. Ricke, Janet R. Donaldson

Faculty Publications

Listeria monocytogenes is a facultative anaerobic foodborne pathogen capable of surviving harsh environments. Recent work has indicated that anaerobic conditions increase the resistance capability of certain strains to environmental stressors. The goal of the study was to conduct a preliminary study to determine whether exposure to anaerobic conditions prior to infection increases the ability to survive in vivo. Gerbils were inoculated with one of five doses of the L. monocytogenes strain F2365 by oral gavage: phosphate-buffered saline (control), 5 × 10⁶ colony forming units aerobic culture (low aerobic), 5 × 10⁸ aerobic culture (high aerobic), 5 × 10 …

Functional Similarity Of Prd-Containing Virulence Regulators In Bacillus Anthracis, Malik Raynor

Dissertations & Theses (Open Access)

Bacillus anthracis produces three regulators, AtxA, AcpA, and AcpB, that control virulence gene expression and are members of an emerging class of regulators termed “PCVRs” (Phosphoenolpyruvate-dependent phosphotransferase regulation Domain-Containing Virulence Regulators). AtxA controls expression of the toxin genes; lef, cya, and pag, and is the master virulence regulator and archetype PCVR. AcpA and AcpB are less well studied. AcpA and AcpB independently positively control transcription of the capsule biosynthetic operon capBCADE, and culture conditions that enhance AtxA activity result in capBCADE transcription in strains lacking acpA and acpB. RNA-Seq was used to assess the regulons of the …

The Characterization Of The Transcription Factor Msab And Its Role In Staphylococcal Virulence, Justin Batte

Dissertations

Staphylococcus aureus is a common human pathogen that is responsible for a wide range of infections, ranging from relative minor skin infections to life-threatening disease such as bacteremia, septicemia, and endocarditis. S. aureus possesses many different virulent factors that aid in its ability to cause this wide array of infections. One major virulence factor includes the production of capsular polysaccharide (CP). The production of CP plays a major role in the virulence response during infection specifically by providing S. aureus an antiphagocytic mechanism that allows the pathogen to evade phagocytosis during an infection. S. aureus has developed complex genetic regulatory …

Characterization Of Two Cyclic Di-Gmp Phosphodiesterases Hypothesized To Regulate Virulence In Streptomyces Scabies, Madison Flasco

Undergraduate Honors Thesis Projects

Common scab is a devastating disease that affects the marketability of potatoes. The disease results in raised and pitted lesions seen on the surface of the potato and is caused by the soil pathogen, Streptomyces scabies. The S. scabies genome has two cyclic-di-GMP phosphodiesterases, RmdA and RmdB, both regulators of morphology and development. These proteins break down the ubiquitous second messenger molecule cyclic di-GMP, a molecule in charge of cell cycle progression and virulence. Both proteins contain diguanylate cyclase domains and phosphodiesterase domains, GGDEF and EAL, respectively, while RmdA contains an additional PAS_4 domain. Despite similar domain configurations, the two …

Secretion Of Tcpf By The Vibrio Cholerae Toxin-Coregulated Pilus Biogenesis Apparatus Requires An N-Terminal Determinant, Christina J. Megli, Ronald K. Taylor

Dartmouth Scholarship

Type IV pili are important for microcolony formation, biofilm formation, twitching motility, and attachment. We and others have shown that type IV pili are important for protein secretion across the outer membrane, similar to type II secretion systems. This study explored the relationship between protein secretion and pilus formation in Vibrio cholerae. The toxin-coregulated pilus (TCP), a type IV pilus required for V. cholerae pathogenesis, is necessary for the secretion of the colonization factor TcpF (T. J. Kirn, N. Bose, and R. K. Taylor, Mol. Microbiol. 49:81–92, 2003). This phenomenon is not unique to V. cholerae; secreted …

Pseudomonas Aeruginosa Ampr Transcriptional Regulatory Network, Deepak Balasubramanian

FIU Electronic Theses and Dissertations

In Enterobacteriaceae, the transcriptional regulator AmpR, a member of the LysR family, regulates the expression of a chromosomal β-lactamase AmpC. The regulatory repertoire of AmpR is broader in Pseudomonas aeruginosa, an opportunistic pathogen responsible for numerous acute and chronic infections including cystic fibrosis. Previous studies showed that in addition to regulating ampC, P. aeruginosa AmpR regulates the sigma factor AlgT/U and production of some quorum sensing (QS)-regulated virulence factors. In order to better understand the ampR regulon, the transcriptional profiles generated using DNA microarrays and RNA-Seq of the prototypic P. aeruginosa PAO1 strain with its isogenic ampR deletion …

Staphylococcus Aureus Response To Long Chain Antimicrobial Fatty Acids, Benjamin Arsic

Electronic Thesis and Dissertation Repository

Staphylococcus aureus is a common pathogen that has the ability to infect virtually every tissue and organ system of the body. Despite its propensity to cause invasive infection, S. aureus is also a commensal organism, asymptomatically colonizing ~25% of the population. Much research has gone into resolving this paradox, focusing on both human and bacterial factors that may contribute to colonization. Antimicrobial fatty acids present on the skin and in the nasal mucosa are important components of the innate immune system, and thus we undertook to further understand how S. aureus responds to these fatty acids, and how this response …

Whole-Genome Sequencing Of Staphylococcus Aureus Strain Rn4220, A Key Laboratory Strain Used In Virulence Research, Identifies Mutations That Affect Not Only Virulence Factors But Also The Fitness Of The Strain, Dhanalakshmi Nair, Guido Memmi, David Hernandez, Jonathan Bard, Marie Beaume, Steven Gill, Patrice Francois, Ambrose L. Cheung

Dartmouth Scholarship

Staphylococcus aureus RN4220, a cloning intermediate, is sometimes used in virulence, resistance, and metabolic studies. Using whole-genome sequencing, we showed that RN4220 differs from NCTC8325 and contains a number of genetic polymorphisms that affect both virulence and general fitness, implying a need for caution in using this strain for such studies.

Crystal Structure Of The Cystic Fibrosis Transmembrane Conductance Regulator Inhibitory Factor Cif Reveals Novel Active-Site Features Of An Epoxide Hydrolase Virulence Factor, Christopher D. Bahl, Christophe Morisseau, Jennifer M. Bomberger, Bruce A. Stanton, Bruce D. Hammock, George A. O'Toole, Dean R. Madden

Dartmouth Scholarship

Cystic fibrosis transmembrane conductance regulator (CFTR) inhibitory factor (Cif) is a virulence factor secreted by Pseudomonas aeruginosa that reduces the quantity of CFTR in the apical membrane of human airway epithelial cells. Initial sequence analysis suggested that Cif is an epoxide hydrolase (EH), but its sequence violates two strictly conserved EH motifs and also is compatible with other alpha/beta hydrolase family members with diverse substrate specificities. To investigate the mechanistic basis of Cif activity, we have determined its structure at 1.8-A resolution by X-ray crystallography. The catalytic triad consists of residues Asp129, His297, and Glu153, which are conserved across the …

The Regulation Of The Icsp Promoter Of Shigella Flexneri By The Virulence Factor Virb, Maria Castellanos, Dustin Harrison, Helen Wing

Undergraduate Research Opportunities Program (UROP)

Shigella flexneri is a pathogenic bacterium that causes severe dysentery in humans commonly known as shigellosis. Shigella encodes an outer membrane protease called IcsP. The regulation of icsP expression is under direct control of a transcriptional factor called VirB, which controls the expression of many virulence genes in Shigella. Previous work has shown through deletion analysis of the icsP promoter region that sequences as far as 1368 base pairs upstream of the transcription starting site are important for the regulation of the icsP gene by VirB. However, it is still unclear whether VirB activation requires sequences within the icsP promoter …

Integration Host Factor Positively Regulates Virulence Gene Expression In Vibrio Cholerae, Emily Stonehouse, Gabriela Kovacikova, Ronald K. Taylor, Karen Skorupski

Dartmouth Scholarship

Virulence gene expression in Vibrio cholerae is dependent upon a complex transcriptional cascade that is influenced by both specific and global regulators in response to environmental stimuli. Here, we report that the global regulator integration host factor (IHF) positively affects virulence gene expression in V. cholerae. Inactivation of ihfA and ihfB, the genes encoding the IHF subunits, decreased the expression levels of the two main virulence factors tcpA and ctx and prevented toxin-coregulated pilus and cholera toxin production. IHF was found to directly bind to and bend the tcpA promoter region at an IHF consensus site centered at position 162 …

Requirements For Vibrio Cholerae Hapr Binding And Transcriptional Repression At The Hapr Promoter Are Distinct From Those At The Apha Promoter, Wei Lin, Gabriela Kovacikova, Karen Skorupski

Dartmouth Scholarship

Virulence gene expression in certain strains of Vibrio cholerae is regulated in response to cell density by a quorum-sensing cascade that influences the levels of the LuxR homolog HapR through small regulatory RNAs that control the stability of its message. At high cell density, HapR represses the expression of the gene encoding the virulence gene activator AphA by binding to a site between −85 and −58 in the aphA promoter. We show here that a second binding site for HapR lies within the hapR promoter from which it functions to repress its own transcription. This site, as determined by gel …

Identification And Characterization Of A Nontypeable Haemophilus Influenzae Putative Toxin-Antitoxin Locus, Dayle A. Daines, Justin Jarisch, Arnold L. Smith

Biological Sciences Faculty Publications

Background: Certain strains of an obligate parasite of the human upper respiratory tract, nontypeable Haemophilus influenzae (NTHi), can cause invasive diseases such as septicemia and meningitis, as well as chronic mucosal infections such as otitis media. To do this, the organism must invade and survive within both epithelial and endothelial cells. We have identified a facilitator of NT(Hi) survival inside human cells, virulence-associated protein D (vapD_Hi, encoded by gene H10450). Both vapD_Hi and a flanking gene, H10451, exhibit the genetic and physical characteristics of a toxin/antitoxin ( TA) locus, with VapD_Hi serving as the …

The Virulence Activator Apha Links Quorum Sensing To Pathogenesis And Physiology In Vibrio Cholerae By Repressing The Expression Of A Penicillin Amidase Gene On The Small Chromosome, Gabriela Kovacikova, Wei Lin, Karen Skorupski

Dartmouth Scholarship

Activation of the tcpPH promoter on the Vibrio pathogenicity island by AphA and AphB initiates the Vibrio cholerae virulence cascade and is regulated by quorum sensing through the repressive action of HapR on aphA expression. To further understand how the chromosomally encoded AphA protein activates tcpPH expression, site-directed mutagenesis was used to identify the base pairs critical for AphA binding and transcriptional activation. This analysis revealed a region of partial dyad symmetry, TATGCA-N6-TNCNNA, that is important for both of these activities. Searching the V. cholerae genome for this binding site permitted the identification of a second one upstream of a …

Mechanism Of Toxt-Dependent Transcriptional Activation At The Vibrio Cholerae Tcpa Promoter, Robin R. Hulbert, Ronald K. Taylor

Dartmouth Scholarship

The AraC homolog ToxT coordinately regulates virulence gene expression in Vibrio cholerae. ToxT is required for transcriptional activation of the genes encoding cholera toxin and the toxin coregulated pilus, among others. In this work we focused on the interaction of ToxT with the tcpA promoter and investigated the mechanism of ToxT-dependent transcriptional activation at tcpA. Deletion analysis showed that a region from −95 to +2 was sufficient for ToxT binding and activation, both of which were simultaneously lost when the deletion was extended to −63. A collection of point mutations generated by error-prone PCR revealed two small regions required …