Microbiology Commons^™

To Mid-Cell And Beyond: Characterizing The Roles Of Gpsb And Ypsa In Cell Division Regulation In Gram-Positive Bacteria, Robert S. Brzozowski

USF Tampa Graduate Theses and Dissertations

The bacterial cell division protein FtsZ is a tubulin homolog that forms a ring-like structure at the site of cell division in most bacterial species. There it acts as a scaffold, aiding in the recruitment of other divisome proteins to the site of cell division. Furthermore, studies focusing on the role of FtsZ treadmilling and septal peptidoglycan synthesis implicates that FtsZ plays a direct role in the ultimate closure of the division septum. Thus, many studies in the field of bacterial cell division have focused on FtsZ in terms of its spatial and temporal regulation as well as its ability …

Biofilm Associated Staphylococcus Aureus Viability Is Altered By Burkholderia Cenocepacia, Bridget Wall, Tiffany J. Brandt, Dr. Deborah Yoder-Himes

Undergraduate Arts and Research Showcase

Respiratory failure caused by chronic and recurrent microbial infections is the most common cause of death for people with cystic fibrosis (CF)¹, a disease causing the formation of thick mucus in the lungs². Most bacteria can form biofilms, collections of sessile cells adhered to a surface by a secreted substance. Biofilm-associated cells develop antibiotic resistance at higher rates³. The thicker mucus in CF lungs is extremely difficult to clear via action of the mucociliary escalator and its presence fosters the formation of bacterial biofilms. Staphylococcus aureus and Burkholderia cenocepacia are two pathogens commonly found …

Mass Spectrometry Based Lipidomic Investigation Of Staphylococcus Aureus Lipid Metabolism And Characterization Of Lipase A, Naren Gajenthra Kumar

Theses and Dissertations

Staphylococcus aureus produces an arsenal of virulence factors and is adept at colonizing multiple hosts by employing specific processes to persist at different sites of infection. Infections caused by this pathogen range from self-limiting infections of the skin like folliculitis to life threatening and chronic conditions like infective endocarditis, non-healing diabetic foot ulcers and atopic dermatitis. As the field continues to expand its understanding of the mechanisms by which this pathogen rapidly develops resistance to new antibiotics, the underlying mechanisms that facilitates niche adaptation and host colonization among healthy individuals remain elusive.

Bacterial and host lipid metabolism presents and area …

Acid Resistance Mechanisms In Staphylococcus Aureus, Chunyi Zhou

Theses & Dissertations

Staphylococcus aureus is a leading cause of opportunistic infections in community and health care settings. To thrive in a great variety of environments, S. aureus has developed the capability of tolerating temporary pH changes, as well as resisting constant acid stress. To evaluate the impact of strong and weak acid stress on S. aureus, growth patterns of JE2 were monitored when cultured in chemically defined media (CDM) at various pH in the presence of hydrochloric acid (HCl), acetic acid, and lactic acid. Our results showed that S. aureus responds to strong and weak acids in different manners. S. aureus …

The Role Of Secreted Proteases In Regulating Disease Progression In Staphylococcus Aureus, Brittney D. Gimza

USF Tampa Graduate Theses and Dissertations

Staphylococcus aureus is a highly successful pathogen capable of producing a wealth of virulence factors in the human host. Of note, ten extracellular proteases are produced alongside these virulence factors and play a multifaceted role during infection. They not only cleave host proteins to promote bacterial invasion, immune evasion and survival, but also control disease progression by modulating the stability of self-derived pathogenic determinants. The importance of the secreted proteases modulating virulence factor stability is evidenced by our groups previous finding that a protease-null strain has a substantially increased infectious capacity in a murine model of sepsis; resulting from the …

Antibiotic Drug Discovery Targeting Bacterial Metabolism, Miranda J. Wallace

Theses and Dissertations (ETD)

Over the last century, the use of antibiotics has enabled many advances in modern medicine, making life as we know it possible. In recent years, however, emerging bacterial resistance to virtually all major antibiotic classes has resulted in a worldwide increase in morbidity, mortality, and financial burden associated with drug resistant infections. The antimicrobial resistance crisis presents an urgent need for new antimicrobials with distinct mechanisms of action from existing drugs. The current pharmaceutical pipeline of new antibiotics is limited due to three obstacles: a lack of understanding of resistance mechanisms, a dearth of novel mechanisms of action among new …

Combating Drug Resistance - Comparison Of The Antibiotic Effect Of Hydrastis Canadensis Extract And Pure Berberine Via Minimum Inhibitory Concentration Assay, William Luke Scott, Timothy D. Trott

Research in Biology

Herbal medicines are a melee of complex organic chemicals, making it difficult to ascertain their direct mechanism of action. In contrast to mainstream pharmaceuticals, it is argued that herbal medicines are effective because of multiple constituents working synergistically. The complexity of herbal medicines may give them advantages over simpler pharmaceuticals in combating antibiotic resistant microbes, but these advantages can be difficult to quantitate. Popular literature frequently espouses the healing properties of herbal medicines, but many of these claims are not scientifically supported. Many gains could be realized in public health and medicine if more research was aimed at validating / …

Staphylococcus Aureus Evasion Of The Innate Immune System, James Paul Grayczyk

Dissertations

Upon entry into the host, pathogens must overcome innate immunity in order to cause disease. The innate immune system represents a fast-acting initial line of defense to prevent infection. In order to withstand innate defenses, bacterial pathogens like the Gram-positive bacterium Staphylococcus aureus, produce a wide array of virulence factors that can inhibit innate immune cell recruitment and antimicrobial activity, or directly target and kill phagocytic leukocytes thereby facilitating pathogenesis. Infection with S. aureus can cause disease in virtually any tissue site and is a significant burden to human health. In this dissertation, we sought to understand how S. aureus …

Biofilm Producing Clinical Staphylococcus Aureus Isolates Augmented Prevalence Of Antibiotic Resistant Cases In Tertiary Care Hospitals Of Nepal, Sarita Manandhar, Anjana Singh, Ajit Varma, Shanti Pandey, Neeraj Shrivastava

Student Publications

Staphylococcus aureus, a notorious human pathogen, is a major cause of the community as well as healthcare associated infections. It can cause a diversity of recalcitrant infections mainly due to the acquisition of resistance to multiple drugs, its diverse range of virulence factors, and the ability to produce biofilm in indwelling medical devices. Such biofilm associated chronic infections often lead to increase in morbidity and mortality posing a high socio-economic burden, especially in developing countries. Since biofilm formation and antibiotic resistance function dependent on each other, detection of biofilm expression in clinical isolates would be advantageous in treatment decision. …

Antibiotic Resistance In Staphylococcus Aureus From Commercially Available Meat, Samuel Schriever, Bradford Berges

Journal of Undergraduate Research

Bacteria can be found everywhere. While some bacteria can help humans to make medicine and clean up environmental disasters, other bacteria can cause horrible disease. After the discovery of antibiotics in the early twentieth century humans have been using them to treat human and animal disease. At first antibiotics very effective at clearing bacterial infections. Bacteria that were able to survive the antibiotics were able to spread and become more common. Bacteria have the ability to share genes and those genes are very favorable for the bacteria that have them. Today, animals are given more antibiotics than humans are. This …

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 …

Hospital And Meat Associated Staphylococcus Aureus And Their Biofilm Characteristics, Trevor Michael Wienclaw

Theses and Dissertations

Biofilm phenotypes were studied in 32 Staphylococcus aureus strains isolated from store-bought meats and 22 from diseased patients in hospitals. Of the meat-associated strains, 21 were methicillin-resistant Staphylococcus aureus (MRSA) and 11 were methicillin-susceptible Staphylococcus aureus (MSSA). The hospital-associated strains included 15 MRSAs and 7 MSSAs. We studied the robustness and composition of the biofilms produced by these strains. We found that on average hospital-associated strains form more robust biofilms than meat associated strains. The model often used to describe S. aureus biofilm composition includes two biofilm types defined by the presence or absence of polysaccharide intercellular adhesin (PIA), PIA-dependent …

Gas Plasma Pre-Treatment Increases Antibiotic Sensitivity And Persister Eradication In Methicillin-Resistant Staphylococcus Aureus, Li Guo, Ruobing Xu, Yiming Zhao, Dingxin Liu, Zhijie Liu, Xiaohua Wang, Hailan Chen, Michael G. Kong

Bioelectrics Publications

Methicillin-resistant Staphylococcus aureus (MRSA) is a major cause of serious nosocomial infections, and recurrent MRSA infections primarily result from the survival of persister cells after antibiotic treatment. Gas plasma, a novel source of ROS (reactive oxygen species) and RNS (reactive nitrogen species) generation, not only inactivates pathogenic microbes but also restore the sensitivity of MRSA to antibiotics. This study further found that sublethal treatment of MRSA with both plasma and plasma-activated saline increased the antibiotic sensitivity and promoted the eradication of persister cells by tetracycline, gentamycin, clindamycin, chloramphenicol, ciprofloxacin, rifampicin, and vancomycin. The short-lived ROS and RNS generated by plasma …

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 …

Anti-Staphylococcal Activity Of Variovorax Paradoxus Eps, Patricia Holt-Torres

Electronic Theses, Projects, and Dissertations

Variovorax paradoxus EPS is a gram-negative rod isolated from the sunflower rhizosphere at CSUSB. Preliminary research has shown that Variovorax paradoxus EPS has anti-staphylococcal activity in liquid and solid co-culture. Anti-staphylococcal activity of Wild type and V. paradoxus EPS 𝚫4519 on 0.5% YE agar with embedded S. aureus AH1710 supports the idea that a soluble molecule is responsible for this activity, as the agar acted as a physical barrier between V. paradoxus EPS and S. aureus colonies. Preliminary genetic analysis of V. paradoxus EPS identified three loci that suitable candidates for the synthesis of a potential anti-staphylococcal small molecule. …

Redirection Of The Immune Response To Staphylococcus Aureus Biofilm Infection, Anna G. Staudacher

Theses & Dissertations

Staphylococcus aureus (S. aureus) is a leading cause of community- and healthcare-associated infections and has a propensity to form biofilms. Biofilm infections are recalcitrant to host immune-mediated clearance as well as antibiotics, making them exceptionally difficult to eradicate. The biofilm environment has been shown to skew the host immune response towards an anti-inflammatory phenotype, characterized by alternatively activated macrophages, recruitment of myeloid-derived suppressor cells (MDSCs), and minimal neutrophil and T cell infiltrates. Our laboratory has attempted to redirect the host immune response towards one that would favor bacterial clearance by employing strategies to augment pro-inflammatory mechanisms. One such …

Molecular Subtyping Of Staphylococcus Aureus Isolates From The U.P. Community For The Presence Of Toxin-Encoding Genes, Carol I. Kessel

All NMU Master's Theses

Staphylococcus aureus is the most common cause of human bacterial infections; however, humans can also be asymptomatically colonized with S. aureus. Asymptomatic carriers can potentially spread S. aureus infection to others. These infections can range from mild to severe. The pathology of a S. aureus infection is often dependent on which toxins are expressed and the virulence factors with which they are associated. One goal of this study was to isolate S. aureus from healthy, consenting adult volunteers who submitted nasal swabs for culture and qRT-PCR analysis to determine which strains are present in the community. This knowledge could …

Cytokine Expression, Cytoskeleton Organization, And Viability Of Sim-A9 Microglia Exposed To Staphylococcus Aureus-Derived Lipoteichoic Acid And Peptidoglycan, Erin Roberts

Browse all Theses and Dissertations

In these experiments, SIM-A9 microglia were exposed to Staphylococcus aureus cell wall components lipoteichoic acid (LTA) and peptidoglycan (PGN) at a concentration of 5 ug/mL for six, twelve, eighteen, and twenty-four hours and the cytokine expression, cytoskeleton organization, and cell viability of the cells were observed. Following LTA and PGN exposure, TNF-a; secretion increased at each time interval and was highest observed at 24 hours. No significant IL-10 secretion was detected. Over the 24 hour period, cell viability and cytotoxicity of LTA and PGN treatment groups were not significantly different from the control, indicating the observed inflammatory cytokine response was …

The Effect Of Gram-Positive Staphylococcus Aureus Cell Wall Components Lipoteichoic Acid And Peptidoglycan On Cytokine Production, Cytoskeletal Arrangement, And Cell Viability On Raw 264.7 Murine Macrophages, Gabrielle String

Browse all Theses and Dissertations

In this study, gram positive Staphylococcus aureus cell wall components such as lipoteichoic acid (LTA) and peptidoglycan (PGN) were used to study the potential inflammatory and anti-inflammatory cytokine response, cytoskeletal arrangement and cell viability on RAW264.7 murine macrophages over 24 hours. The effect of S.aureus LTA and PGN (5 [mu]g/mL) on RAW 264.7 macrophages was evaluated every six hours for twenty-four hours. Inflammatory cytokine (TNF-a) production peaked at 6 hours before decreasing over time. Anti-inflammatory cytokine (IL-10) production peaked between 12 and 18 hours. During the first twenty-four hours, cytotoxicity of treated macrophages, as defined as the release of lactate …

Amino Acid Catabolism In Staphylococcus Aureus, Cortney Halsey

Theses & Dissertations

Staphylococcus aureus must rapidly adapt to a variety of carbon and nitrogen sources during invasion of a host. Within a staphylococcal abscess, preferred carbon sources such as glucose are limiting, suggesting S. aureus survives through the catabolism of secondary carbon sources. S. aureus encodes pathways to catabolize multiple amino acids including those that generate pyruvate, 2-oxoglutarate, and oxaloacetate. To assess amino acid catabolism, S. aureus JE2 and mutants were grown in complete defined medium containing 18 amino acids but lacking glucose (CDM). A mutation in glutamate dehydrogenase (gudB), which generates 2-oxoglutarate from glutamate, abrogated growth in CDM suggesting …

Fluid Dynamic Factors As A Cause And Effect Of Biofilm Formation Of Staphylococcus Aureus Biofilms, Erica Sherman

Department of Mechanical and Materials Engineering: Dissertations, Theses, and Student Research

Staphylococcus aureus bacteria are able to form biofilms and distinctive tower structures that facilitate their ability to tolerate treatment and to spread within the human body. The formation of towers, which break off, get carried downstream and serve to initiate biofilms in other parts of the body are of particular interest here. It is known that flow conditions play a role in the development, dispersion and propagation of biofilms. The influence of flow on tower formation and what factors lead to tower formation is not at all understood The hypothesis being examined is that tower structures form within a specific …

Spatiotemporal Control Of Staphylococcus Aureus Biofilm Development, Derek E. Moormeier

Theses & Dissertations

Chronic biofilm-related infections caused by the human pathogen Staphylococcus aureus often lead to significant increases in morbidity and mortality in both hospital- and community-associated settings. Typically, S. aureus biofilm development occurs in three stages: 1) attachment, 2) tower maturation, and 3) dispersal. Here, utilizing BioFlux1000 time-lapse microscopy we have expanded upon these fundamental stages of biofilm development and also unveiled and characterized two additional stages (multiplication and exodus).

The attachment and multiplication stages were shown to be protease sensitive but independent of most cell surface-associated proteins. Following multiplication, an exodus of the biofilm population that followed the transition of the …

A Novel And Rapid Staphylococcus Aureus Bacterial Identification Method Utilizing Immunomagnetic Beads And Single Cell Laser-Light Scattering, Kaylagh Hollen

All NMU Master's Theses

Staphylococcus aureus is the most commonly isolated human associated bacterial pathogen. It plays an important role in skin and soft-tissue infections, pneumonia, endocarditis, osteomyelitis, foreign-body infections, and sepsis. S. aureus diagnosis and treatment requires a minimum of 24-48. With this in mind, previous studies suggest that faster pathogen identification has been linked to improved patient outcomes. Improved patient outcomes including a reduction in hospitalization time, decreased risk of nosocomial infections, and decreased in medical costs. The impact of faster identification on patient outcome has led us to develop an alternative method of S. aureus identification via ImmunoMagnetic Separation (IMS) and …

Discovery And Characterization Of Bacteriophage To Control Mrsa, Jacob B. Hatch, Bradford Berges

Journal of Undergraduate Research

About 20% of humans are carriers of Staphylococcus aureus (SA). There were an estimated 11,000 deaths in the United States in 2005 attributed to SA, with the majority caused by MRSA (Methicillin Resistant Staphylococcus Aureus) isolates [1]. Many MRSA isolates have developed resistance to all but one antibiotic drug: vancomycin. However, other bacteria have developed resistance to vancomycin, suggesting that in time MRSA will likewise become non-responsive to this last available drug and MRSA infections will be untreatable. This project looks to find an alternate method of MRSA treatment. Bacteriophage (phage) are viruses that infect bacterial cells in order …

Isolation And Characterization Of Novel Lytic Phage To Treat Mrsa, Tyler White, Bradford Berges

Journal of Undergraduate Research

The bacterium Staphylococcus Aureus is the cause of a serious skin disease that is known to cause life-threatening infections such as pneumonia, endocarditis, meningitis, sepsis, bacteremia, and toxic shock syndrome [1]. Nonetheless, most SA infections are readily remedied via antibiotic treatment with methicillin. However, over the course of frequent exposure to various antibiotics, the bacteria have evolved a mutant strain (MRSA) that is completely resistant to the drug. In 2005 an estimated 11,000 deaths occurred that can be attributed to SA, with the majority caused by MRSA isolates [2]. Albeit the mutant is named MRSA, isolates of this strain are …

Identification Of Novel Cyclic Lipopeptides From A Positional Scanning Combinatorial Library With Enhanced Antibacterial And Antibiofilm Activities, Nina Bionda, Renee M. Fleeman, César De La Fuente-Núñez, Maria C. Rodriguez, Fany Reffuveille, Lindsey N. Shaw, Irena Pastar, Stephen C Davis, Robert E W Hancock, Predrag Cudic

Molecular Biosciences Faculty Publications

Treating bacterial infections can be difficult due to innate or acquired resistance mechanisms, and the formation of biofilms. Cyclic lipopeptides derived from fusaricidin/LI-F natural products represent particularly attractive candidates for the development of new antibacterial and antibiofilm agents, with the potential to meet the challenge of bacterial resistance to antibiotics. A positional-scanning combinatorial approach was used to identify the amino acid residues responsible for driving antibacterial activity, and increase the potency of these cyclic lipopeptides. Screening against the antibiotic resistant ESKAPE pathogens revealed the importance of hydrophobic as well as positively charged amino acid residues for activity of this class …

Transcriptomic Analysis Of Staphylococcal Srnas: Insights Into Species-Specific Adaption And The Evolution Of Pathogenesis, William H. Broach, Andy Weiss, Lindsey N. Shaw

Molecular Biosciences Faculty Publications

Next-generation sequencing technologies have dramatically increased the rate at which new genomes are sequenced. Accordingly, automated annotation programs have become adept at identifying and annotating protein coding regions, as well as common and conserved RNAs. Additionally, RNAseq techniques have advanced our ability to identify and annotate regulatory RNAs (sRNAs), which remain significantly understudied. Recently, our group catalogued and annotated all previously known and newly identified sRNAs in several Staphylococcus aureus strains. These complete annotation files now serve as tools to compare the sRNA content of S. aureus with other bacterial strains to investigate the conservation of their sRNomes. Accordingly, in …

Regulation Of Vancomycin Resistance And Stress Response By The Msaabcr Operon In Staphylococcus Aureus, Dhritiman Samanta

Dissertations

Staphylococcus aureus is the predominant cause of public health problems around the world. Vancomycin has been an important antibiotic against Methicillin Resistant Staphylococcus aureus (MRSA) infections. However, Vancomycin Intermediate S. aureus (VISA) strains have been reported. These strains are characterized by thick cell walls, reduced autolytic rate, reduced PBP4 activity, and increased amount of D-Ala-D-Ala termini in the cell wall. In this study, we show that the msaABCR operon regulates vancomycin resistance in two clinical VISA strains. Deletion of the msaABCR operon in strains Mu50 and HIP6297 resulted in a significant decrease in the minimum inhibitory concentration (MIC) for vancomycin. …

Isolation And Host Range Of Staphylococcus Aureus Bacteriophages And Use For Decontamination Of Fomites, Kyle C. Jensen

Theses and Dissertations

Staphylococcus aureus is a common bacterium found on the skin and mucosal membranes of about 20% of the population. S. aureus growth on the skin is harmless, but if it bypasses the skin it can causes life-threatening diseases such as pneumonia, meningitis, bacteremia, and sepsis. Antibiotic-resistant strains of S. aureus, called Methicillin Resistant S. aureus (MRSA), are resistant to most antibiotics except vancomycin. However, vancomycin resistant strains of MRSA are becoming more common. In this study, 12 phages were isolated capable of infecting human S. aureus and/or MRSA strains. Five phages were discovered through mitomycin C induction of prophages and …

Discovery And Characterization Of Bacteriophage To Control Mrsa, Jacob B. Hatch, Bradford Berges

Journal of Undergraduate Research

About 20% of humans are carriers of Staphylococcus aureus (SA). There were an estimated 11,000 deaths in the United States in 2005 attributed to SA, with the majority caused by MRSA (Methicillin Resistant Staphylococcus Aureus) isolates [1]. Many MRSA isolates have developed resistance to all but one antibiotic drug: vancomycin. However, other bacteria have developed resistance to vancomycin, suggesting that in time MRSA will likewise become non-responsive to this last available drug and MRSA infections will be untreatable. This project looks to find an alternate method of MRSA treatment. Bacteriophage (phage) are viruses that infect bacterial cells in order …