Pathogenic Microbiology Commons^™

Development, Expansion And Role Of Myeloid-Derived Suppressor Cells In Post-Sepsis Immune Suppression, Tuqa Alkhateeb

Electronic Theses and Dissertations

Myeloid-derived suppressor cells (MDSCs) numbers increase significantly in sepsis and are associated with high mortality rates. These myeloid cell precursors promote immunosuppression, especially in the late (post sepsis) stage. However, the mechanisms that underlie MDSC expansion and programming are not completely understood. To investigate these mechanisms, we used a cecal-ligation and puncture (CLP) mouse model of polymicrobial sepsis that progresses from an early/acute proinflammatory phase to a late/chronic immunosuppressive phase. Previous studies in our laboratory showed that microRNA (miR)-21 and miR-181b elevate levels of the transcription factor nuclear factor 1 (NFI-A) that promotes MDSC expansion. We report here that miR-21 …

Utility Of Molecular Tools For The Identification And Characterization Of Infectious Agents, Shih Keng Loong

Student Works (2020-2029)

Molecular tools involving nucleic acid sequence-based detection offer rapid, sensitive and specific identification of bacterial infections, all together instrumental in timely patient management and infection control. Analyses of DNA sequences played a pivotal role in the discovery of novel or uncommon bacterial pathogens. This method can be applied to detect and identify rare and potentially pathogenic bacterial infectious diseases emerging from the interactions between human, animal and the environment. In this study, the 16S rDNA sequencing demonstrated superior utility against the conventional phenotypic identification systems for the identification of rare pathogenic bacteria. Its inability to differentiate between certain bacterial species …

Repression Of Tick Microrna-133 Induces Organic Anion Transporting Polypeptide Expression Critical For Anaplasma Phagocytophilumsurvival In The Vector And Transmission To The Vertebrate Host, Ellango Ramasamy, Vikas Taank, John F. Anderson, Hameeda Sultana, Girish Neelakanta

Biological Sciences Faculty Publications

The microRNAs (miRNAs) are important regulators of gene expression. In this study, we provide evidence for the first time to show that rickettsial pathogen Anaplasma phagocytophilum infection results in the down-regulation of tick microRNA-133 (miR-133), to induce Ixodes scapularis organic anion transporting polypeptide (isoatp4056) gene expression critical for this bacterial survival in the vector and for its transmission to the vertebrate host. Transfection studies with recombinant constructs containing transcriptional fusions confirmed binding of miR-133 to isoatp4056 mRNA. Treatment with miR-133 inhibitor resulted in increased bacterial burden and isoatp4056 expression in ticks and tick cells. In contrast, treatment with …

Plant Defensin Antibacterial Mode Of Action Against Pseudomonas Species, Andrew E. Sathoff, Shawn Lewenza, Deborah A. Samac

Research & Publications

Background: Though many plant defensins exhibit antibacterial activity, little is known about their antibacterial mode of action (MOA). Antimicrobial peptides with a characterized MOA induce the expression of multiple bacterial outer membrane modifications, which are required for resistance to these membrane-targeting peptides. Mini-Tn5- lux mutant strains of Pseudomonas aeruginosa with Tn insertions disrupting outer membrane protective modifications were assessed for sensitivity against plant defensin peptides. These transcriptional lux reporter strains were also evaluated for lux gene expression in response to sublethal plant defensin exposure. Also, a plant pathogen, Pseudomonas syringae pv. syringae was modified through transposon mutagenesis to …

Contributions Of Gyra And Parc Mutations And Qnrs2 Acquisition To Ciprofloxacin Resistance In Aeromonas Veronii Hm21, Daniel J. Silverstein

Honors Scholar Theses

In recent years, ciprofloxacin resistant (CpR) Aeromonas veronii and A. hydrophila strains have been isolated from the wounds of patients receiving leech therapy. Genome comparisons of these CpR isolates revealed the presence of chromosomal mutations in gyrA and parC as well as the gain of qnrS2 on either a large, 34 kb, conjugatable, low-copy plasmid, pAv42, or on a small, 6.8 kb, high-copy plasmid, pAh1471. The minimum inhibitory concentration, MIC, for Cp of these clinical isolates ranged from 1 to ≥32 µg/mL and some harbored a qnrS2 containing plasmid. We wanted to assess the contributions of these factors in an …

Analysis Of Biofilm Remediation Capacity For Octenyl Succinic Anhydride (Osa), A Bioactive Food Starch Modifier Compound, Matthew R. Borglin

Master's Theses

Matthew R. Borglin

This thesis demonstrates efficacy of Octenyl Succinic Anhydride (OSA), as a biofilm sanitizer. Biofilms allow bacteria to adhere to solid surfaces with the use of excreted polymeric compounds. For example, surfaces found in food production or processing facilities such as the interior of a raw milk holding tank, are some of the most susceptible to biofilm contamination. When present, biofilms can cause a variety of negative effects, which include; reduction of product shelf life, corrosion, and outbreaks of foodborne illnesses. The close association of biofilms with the majority of foodborne illness cases led the US Environmental Protection …

Insights Into The Chlamydial Niche: The Dynamic Roles Of Inclusion Membrane (Inc) Proteins In Chlamydia Trachomatis Development, Macy G. Wood

Theses & Dissertations

Chlamydia trachomatis (Ctr) is the leading cause of bacterial sexually transmitted infections. Ctr, an obligate intracellular bacterium, develops within a membrane-bound vacuole called an inclusion. The inclusion membrane is modified by chlamydial inclusion membrane (Inc) proteins, the functions of which are poorly characterized. Bacterial two-hybrid analyses found some Incs (e.g., IncF) interacted with numerous Incs while others (e.g., IncA) did not. We hypothesize that some Incs organize the inclusion through Inc-Inc interactions whereas other Incs promote chlamydial-host interactions by binding eukaryotic proteins. To test our hypothesis, we implemented the ascorbate peroxidase proximity labeling system (APEX2), which labels proximal proteins with …

Investigation Of Rapid Diagnostic Tests For Characterization Of Mycobacterium Avium Complex (Mac) From Various Isolates And Identification Of Virulence Factors Of Porcine Reproductive And Respiratory Syndrome Virus (Prrsv) In Vitro, Claudia Antonika

UCARE: Research Products

Chapter 1

Mycobacterium avium complex (MAC) has become one of the major problems in public health and livestock. Members of MAC, such as M. avium subsp paratuberculosis (MAP) and M. avium subsp hominissuis (MAH), are responsible for many opportunistic infections and the loss of livestock. MAP is economically significant to the beef and dairy industries because it is the etiologic agent of Johnes’s disease, a chronic and fatal enteritis in ruminants. Tracing the infection sources of MAC could be difficult since it infects many types of hosts in the natural environment. Furthermore, there is less information known about MAP pathogenicity …

The Effects Of Farnesol, A Quorum Sensing Molecule From Candida Albicans, On Alcaligenes Faecalis, Savannah Hutson

Undergraduate Honors Theses

Quorum sensing molecules have become a recent focus of study to learn if and how they can be used, both on their own and in conjecture with current antimicrobial methods, as a means of bacterial control. One such quorum sensing molecule is the sesquiterpene alcohol, Farnesol, which is synthesized and released by the fungus, Candida albicans. In most in-vivo cases, our laboratory has shown that Alcaligenes faecalis overtakes C. albicans, preventing its growth. However, as a way to counteract this inhibitory effect, Farnesol may be one way that Candida has found to fight back. In this study, we …

A Study Of The Polymicrobial Inhibitory Interactions Between Alcaligenes Faecalis And Staphylococcus Aureus, Blakeley Griffin

Undergraduate Honors Theses

Members of the Staphylococcus genus are found as a part of normal microflora in humans and can commonly be found on the skin or in the nasal cavity. However, these microorganisms can cause serious and life-threatening opportunistic infections when there is a break in the physical barrier of skin. These infections have become difficult to treat as resistant strains emerge, particularly Methicillin Resistant Staphylococcus aureus (MRSA). MRSA has become a commonly acquired nosocomial infection which is difficult to treat with conventional antibiotics of the blactam class. Even Vancomycin, a last resort antibiotic, has been ineffective on some infections. Furthermore, …

A Putative Cystathionine Beta-Synthase Homolog Of Mycolicibacterium Smegmatis Is Involved In De Novo Cysteine Biosynthesis, Saroj Kumar Mahato

Graduate Theses and Dissertations

Mycobacteria include serious pathogens of humans and animals. Mycolicibacterium smegmatis is a non-pathogenic model that is widely used to study core mycobacterial metabolism. This thesis explores mycobacterial pathways of cysteine biosynthesis by generating and study of genetic mutants of M. smegmatis. Published in vitro biochemical studies had revealed three independent routes to cysteine synthesis in mycobacteria involving separate homologs of cysteine synthase, namely CysK1, CysK2, and CysM. However, in vivo data were lacking. The M. smegmatis genome encodes only a CysM homolog and lacks orthologs for CysK1 or CysK2. The gene that codes for CysM is a part of an …

Second Messenger Cyclic-Di-Gmp Regulation In Acinetobacter Baumannii, Justin Deal

Undergraduate Honors Theses

Over time, “superbugs,” or bacteria that have become resistant to antibiotics, have become a great concern in modern medicine. Viable alternates are currently being looked into as effective and safe ways to prevent or treat infections caused by these superbugs. One such method is through the utilization of the second messenger molecule cyclic-di-GMP (c-di-GMP) that has been shown to regulate phenotypes within other bacteria that may control surface colonization in Acinetobacter baumannii. Through a series of experiments, the active enzymes that create c-di-GMP - diguanylate cyclases - and break down c-di- GMP - phosphodiesterases - have been inactivated in …

Regulation Of Weak Acid-Dependent Cell Death By The Msaabcr Operon During Biofilm Formation Of Staphylococcus Aureus, Sarah-Elizabeth M. Polley

Honors Theses

Staphylococcus aureus is a dangerous human pathogen and a leading cause of both nosocomial and community-associated infections. Infection by S. aureus is a growing health concern due to the emergence of antibiotic resistance and the capability to form robust biofilms. Major constituents of the biofilm extracellular matrix are demonstrated to be released via controlled autolysis of a subpopulation of the biofilm. As indicated in past studies, the programmed cell death (PCD) of S. aureus grown in a biofilm microenvironment is potentiated by the buildup of acetate, a byproduct of glucose metabolism, which causes cytoplasmic acidification. Furthermore, it has been shown …

Development Of In Vitro Models To Study The Rapid Extraintestinal Dissemination Of Salmonella., Adarsh Gopinath

Electronic Theses and Dissertations

Salmonella appears in the bloodstream of mice in as little as 15 minutes after oral inoculation and establishes persistent colonies in the spleen and liver. While its pathway to blood is undetermined, this phenomenon is dependent on the activity of Salmonella pathogenicity island 2 (SPI-2) coded type III secretion system (T3SS) and CD18+ phagocytes. We hypothesize that dendritic cells associated with the basal face of the gut epithelium, that are naturally migratory and known to sample for luminal antigens directly transport Salmonella to the bloodstream. This process comprises of at least two phases, dissociation and reverse transmigration. We define dissociation …

Transcriptomic Responses To Coaggregation Between Oral Bacteria, V R Mutha Naresh

Student Works (2020-2029)

Cell-cell interactions between genetically distinct oral bacteria form macroscopic clumps known as coaggregates. These interactions, termed coaggregation contribute to the formation of highly structured multispecies communities in oral cavity, known as oral biofilms. Individual species also compete and collaborate with other neighboring species through metabolic interactions allowing growth of other species by coaggregation. This study involves the mixing of selected pairs of oral bacteria to form coaggregates and quantifying differential gene expression levels in each pair in comparison with mono-species coaggregation. The dual RNA-Seq approach made it possible to see how bacteria respond to one another leading to different metabolic …

Modulation Of Host Innate Immune Cells By Yersinia Pestis To Create A Permissive Environment For Replication., Amanda Rose Pulsifer

Electronic Theses and Dissertations

Yersinia pestis has gained widespread infamy due to the historic outbreak during the middle ages, referred to as The Black Death. Infection with Y. pestis typically begins with deposition of Y. pestis into the dermis (bubonic plague) or respiratory tract (pneumonic plague). Tissue resident macrophages are the first innate immune cell encountered by Y. pestis. Macrophages are likely a way for Y. pestis to avoid neutrophils early in infection when the neutrophil neutralizing Type Three Secretion System is not expressed. This work focuses on which Rab host proteins are manipulated by Y. pestis, and how neutrophils are forced to …

Evaluation Of Microbiome On Chicken Necrotic Enteritis And Growth Performance, Mussie Abraha

Graduate Theses and Dissertations

Necrotic enteritis (NE) has re-emerged following restriction of antimicrobial usage and costs $6 billion every year worldwide. The primary objective of the studies was to evaluate prevention and treatment of NE using a microbiota metabolic product, secondary bile acid deoxycholic acid (DCA), in drinking water. Day-old birds were tagged and placed in floor pens. In experiments 1 and 2, the birds were infected with Eimeria maxima (Em) at d 18 and C. perfringens at d 23 and 24 and the birds were euthanized at d 26. In experiment 3, birds were infected with Eimeria at d 20 and C. perfringens …

A Novel Mode Of Action Of C-Reactive Protein In Protecting Against Streptococcus Pneumoniae Infection And Synergy With Antibiotics, Donald Ngwa

Electronic Theses and Dissertations

C-reactive protein (CRP) is a part of the innate immune system, is synthesized in the liver, its blood level increases in inflammatory states, and it binds to Streptococcus pneumoniae. The conformation of CRP is altered under conditions mimicking an inflammatory milieu and this non-native CRP also binds to immobilized/aggregated/pathogenic proteins. Experiments in mice have revealed that one of the functions of CRP is to protect against pneumococcal infection. For protection, CRP must be injected into mice within two hours of administering pneumococci, thus, CRP is protective against early-stage infection but not against late-stage infection. It is unknown how CRP protects …

Microbiota Accessible Carbohydrates And Susceptibility To Clostridioides Difficile Infection, Keegan Schuchart, Jennifer Auchtung, Thomas Auchtung

UCARE: Research Products

This experiment looked into the effects of dietary microbiota accessible carbohydrates (MACs)on the composition of human GI microbe colonies and their susceptibility to pathogen infection. Three different concentrations of MAC ingrained media were tested in mini bioreactor arrays, treated with antibiotics, infected with Clostridioides difficile, then allowed to recover. Microbe community was examined for strain composition and diversity before and after the antibiotic treatment and Clostridioides difficile infection. After the initial mini bioreactor run, individual isolates were investigated in their ability to respond to specific MACs Arabinoglactina, Inulin, and Soluble starch at experimental concentrations.

Biological Sex Influences Susceptibility To Acinetobacter Baumannii Pneumonia In Mice, Sílvia Pires, Adeline Peignier, Jeremy Seto, Davida S. Smyth, Dane Parker

Publications and Research

Acinetobacter baumannii (A. baumannii) is an extremely versatile multidrug-resistant pathogen with a very high mortality rate; therefore, it has become crucial to understand the host response during its infection. Given the importance of mice for modeling infection and their role in preclinical drug development, equal emphasis should be placed on the use of both sexes. Through our studies using a murine model of acute pneumonia with A. baumannii, we observed that female mice were more susceptible to infection. Likewise, treatment of male mice with estradiol increased their susceptibility to infection. Analysis of the airway compartment revealed enhanced inflammation and reduced …