Microbiology Commons^™

Introduction Of Plasmid Dna Into Sneathia Vaginalis; The First Step To Genetic Manipulation, Rea Mansukhani, Cathyrn O'Brien, Zion Mccoy, Kimberly K. Jefferson

UROP Posters

Many bacterial pathogens are capable of secreting one or more toxins, all of which can be categorized based on their function, target, or mechanism of action. Pore-forming toxins, for instance, are characterized based on their ability to perforate the host membrane. This may result in the delivery of bacterial substances into the host cell, release of compounds from the host cell, and/or death of the host cell. Research from our lab has demonstrated that Sneathia vaginalis, a gram-negative anaerobic bacterial species that is associated with bacterial vaginosis and preterm birth, produces a pore-forming toxin referred to as cytopathogenic toxin …

Transitions In Bacterial Communities Across Two Fermentation-Based Virgin Coconut Oil (Vco) Production Processes, Zomesh A N Maini, Crisanto M. Lopez

Biology Faculty Publications

Despite being one of the most used methods of virgin coconut oil (VCO) production, there is no metagenomic study that details the bacterial community shifts during fermentation-based VCO production. The identification and quantification of bacteria associated with coconut milk fermentation is useful for detecting the dominant microbial genera actively involved in VCO production which remains largely undescribed. Describing the constitutive microbial genera involved in this traditional fermentation practice can be used as a preliminary basis for improving industrial practices and developing better fermentation procedures. In this study, we utilized 16S rRNA metagenomic sequencing to trace the transitions in microbial community …

Antibacterial Activity Of Commiphora Molmol (Myrrha) Against The Periodontal Pathogen, Aggregatibacter Actinomycetemcomitans, Khalid Nafea Alharbi

Selected Full-Text Master Theses 2021-

The exogenous pathogen Aggregatibacter actinomycetemcomitans (A. actinomycetemcomitans) is most frequently involved in periodontitis and other systemic diseases. The treatment of these infections involves antibiotic therapy with Amoxicillin being the most popular antibiotic against A. actinomycetemcomitans but now, the drug is not 100% effective due to the development of 0-84% antibiotic resistance. The present study aimed to determine antibacterial activity of Commiphora molmol (Myrrha) against the periodontal pathogen, Aggregatibacter actinomycetemcomitans. In detail, the strain LIU1239 of Aggregatibacter actinomycetemcomitans was used where the plasmid pVJT128 is used for transposon mutagenesis and the Escherichia coli strain MV10Nal (stock # LIU4). The …

Phip Variants Associated With Chung–Jansen Syndrome Disrupt Replication Fork Stability And Genome Integrity, Neysha Tirado-Class, Caitlin Hathaway, Wendy K. Chung, Huzefa Dungrawala

Molecular Biosciences Faculty Publications

Chung–Jansen syndrome (CJS) is a rare, autosomal dominant disorder characterized by developmental delay, intellectual disability/cognitive impairment, behavioral challenges, obesity, and dysmorphic features. CJS is associated with heterozygous variants in PHIP (Pleckstrin-Homology Interacting Protein), a gene that encodes one of several substrate receptors for Cullin4-RING (CRL4) E3 ubiquitin ligase complex. Full-length PHIP, also called DCAF14, was recently identified to function as a replication stress response protein. Herein, we report the identification of two PHIP missense variants identified by exome sequencing in unrelated individuals with CJS. The variants p.D488V and p.E963G occur in different functional elements of DCAF14-WD40 repeat domain and pleckstrin …

Sequence Properties Of An Intramolecular Interaction That Inhibits P53 Dna Binding, Emily Gregory, Gary W. Daughdrill

Molecular Biosciences Faculty Publications

An intramolecular interaction between the p53 transactivation and DNA binding domains inhibits DNA binding. To study this autoinhibition, we used a fragment of p53, referred to as ND WT, containing the N-terminal transactivation domains (TAD1 and TAD2), a proline rich region (PRR), and the DNA binding domain (DBD). We mutated acidic, nonpolar, and aromatic amino acids in TAD2 to disrupt the interaction with DBD and measured the effects on DNA binding affinity at different ionic strengths using fluorescence anisotropy. We observed a large increase in DNA binding affinity for the mutants consistent with reduced autoinhibition. The ΔΔG between DBD and …

Isolation And Characterization Of Novel B. Subtilis Bacteriophages, Johnson Phung, Rebecca Adams

Science University Research Symposium (SURS)

Bacteriophages are viruses that infect bacteria cells. Bacteria, being the most abundant form of life on the planet, have an estimated ten to a hundred different types of bacteriophages that infect each species. Bacteriophages are everywhere on Earth, but the number of phages that have been identified is only a fraction of what are thought to exist. This project focuses on isolating and identifying phages that infect the bacterium Bacillus subtilis. B subtilis is a decomposition microbe commonly found in the soil. Our first goal was to determine whether the 3 samples of Bacillus subtilis phages that were previously …

Adenoviral-Vectored Centralized Consensus Hemagglutinin Vaccine Provides Broad Protection Against H2 Influenza A Virus, Erika M. Petro-Turnquist, Brianna L. Bullard, Matthew J. Pekarek, Eric A. Weaver

Nebraska Center for Virology: Faculty Publications

Several influenza pandemics have occurred in the past century, one of which emerged in 1957 from a zoonotic transmission of H2N2 from an avian reservoir into humans. This pandemic caused 2–4 million deaths and circulated until 1968. Since the disappearance of H2N2 from human populations, there has been waning immunity against H2, and this subtype is not currently incorporated into seasonal vaccines. However, H2 influenza remains a pandemic threat due to consistent circulation in avian reservoirs. Here, we describe a method of pandemic preparedness by creating an adenoviral-vectored centralized consensus vaccine design against human H2 influenza. We also assessed the …

Computational Strain Analysis In Microbiome Datasets With Application To Skin Disease, Minerva Fatimae Ventolero

Electronic Theses and Dissertations, 2020-2023

Identifying bacterial strains is important not only for human health research but also for various environmental and industrial applications. The plethora of available shotgun metagenomic datasets provide an unprecedented opportunity to discover novel bacterial strains in different environmental niches. In this thesis, we evaluated the existing novel-strain-based tools and showed that their performance is still suboptimal. Due to the difficulty in distinguishing strains of similar abundance by available tools, the SMS (strains in multiple samples) tool was developed to de novo identify bacterial strains in multiple shotgun metagenomic samples. We showed that SMS distinguishes strains of similar abundance well and …

Elucidation Of Novel And Established Campylobacter Species With Clinical And Agricultural Significance Through Phenotypic, Genotypic, And Taxonomic Investigation, Caoimhe Lynch

Theses

Campylobacter spp. are well-established human, veterinary and economic pathogens, with a broad host range spanning from terrestrial and marine mammalian, avian and reptilian hosts. The scope of the study includes novel and notorious species within the genus, with reference to zoonotic agents Campylobacter jejuni and Campylobacter coli - the leading cause of human bacterial gastroenteritis in the EU and Ireland, Campylobacter fetus that represents the most common campylobacter causing bacteraemia and two novel species isolated from pigs not described in the literature previously. C. jejuni and C. coli isolates (n = 350) recovered from broiler caecal and neck skin samples, …

Investigating The Development Of Alcohol Tolerance By The Opportunistic Pathogen Staphylococcus Aureus, Vaughn Perveiler

West Chester University Master’s Theses

With the ongoing COVID-19 pandemic, the use of alcohol-based antiseptics by the general public has greatly increased. While this form of antimicrobial is effective in stopping the spread of both bacterial and viral infections, little research has been conducted in the development of alcohol tolerance by microorganisms or its mechanisms. In this study, experimental evolution was conducted on four separate strains of Staphylococcus aureus to determine if tolerance can develop towards moderate to high concentrations of ethanol. Once ethanol tolerance was established in the evolved populations, whole genome sequencing was performed to identify the potential mutations responsible for the observed …

Evaluation Of Oasl And Herc5’S Role In The Non-Lytic Clearance Of Influenza A Virus From Club Cells, Steve Crisafulli

West Chester University Master’s Theses

Influenza A virus (IAV) is a highly infectious pathogen responsible for causing severe respiratory illness and death in humans and animals worldwide. Due to highly effective strategies to negate host antiviral defenses, IAV leads to the death of nearly all infected cells. Furthermore, IAV induces high levels of genome-damaging oxidative stress within infected cells and suppresses the cellular mismatch repair (MMR) mechanism, thereby inhibiting expression of key antiviral genes, which further contributes to cell death. However, recent studies have demonstrated that a subset of respiratory epithelial cells, called club cells, are able to non-lytically clear IAV and continue to survive …

Effect Of Metal-Containing Nanoparticles On Bacterial Biofilms And On The Microbiome Of Girardia Tigrina, Azar Saikali

West Chester University Master’s Theses

Anthropogenically-produced nanoparticles are a form of nanotechnology being used in industries including food and textiles. Humans and livestock are frequently exposed to metal-containing nanoparticles (MCNPs), that have been washed into streams and rivers, have been deliberately used in food packaging as antimicrobials, preservatives or for supplementation. The animal microbiome, which consists of a diverse community of microorganisms, provides a number of benefits to the host in terms of nutrition availability, immune support, and can influence behavior. Biofilms of diverse microbes may cause detrimental effects, for instance by causing dental diseases in humans. However, the scientific community has not reached consensus …

Unique Features Of Alarmone Metabolism In Clostridioides Difficile, Asia Poudel, Astha Pokhrel, Adenrele Oludiran, Estevan J. Coronado, Kwincy Alleyne, Marrett M. Gilfus, Raj K. Gurung, Surya B. Adhikari, Erin B. Purcell

Chemistry & Biochemistry Faculty Publications

The “magic spot” alarmones (pp)pGpp, previously implicated in Clostridioides difficile antibiotic survival, are synthesized by the RelA-SpoT homolog (RSH) of C. difficile (RSH_Cd) and RelQ_Cd. These enzymes are transcriptionally activated by diverse environmental stresses. RSH_Cd has previously been reported to synthesize ppGpp, but in this study, we found that both clostridial enzymes exclusively synthesize pGpp. While direct synthesis of pGpp from a GMP substrate, and (p)ppGpp hydrolysis into pGpp by NUDIX hydrolases, have previously been reported, there is no precedent for a bacterium synthesizing pGpp exclusively. Hydrolysis of the 5′ phosphate or pyrophosphate from GDP …

Copper(Ii) And Silver(I)‑1,10‑Phenanthroline‑5,6‑Dione Complexes Interact With Double‑Stranded Dna: Further Evidence Of Their Apparent Multi‑Modal Activity Towards Pseudomonas Aeruginosa, Anna Clara Milesi Galdino, Lívia Viganor, Matheus Mendonça Pereira, Michael Devereux, Malachy Mccann, Marta Helena Branquinha, Zara Molphy, Sinéad O'Carroll, Conor Bain, Georgia Menounou, Andrew Kellett, André Luis Souza Dos Santos

Articles

Tackling microbial resistance requires continuous efforts for the development of new molecules with novel mechanisms of action and potent antimicrobial activity. Our group has previously identified metal-based compounds, [Ag(1,10-phenanthroline-5,6-dione)2]ClO4 (Ag-phendione) and [Cu(1,10-phenanthroline-5,6-dione)3](ClO4)2.4H2O (Cu-phendione), with efficient antimicrobial action against multidrug-resistant species. Herein, we investigated the ability of Ag-phendione and Cu-phendione to bind with double-stranded DNA using a combination of in silico and in vitro approaches. Molecular docking revealed that both phendione derivatives can interact with the DNA by hydrogen bonding, hydrophobic and electrostatic interactions. Cu-phendione exhibited the highest binding affinity to either major (− 7.9 kcal/mol) or minor (− 7.2 kcal/mol) …

Role Of Host Restriction Factors On Porcine Reproductive And Respiratory Syndrome Virus (Prrsv) Replication, Pratik Katwal

Electronic Theses and Dissertations

In this study, the role of IFITM3 on PRRSV replication was studied in vitro by expressing exogenous IFITM3 in MARC-145 cells. An average of 31% reduction in PRRSV N protein expression and an average of 5.4 fold decrease in virus titer in the supernatant were observed in IFITM3 overexpressing cells as compared to vector control cells at 24 hours post infection (hpi). Moreover, there was a positive correlation between interferon- induced IFITM3 up-regulation and reduced PRRSV replication. To determine the role of endogenous IFITM3 in PRRSV replication, siRNA induced knockdown of IFITM3 was employed. RT-PCR validated the successful silencing of …

Decoding The Mystery Of Antibiotic Persistence, Tahmina Hossain

Electronic Theses and Dissertations

This work combines microbiology, molecular biology, Next-Generation Sequencing and system biology approaches to explore the mechanism of antibiotic persistence: a multi-drug tolerant, non-dividing, and metabolically altered state present in a subpopulation of cells due to phenotypic diversity rather than genetic variation (i.e. mutations). Persister can survive lethal antibiotic state and resuscitate after the treatment period is over. They are considered as the major contributing factor behind recurring infections. They also have a high mutation rate, which increases the chances of bacteria gaining antibiotic resistance. The formation of this phenotypic variant (persister) threatens the therapeutic effectiveness of antibiotics and understanding how …

The Bgs 13 Mutant Of Pichia Pastoris And Its Effect On Structural Changes Of The Reporter Protein B-Lactogolbulin, Bushra Irshad

University of the Pacific Theses and Dissertations

Pichia pastoris, a methylotrophic yeast, is an ideal host for recombinant protein expression. It has the capability of performing many eukaryotic post-translational modifications and grows to high cell densities. However, P. pastoris’s secretion properties are not always efficient, and its secretory pathway mechanisms have not been thoroughly elucidated. A previously identified mutant strain, bgs13, was found to efficiently secrete most recombinant proteins tested, raising the possibility that this bgs13 mutant is a universal super secreter and understanding its secretion process is needed. In this study, we used a reporter protein, ?-lactoglobulin (b-LG), to perform structural analysis and comparisons of protein …

Leveraging Queueing Theory To Develop Advanced Synthetic Biological Circuits, Prajakta K. Jadhav

Electronic Theses and Dissertations

The complexity and redundancy in the network topology of the cell hinder our understanding of the natural system, making it challenging to engineer living organisms with the desired functionality. The long-term goal of synthetic biology and bioengineering is to engineer cells to perform specific functions with increased robustness. The robust engineered systems can be easily applied to different species with the same outcome. An effective approach is a bottom-up approach where we tease apart the biological pathways and study them independently. Understanding the underlying mechanisms of biological pathways can be further applied to construct complex biological circuits that function similarly …

Characterizing The Lir Domain Of Abin1 And Identifying Its Role As A Regulator Of Mitophagy, Andrew Rhiner

Electronic Theses and Dissertations

A20 Inhibitor of NF-κB (ABIN1/TNIP1) is a known regulator of TNFα signaling induced cell death and inflammation. The regulatory activity has been attributed to ABIN1’s recruitment of the ubiquitin editing enzyme TNF-α-induced protein 3 (TNFAIP3/A20) to Receptor-interacting serine/threonine-protein kinase 1 (RIPK1). The regulation of RIPK1 by ABIN1 and A20 relies on a third player, linear ubiquitin chain assembly complex (LUBAC), which is involved in the recruitment of ABIN1 to RIPK1. Loss of LUBAC or ABIN1 is embryonic lethal, but loss of A20 is not embryonic lethal. The embryonic lethality due to loss of ABIN1, paired with the lack of lethality …

Characterization Of Porcine Respiratory Epithelial Cells And Their Innate Immune Responses To Bacterial And Viral Ligands, Yam Prasad Gautam

Electronic Theses and Dissertations

In response to a pathogenic attack, the host produces a series of defense mechanisms through various intracellular signaling pathways. The byproduct of these signaling pathways helps tackle the invading pathogen and protects the body from getting into a diseased state. This system is called the immune system. The immune system can be divided into two branches namely the innate immune system and adaptive immune system. The groups of immune cells that provide protection regardless of the pathogen specificity constitute the innate immune system. The system that acts according to the pathogen specificity is called the adaptive immune response. The production …

Spatial Localization Of Markers And 3d-Cell Model For Study Of The Human Cytomegalovirus Cytoplasmic Virion Assembly Compartment, Taylor Alan Vensko

Wayne State University Theses

Human cytomegalovirus (HCMV) causes severe disease in immunocompromised individuals and is a leading cause of congenital disease. Efficient assembly of virions (viral particles) is an intricate process that requires modulation and modification of host systems. HCMV induces an extensive rearrangement of the cellular endocytic recycling compartment into the site of virion maturation and egress: the cytoplasmic virion assembly compartment (cVAC). The HCMV cVAC is a distinguishing characteristic of infection, although studies have only been performed in traditional cell culture with limited observations in infectedhuman tissues. To study the cVAC, viral and cellular markers that are discrete to their respective compartments …

Inhibition Of Lysine Acetyltransferases Kat 3a/3b And Its Effect On Poliovirus Proliferation, Eduards Norkvests

Legacy Theses & Dissertations (2009 - 2024)

Ribosomes have been traditionally viewed as invariable machinery, but in recent years more and more evidence for their heterogeneity has emerged. Core ribosomal proteins, ribosomal-associated proteins, and post-translational modifications all contribute to the variations found within these crucial subcellular components.In our research, we focused on lysine acetylation – a post translational modification found on many ribosomal proteins. We used a chemical inhibitor of acetyltransferases KAT 3A and KAT 3B to observe the effect that inhibiting acetylation may have in the context of translation. By using poliovirus to shut off conventional cellular translation, we were able to look at a narrow …

Mechanisms By Which Protective Human Outer Surface Protein A Antibodies Block Transmission Of Borrelia Burgdorferi, Amber M. Frye

Legacy Theses & Dissertations (2009 - 2024)

Borrelia burgdorferi (B. burgdorferi) is the causative agent of Lyme disease in the United States and Europe, which remains a major threat to public health. While currently no human Lyme disease vaccine is available, a previously licensed OspA-based vaccine was shown to effectively prevent transmission of B. burgdorferi from ticks to mammalian hosts. Concerning the vaccines mechanism of action, OspA antibodies enter the bloodmeal of a feeding tick and engage with B. burgdorferi producing OspA on their outer surface in a manner that impairs migration of the spirochete to the host; however exact antibody-spirochete interaction is unknown, despite the importance …

Genomic Epidemiology Of Clinical Salmonella Enterica In New Hampshire, 2017-2020, Madison R. Turcotte

Legacy Theses & Dissertations (2009 - 2024)

Antimicrobial resistance in Salmonella enterica remains a challenging public health threat. We aim to characterize the population structure and evolutionary history of 394 genomes of S. enterica isolates recovered from human clinical cases in New Hampshire, USA from 2017 to 2020. The population is phylogenetically diverse, consisting of 78 STs and 67 serotypes. We detected 61 genes associated with resistance to14 different antimicrobial classes. Rather than a single multidrug resistant clone expanding in the state, we found multiple lineages carrying different combinations of independently acquired resistance determinants. New or emerging lineages can therefore rapidly spread over relatively short timeframes. In …

Isolating Bacteriophage For Potential Treatment Of Chronic Multi-Drug Resistant Escherichia Coli Infections, Leila Oswalt

Honors Projects

The misuse and overuse of antibiotics has led to the intense rise in antibiotic resistance. As society transitions into the post antibiotic era, there will be a great need for new therapeutic strategies to address multiple drug resistant bacterial infections. One such method, called bacteriophage therapy, allows for specific targeting of certain pathogenic bacteria through the use of viruses that attack bacteria; termed “bacteriophage” or simply “phage”. Urinary tract infections are among the most common pathological human infections that rely heavily on the use of antibiotics, the major cause of which is the bacterium Escherichia coli. During the Spring 2021-Spring …

Expanding Mouse-Adapted Yamagata-Like Influenza B Viruses In Eggs Enhances In Vivo Lethality In Balb/C Mice, Matthew J. Pekarek, Erika M. Petro-Turnquist, Adam Rubrum, Richard J. Webby, Eric A. Weaver

Nebraska Center for Virology: Faculty Publications

Despite the yearly global impact of influenza B viruses (IBVs), limited host range has been a hurdle to developing a readily accessible small animal disease model for vaccine studies. Mouseadapting IBV can produce highly pathogenic viruses through serial lung passaging in mice. Previous studies have highlighted amino acid changes throughout the viral genome correlating with increased pathogenicity, but no consensus mutations have been determined. We aimed to show that growth system can play a role in mouse-adapted IBV lethality. Two Yamagata-lineage IBVs were serially passaged 10 times in mouse lungs before expansion in embryonated eggs or Madin–Darby canine kidney cells …

Evaluate The Efficacy Of A Mixture Of Peroxyacetic Acid And H2o2 Against The Survival And Cross-Contamination Of The Salmonella Surrogate Enterococcus Faecium On Tomatoes During Triple-Wash, Corey Waitman Coe

Graduate Theses, Dissertations, and Problem Reports (ETD)

Triple-wash with a mixture of peroxyacetic acid and H2O2 (SaniDate-5.0) during post-harvest processing of fresh produce has been recommended by West Virginia Small Farm Center to improve microbial safety. It has been well recognized that the washing of produce is more important for preventing cross-contamination than reducing foodborne pathogens. Furthermore, it may help improve public confidence in that the produce they obtain from locally grown farmers is safe for their consumption. determine the efficacy of SaniDate-5.0 for reducing the survival and preventing cross-contamination of the Salmonella surrogate Enterococcus faecium on tomatoes during triple-wash.

E. faecium ATCC-8459 (resistant to 100-ppm nalidixic-acid) …

College Of Natural Sciences Newsletter, January 2022, College Of Natural Sciences

College of Natural Sciences Newsletters and Reports

Volume 3, Issue 1

Page 1 Dean's Message
Page 2 Awards & Recognition
Page 3 Outreach in Harrisburg, SD
Page 4 Geography Alumni Named Among 'Highly Cited Researchers', 53rd Annual Geography Convention
Page 5 Geography Convention Speakers, Brookings/SDSU Day
Page 6 Media Coverage of Natural Sciences
Page 7 Open PRAIRIE Data
Page 8 Fall 2021 Dean's List
Page 9 Fall 2021 Dean's List, cont.

A Review Of Virus-Vectored Vaccines: Current Production Methods, Uses, Issues, And Future Perspectives, Jaden Marks

Biology and Microbiology Graduate Students Plan B Research Projects

Viral vector vaccines have recently risen to the forefront of vaccinology, but there are multiple factors that must be addressed before they achieve widespread approval and use. The concept of using viruses as vectors is not new, and numerous attempts to create vaccines that utilize virus vectors have been made in the past 25 years. Despite this, only one viral vector vaccine has gained full approval by the FDA. The goal of this review is to provide a comprehensive overview of the types of viral vector vaccines, production and purification methods, current and future viral vector vaccines, the issues and …

Review Of A Chimeric Hemagglutinin-Based Influenza Vaccine Approach, Whitney Muck

Biology and Microbiology Graduate Students Plan B Research Projects

Influenza virus has affected human populations for centuries, causing an acute respiratory effect on the body. Even with scientific advancements in vaccine research, there are still serious limitations to the influenza vaccine. Varying strains and the constant recombination of the influenza virus has caused annual influenza circulation to be difficult to predict. The ongoing 40 race to develop a more efficient vaccine to broadly protect against the variety of influenza strains remains a public health priority. Many strategies have been studied to improve the current vaccine. The use of a chimeric hemagglutinin-based vaccine is in the first stages of human …