Microbiology Commons^™

Understanding The Molecular Mechanism Of Ilv1 Stress Sensitivity In Yeast By Examination Of Differential Patterns Of Gene Expression Between Wild-Type And Ilv1 Knockout Strains Of Saccharomyces Cerevisiae, Emily Schuster

Honors Theses

As arguably the most widely used industrial eukaryote, Saccharomyces cerevisiae has become one of the many model organisms used to study molecular genetics. A recent unpublished study at the University of Tennessee - Chattanooga showed that yeast cells that had the ILV1 gene knocked out had reduced survivability under stress conditions. This study then focused on the role of ILV1 in the stress response of cells by measuring the expression levels of 11 candidate genes that are all involved in some critical pathway in the yeast to see how gene expression was influenced as a result of the loss of …

Analyzing Staphylococcal Contamination On Surfaces And Bedside Areas Of A Neonatal Intensive Care Unit Of A Children's Hospital, Daniel See

Honors Theses

Staphylococci species are known to be a cause of Healthcare-Associated Infections (HAIs) in neonatal intensive care units (NICU). There is limited research about the surveillance and identification of staphylococci bacteria from NICUs. Surveillance of bacteria within the NICU helps to identify areas acting as reservoirs for bacteria so that new cleaning policies and techniques can be put in place to stop the spread of HAIs. The objective of this study was to swab sample sites in a local level IV hospital NICU and identify locations of staphylococci presence throughout the NICU. Forty-one swabs were selected from over 900 swabs collected …

Invasion Properties Of Listeria Monocytogenes In Aged Cells, Mina Lane Chasteen Burton

Honors Theses

Listeriosis is a bacterial infection caused by the gram-positive pathogen Listeria monocytogenes. When compared to other foodborne illnesses, listeriosis has a higher death rate due to an increased incidence of complications such as meningitis, hydrocephalus, and sepsis in immunocompromised populations. Elderly individuals experience a condition known as immunosenescence, which is a gradual compromisation of the immune system brought on by natural aging. Hutchinson-Gilford progeria syndrome (HGPS) is a rare genetic disorder causing premature aging, which allows HGPS cell lines to be used as models to further research in the field of biogerontology. This study utilizes the F2365 strain of …

Antibiotic-Resistant Bacteria In Freshwater Crayfish, Colby Finch

Honors Theses

The presence of antibiotic-resistant bacteria is increasing in natural aquatic environments. Alongside this, organisms that live in these ecosystems are increasingly harboring antibiotic-resistant bacteria. In this study, I analyzed the capacity for the crayfish species Procambarus vioscai paynei to harbor antibiotic-resistant bacteria. Crayfish, as well as water and sediment, were sampled from a pond at the University of Mississippi Field Station. The guts of crayfish were plated on TSA agar, as well as agar containing vancomycin, erythromycin, penicillin, tetracycline, or ciprofloxacin. Following incubation, counts of bacteria were determined. Selected bacterial isolates were tested for multiple antibiotic-resistance. Bacterial isolates were also …

A Comparison Of Heterotroph Isolation And Sequencing Methods From Various Cyanobacterial And Algal Microbiomes, Victoria Starks

Honors Theses

Cyanobacteria have provided a vast, new source of natural products to be utilized in drug development. Because of their non-axenic nature, cyanobacteria typically have an abundance of symbiotic heterotrophs living in association with them. These bacteria can play significant roles in the survival of its cyanobacterial host as well as provide the potential production of unique compounds. The possibility of unknown natural products is only increased by the flexible nature of these bacteria, as altering its environmental state can change the activity of biosynthetic pathways and even activate novel production. Our research team’s intent is to isolate cyanobacterial strains from …

Detecting Potentially Neurotoxic Pseudo-Nitzschia Species In The Grand Strand Area, Chesney Price

Honors Theses

Pseudo-nitzschia is a globally distributed diatom genus which contains 26 neurotoxin (domoic-acid) producing species. In shallow coastal waters these diatoms are associated with both the development of harmful algal blooms and the bio-accumulation of toxins in shellfish that can impact human health. Pilot data recovered in 2018 indicated the presence of multiple species of Pseudo-nitzschia, including domoic-acid producing taxa. We hypothesize that we will recover a similar species diversity using multiple sampling dates during the summer of 2021. Environmental DNA (eDNA) samples taken from the public shellfish harvesting area waters at Huntington Beach State Park (SC) over a two month …

Composting During Winter Versus Summer: A Microbial Analysis, Madison Ochs

Honors Theses

Pre-consumer food waste, hay, and woodchips were composted using two different methods of static composting: forced aeration (FA) and natural aeration (NA). The composting process was performed during the winter and summer for 106 and 124 days, respectively. The composting process for both winter and summer processes did not reach the temperature above the 40°C threshold of the thermophilic stage in composting. The microbial communities present during different timepoints (start, peak, and final) along the composting process were analyzed with DNA sequencing techniques and visualized using QIIME 2, an open-source bioinformatics platform. The microbial communities of the winter and summer …

The Role Of A-Layer In Polyunsaturated Fatty Acid (Pufa)-Mediated Effects On Aeromonas Salmonicida Subsp. Salmonicida, Allen Lin

Honors Theses

Aeromonas salmonicida subsp. salmonicida is a Gram-negative bacterium that infects salmonids and non-salmonids worldwide leading to an infection known as furunculosis, which is characterized by skin lesions and hemorrhages of the fish epidermis. This infection is carried out by a A+ (virulent strain) of A. salmonicida containing an important virulent factor known as the A-layer, which is a 2D paracrystalline structure that binds to the basement membrane and functions to promote adherence to host membranes and resistance to host defense. The purpose of this study was to examine the ability of the A+ A. salmonicida to incorporate exogenous fatty acids …

Calibration Study Of A Bacterial Quantification Technique Using Transport Swabs And Line Inoculations On Agar Growth Medium, Seth Larue

Honors Theses

Enumeration of bacteria has been the subject of research for over a century. Some of the techniques that have been developed include viable plate counts, the most probable number assessment, and more modern molecular techniques. Many of the newer molecular-based techniques provide little or no information about the viability of the cells being counted, and are generally not quantitative. Having quantitative data for potentially pathogenic bacteria on surfaces can be very useful in many settings, particularly in healthcare facilities. Knowing the numbers of bacterial cells present on surfaces within healthcare facilities may be important to attempt to study the cause …

Diagnostic Approaches To Combat Antibiotic Resistance In Bacteria, Alexander Oliveri, Shade Smith, Anshul Anugu

Honors Theses

The threat of antibiotic resistance is a major problem faced by the healthcare field affecting millions of people and costing tens of thousands of lives annually. Of the potential ways to mitigate this issue the field of antibiotic resistance testing presents an opportunity for significant improvement and benefits. Several methods of such diagnostic processes can yield more informative results than the current commonly used Kirby-Bauer test. However, there are benefits and limitations to each method. In the context of a clinically relevant diagnostic for antibiotic resistance, the microarray platform exhibits the necessary breadth with opportunities to overcome some limitations with …

Purification Of Pcbp2 And Its Effect On The Formation Of Dna I-Motifs And Secondary Structures, Nathan Redden

Honors Theses

Several secondary, non-B forms of DNA are associated with the promoter regions of many oncogenes, including the intensely studied c-MYC gene that codes for a transcription factor. Two such secondary structures, G-quadruplexes, and i-motifs, have become topics of interest due to their prevalence within oncogenic promoters, as well as their potential accessibility for drug targeting. B-form DNA can adopt alternative structures under certain conditions. These alternative structures are favored when there is an asymmetric distribution of guanosine and cytosine on complimentary DNA strands. The strand rich in guanosine is able to form a quadruple stranded DNA complex termed a G-quadruplex. …

Construction And Analysis Of Three Multi-Partite Synthetic Microbial Communities, Alexander J. Lazzara, Jacob K. Fanning

Honors Theses

Microbial Communities are of interest to molecular biologists hoping to understand the nature of metabolic interactions between co-existing, or possibly mutualistic, organisms. These interactions are ubiquitous in nature, but understanding the molecular mechanisms involved remains challenging and not well understood. Here, we design three tri-partite microbial circuits based on possible interactions among involved microbes, which are discussed and may suggest mutualistic interactions. Carbon and nitrogen molecular pathways and the intracellular metabolism of each microbe is discussed. We present minimal growth media that will ensure that organisms utilize available resources, which may originate from metabolic processes in neighboring microbes, simulating a …

The Impact Of Oxygen Availability And Oxidative Stress On Regulation Of Cyclic-Dimeric-Gmp In Listeria Monocytogenes Strain F2365, Anna G. Welch

Honors Theses

Listeria monocytogenes is a gram-positive foodborne pathogen that results in the infectious disease listeriosis. The second messenger molecule cyclic-dimeric-GMP has been found to be responsible for the regulation of expression of many of its virulence factors with diguanylate cyclases and phosphodiesterases regulating the intracellular concentration of cyclic-di-GMP through synthesis and degradation, respectively. This study investigated the possibility that the availability of oxygen is the environmental signal crucial to the regulation of these enzymes. Prior research in our laboratory has demonstrated that the intracellular concentration of cyclic-di-GMP is elevated when exposed to anaerobic conditions. Due to this, it was hypothesized that …

Study Of The Role Of The Msaabcr Operon In Tricarboxylic Acid Cycle Activity And Persister Cell Formation In Staphylococcus Aureus, Karsen Motter

Honors Theses

Staphylococcus aureus is an important human pathogen that causes wide arrays of infections ranging from minor skin infections to lethal systemic conditions such as infective endocarditis, osteomyelitis, sepsis and pneumonia. These systemic diseases are often difficult to treat due to the presence of persister cells. Persister cells are a phenotypic variant of the bacterial population that exhibit extreme and transient antibiotic tolerance accompanied by a transient halt in growth. Upon cessation of antibiotic treatment, however, persisters resume growth which results in recurrence of infections. This characteristic of persister cells therefore displays high clinical significance. In this study, we show the …

Interregulation Between Msaabcr Operon And Ccpe To Determine Staphylococcal Metabolism And Virulence, Erin R. Cox

Honors Theses

Staphylococcus aureus is a complex human pathogen that causes problems in both healthcare and community settings. Staphylococcus aureus is a gram-positive, sphere shaped bacterium that usually colonizes in the nasal cavity of healthy individuals. Staphylococcus aureus infections are a growing health concern due to its ability to produce virulence factors, such as pigmentation, protease production, and capsule formation. Two regulators in S. aureus virulence factors are msaABCR, which is a newly characterized operon, and ccpE. In order to determine the interaction between these two regulators in regulating virulence and metabolism in S. aureus, ccpE and ccpE/ …

Phosphodiesterase D Is Involved In Bile Resistance In Listeria Monocytogenes, Sophia A. Ali

Honors Theses

Listeria monocytogenes is a deadly foodborne bacterium that is responsible for almost 20% of food-related deaths in the United States. Listeria monocytogenes contaminates ready-to-eat products such as cheese, deli meat, and ice cream. Once ingested, it invades the intestinal lining and can enter the bloodstream, causing listeriosis. There is a gap in the knowledge of the pathogenesis of L. monocytogenes in how it is able to survive in the gastrointestinal tract in the presence of bile, which has bactericidal properties. Previous studies have suggested that the second messenger cyclic-dimeric-GMP may be involved in the regulation of virulence factors of Listeria. …

Phenotypic Characterizations Of Msaabcr Operon Deletion In Staphylococcus Epidermidis Rp62a, Raelyn Williams

Honors Theses

Staphylococcus epidermidis is a human pathogen that is increasingly known for its role in hospital infections associated with implantable medical devices. Antibiotic resistance has become a concerning issue for these infections as this bacteria have various virulence traits that help to evade immune response and antibiotic treatment. Currently, the most effective way to treat S. epidermidis infection is removal of the implant and long-term antibiotic treatment. S. epidermidis causes infection by expressing several protein factors that induce biofilm formation, the bacteria’s primary virulence mechanism. The purpose of this study was to perform the phenotypic characterizations of the msaABCR operon in …

The Role Of Gac/Rsm Signal Transduction Pathway In The Biology Of Antagonistic Marine Bacterium Gynuella Sunshinyii 449, Anhthu Truong

Honors Theses

Gynuella sunshinyii is a newly characterized bacterium with a remarkable ability to produce multiple secondary metabolites that act as potential antibiotics for medical applications. Currently, little is known about gene regulation in this unusual microorganism. In 2018, studies conducted of rhizosphere microbiomes of tidal marsh grasses on Deer Island, MS, resulted in multiple isolates of halophilic, aerobic, Gram-negative bacteria with antagonistic properties. One of the most active antagonists was identified as Gynuella sunshinyii 449. The genome of strain 449 was sequenced with Illumina MiSeq, assembled with Unicycler and SPAdes, and annotated with RASTtk. The annotation revealed a 6.09 Mb genome …

A Metabolomics-Based Approach To The Screening Of Endometrial Cancer: Development Of A Gas Chromatography-Ion Trap Mass Spectrometry-Based Method, Allycia Lee

Honors Theses

Endometrial Cancer (EC) is the most common malignant tumor of the female reproductive tract. Currently there are no methods for effectively screening EC, but one method that is thought to be useful in screening EC is metabolomics. Metabolomics involves the analysis of metabolites which are low molecular weight inorganic and organic chemicals that are substrates, intermediates, and by-products of enzyme-mediated biochemical reactions in the cell. Previous research shows that the metabolic signature of EC patients are discernable from those of healthy patients. To aid in the development of an effective screening method for EC, blood serum, urine, and saliva samples …

Increasing Stress Tolerance In Saccharomyces Cerevisiae Using Lipocalin Genes, Ximena Leon

Honors Theses

Saccharomyces cerevisiae is a microorganism that is commonly used in industries, but its productivity is significantly hindered by its inability to resist high stress levels found in industrial processes. By increasing stress tolerance in S. cerevisiae, its role in industrial processes could be maximized. The objective of this study is to determine if complementation of lipocalin genes into the wildtype and knockout strains of S. cerevisiae can increase stress tolerance against a variety of stressors. We identified the knockout yeast strains, ALD3, ALD4, PDX3, and ILV1, as being sensitive to various stressors including salinity, heat, oxidative, and osmotic stresses. Arabidopsis …

Characterization Of A Plasmid-Based Dna Vaccine For Simian Immunodeficiency Virus, Priya Sanipara

Honors Theses

Described as one of the world’s worst pandemics, HIV (Human Immunodeficiency Virus) infects millions of people each year and is the cause for AIDS (Acquired Immunodeficiency Syndrome). Despite the development of vaccines for numerous infectious diseases such as polio, small pox, and influenza, a vaccine for HIV remains elusive due to the virus’s high mutation rate and ability to evade the immune system. HIV causes depletion of CD4+ lymphocytes, resulting in a weakened immune system. However, the development of a plasmid-based DNA vaccine approach may help revolutionize vaccine development for HIV due to its ability to confer cellular and humoral …

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 …

The Effects Of Light On The Bacterial Biomass Of Decomposing Leaf Litter In Freshwater, Savannah L. Underwood

Honors Theses

Recent evidence has suggested that the presence of light (and in conjuction, periphytic algae) stimulates the growth and production of bacteria on decaying plant litter. However, the current method of measuring bacterial biomass, flow cytometry, drastically underestimates the bacterial biomass levels associated with litter samples. To determine and correct for this underestimation, the present study determined conversion factors using direct counting methods via epifluorescence microscopy. These conversion factors were then applied to flow cytometry counts for seven different studies. The studies analyzed the effects of light and other covarying factors on the bacterial biomass associated with various samples of leaf …

Phospholipid Remodeling Via Exogenous Polyunsaturated Fatty Acid Uptake Modulates Stress Resistance In Vibrio Cholerae, William Strike

Honors Theses

Antibiotic-resistant pathogens represent an escalating threat to public health worldwide, substantially increasing the burden of healthcare and community-acquired infections. Several factors contribute to the emergence and spread of this threat, including but not limited to improper antibiotic use and prescriptions in health-care settings and the community, increasing global travel and migration from countries that have higher levels of antibiotic-resistant pathogens, and a lack of new antibiotics under development. According to the World Health Organization, rising rates of resistance among Gram-negative bacteria (such as Vibrio cholerae) are of particular concern. These bacteria have evolved a number of endogenous membrane remodeling strategies …

Investigation Of Origin And Growth Of Fungi In Coastal Beach Sand, Ruth L. Wright

Honors Theses

In a previous study from this lab (Stevens, Evans, & Aguirre, 2012) yeast colonies were isolated and grown grown from sand collection at pristine, medium use and high use beaches along the grand strand. In that study greater yeast abundance and greater diversity both correlated with higher census of human use, suggesting that some yeast growth of sites was anthropogenic. However the sand matrix itself also varied from very fine, uniform, dark sand at the pristine beach to coarser, color varying, less tightly packed sand at residential and commercial beaches. This suggested an alternative hypothesis to differential colonization and growth …

The Importance Of Rnd-Type Efflux Pumps In The Interactions Of Pseudomonas Aeruginosa With 4-Methoxybenzaldehyde, Sanchirmaa Namjilsuren

Honors Theses

Pseudomonas aeruginosa is a common hospital-acquired pathogen and is often associated with high mortality rates due to the development of multidrug-resistance. Antimicrobial plant-derived (phyto-) aldehydes present a promising alternative to antibiotics due to their broad-spectrum antimicrobial activity and low propensity to trigger resistance. However, two main problems preclude the widespread application of these compounds: chemical instability and low antimicrobial efficacy. The ongoing collaboration between the Mavrodi and Patton labs at USM has recently addressed the problem of chemical instability by incorporating plant aldehydes into polymer materials called Pro-Antimicrobial Networks via Degradable Acetals (PANDAs). Thus, this study aimed to address the …

The Impact Of Oxygen On The Intracellular Survival Of Listeria Monocytogenes, Amber N. Coats

Honors Theses

Listeria monocytogenes is an intracellular pathogen that is responsible for the foodborne disease listeriosis. In order to invade host cells and establish an infection, this pathogen must travel through the gastrointestinal tract where it is exposed to a variety of stressors, including low pH, bile, and variations in oxygen concentrations. These stressors necessitate this organism’s ability to sense and respond to its environment. Previous research has shown that bacteria use two-component systems to sense and regulate genes needed for improved survival in stressful environmental conditions. The goal of this study was to determine if the expression of the putative oxygen …

The Effects Of Nutrient Limitation On Toxin Production For The Harmful Algal Bloom Species, Pseudo-Nitzschia, In The Gulf Of Maine, Julia M. Park

Honors Theses

Pseudo-nitzschia is a pennate marine diatom that produces the neurotoxin domoic acid (DA). Domoic acid accumulates in the digestive tract of filter feeders and becomes concentrated at higher trophic levels. When DA concentrations reach critical levels, consumption of toxic shellfish can induce a condition known as Amnesic Shellfish Poisoning (ASP), which has been responsible for the deaths of marine mammals, seabirds and humans. This study worked to respond to a toxic Pseudo-nitzschia bloom in the Gulf of Maine during the fall of 2016, with the goal of quantifying and identifying Pseudo-nitzschia spp. in the bloom region and to determine the …

Haloferax Volcanii For Carotenoid Production, Erika Smith

Honors Theses

Carotenoids are used as natural colorants and have been shown to have numerous health benefits, making these molecules highly desirable in the aquaculture, pharmaceutical, and nutraceutical markets. There is increasing consumer demand for the natural synthesis of carotenoids, but current methods are limited by the cost and difficulty of extraction from organisms. The halophilic archaeon Haloferax volcanii is amenable to extraction and has an endogenous carotenoid biosynthetic pathway that culminates in the accumulation of the carotenoid bacterioruberin; however, H. volcanii also synthesizes important precursors that can be converted into more highly-desired carotenoids, such as β-carotene and astaxanthin, in the presence …

Vibrio Vulnificus Monitoring Along The Coastal Bend And The Rio Grande Valley, Texas, Louay Bachnak

Honors Theses

Vibrio vulnificus is a common opportunistic pathogen found in marine, coastal, and estuarine waters. This one micrometer microorganism is the leading cause of deaths in the United States due to food poisoning after consumption of raw or undercooked seafood. The optimal temperature for survival is between 15.6°C and 35.0°C with moderate salinities – conditions frequently found in the Gulf of Mexico (Lin, Payne, & Schwarz, 2003).

In the United States, V. vulnificus infections are more common in states bordering the Gulf of Mexico, including Texas. However, infections can still occur in individuals consuming infected food more inland or sustain wounds …