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A Forensic Assessment Of Current Water Quality Using Idexx Techniques In The Himmarshee Canal And New River In Fort Lauderdale, Florida, Catherine Margaret Bilodeau

All HCAS Student Capstones, Theses, and Dissertations

Fecal contamination has continued to plague local communities around the United States and the world, especially in highly populated areas like Southern Florida. Newer techniques are beginning to be used to track microbes properly as they are found in waterways. Some more recent techniques in Microbial Source Tracking (MST) use IDEXX kits to detect fecal indicator bacteria (FIB). Two IDEXX kits Colilert-18^® which detects total coliforms and Escherichia coli in water or fecal coliforms in wastewater, and Enterolert^®, which targets enterococci, were used in the waterways of the Himmarshee Canal and South Fork of the New River …

Investigating The Impact Of Nonenzymatic Lysine Acetylation On The Function Of The Bacterial Ribosome, Sarah Caldwell Feid

Dissertations

An under-investigated target of lysine acetylation is the bacterial ribosome. Although lysine acetylations on the ribosome are common and conversed in diverse bacterial species, little work has been done to understand how lysine acetylation affects the bacterial ribosome. The goal of this work is to determine if lysine acetylation has functional impact on the bacterial ribosome.I have identified in vitro and in vivo effects of nonenzymatic, AcP-dependent lysine acetylation on translation and the ribosome. In vitro acetylation of transcription-translation reactions causes a translation-specific defect that is unaffected by the addition of the CobB deacetylase. This suggests certain AcP-sensitive residues that …

Metabolic Engineering Of Escherichia Coli For N-Acetyl-Glucosamine Production, Anica Tasnim Protity

Graduate Research Theses & Dissertations

Glucosamine (GlcN) and GlcN-based supplements, such as N-acetyl-glucosamine (GlcNAc), are commonly used by osteoarthritis patients to relieve pain and maintain the health of bone joints. However, the current production methods for GlcN-based products are not eco-friendly and pose risks to individuals with shrimp allergies. Microbial cell-based systems have been explored to address these issues as an eco-friendly alternative for GlcN and GlcNAc production. This study focused on developing an Escherichia coli (E. coli) strain for GlcNAc fermentation. E. coli has an endogenous pathway for peptidoglycan biosynthesis involving GlcNAc-1-P. However, altering this pathway could affect cell growth and integrity. Therefore, two …

The Sos Response In Escherichia Coli K12: An Exploration Of Mutations In Lexa And Reca Using Fluorescence Microscopy, Steven Van Alstine

Doctoral Dissertations

Faithful replication of the genome is paramount for maintaining the fitness of an organism. Therefore, life has evolved inducible mechanisms to be able to repair damaged DNA and maintain evolutionary fitness. The SOS response is a highly conserved DNA damage inducible response that is tightly regulated. Multiple factors contribute to the ability of the cell to perform proper DNA repair and induction of the SOS response including the amount of RecA, mutations in RecA that affect competition for DNA, and other proteins that interact with the RecA filament. The complex relationship between RecA and LexA is the subject of this …

Implications Of Antibiotic And Bacteriophage Resistance In Environmentally Isolated E. Coli, Michael Connolly

Honors Theses

The increasing prevalence of antibiotic-resistant bacteria is an emerging problem for humans. Clinical misuse, overuse in agricultural and food settings, and limited numbers of new antibiotics have accelerated the proliferation of antibiotic-resistant bacteria. To confront this threat, scientists must develop new therapeutics that kill these antibiotic-resistant bacteria. In this study, we used Escherichia coli to analyze antibiotic and bacteriophage susceptibility. E. coli is a common, mostly benign, enteric, gram-negative bacteria. We isolated three E. coli strains from the Hans Groot Kill, a stream that runs through Union College’s campus. We sought to assess various E. coli strains’ antibiotic resistance, susceptibility …

Influence Of Nutrients And The Native On E. Coli Survival In The Beach Environment, Brigid Meyers

Theses and Dissertations

E. coli is used as an indicator for water quality to determine if water poses a health risk for pathogens. Past research has shown that E. coli is present in high numbers in freshwater beach sands distinct from fecal pollution events, yet the precise mechanism for their persistence in not well understood. Persistent E. coli populations in sand can resuspend into adjacent water and lead to increased beach closures when no threat is present. This work identifies factors that influence the survival of E. coli in sand using laboratory microcosms to replicate beach conditions. Microcosms were deployed to examine the …

Studying The Lysine Acetylation Of Aconitase Isozymes In E. Coli, Sara Ottinger

Chemistry & Biochemistry Undergraduate Honors Theses

The contents of this thesis have been modified from the publication “Araujo J, Ottinger S, Venkat S, Gan Q and Fan C (2022) Studying Acetylation of Aconitase Isozymes by Genetic Code Expansion. Front. Chem. 10:862483”. Though studies have found multiple lysine sites in which acetylation takes place in Escherichia Coli aconitase, acetylation’s effects on the enzyme’s activity have yet to be studied. Aconitase is the dehydratase-hydratase found in the citric acid and glyoxylate cycles responsible for the reversible isomerization of citrate to isocitrate via cis-aconitate intermediate. There are two isoforms of aconitase in E. coli: AcnA and AcnB. In …

Examining Bacteriophage Reliance On The Tola Protein Of Escherichia Coli, Ellen Davis

Honors Projects

With the ever-growing crisis of antibiotic resistant bacteria, bacteriophage provide an important potential alternative therapy to treat infections when antibiotics are ineffective. For this reason, it is important to understand the mechanisms bacteriophage use to enter their bacterial host cell. The cellular envelope of the gram-negative bacterium E. coli contains a protein system known as Tol-Pal, which utilizes energy from the Proton Motive Force to carry out several cellular functions, including some associated with cellular division and outer-membrane stability. Certain bacteriophage take advantage of the energy produced by this system to cross the bacterial cell membrane. One important protein of …

Evaluating The Effects Of Pine And Miscanthus Biochar On Water Activity And Escherichia Coli Populations In Commercial Broiler Litter, Christopher Austin Marty

Theses and Dissertations

The decrease in subtherapeutic antibiotic administration in poultry has increased the need to address production challenges caused by pathogens, such as E. coli. One potential way to improve bird health and reduce bacterial infection is through the addition of litter amendments that absorb moisture. Biochar (BC) has previously been shown to increase water holding capacity in poultry litter, but its effects on E. coli mitigation are unknown. The objectives of this research were to 1) evaluate water activity of poultry litter amended with pine and miscanthus BC, and 2) determine the effects of different BC inclusion rates on litter …

Chga-Depleted Urothelial Amp And Immune Response During Urinary Tract Infection, Theadora Jane Ceccarelli

Master's Theses

Women are 8 times more likely than men to experience a urinary tract infection (UTI) with up to 60% of women reporting a UTI in their lifetime. This significant healthcare burden is caused by the infection of uropathogenic Escherichia coli in the urinary tract. At this time, treatment is limited to antibiotic therapy, which is challenged by antibiotic-resistance. These factors make UTIs an NIH priority and are motivation for our research.CgA is a protein of the granin family co-released with catecholamines from neuroendocrine cells throughout the body. Pro-hormone convertases post-translationally cleave CgA into peptides, like catestatin, which can have antimicrobial …

Molecular Roles Of Small Inner Membrane Proteins In Escherichia Coli Cell Envelope Integrity, Aaron Mychack

Dissertations, Theses, and Capstone Projects

The biological membrane is an essential, defining feature of all cells. Biological membranes comprise phospholipid bilayers as well as a complement of proteins which are unique to a given organism. These proteins play a central role in dictating the biochemical state of the cell’s internal cytoplasm by controlling selective passage of solutes in and out of the cell, transducing signals in response to extracellular stimuli, and controlling the biogenesis of the bilayer itself which is critical towards barrier function. For most bacteria, the periphery of the cell is multi-layered, including both a biological membrane as well as a peptidoglycan cell …

Analysis And Optimization Of Colorimetric Nanosensors For Rapid Detection Of Microbes In Water, Ruby A. Lang

Master's Theses

Access to safe water is a basic human right recognized by the United Nations General Assembly in 2010 (WHO, 2020). However, a least 2.2 billion people globally still are without safely managed water services meaning they use a drinking water source that can be contaminated with faeces (WHO, 2020). With such a pressing global health issue, it is clear that improvement to water systems is important and required in the Agenda 2030 Sustainable Development Goals (SDGs). However, to improve water systems and prove they are safe water sources, water quality testing must occur. A solution to this issue is the …

Characterization Of The Physical And Chemical Effect Of Membrane Disruption And Protein Inhibiting Treatments On E. Coli, Khadijah Wright

Honors Undergraduate Theses

The increase in antibacterial resistance has placed the issue of microbial multi-drug resistance on a global stage (Gurunathan, 2019). This issue poses a threat to human and animal health as well as to the environment (Aslam et al., 2018). It affects not only the efficacy of treatment but also how those treatments are conducted (Friedman, Temkin, & Carmeli, 2016). As a result of this ongoing threat, new treatments that have potent effects on bacteria are necessary. One scientific response to this issue has been the development of multifunctional nanoparticles (NPs)(H. Wang et al., 2018). NPs have the ability to be …

16s Rrna Analysis And Toxin Gene Presence In Escherichia Coli Isolated From Beach Water And Sand At A Public Beach (Erie County, Ny), Jennifer D. Jackson

Biology Theses

Every year, thousands of people utilize beaches for recreation, but most are unaware of Escherichia coli (E. coli) contamination and the possibility of acquiring an infection. In this study, 173 strains of E. coli were isolated from sand and adjacent waters from a public beach in Erie County, NY and analyzed for genetic relatedness based on sequence differences in the variable regions of the 16S rRNA gene. Some of the variable regions (V1 and V6) proved useful in constructing phylogenetic trees but the discriminatory power of these regions was inadequate to resolve intraspecies differences. Therefore, whether extant populations …

The Effect Of Proteolytic Queues On Antibiotic Tolerance And Persistence Cells Population In Escherichia Coli, Alawiah Abualrahi

Electronic Theses and Dissertations

A major contributing factor to the abundance of antibiotic-resistant microorganisms and failed antibiotic treatment is survival due to antibiotic tolerance and persistence. Antibiotic tolerance is a widespread phenomenon that enables cells to survive treatment without carrying a resistance gene. This phenomenon renders antibiotic treatments less effective and facilitates antibiotic resistance. We are particularly interested in proteases, responsible for degradation of proteins, because of their known relationship to tolerance and persistence. Here, we examine the effects of proteases and antibiotic survival using queueing theory, in which one type of customer competes for processing by servers, that has traditionally been applied to …

Post-Translational Modifications And Functional Studies Of Dksa In Escherichia Coli, Andrew Charles Isidoridy

Legacy Theses & Dissertations (2009 - 2024)

DksA is a bacterial gene regulator that functions synergistically with the stress alarmone ppGpp to mediate the stringent response. DksA also functions independently of ppGpp to regulate transcription of a number of genes. DksA function is dependent on its binding affinity to RNA polymerase and requires specific interactions between RNAP and catalytic amino acids located on the coiled coil tip, D74 and A76. While much of the previous work on DksA has focused on understanding the mechanisms of action and the numerous gene targets for transcriptional regulation, little is known about the mechanisms by which DksA expression and function may …

Evaluating Human-Associated Escherichia Coli Marker Genes For Microbial Source Tracking Applications In Florida, Jacob K. Senkbeil

USF Tampa Graduate Theses and Dissertations

Fecal pollution of surface water can lead to human health issues because waterborne transmission of pathogens is a common cause of gastroenteritis. Genetic markers developed for human-associated Escherichia coli (H8, H12, H14, and H24) are promising tools for microbial source tracking (MST) of sewage in environmental waterbodies but are untested in the US. H gene marker performance characteristics (specificity, sensitivity, and prevalence) were assessed, with the goal of developing a quantitative PCR assay for the markers with the best performance. Individual E. coli isolates from reference samples were tested to determine end-point PCR assay performance. Three hundred E. coli strains …

Clpxp-Regulated Proteins Suppress Requirement For Reca In Dam Mutants Of Escherichia Coli K-12, Amie Savakis

Masters Theses

Double strand breaks (DSB) are a common source of DNA damage in both prokaryotes and eukaryotes. If they are not repaired or are repaired incorrectly, they can lead to cell death (bacteria) or cancer (humans). In Escherichia coli, repair of DSB are typically accomplished via homologous recombination and mediated by RecA. This repair pathway, among others, is associated with activation of the SOS response. DNA adenine methyltransferase (dam) mutants have an increased number of DSB and, therefore, are notorious for being RecA-dependent for viability. Here, we show that the synthetic lethality of Δdam/ΔrecA is suppressed when clpP is removed, suggesting …

Regulation Of The Tubulin Homolog Ftsz In Escherichia Coli, Monika S. Buczek

Dissertations, Theses, and Capstone Projects

Escherichia coli is a well-known pathogen, and importantly, a widely used model organism in all fields of biological sciences for cloning, protein purification, and as a model for Gram-negative bacterial species. And yet, researchers do not fully understand how this bacterium replicates and divides. Every year additional division proteins are discovered, which adds complexity to how we understand E. coli undergoes cell division. Due to their specific roles in cytokinesis, some of these proteins may be potential targets for development of antibacterials or bacteriostatics, which are much needed for fighting the current global antibacterial deficit. My thesis work focuses on …

Bacteria That Resist Centrifugal Force, Nickolas Kessler

Honors Undergraduate Theses

Our lab discovered that approximately 1 in 10,000 Escherichia coli cells in stationary phase remain in suspension after a high g-force centrifuge event. To establish the mechanism behind this curious phenotype, multiple mutant strains of E. coli were independently evolved such that the majority of their populations resisted migration when exposed to high centrifugal forces. Genomic DNA sequencing of the mutants' revealed unique, isolated mutations in genes involved in capsule synthesis and exopolysaccharide (EPS) production. Each mutant exhibits a novel mechanism that allows them to remain in suspension. The mutants were further characterized by determining their growth rates, strengths …

Molecular Analysis Of Β-Lactamase Genes In Antibiotic Resistant Bacteria, Neisha Medina Candelaria

Honors Projects

The wide overuse of antibiotics in both the medicine as well as agricultural has lead to a problem with antibiotic resistance. This present research experiment aims to identify various environmental bacterial samples to find antibiotic resistant strains. By looking at these strains we are then able to develop a system to monitor how antibiotic resistance spreads and develops over time. We can then compare different bacteria species to identify any similarities and the prevalence of the resistant strains.

Colicins - A Sound Antimicrobial Approach For The Prevention Of Catheter-Associated Urinary Tract Infections, Sandra M. Roy

Doctoral Dissertations

The emergence and spread of antibiotic resistance has created one of the greatest challenges in fighting infectious disease. We address the rise of antibiotic-resistant pathogens by examining the evolutionary history of a class of resistance determinants, the SHV b-lactamases. We isolated the genes that encode the SHV beta-lactamases (bla_SHV genes) from clinical settings and from an environment essentially devoid of antibiotic use. Our data suggests that, counter to current dogma, the use of antibiotics in the clinic is not creating these resistance genes; genes for antibiotic resistance already exist in nature and our use of antibiotics in clinical …

Investigating The Sources And Dynamics Of Escherichia Coli In A Missouri Ozarks Watershed, Kaleb Bassett

Masters Theses

"Escherichia coli, an intestinal bacterium, can serve as an indication of the presence of pathogenic microorganisms in water systems used by humans for recreation, agriculture, or drinking water. Many aquatic systems in the United States exceed the E. coli standard, set by the Environmental Protection Agency, for safe drinking water and recreational use. During 2016, a water sampling program was established in the Mill Creek watershed, a rural watershed located near the city of Newburg in Phelps County, Missouri. Water samples were collected before, during, and after storms throughout the year to examine the relationship between E. coli …

The Selective Survival Of Escherichia Coli In Freshwater Beach Sand, Natalie Ann Rumball

Theses and Dissertations

The quantification of Escherichia coli or E. coli is the most common method used to detect recent fecal pollution in recreational water, as this species is known for its high abundance in fecal matter and assumed host-associated nature. However, it has been determined that some strains are capable of long-term survival and potential propagation in non-host environments, such as the beach sand. These long-term environmental survivors are host-independent and are not associated with the same health risks as those E. coli from recent fecal pollution. However, they have been shown to impact how water quality is perceived as they are …

Inhibition Of Bacterial Growth And Prevention Of Bacterial Adhesion With Localized Nitric Oxide Delivery, Julia Osborne

Dissertations, Master's Theses and Master's Reports

Bacterial infections continue to be a problem at the site of an indwelling medical device, and over the years, various bacterial strains have become more resistant to current antibiotic treatments. Bacterial infection at an indwelling medical device can be dangerous and affect the performance of the medical device which can ultimately lead to the failure of the device due to bacterial resistance to treatment.

Nitric Oxide (NO) has been shown to possess antibacterial properties to prevent and inhibit bacterial growth. NO releasing coatings on indwelling medical devices could provide a reduction in bacterial infections that occur at the device site …

Ecological Dynamics In Compost-Amended Soils And The Resulting Effects On Escherichia Coli Survival, Anya Cutler

Graduate College Dissertations and Theses

Escherichia coli (E. coli) are common and typically innocuous copiotrophic bacteria found in the mammalian gut microbiome. However, over the past 30 years, pathogenic E. coli have been responsible for several outbreaks of foodborne illness linked to contaminated produce. The introduction of Escherichia coli to an agricultural soil, via contaminated water, compost, or raw manure, exposes the bacterium to a medley of ecological forces not found in a mammalian gut environment. This study assesses a variety of abiotic and biotic soil factors that influence the ability of an "invasive" copiotrophic coliform bacterium to survive in compost-amended agricultural soil. The study …

Demographics And Transfer Of Escherichia Coli Within Bos Taurus Populations, Joshua Ryan Dillard

Master's Theses

In the United States, symptoms caused by pathogenic strains of Escherichia coli are on the rise. A major source of these pathogenic strains is the E. coli in the digestive tract of cattle. The purpose of this project was to determine if E. coli are transferred between individuals of the same species and if interspecies transmission is possible. Proximity of cattle was also studied as a contributing factor to the transfer of E. coli. To accomplish this goal, E. coli isolates from cattle and cohabitating ground squirrels were compared through a new method of bacterial strain typing called pyroprinting. …

Impedance Biosensors For The Rapid Detection Of Viral And Bacterial Pathogens Using Avian Influenza Virus Subtypes H5n1 And H7n2 And Escherichia Coli O157:H7 As Model Targets, Jacob David Lum

Graduate Theses and Dissertations

This research investigated impedance biosensors for the rapid detection of viral and bacterial pathogens using avian influenza virus (AIV) subtypes H5N1 and H7N2 and Escherichia coli O157:H7 as the model targets, which were chosen due to their impact on the agricultural and food industries. For the detection of AIV H7N2, a single stranded DNA aptamer was selected using systematic evolution of ligands by exponential enrichment (SELEX). The selected aptamer and a previously selected aptamer against AIV H5N1 were used in a microfluidics chip with an embedded interdigitated array microelectrode to fabricate an impedance biosensor for specific detection of AIV H7N2 …

The Movement Of Escherichia Coli And Enterococci Among Beach Sand, Lyngbya Wollei, And The Water Column: Implications For Human Health, Krystal Ann Bakkila

Wayne State University Theses

Occurrence of the filamentous cyanobacteria Lyngbya wollei has become an increasing concern in the Great Lakes region. Prior to the early 1900's, L. wollei had been occasionally documented in the Great Lakes region, but in the last decade it has been observed with increasing frequency. In the Great Lakes L. wollei has been proliferating, fragmenting, and accumulating ashore, where it decays forming thick detrital mats harboring potentially harmful bacteria such as Escherichia coli and enterococci. While the filamentous green algae species Cladophora glomerata has been well studied in this region, very little research has been done on the cyanobacteria L. …

The Pleiotropic Effects Of Beneficial Mutations Of Adapted Escherichia Coli Populations, Brian Scott Van Dam

Honors Theses and Capstones

Mutations that improve fitness in one environment can often be beneficial, deleterious, or neutral in alternative environments. When a single mutation effects fitness in multiple environments, it is said to be a pleiotropic, which can have important consequences for niche specialization, niche expansion, speciation, and even extinction in the face of environmental change. While previous studies have revealed that pleiotropy is nearly universal, the role of adaptive history in the spectrum of pleiotropic effects has yet to undergo detailed experimental observation. Using experimental evolution we gathered beneficial mutations in a previously adapted strain of Escherichia coli growing in the same …