Pathogenic Microbiology Commons^™

Phosphatase Activity And Structure Of The Spoiie Protein In Clostridioides Difficile, Kathryn Wootten

Biological Sciences Undergraduate Honors Theses

Clostridioides difficile is a major cause of severe gastrointestinal infections and diseases, particularly in healthcare settings. Because of its ability to produce infectious endospores by sporulation, the protein SpoIIE, which is known to play a role in the regulation of this process, is being explored as a potential therapeutic target. In order to gain better insight into the structure and function of this protein, a concentrated and purified sample of the protein must be obtained. The goal of this study was to determine the conditions that would yield the highest amount of SpoIIE. The density of the cultures of bacteria …

The Impact Of Reactive Oxygen Species On Bactericidal Activity And Drug Resistance In Salmonella Typhimurium Upon Chlorine And Eugenol Exposure, Mohammed Aljuwayd

Graduate Theses and Dissertations

Salmonella Typhimurium (S.T) is a foodborne pathogenic bacterium which causes salmonellosis in human beings. Chicken is the most common source of S.T contamination. In chapter I, I discuss the growth of the poultry industry, the role chickens play as Salmonella reservoirs, the spread of drug-resistant strains caused by chicken husbandry and antibiotic use, and how eugenol can be used in poultry feed to decrease Salmonella infections and improve intestinal health. In chapter Ⅱ, I summarize the role of reactive oxygen species (ROS) in bactericidal activity. In chapter Ⅲ, I describe the killing mechanism of sodium hypochlorite (NaOCl) and identify the …

Persistence Of Salmonella Enterica Subsp. Enterica Ser. Javiana, Listeria Monocytogenes, And Listeria Innocua In Hydroponic Nutrient Solution, Gayatri R. Dhulappanavar, Kristen E. Gibson

Food Science Faculty Publications and Presentations

This study aimed to determine the persistence of Salmonella Javiana, Listeria monocytogenes, and Listeria innocua in nonsterile, hydroponic nutrient solution (NS) at 15, 25, 30, and 37°C over a 21-day period to mimic time from seedling to mature lettuce. Bacteria were inoculated in modified Hoagland's NS at 106 CFU/mL and maintained at 15, 25, 30, and 37°C. Samples were collected at various time points, and bacteria were quantified. A mixed model was used to determine the effect of bacteria type, time (day), and temperature on bacteria concentration (log CFU/mL). The least-squares means were calculated to compare the mean log …

The Bactericidal Activity Of A Novel Aneurinibacillus Aneurinilyticus Isolate Effectively Controls Foodborne Pathogens Campylobacter Jejuni And Listeria Monocytogenes, Peter M. Rubinelli, Rohana Liyanage, Jackson Lay, Jennifer C. Acuff

Chemistry & Biochemistry Faculty Publications and Presentations

Human zoonotic infection with Campylobacter is a major cause of gastroenteritis in the United States and worldwide. Listeria monocytogenes causes a potentially fatal infection in humans and is often attributed to contaminated food. Genetic typing has demonstrated that Campylobacter infection is often associated with the consumption of contaminated poultry products, with Campylobacter often colonizing the poultry gastrointestinal tract, while listeriosis is commonly associated with the consumption of contaminated ready-to-eat (RTE) foods. In this study, a strain of endospore-forming bacterium (strain NH) that is bactericidal towards the human food pathogens Campylobacter jejuni and Listeria monocytogenes was identified and characterized. Transwell experiments …

Horizontal Transfer Of Probable Chicken-Pathogenicity Chromosomal Islands Between Staphylococcus Aureus And Staphylococcus Agnetis, Douglas D. Rhoads, Jeff Pummil, Nnamdi S. Ekesi, Adnan A.K. Alrubaye

Cell and Molecular Biology Faculty Publications and Presentations

Staphylococcus agnetis is an emerging pathogen in chickens but has been most commonly isolated from sub-clinical mastitis in bovines. Previous whole-genome analyses for known virulence genes failed to identify determinants for the switch from mild ductal infections in cattle to severe infections in poultry. We now report identification of a family of 15 kbp, 17–19 gene mobile genetic elements (MGEs) specific to chicken osteomyelitis and dermatitis isolates of S. agnetis. These MGEs can be present in multiple copies per genome. The MGE has been vectored on a Staphylococcus phage that separately lysogenized two S. agnetis osteomyelitis strains. …

In Silico Analyses Of Diversity And Dissemination Of Antimicrobial Resistance Genes And Mobile Genetics Elements, For Plasmids Of Enteric Pathogens, Suad Algarni, Jing Han, Dereje D. Gudeta, Bijay K. Khajanchi, Steven C. Ricke, Young Min Kwon, Douglas D. Rhoads, Steven L. Foley

Animal Science Faculty Publications and Presentations

Introduction: The antimicrobial resistance (AMR) mobilome plays a key role in the dissemination of resistance genes encoded by mobile genetics elements (MGEs) including plasmids, transposons (Tns), and insertion sequences (ISs). These MGEs contribute to the dissemination of multidrug resistance (MDR) in enteric bacterial pathogens which have been considered as a global public health risk.

Methods: To further understand the diversity and distribution of AMR genes and MGEs across different plasmid types, we utilized multiple sequence-based computational approaches to evaluate AMR-associated plasmid genetics. A collection of 1,309 complete plasmid sequences from Gammaproteobacterial species, including 100 plasmids from each of the following …

The Role Of Microbiota-Biotransformed Bile Acid Metabolites On Campylobacter Jejuni- And Clostridium Perfringens-Infection, Ying Fu

Graduate Theses and Dissertations

Campylobacter jejuni (C. jejuni) and Clostridium perfringens (C. perfringens) are two leading pathogenic bacteria responsible for both human and animal diseases. In chapters Ⅱ to Ⅳ, I presented an overview of current knowledge on the role of immune response, microbiota, bile acids and their microbial metabolites on campylobacteriosis, and C. perfringens induced necrotic diseases.In chapter Ⅴ, I investigated effects of deoxycholic acid-modulated anaerobes (DCA-Anaero) on attenuating chicken transmission-exacerbated campylobacteriosis. SPF Il10-/- mice were gavaged with clindamycin for 7 days followed by infected with three C. jejuni strains, Cj-P0 (C. jejuni AR101), Cj-P1 (from chicken pre-infected with C. jejuni AR101), and …

The Antimicrobial Resistance Plasmid Mobilome Of Salmonella Enterica And Related Enteric Species And Factors That Influence The Transfer Efficiency., Suad Algarni

Graduate Theses and Dissertations

The dynamic distribution of antimicrobial resistance genes in Salmonella enterica is considered a public health risk. S. enterica is one of the most important etiological agents of foodborne illness and has a critical impact on global human health. In S. enterica and related species, mobile genetic elements (MGEs) serve as primary vehicles for the dissemination of antibiotic resistance genes in the bacterial evolution. This dissemination can be impacted by different selective pressures that leads to diverse antibiotic response phenotypes. This project focusses on the dynamics of antimicrobial resistance genes, particularly exploring the transfer efficiency of multidrug resistance plasmids in S. …

Characterization Of Genetic Pathways Involved In Resistance To A Novel Antifungal Peptide, Kayla L. Haberman

Graduate Theses and Dissertations

Antibiotic resistance is increasing prevalence, particularly in Candida glabrata. This opportunistic pathogen is closely phylogenetically related to Saccharomyces cerevisiae; however, its characterization is limited. C. glabrata is only second to Candida albicans as a fungal pathogen in immunocompromised patients. Commonly resistant to azoles, the most common fungal therapy, it has become costly and challenging to treat. A histatin 5 derived antifungal peptide, KM29, has a high degree of efficacy in Candida species and S. cerevisiae. The objective of this work is to advance our understanding of the mechanism of action of KM29 against C. glabrata. Previous work in the lab …

Bacterial Chondronecrosis With Osteomyelitis In Broilers: Genomics, Phylogenomics, And Methods To Detect Specific Pathogens During Outbreaks., Abdulkarim Abdulaziz A. Shwani

Graduate Theses and Dissertations

Lameness is a major issue in animal welfare and the broiler industry. Bacterial chondronecrosis with osteomyelitis (BCO) is one of the main causes of lameness. Many staphylococcal species, including Staphylococcus agnetis isolate 908, have been isolated from the bones and blood of lame broilers at the University of Arkansas. Staphylococcus agnetis is a coagulase-variable, Gram-positive bacterial species that has been previously associated with subclinical or mild clinical cases of mastitis in dairy cattle. The annotated complete genome of hypervirulent strain 908 was published at NCBI. In this study, it has been compared to nine genomes we assembled for hypervirulent isolates …

Diffusive Dynamics Of Biological Macromolecules In The Cytoplasm Of Live Bacteria, Asmaa Ali Sadoon

Graduate Theses and Dissertations

Bacteria are small organisms that play important roles in our bodies and in maintaining our surrounding environment. However, bacterial infections are considered as one of the health diseases that threatens the lives of humans. Resistance of bacteria to antibiotics has made treating bacterial infections more challenging. With the emergence of many developed optical techniques, it has become possible to study various biological mechanisms of molecules inside these small organisms with high resolution and precise localization. In this research, single particle tracking photoactivation localization microscopy (sptPALM) technique was used to investigate the dynamics of fluorescent histonelike nucleoid-structuring proteins (H-NS) in live …

Enzymatic Degradation Of Microcystin-Lr By Microcystinase (Mlra), Faisal Alqhtani

Graduate Theses and Dissertations

Microcystin-LR (MC-LR) is affecting the water supply worldwide. Hence, a way to eliminate this toxin is an essential target. In this study, successful cloning of the mlrA gene and producing MlrA enzyme that can degrade the cyclic MC-LR to linearized MC-LR was done. MlrA protein was expressed in Escherichia coli BL-21 (E. coli). Also, enhancing the MlrA yield by adding nickel to LB media was a success in producing more MlrA enzyme from the same volume. Even though the enzyme showed no activity after adding Ni, the enzyme was expressed at a higher yield. Furthermore, it was to investigate adding …

Directed Genome Evolution To Identify Genes For Macrophage Survival By Staphylococcus Agnetis, Sonali Lakshika Anne Lenaduwe Lokuge

Graduate Theses and Dissertations

Bacterial Chondronecrosis with Osteomyelitis (BCO) is a debilitating infection that negatively impacts animal welfare and costs the broiler industry billions of dollars annually. We have previously isolated Staphylococcus agnetis 908 from BCO samples obtained from broilers at the University of Arkansas research farm. This isolate can induce BCO lameness at greater than 50% in broilers exposed to the pathogen in drinking water. We found that S. agnetis 908 is capable of surviving and escaping macrophages compared to a closely related cattle isolate,1379. Through Directed Genome Evolution (DGE) we identified that this difference is at least partially associated with an alanine …

Developing A Microdialysis Sampling-Based Biofilm/Macrophage Co-Culture Model, Alda Diaz Perez

Graduate Theses and Dissertations

The host immune system and bacterial cells are known to interact during the human lifetime. Bacteria secrete a wide variety of signaling molecules, known as quorum sensing (QSC) molecules, that modulate the host immune system. While immune-biofilm interactions involve this chemical signaling network, the mechanisms through which this occurs are not well understood. This work aimed to develop a new method that can be used not only in vitro settings but also in vivo. The microdialysis sampling technique has widely been used in in vitro and in vivo settings in humans, mice, and rats for the collection of neuropeptides, cytokines, …

Pathogenicity Of Staphylococcus Agnetis Associated With Lame Broilers With Osteomyelitis, Sura Faris Salih Zaki

Graduate Theses and Dissertations

We developed High Resolution Melt Analysis (HERMA) as a rapid and reliable molecular diagnostic assay for the detection and identification of the main bacterial species recovered from the blood and lesions of bacterial chondronecrosis with osteomyelitis (BCO) from the lame broilers at the University of Arkansas poultry research farm. Also, the present study confirms that raising young birds on suspended wire flooring has been proved to successfully induce lameness attributable to BCO with birds being more susceptible to bacteraemia than those that were raised on litter flooring. The newly described pathogen, Staphylococcus agnetis, has been reported to be overrepresented in …

An Impedimetric Aptasensing System For The Rapid Detection Of Salmonella Typhimurium, America Sotero

Graduate Theses and Dissertations

Salmonella Typhimurium is a foodborne pathogen associated with raw and undercooked eggs, poultry, beef, fruits, and vegetables. In the United States, Salmonella is responsible for approximately 1.2 million illnesses, 23,000 hospitalizations, and 450 deaths annually. For many years, conventional detection methods such as culture-dependent and PCR-based methods have been the “golden standards” for the detection of this pathogen due to their high sensitivity and reliability. However, they still have some disadvantages such as long enrichment steps and high costs that need to be overcome. The development of a rapid and reliable method for the detection of S. Typhimurium is needed …

Examining Pathogenesis And Preventatives In Spontaneous And Staphylococcus-Induced Bacterial Chondronecrosis With Osteomyelitis In Broilers, Nnamdi Simon Ekesi

Graduate Theses and Dissertations

Bacterial chondronecrosis with osteomyelitis is the most important cause of lameness in broilers. This is important to poultry production, as it poses animal welfare issues, and causes a significant loss in revenue. The remediation of this disease requires the study of its etiology with fitting models and evaluating preventatives. The research reported herein covers genomic virulence analysis of BCO isolates, mainly Staphylococcus aureus, and Escherichia coli retrieved from lame birds. We found that S. aureus isolates were closest to chicken strains in Europe but may have been in the Arkansas area for a decade. Phylogenomics suggest our S. aureus is …

Investigating The Potential Of Microbial Metabolites And Mtor Signaling On Chicken Necrotic Enteritis, Mohit Bansal

Graduate Theses and Dissertations

Necrotic enteritis (NE), caused by C. perfringens and coccidiosis, is responsible for substantial economic loss annually. In the first chapter, we investigated the effect of secondary bile acid on necrotic enteritis. Day-old broiler chicks were randomly assigned to 5 groups of diets supplemented with 0 (basal diet), 0.8, 1.0, and 1.5 g/kg (on top of basal diet) deoxycholic acid (DCA). The birds were challenged with Eimeria maxima (20,000 oocysts/bird) at d 18 and C. perfringens (109CFU/bird/day) at d 23 and d 24 to induce NE. Birds were sacrificed at d 26, and ileal tissue and digesta samples were collected. DCA …

Biogeography Of The Bovine Respiratory Microbiome And Its Relationship With Bovine Respiratory Disease, Jianmin Chai

Graduate Theses and Dissertations

Bovine respiratory disease (BRD) is the most common and costly disease in the beef cattle industry, leading to high morbidity, mortality and huge economic loss. Despite the recent advances in vaccination and antimicrobial techniques, no significant health-improved outcomes have developed. Due to a deep investigation of the microbiome, respiratory microbiotas are known to have important roles for host health and disease. However, BRD specific pathogens have not yet been identified since they are found in both healthy and diseased animals. A systemic and comprehensive study of the biogeography of the bovine respiratory microbiome and its relationship with BRD is lacking …

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 …

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 …

Understanding The Molecular Strategies Of Campylobacter Jejuni For Survival In Amoeba And Chicken., Deepti Pranay Samarth

Graduate Theses and Dissertations

Campylobacter jejuni endure to be major cause of gastroenteritis in humans worldwide. C. jejuni is fastidious in laboratory setup but can cause waterborne infection through contaminated water where none of these fastidious conditions are met. This dissertation presents an assortment of studies focused in reviewing three major factors which could present a helping hand to C. jejuni in its environmental survival viz. i) association with free-living amoebae (FLA) ii) horizontal gene transfer (HGT) contributing towards its genetic diversity iii). Viable but non-culturable (VBNC) state.

Acanthamoeba is a FLA linked to environmental survival of many intracellular pathogens, including C. jejuni. In …

Aqueous Ozone Inactivation Of Viruses And Bacteria On Biotic And Abiotic Surfaces, Cailin Dawley

Graduate Theses and Dissertations

Produce is susceptible to contamination by foodborne pathogens. Food service establishments utilize sanitizing agents to reduce microbes on produce surfaces. The research objectives were to evaluate the efficacy of aqueous ozone 1) on the inactivation of viruses and bacteria on produce; 2) on the inactivation of viruses on stainless steel; and 3) against viruses in association with bacteria on produce surfaces. For objective 1, Boston bibb lettuce and cherry tomatoes were spot inoculated with a cocktail of viruses (murine norovirus (MNV) and MS2 bacteriophage) or bacteria (Enterobacter cloacae and Bacillus cereus) and washed for 40 min with samples taken every …

Role Of Incompatibility Group 1 (Inci1) Plasmid-Encoded Factors On Salmonella Enterica Antimicrobial Resistance And Virulence, Pravin Raghunath Kaldhone

Graduate Theses and Dissertations

Foodborne illnesses are a leading cause of infectious diseases in the world. Among enteric organisms Salmonella is a key pathogen. It’s high prevalence in poultry and other food-animal sources make it imperative to study. Salmonella has the ability to modify its genetic content with help of mobile genetic elements such as plasmids. Incompatibiltiy group 1 (IncI1) plasmids are commonly reported in Salmonella. This study evaluates role on IncI1 plasmids in antimicrobial resistance and virulence in Salmonella. Genetic determinants of resistance and virulence are noted among our IncI1-containing Salmonella isolates. These genetic elements are also transferable and reported to carry respective …

Experimental Approaches To Understand And Control Salmonella Infection In Poultry, Yichao Yang

Graduate Theses and Dissertations

Salmonella is a major foodborne pathogen around the world and chickens are the major reservoir to transmit Salmonella into the human food chain. For decreasing the infection of Salmonella, we developed six attenuated live vaccines based on Salmonella Enteritidis (SE) and Typhimurium (ST) for testing the cross-serovar and cross-serogroup protection from the challenge of Salmonella Heidelberg and Campylobacter jejuni. One of the constructed vaccine strain showed ability to protect against challenge from Salmonella Heidelberg. Even though some preventive approaches are able to decrease Salmonella colonization in the gastrointestinal tract of chickens or other farm animals, Salmonella transmission mechanisms remain unclear. …

Genetic Determinants Of Salmonella And Campylobacter Required For In Vitro Fitness, Rabindra Kumar Mandal

Graduate Theses and Dissertations

Non-typhoidal Salmonella (NTS) and Campylobacter play a major role in foodborne illness caused by the consumption of food contaminated by pathogens worldwide. A comprehensive understanding of the genetic factors that increase the survival fitness of these foodborne pathogens will effectively help us formulate mitigation strategies without affecting the nutrition ecology. The objective of this study was to identify the genetic determinants of Salmonella and Campylobacter that are required for fitness under various in vitro conditions. For the purpose, we used a high throughput Transposon sequencing (Tn-seq) that utilizes next generation sequencing (NGS) to screen hundreds of thousands of mutants simultaneously. …

A Portable And Automatic Biosensing Instrument For Detection Of Foodborne Pathogenic Bacteria In Food Samples, Zhuo Zhao

Graduate Theses and Dissertations

Foodborne diseases are a growing public health problem. In recent years, many rapid detection methods have been reported, but most of them are still in lab research and not practical for use in the field. In this study, a portable and automatic biosensing instrument was designed and constructed for separation and detection of target pathogens in food samples using nanobead-based magnetic separation and quantum dots (QDs)-labeled fluorescence measurement. The instrument consisted of a laptop with LabVIEW software, a data acquisition card (DAQ), a fluorescent detector, micro-pumps, stepper motors, and 3D printed tube holders. First, a sample in a syringe was …

Exploring The Mechanisms Of Action Of Antifungal Peptides Using Saccharomyces Cerevisiae., Michelle L. Mason

Biological Sciences Undergraduate Honors Theses

Candida albicans is a normal inhabitant of the skin and mucosal membranes of humans, however, in individuals with depressed immune systems or disrupted cutaneous flora, Candida can overgrow and cause serious infection. Candida infection is the fourth leading cause of nosocomial infection in the United States. These infections are often associated with longer hospital stays and higher mortality. Current drug therapies for this infection are largely ineffective due to the increased drug resistance of Candida species, and for some therapeutics, high levels of drug toxicity to humans. Histatin 5 is a naturally occurring salivary peptide that has strong antifungal properties. …

The Ccaat-Binding Factor Dependent Regulation Of The Oxidative Stress Response In Candida Albicans, Ananya Chakravarti

Graduate Theses and Dissertations

The success of Candida albicans as an opportunistic human pathogen has been attributed to several factors, including the ability to survive in limiting iron environments and the ability to evade the respiratory burst of human macrophages and neutrophils. The goal of this research is to elucidate the role of the CCAAT-binding factor in the oxidative stress response of Candida albicans. Prior whole genome microarray studies performed in our lab compared the gene expression of a wild type Candida albicans strain versus a hap5Δ strain under iron-limiting growth conditions. Among the differentially regulated genes, CTA1, encoding catalase, had a four-fold higher …

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 …