Life Sciences Commons^™

Genetic And Genomic Analyses Of Bacterial Responses To Different Stress Environments, Sakshi Rampal

Graduate Theses and Dissertations

Bacteria encounter a plethora of environmental stresses and have evolved different mechanisms to recognize and respond to various harmful conditions. Understanding and elucidating common themes as well as unique aspects of the molecular mechanisms underlying stress adaptation is important and can provide valuable strategies for applications. This study focuses on stress responses in three different bacteria, namely, Acidothermus cellulolyticus, Mycobacterium smegmatis and Escherichia coli. The thermophilic and acidophilic organism A. cellulolyticus was used as a model system to understand the effects of lignin phenolic acids on cellulolytic bacteria. Lignin phenolic acids pose a significant challenge to microbial deconstruction of lignocellulosic …

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 …

Effects Of Acute Heat And Oxidative Stress On The Hepatic Expression Of Orexin And Its Related Receptors, Stephanie Khaldi

Graduate Theses and Dissertations

It is widely known that orexin A and B peptides as well as their receptors are expressed in the hypothalamus and distributed throughout the central nervous system, but there have been few studies regarding its presences in other parts of the body. There is now evidence that orexin (ORX) and its receptors (ORXR1/2) are present in the avian liver; however, their regulation under different environmental conditions is still unknown. In the current study, we sought to determine the effects of heat and oxidative stress using hydrogen peroxide (H2O2) and 4-hydroxynonenal (4-HNE) on the hepatic expression of ORX and ORXR1/2 in …

Listeria Monocytogenes Strain Variation: Starvation, Metabolism, And Macrophage Survival, Nathan A. Jarvis

Graduate Theses and Dissertations

Listeria spp. are often isolated from raw and processed food products. Listeria monocytogenes is responsible for death rates of 10 to 30% in immune compromised people; it can also infect a vast range of animals. There are numerous reports of poultry harboring or becoming infected with L. monocytogenes. First, this research assessed the ability of L. monocytogenes to replicate within chicken macrophages. Using the gentamicin-killing assay, a cell culture of chicken macrophage-like cells (HD11) were infected with two strains of L. monocytogenes individually (EGD-e and Scott A). Results show that unlike data on mouse macrophages, intracellular L. monocytogenes do not …

Shifts In Myxomycete Community Structure In Selected Microhabitats Across Nutrient Treatments In A Lowland Tropical Forest Of Panama, Laura M. Walker

Graduate Theses and Dissertations

Myxomycetes (plasmodial slime molds) are abundant amoeboid predators of bacteria and other microorganisms. They are found worldwide, largely in association with decaying plant material in terrestrial habitats. Their consumption of bacterial prey puts microbial predators such as myxomycetes in a key position in various ecosystem processes wherein they help regulate the flow of nutrients (e.g., N and C) through the ecosystem. The importance of microbial predators in nutrient cycling and plant productivity is well established. Given the extent to which tropical ecosystems influence global nutrient fluxes, along with the ecologically significant role that myxomycetes play in these processes, there is …

Characterization Of Histone Mutants Associated With Mitotic Defects In Saccharomyces Cerevisiae, Md Riajul Hossain

Graduate Theses and Dissertations

Nucleosomes, the basic unit and the building blocks of chromatin have an essential role in the tight packaging of DNA into higher order chromatin. Work from our lab and others have provided information on the contributions of different histone proteins and specific domains within the nucleosome made to create the centromeric chromatin structure required for normal chromosome segregation during mitosis. The DNA entry/exit site is a particular region of the nucleosome where histone H2A, H3 and H4 form critical interactions that appear to be essential for the association of Sgo1, a tension sensing protein that monitors kinetochore-microtubule attachment. In our …

Dissemination And Persistence Of Plasmid Located, Integron Associated Antibiotic Resistant Genes In Wastewater Treatment Plant Effluent And Stream Water Bacteria, _ Suhartono

Graduate Theses and Dissertations

Transmissible plasmid-mediated integrons play important role in the persistence and dissemination of antibiotic resistance throughout the environment. Plasmids from 139 multi-antibiotic resistant Escherichia coli recovered from wastewater treatment plant effluent and upstream and downstream receiving stream water in Northwest Arkansas were extracted and profiled. Genes of class 1 and class 2 integrase (intI), mobilization (mob), sulfamethoxazole resistance (sul), and trimethoprim resistance (dfr) were detected using PCR and confirmed through DNA sequencing. Plasmids from almost half of the isolates (47%) were transmissible with mobF12 gene as the most frequently detected mobilization gene. Plasmid-borne class 1 with and without class 2 integrons …

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 …

Oxidative Stress Response In Archaea: Elucidation Of Oxidant Sensing And Tolerance Mechanisms In Methanosarcina Acetivorans, Matthew Edward Jennings

Graduate Theses and Dissertations

Methanogens are archaea possessing a conserved metabolic pathway which produces methane. Many of the enzymes in the methanogenesis pathway are Fe-S proteins, meaning methanogens are sensitive to conditions which disrupt Fe-S clusters. Molecular oxygen is capable of disrupting Fe-S clusters through oxidation of the iron atoms. Furthermore, reduced iron can facilitate the production of reactive oxygen species (ROS), meaning methanogens must possess antioxidant mechanisms. Detection and eradication of ROS is important for all cells, due to the potentially fatal consequences of unchecked oxidation. This dissertation presents two separate projects investigating mechanisms the model methanogen Methanosarcina acetivorans possess for dealing with …

Reduction Of Campylobacter Jejuni On Chicken Wingettes By Treatment With Caprylic Acid, Chitosan Or Protective Cultures Of Lactobacillus Spp., Ann Nicole Woo-Ming

Graduate Theses and Dissertations

Poultry is one of the main sources of protein in the United States and in 2014 Americans ate approximately 100 lbs per person. However, consumption of poultry products is strongly associated with foodborne illness from Campylobacter. In the first study, chitosan, caprylic acid and their combination were evaluated as a coating treatment for the reduction of Campylobacter jejuni on poultry products. For the initial screening trials, chitosan of three different molecular weights (15-50 kDa, 190-310 kDa and 400-600 kDa) was evaluated at three concentrations (0.5%, 1.0%, 2.0%) and separately, caprylic acid was tested at 0.5%, 1.0% and 2.0% using chicken …

Understanding The Causal Agent Of Rose Rosette Disease, Patrick Louis Di Bello

Graduate Theses and Dissertations

A number viruses are known to infect roses, ranging from those in the genera Nepovirus, and Ilarvirus, which have been reported since the inception of rose virology, to recently discovered viruses in the genera Carmovirus, Closterovirus, Emaravirus, Luteovirus, Rosadnavirus, and Potyvirus. Of the viral diseases in rose, arguably the most damaging is Rose rosette (RRD), which is associated with the Emaravirus, Rose rosette virus (RRV). The objective of this thesis is to fill in the gaps in knowledge on the epidemiological aspects of RRD and RRV. There has been significant progress in the epidemiology of the RRD agent prior to …

The Functional And Distributional Ecology Of Mycetozoans Under Changing Edaphic And Climatic Dynamics, Geoffrey Lloyd Zahn

Graduate Theses and Dissertations

Investigations into the distribution and ecosystem functions of fruiting amoebae revealed that local-scale environmental conditions can largely explain broad biogeographical patterns in species assemblage, the way in which amoeboid predators shape bacterial communities and how this top-down influence may affect global biogeochemical processes in a changing climate. The distribution and assemblage of protosteloid amoebae on the islands of New Zealand and Hawaii did not yield any expected patterns of island biogeography, and conformed to other global regions studied. The strongest predictor of species richness in a given region was sampling effort and these species do not appear to have any …

Understanding The Evolution Of Aggregative Multicellularity: A Molecular Phylogenetic Study Of The Cellular Slime Mold Genera Sorodiplophrys And Pocheina, Alexander Tice

Graduate Theses and Dissertations

Cellular slime molds are amoeboid protists that have a unicellular trophic phase and multicellular dispersal stage formed through the aggregation of individuals in their life cycles. These organisms were once thought to form a monophyletic group in the Mycetozoa. After careful morphological, ultrastructural, and molecular studies, cellular slime molds are now thought to be distantly related organisms that have all converged on the cellular slime mold habit. The following thesis consists of two molecular phylogenetic studies on two named genera of cellular slime mold for which little or no molecular data were publically available. In the first study, gene sequence …

Evaluation Of Tulane Virus As A Surrogate For The Study Of Human Norovirus, Sabastine Eugene Arthur

Graduate Theses and Dissertations

Among all known causes of acute gastroenteritis, human noroviruses (HuNoV) are the primary cause (68%) of outbreaks and are associated with 78% of illnesses, 46% of hospitalizations, and 86% of deaths. The main obstacle to studying the pathogenesis of HuNoV is the lack of cell culture system and small animal model. Murine norovirus (MNV) and feline calicivirus (FCV) have been utilized as model surrogate viruses to study HuNoV. In this research, a more recent surrogate virus, Tulane virus (TV), was evaluated for physicochemical stability and environmental persistence. The primary goal was to determine the suitability of TV as a surrogate …

Methane Emissions From Direct-Seeded, Delayed-Flood Rice Grown On A Clay Soil, Alden Daniel Smartt

Graduate Theses and Dissertations

Due to the production of methane (CH4) under flooded-soil conditions, rice (Oryza sativa L.) cultivation is a major contributor to agricultural CH4 emissions. Studies examining CH4 emissions from rice have only recently been initiated in Arkansas and no data have been collected from rice produced on clay soils in Arkansas. Therefore, research was conducted in 2012 and 2013 at the Northeast Research and Extension Center in Keiser, Arkansas to examine the factors affecting CH4 emissions from rice produced on a Sharkey clay (very-fine, smectitic, thermic Chromic Epiaquerts). The objectives of this study were to determine: 1) the effect of vegetation …

Management Of A Primordial Problem: Redox-Sensitive Transcriptional Regulation In Methanosarcina Acetivorans, Ryan Christopher Sheehan

Graduate Theses and Dissertations

The primordial Earth which hosted the first forms of life was an environment free of oxygen. Early organisms utilized metabolisms dependent upon anaerobic conditions and incorporated systems to which oxygen is deleterious. As the content of oxygen in Earth's atmosphere increased, anaerobic organisms had to acquire methods to sense and combat oxygen and reactive oxygen species. Several mechanisms were advantageous to such anaerobic organisms which correlated transcriptional processes with the redox state of the cell so that energy may be conserved and oxygen stress recovery genes activated during periods of oxidative stress. Iron sulfur (Fe-S) cluster cofactors incorporated within RNA …

A Phylogenetic Analysis Of Dictyostelium Purpureum Based On Nuclear Rdna Sequences, Mahmoud Suliman

Graduate Theses and Dissertations

Dictyostelids (cellular slime molds) are eukaryotic microorganisms that have both unicellular and multicellular stages during their life cycle. In this study, a molecular phylogenetic analysis was conducted for isolates of one species (Dictyostelium purpureum) based DNA sequences of the ITS, 5.8S and SSU regions of nuclear rDNA. Moreover, a detailed morphological study was carried out using images obtained with both dissecting and compound microscopes. Mating experiments were carried out to assess macrocysts formation between each pair of isolates. The constructed molecular phylogenetic trees indicate that (1) D. purpureum isolates are more closely related to each other than to other species …

Bacterially-Mediated Formation Of Rock Coatings In Kärkevagge, Swedish Lapland: A Mineralogical And Micro-Environmental Analog For Mars, Cassandra L. Marnocha

Graduate Theses and Dissertations

The search for past or present life on Mars is, for now, limited to surface environments. An often neglected surface environment that could have served as an abode for life and could presently preserve evidence of that life is that of rock coatings. Rock coatings are mineral accretions on rock surfaces. On Earth, they are widespread and occur with considerable chemical diversity. There is growing evidence for a biotic role in their formation on Earth, particularly with respect to rock varnish. As a result, rock varnish has become a target of astrobiological interest on Mars, where varnish-like coatings have been …

Proteomic Studies Of The Influenza Virus-Human Cell Interactions: The Responses Of Host Cell Protein Expression To Viral Infection And The Novel Host Proteins That Interact With Virus Protein Ns1, Yimeng Wang

Graduate Theses and Dissertations

Influenza A viruses (IAVs) continue to be a threat to human health. Despite extensive studies, the mechanisms underlying the IAVs-host interactions during IAV infection remain elusive. We employed quantitative proteomic methods to systematically explore the host cell protein expression responses to IAV infection and examine the function of a critical IAV protein called NS1 by identifying its host binding partners. Specifically, we used a 2-dimentional gel electrophoresis (2-DE) based proteomic method to screen host proteins whose expression was substantially altered by IAV. One critical protein named IκB kinase-gamma (IKKγ) was found to be significantly down-regulated during IAV infection. Functional studies …

Qcm Aptasensor For Rapid And Specific Detection Of Avian Influenza Virus, Luke Brockman

Graduate Theses and Dissertations

There has been a need for rapid detection of avian influenza virus (AIV) H5N1 due to it being a potential pandemic threat. Most of the current methods, including culture isolation and PCR, are very sensitive and specific but require specialized laboratories and trained personnel in order to complete the tests and are time-consuming. The goal of this study was to design a biosensor that would be able to rapidly detect AIV H5N1 using aptamers as biosensing material and a quartz crystal microbalance (QCM) for transducing method. Specific DNA aptamers against AIV H5N1 were immobilized, through biotin and streptavidin conjugation, onto …

Cell Bioenergetics In Leghorn Male Hepatoma Cells And Immortalized Chicken Liver Cells In Response To 4-Hydroxynonenal Induced Oxidative Stress, Alissa Laura Piekarski

Graduate Theses and Dissertations

Bioenergetic mechanisms responsible for ATP production are essential in carrying out maintenance and cell-specific functions. In this study, hepatocytes (liver cells) were used to test both endogenous and exogenous stress on cellular respiration. The secondary lipid peroxide, 4-hydroxynonenal (HNE), was used because it alters bioenergetics by increasing mitochondrial proton leak that attenuates mitochondrial radical production and, therefore, endogenous oxidative stress. The major objective of this study was to demonstrate effects of HNE-induced oxidative stress on avian hepatocyte bioenergetics. Various chemical which help enable the determination of oxygen (O2) consumption linked to ATP synthesis (oligomycin), maximal O2 consumption (FCCP), and proton …

The Use Of Diatoms To Infer Environmental Change Of The Lower White River, Southeastern Arkansas, Ruchi Bhattacharya

Graduate Theses and Dissertations

This dissertation describes the limnology of the lower White River oxbow lakes in southeast Arkansas, inundated by the major 2008 spring flood. The flood water and sediment deposited in oxbow lakes were analyzed for water chemistry, sediment geochemistry and diatom assemblages and used as an analogue for past floods. The 2008 spring flood inundated and homogenized the entire floodplain; also represented by the predominantly planktonic diatom assemblage suggesting riverine input. Indicator species for eutrophication were also recorded consistent with the high nutrient content in the lakes. However, the variance in the surface diatom assemblage was best explained by the lake …

Genome-Wide Identification Of Conditionally Essential Genes In Salmonella Typhimurium Using Tn-Seq Method, Anita Khatiwara

Graduate Theses and Dissertations

ABSTRACT

As more whole genome sequences become available, there is an increasing need for high-throughput methods that link genes to phenotypes and facilitate discovery of new gene functions. The objective of this study was to develop a high-throughput method to study gene functions in bacteria and use this method to study gene functions of S. enterica serotype Typhimurium (S. Typhimurium) under various environmental conditions encountered during its life cycle. Chapter I of this dissertation reviews the history and evolution of functional genomics in bacteria with focus on Salmonella, along with the recent techniques available. Chapter II, deals with the development …

Utilization Of Natural Green Tea And Grape Seed Extracts And Nisin To Reduce Conventional Chemical Preservatives And To Inhibit The The Growth Of Listeria Monocytogenes In Ready To Eat Low And High Fat Chicken And Turkey Hotdogs, Amara Venkata Sunil Perumalla

Graduate Theses and Dissertations

Ready-to-eat meat (RTE) products such as hotdogs represent a popular segment in convenience food purchases. Increased demand has led the processors to extend the shelf life by minimizing lipid oxidation and post-processing contaminants such as Listeria monocytogenes. There is a growing interest in the food processors and consumers regarding the use of natural alternatives in place of synthetic food additives to control the growth of foodborne pathogens and (or) lipid oxidation. In recent years, green tea (GTE) and grape seed extracts (GSE) are increasing choices as they have demonstrated antioxidant as well as antimicrobial properties in various food applications. …

Role Of Lactic Acid Bacteria As A Bio-Sanitizer To Prevent Attachment Of Listeria Monocytogenes On Deli Slicer Contact Surfaces, And The Influence Of Listeria Innocua On The Attachment Of L. Monocytogenes On Selected Materials Located In The Food-Processing Environment, Jean Baptiste Ndahetuye

Graduate Theses and Dissertations

Listeria monocytogenes an important foodborne pathogen that continues to be a serious problem for the food industry. This pathogen contaminates food primarily during post-harvest in the food-processing environment. Its ecology in the food-processing environment is not well understood but previous research has demonstrated the ability of L. monocytogenes to survive on food contact surfaces after cleaning and disinfection. The current study explored the attachment of three lactic acid bacteria (LAB) strains, Lactobacillus animalis , Lb. amylovorus and Pediococcus acidilactici , their combination in a cocktail, and their influence to prevent the attachment of L. monocytogenes at room temperature by examining …

In Vitro Selection Of Aptamers Against Avian Influenza Virus H5n1, Jingjing Zhao

Graduate Theses and Dissertations

Over $10 billion losses in the poultry industry were caused by avian influenza (AI) so far. Rapid and specific detection of avian influenza virus is urgently needed with the concerns over the outbreaks of highly pathogenic H5N1 influenza virus and cases of animal and human infection. Aptamers are oligonucleic acid or peptide molecules that bind a specific target molecule with good affinity. They show better thermal stability than antibodies. The goal of this research was to select DNA-aptamers as the specific recognition element of AI H5N1virus to be used in detection assays specific for field application. In this study, Systematic …

Detection Of Two Pathogens Of High Importance To The National Poultry Improvement Plan: Salmonella Spp. And Mycoplasma Spp., Robin Levi Jarquin

Graduate Theses and Dissertations

In 1935, the National Poultry Improvement Plan (NPIP) was created to control Salmonella gallinarum and Salmonella pullorum . These two pathogens were devastating economically for poultry producers. Through cooperative efforts using vaccination and strict biosecurity, these two pathogens were eradicated from the United States. Currently, the NPIP program is targeting two other poultry pathogens, Salmonella enterica serovar Enteritidis and Mycoplasma . In the broiler industry it targets 2 specific Mycoplasma species (synoviae, gallisepticum). Vaccinations for these bacteria are available, but are not fully effective at controlling all strains and serovars. Thus, constant monitoring systems and strict biosecurity measures are necessary …

Host-Virus Interactions Of Infectious Laryngotracheitis Virus Infection In Cultured Cells, Jeong Yoon Lee

Graduate Theses and Dissertations

Infectious laryngotracheitis virus (ILTV; Gallid herpesvirus 1) causes upper respiratory diseases in mainly chickens and exhibits 90-100% of high morbidity and up to 70% of mortality, resulting in huge economic losses in the poultry industry worldwide.

To study host-ILTV interactions, the changes in genome-wide gene expressions in response to wild-type and vaccine ILTV infections in primary chicken embryo lung cells were investigated using microarray analysis. Results provide crucial insights into host cell pathogenic and immunogenic responses against wild-type and vaccine ILTV infections. Using microarray method and Ingenuity Pathway Analysis (IPA) bioinformatics tool, 273 and 306 differentially expressed genes were identified …

Systematics Of Protosteloid Amoebae, Lora Lindley Shadwick

Graduate Theses and Dissertations

Because of their simple fruiting bodies consisting of one to a few spores atop a finely tapering stalk, protosteloid amoebae, previously called protostelids, were thought of as primitive members of the Eumycetozoa sensu Olive 1975. The studies presented here have precipitated a change in the way protosteloid amoebae are perceived in two ways: (1) by expanding their known habitat range and (2) by forcing us to think of them as amoebae that occasionally form fruiting bodies rather than as primitive fungus-like organisms. Prior to this work protosteloid amoebae were thought of as terrestrial organisms. Collection of substrates from aquatic habitats …