Microbiology Commons^™

Vertical Transmission Of Zika Virus In Monodelphis Domestica, Mohini Moulick

Theses and Dissertations

Zika Viral infection in pregnant women may lead to infants born with microcephaly and other neurological complications, making Zika viral research imperative. Prior research demonstrated Monodelphis domestica to be a unique animal model for Zika virus studies, and viral proteins were detected in brain tissue of juveniles after intracerebral inoculation of infants. We wanted to determine if we can detect a viral protein, non-structural protein 1 (NS1), in brain tissue after intraperitoneal inoculation and we found it to be possible through immunohistochemistry (IHC) analysis. We also investigated if inoculated dams (mothers) would give rise to Zika positive pups (natural inoculation), …

The Kinetics Of Biocontaminant Inactivation: A Comparative Study With Bacillus Globigii And Bacillus Anthracis, Leigh M. Durden

Theses and Dissertations

Bacillus anthracis has been a biological threat for decades and after the 2001 terrorist attack via the U.S. Postal Service where five people died, there was heightened awareness and inquiry on the subject (Goel, 2015; Greenberg et al., 2010; Nicholson & Galeano, 2003; Spotts Whitney et al., 2003). The health risk to humans makes this biological agent, a Tier 1 agent and one of the most likely to be used in an attack (Anthrax as a Bioterrorism Weapon | CDC, n.d.; Driks, 2009; Rose & Rice, 2014). B. anthracis is one of most resistant and potent biological warfare agents as …

Development Of Whole-Cell Biosensors For Early Detection Of Oral Squamous Cell Carcinoma, Sharona Adina Priyev

Theses and Dissertations

Accurate and timely diagnosis is crucial in preventing progression to advanced disease stages and in improving survival rates of oral squamous cell carcinoma (OSCC). Using synthetic biology tools, we aim to integrate synthetic biological circuits into the construction of a novel, whole-cell biosensor (WCB) that will detect salivary biomarkers of OSCC. In treating biological parts and genetic circuits as modular pieces, we can intentionally rearrange selected domains of interest to create new transcription factors that will enable the detection of molecules of interest. Our end-goal is the creation of portable devices carrying living cells that express our synthetic genetic circuits …

Probing The Porphyromonas Gingivalis Nitrosative Stress Response Network, Alexandra Galina

Theses and Dissertations

Objective: To persist in the oral cavity, the perio-pathogen Porphyromonas gingivalis must have an efficient nitrosative stress response mechanism. To better understand how P. gingivalis responds to nitrite (NO₂^-) we must determine what genes are involved in the nitrosative stress response network.

Methods: Through genomic screening, a list of genes was generated that were hypothesized to be part of the nitrosative stress response network and believed to be involved in nitrate/nitrite metabolism or redox state of the cells. Protein coding regions were replaced with the ermF sequence by allelic exchange mutagenesis. 15 mutants were created …

The Type Iv Pilus Secretin Bfpb: Structural Analysis And Binding Interactions, Janay I. Little

Theses and Dissertations

Enteropathogenic Escherichia coli (EPEC) causes severe diarrhea in young children. The type IV pilus (T4P) of EPEC, known as the bundle-forming pilus (BFP), plays an important role in EPEC pathogenesis. T4Ps are a family of surface appendages that are important for adhesion, colonization, biofilm formation, virulence, twitching motility and many other functions. One essential component of the BFP system is the secretin, BfpB. Secretins are a large family of integral outer membrane proteins found in T4Ps as well as type II and type III secretion systems, and filamentous phages. Details of the secretin structure have been limited to the overall …

Treponema Denticola Synthesizes C-Di-Amp And Encodes The Cdaa-Type Diadenylate Cyclase Cdaa, Claire R. O'Brien

Theses and Dissertations

Periodontitis is a form of oral disease characterized by dysbiosis of the oral microbiome, leading to inflammation, bone resorption, and in severe cases, entire tooth loss, affecting 42% of adults in the US. One of the bacteria most associated with periodontal disease progression is Treponema denticola (Td), an oral spirochete which inhabits the mouth in small quantities during health but which can dominate the biofilms that form during periodontal disease. The ability of Td to survive in a disease environment and contribute to the progression of disease requires the use of robust signaling networks. Analysis of Td cultures …

Evaluation Of Novel Litter Amendments For Use In The Commercial Broiler Industry, Jorge A. Urrutia

Theses and Dissertations

Poultry litter amendments (LA) are commonly used to maintain good litter quality; the most common ones; are acidifiers. However, concern over the lifespan of LA acidifiers (d) and corrosive nature revealed a need for alternative LA. Products of interest include biochar (BC), superabsorbent polymers (SAPs), and alum mud (AM). Experiment 1 was a screening study (n=17 treatments, no birds) conducted to determine the optimal application rate/method of BC, SAPs, and AM vs.commercial LA and a control. Most SAPs treatments measured metrics may be similar (i.e., bacteria, water activity) or better (pH) vs. commercial LA (PLT). High inclusions of novel LA …

Survival, Regrowth And Morphotype Formation Of Salmonella Strains After Exposure To High Or Low Concentrations Of First Generation Qac In Water, Grace Umutesi

Theses and Dissertations

This study determines the differences in survival of eight Salmonella strains concentration of BAC in water, followed by regrowth in high and low nutrition conditions containing sublethal concentration of BAC. Our results show strain difference in survival, persistence of Salmonella in BAC at different concentrations. All eight strains of Salmonella were non-recoverable when exposed to 40 to 48 µg/ml BAC for 1h in water. When exposed to 24 µg/ml of BAC for 1 h in water, S. Typhimurium ATCC 14028 and S. Heidelberg ATCC 8326 cells were non-detectable from the initial 7 log CFU/ml. S. Blockley 7175, S. Virchow 7207, …

Metagenomic Analysis Of Root-Associated Microbiome Of Healthy And Taproot Decline-Affected Soybeans And Identification Of Healthy Soybean Root Endophytes With Protective Activity Against The Causal Agent, Xylaria Necrophora, Uyen Wesser

Theses and Dissertations

Plant roots are associated with a diverse microbial community of bacteria and fungi. The root microbiome communities associate with the root surface or penetrate the superficial layers of root tissues. It is of high agronomic interest to characterize root microbiomes and identify microbes with beneficial effects on plant defense against pathogens. Soybean taproot decline (TRD) is an emerging soil-borne disease caused by a fungus classified as Xylaria necrophora, challenging to control and with devastating effects on yield. Our group has initiated a study of the soybean root microbiome with three primary experiments: (I) perform a comparative study of root …

Effects Of Microcystin-Lr On Channel Catfish (Ictalurus Punctatus) Susceptibility To Microbial Pathogens (Aeromonas Hydrophila And Edwardsiella Piscicida), Alison Marchant

Theses and Dissertations

Microcystin-LR is a hepatotoxin produced by cyanobacteria. Aeromonas hydrophila and Edwardsiella piscicida infections are leading causes of losses in market-sized channel catfish (Ictalurus punctatus). These older fish should have natural immunity in place and a predisposing factor is likely a prerequisite for these disease outbreaks. While microcystin-LR rarely causes mortality in warm-water aquaculture, we believe it may be a predisposing factor that leads to bacterial disease outbreaks during the summer months due to its ability to damage the liver. Our study investigated microcystin-LR’s effects on channel catfish susceptibility to these pathogens. We found that a sublethal dose of microcystin-LR induced …

Molecular And Phenotypic Characterization Of Escherichia Coli Isolated From Broiler Chicken Flocks In Mississippi, Priyanka Devkota

Theses and Dissertations

Avian pathogenic Escherichia coli (APEC) is the causative agent of colibacillosis. APEC causes significant economic losses to the poultry industry. In this study, 66 E. coli isolates were collected from broiler flocks across Mississippi and categorized as clinical and non-clinical isolates. Genotypic and phenotypic characterization of virulence factors and antimicrobial resistance profiles of these E. coli were examined. The data disclosed a higher prevalence of virulence genes in clinical isolates than in non-clinical isolates. High differences on the genotypic and phenotypic antimicrobial resistance among clinical and non-clinical isolates were observed. Whole genome sequences of avian E. coli elaborated a diverse …

Effects Of Mass Death On Community Structure And Ecosystem Function, Abby Kimpton Jones

Theses and Dissertations

Death and decomposition are natural processes that are generally well-understood. However, large events of death, such as mass mortality events (MMEs) are increasing in frequency and their impacts on the ecosystem are largely unknown. These events may have both bottom-up effects from increased nutrient input as well as top-down effects from loss of an ecological functional group by the affected population. Different functional MMEs may result in different top-down effects, creating cascading effects. In Chapter 1, I test the hypothesis that scavenger and herbivore simulated MMEs generate novel bottom-up and top-down effects. Results indicate that MMEs have a significant effect …

Evaluating Soil Health Changes Following Cover Crop And No-Till Integration Into A Soybean (Glycine Max) Cropping System In The Mississippi Alluvial Valley, Alexandra Gwin Firth

Theses and Dissertations

The transition of natural landscapes to intensive agricultural uses has resulted in severe loss of soil organic carbon (SOC), increased CO₂ emissions, river depletion, and groundwater overdraft. Despite negative documented effects of agricultural land use (i.e., soil erosion, nutrient runoff) on critical natural resources (i.e., water, soil), food production must increase to meet the demands of a rising human population. Given the environmental and agricultural productivity concerns of intensely managed soils, it is critical to implement conservation practices that mitigate the negative effects of crop production and enhance environmental integrity. In the Mississippi Alluvial Valley (MAV) region of Mississippi, USA, …

Optimized Selenium Status, Gut Microbiota, And Type 2 Diabetes, Ying-Chen Huang

Theses and Dissertations

We have previously demonstrated that long-term dietary Se deficiency in old Terc^-/- mice with humanized telomeres induces type-2 diabetes and exacerbates age-dependent increases in the abundance of A. muciniphila and Lachnospiraceae, which are related to obesity and metabolic syndromes. The objectives of this dissertation are: 1) to determine the minimum intake of Se required for type 2 diabetes prevention in middle-aged mice; 2) to evaluate the efficacy of A. muciniphila and R. torques (a Lachnospiraceae family member) to intervene dietary Se deficiency-induced type 2 diabetes and the underlying mechanisms; 3) to assess sex differences in the responses to …

Identification Of Pneumococcal Membrane Proteins Involved In Colonization/Biofilm Formation And Cognate Host Cellular Receptors, Yoonsung Hu

Theses and Dissertations

Colonization is prerequisite for infection and transmission of Streptococcus pneumoniae, or pneumococcus. Currently available pneumococcal conjugate and pneumococcal polysaccharide vaccines can provide protection against a limited number of capsular serotypes. Implementation of vaccines has decreased the frequency of invasive pneumococcal disease and their colonization rates, but only in a serotype-dependent manner. This has led to serotype replacement in pneumococcal ecology and increased invasive disease caused by non-vaccine serotypes. Development of conserved protein-based vaccine that can provide protection against all pneumococcal serotypes is needed. Numerous surface proteins are conserved in all serotypes, and some are known to be involved in …

An Analysis Of The Effects Of Temperature And Other Environmental Factors On Microorganismal Productivity Within Aquatic Ecosystems Using Long-Term Data From Various Lter And Ilter Sites, Andrew Dorbu

Theses and Dissertations

Global climate change is a phenomenon resulting in more extreme weather patterns and species diversity loss. In this study, I explore the impacts of climate change on regional patterns of microbial productivity. Variation in microbial productivity was explained using temperature, salinity, and dissolved oxygen as predictors in regression models with data from the LTER and iLTER network of sites that ranged from the poles to the equator. I found a positive relationship between temperature and productivity. Antarctica and Arctic sites exhibited the strongest positive relationships supporting prior research demonstrating temperature as one of the driving forces of productivity change in …

Development Of A Methodology For The Quantification Of Reaerosolization Of A Biological Contaminant Surrogate Particle From Military Uniform Fabric, George D. Cooksey

Theses and Dissertations

During a mass casualty medical evacuation after a bioaerosol attack, a decontamination method is needed that is effective at both decontamination and preventing the secondary hazard of biological particles reaerosolizing from contaminated clothing. However, neither the efficacy of current decontamination methods nor the risk of biological particle reaerosolization is significantly explored in existing literature. The goals of this thesis were to develop a repeatable methodology to quantify the reaerosolization of a biological contaminate off Airman Battle Uniform (ABU) fabric swatches, and to test the efficacy of one decontamination method (high-volume, low-pressure water) using 1 mpolystyrene latex (PSL) spheres as a …

Heterologous Production Of Plant Secondary Metabolites Resveratrol And Caffeic Acid In E.Coli And Synechocystis Sp. Pcc 6803, Yong Xue

Theses and Dissertations

Caffeic acid and resveratrol are polyphenol compounds produced by plants as secondary metabolites to protect against biotic and abiotic stresses. In the last decades, numerous studies have demonstrated the beneficial effects of these compounds on human health as anticancer, antioxidant, anti-virus and anti-inflammatory agents. In plants, both caffeic acid and resveratrol are synthesized either from phenylalanine or tyrosine. Recently, heterologous production of phenylpropanoids using genetically engineered bacteria is gaining more and more attentions from industry because it can dramatically improve the yield with reduced costs. The major aim of the work is to explore the possibility of using Synechocystis or …

Design Of A Synthetic Temperature-Activated Epigenetic Biocontainment System, Owen Rivers Page

Theses and Dissertations

The need for an internal control to govern engineered strain survival is becoming apparent when considering the biosafety and ethics within biotechnological research. Using understudied regulatory elements known as RNA thermometers (RNATs) that can modulate protein translation based on temperature changes and a Type II restriction modification system, we posit the creation of a synthetic temperature-activated epigenetic biocontainment system, termed the ‘sentinel switch’. We have mined a wide variety of bacteria from Yersinia pestis to Synechocystis sp. looking for novel RNATs. A complementary synthetic biology approach based on library generation and screening of chimeric RNATs (ChRNATs) was also conducted. RNATs …

The Influence Of Salinity On The Structure And Function Of Soil Prokaryotic Communities In Coastal Freshwater Wetlands, Joseph C. Morina Ii

Theses and Dissertations

Microbial communities are critical biological components of the world’s ecosystems, and their respiratory pathways are directly involved in the biogeochemical cycling of essential nutrients. As genomic technologies advance, allowing for more detailed profiling of microbial communities, efforts have successfully linked microbial community composition to ecosystem-level functions and have shown microbial communities are susceptible and resistant to disturbance events. The goal of this dissertation is to address the temporal scales in which microbial communities respond to the disturbance of salinization, and the repercussions this has on microbially-meditated carbon and nitrogen cycling. Coastal freshwater wetlands are an excellent study system to investigate …

Antiviral Effects Of Metalloshielding: Differential Antiviral Activity Of Polynuclear Platinum And Cobalt Compounds, Mary Zoepfl

Theses and Dissertations

The majority of antiviral drug development has focused on virus-specific discovery targeting discrete steps in the individual life cycles. Although great strides have been made for a number of clinically relevant diseases such as human immunodeficiency virus, influenza virus, and hepatitis B, broad spectrum antivirals do not exist. Broad spectrum antivirals would offer (1) treatment for viruses without specifically-targeted antivirals, (2) treatment for viruses which have developed resistance to their available treatments, and (3) a rapidly deployable treatment option in viral epidemics. Many viruses including human cytomegalovirus (HCMV), HIV, and SARS-CoV-2. rely on heparan sulfate (HS), a highly sulfated glycosaminoglycan …

Human Papillomavirus 16 E2 Regulates Host Cell Pathways Important For Cancer Progression And Treatment Sensitivity Which May Contribute To Cancer Outcomes, Christian Fontan

Theses and Dissertations

Human papillomaviruses (HPVs) are causative agents in around 5% of all cancers, including cervical and oropharyngeal. A feature of HPV cancers is their better clinical outcome compared with non-HPV anatomical counterparts. In turn, the presence of E2 predicts a better clinical outcome in HPV-positive cancers; the reason(s) for the better outcome of E2-positive patients is not fully understood.

Previously, we demonstrated that HPV16 E2 regulates host gene transcription that is relevant to the HPV16 lifecycle in N/Tert-1 cells. One of the genes repressed by E2 and the entire HPV16 genome in N/Tert-1 cells is TWIST1. In these studies, we demonstrate …

Modification Of Culture Medium And Identification Of Microbial Contaminants For Improved In Vitro Propagation Of Freshwater Mussels, Raquel M. Wetzell

Theses and Dissertations

In vitro propagation efforts play an essential role in conserving and restoring threatened freshwater mussel populations by circumventing the need for a fish host. Across a broad range of taxa, transformation is induced with an artificial M199 medium and rabbit serum. However, such formulation may not be sufficient in culturing critical species with more specific physiological requirements. In this study, multiple serum mixtures were tested to improve in vitro transformation of two freshwater mussel species: yellow lampmussel (Lampsilis cariosa) and tidewater mucket (Atlantaconcha ochracea). These species were selected because they parasitize similar fish host species but …

Bioinformatic Pipeline For Determining Terminal Repeats In The Human Cytomegalovirus Genome Assembled With Pacbio Long Read Sequences, Ahmed Al Qaffas

Theses and Dissertations

Human Cytomegalovirus (HCMV) is a member of the betaherpesvirinae subfamily of the Herpesvirus family. HCMV infection is common among adults worldwide, with an estimated seroprevalence of 66 to 95%, depending on the geographic region (Zuhair et al., 2019). Although most of the virus genomic content has been studied extensively, the terminal repeating region sequences remain understudied. Two main challenges hindered the study of the region: a) limitations of sequencing technologies; and b) misassembly of the repeats due to its complex nature. Here I show a novel bioinformatics pipeline that takes advantage of PacBio's long reads to resolve the challenges mentioned …

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 …

Genetic Characterization Of Antimicrobial Activities Of Endophytic Bacteria Burkholderia Strains Ms455 And Ms389, Jiayuan Jia

Theses and Dissertations

Strains MS455 and MS389, endophytic bacteria, were isolated from healthy soybean plant growing adjacent to a patch of plants affected by charcoal rot disease, caused by the fungal pathogen Macrophomina phaseolina. The complete genomes of both strains were sequenced and identified as Burkholderia species Strain MS455 exhibits broad-spectrum antifungal activities against economically important pathogens, including Aspergillus flavus. Random and site-specific mutations were employed in discovery of the genes that share high homology to the ocf gene cluster of Burkholderia contaminans strain MS14, which is responsible for production of the antifungal compound occidiofungin. RNA-seq analysis demonstrated ORF1, a …

Prospecting For And Isolation Of Microalgae In South Texas, Mauricio Alejandro Peña

Theses and Dissertations

Microalgae is a very important microorganism that can be used for beneficial purposes for humans. One way to show the importance of algae in this planet is the contribution of Oxygen it provides to the atmosphere, which many organisms rely on (Medeiros et al., 2015). Carbon footprint has been one of the greatest causes for global warming and it is due to the use of fossils to create diesel. One of the ways to avoid contamination is to replace those fuels that are used on daily basis, such as for transportation. The microalgae are able to produce lipids that can …

Insights Into Halophilic Microbial Adaptation: Analysis Of Integrons And Associated Genomic Structures And Characterization Of A Nitrilase In Hypersaline Environments, Sarah Sonbol

Theses and Dissertations

Hypersaline environments are extreme habitats that can be exploited as biotechnological resources. Here, we characterized a nitrilase (NitraS-ATII) isolated from Atlantis II Deep brine pool. It showed higher thermal stability and heavy metal tolerance compared to a closely related nitrilase.

We also studied integrons in halophiles and hypersaline environments. Integrons are genetic platforms in which an integron integrase (IntI) mediates the excision and integration of gene cassettes at specific recombination sites. In order to search for integrons in halophiles and hypersaline metagenomes, we used a PCR-based approach, in addition to different bioinformatics tools, mainly IntegronFinder.

We found that integrons and …

Uncovering Potential Biofilm Modulators, Liliana Margent

Theses and Dissertations

Biofilms have negative and positive impacts depending on the context they develop. In this work, I optimized a biofilm assay to evaluate the effect of three different types of compounds indole derivatives, 2,5 Diketopiperazines, and 12-hydroxystearic acid on the biofilm formation of E. coli.

Mining Selected Metagenomes/Metatranscriptomes For Biosynthetic Gene Clusters And Antimicrobial Resistance Genes, Ahmed Yamany

Theses and Dissertations

Antimicrobial resistance is one of the serious global challenges in the current century. The fact that resistance genes transfer between bacteria, coupled with the fact that the world is connected through complex dynamics. Studying microbial behavior and understanding the different factors coffering microbial resistance to a broad spectrum of the available drug classes, parallel with a comprehensive analysis of the natural microbial products as the primary source of the novel antibiotics, might shed some light on solutions for this problem. Microbial environments harbor a wide range of secondary metabolites (SM) with different functional groups. SMs are not directly involved in …