Microbiology Commons^™

Relationships Between Fecal Indicator Bacteria And Environmental Factors At Edgewater Beach, Anna M. King

Williams Honors College, Honors Research Projects

To provide more timely estimates of indicator bacteria concentrations in recreational waters, environmental agencies are using predictive models to supplement conventional bacteria enumeration methods. As a tool to develop predictive models, correlational relationships between variables can be examined, to determine the statistical significance of explanatory variables. Prior research at Lake Erie beaches suggests that environmental conditions such as average wave height, rainfall, turbidity, and water temperature may demonstrate a strong relationship to concentrations of fecal indicator bacteria Escherichia coli and Enterococcus faecalis (enterococci). These relationships were examined at Edgewater Beach in Cleveland, Ohio, using statistical correlation tests on data collected …

Metabolism Of Herpes Simplex Virus 1 Infected Raw 264.7 Macrophages, Mary K. Jenkins

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Macrophages are immune cells that phagocytize pathogens and act as antigen presenting cells. Macrophages are critical in regulating the adaptive immune response and play a key role in neutralizing infection. Macrophages exhibit a variety of phenotypes which have different energy requirements. In the case of HSV-1 infection the virus has been shown to alter metabolic processes of its host. Utilizing a XF24 Extracellular Flux Analyzer the metabolic activity of M0, M1, and M2 (IL-4, IL-13, and IL-10) RAW 264.7 macrophages was quantified for both uninfected cells and in response to HSV-1 infection. The analysis showed uninfected M0 and M2 (IL-4, …

The Anti-Apoptotic Effect Of Hsv-1 On Murine Macrophages: Raw 246.7 Murine Macrophage Cell Line, Mofeda Abdussalam Alhanghari

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Herpes simplex virus type 1 (HSV-1) is a worldwide pathogen that affects humans and has the ability to establish a latent state of infection in the sensory nerve ganglia after primary infection of epithelial cells (Boutell and Everett, 2003). HSV-1 is a very contagious virus, which can be transmitted from person to person and cause cold sores in the infected person. Rarely, infection can lead to more serious complications, such as encephalitis. Most HSV-1 infections usually occur in childhood with lifelong potential for symptomatic or asymptomatic viral shedding episodes (Looker et al., 2015). HSV- 1 infects 60%-80% of people throughout …

Clinical Coagulase-Negative Staphylococci; Identification, Characterisation And Treatment, Emma Gabriel

Theses

The work presented in this thesis focuses on clinically significant staphylococci (with an emphasis on coagulase-negative staphylococci) in terms of their isolation, identification, characterisation, and biocontrol.

CoNS are increasingly being recognised as true nosocomial pathogens yet remain under-characterised, particularly from an Irish clinical perspective. We examined a large cohort of clinically-derived CoNS, isolated from blood (n=l35), identifying them to species level. Extensive antibiotic resistance against nine clinically-relevant antibiotics was recorded. Additionally, widespread biofilm forming capabilities associated with increased virulence was detected both phenotypically and genotypically. We identified multiple species of CoNS in circulation in an Irish healthcare setting, and determined …

An Investigation Of The Use Of Natural Agents To Control Mycobacteria, Olga Lesniak

Theses

Economic losses that are caused by Mycobacterium avium paratuberculosis (MAP) infections represent a major obstacle for farmers and also for the dairy industry. In order to control the population of MAP, alternative control agents must be investigated. One of the studies described in this thesis focused on using a bacteriophage therapy based approach to assess its efficacy on simulated industrial environments and food models. Another study described in this thesis investigated the antimicrobial effects of parsley and garlic on a range of mycobacteria species. Both studies used conventional microbiological methods such as plaque assays, AlamarBlue assay (MABA) and growth curves, …

Efficiency Of The Targetron Gene Knockout System As A Transformative Protocol For The Mutagenesis Of Listeria Monocytogenes, Hilaree Noele Frazier

Online Theses and Dissertations

The process of using electroporation to introduce plasmid DNA into host cells is a valuable molecular technique that is increasingly employed in labs worldwide. Electroporators are generally small and relatively inexpensive, making them attractive systems to use for a variety of purposes. Electroporation protocols are numerous in the published literature and encompass all cell types, from prokaryotic bacterial cells to eukaryotic human cells. The TargeTron Gene Knockout System by Sigma-Aldrich is an affordable option for the electroporation of numerous bacterial species. However, its use in Listeria monocytogenes has not been extensively characterized. Here we sought to discuss the effectiveness of …

Late Events Of Human Metapneumovirus Infection: Insights Into Viral Spread Within The Respiratory Epithelium, Farah El Najjar

Theses and Dissertations--Molecular and Cellular Biochemistry

Human metapneumovirus (HMPV) is a leading cause of respiratory tract infections worldwide across all age groups, and is particularly devastating in the pediatric, elderly and immunocompromised populations. Despite its high prevalence and burden on human health, there are currently no treatments or vaccines against HMPV infections. HMPV is an enveloped virus that belongs to the paramyxovirus family. Paramyxoviruses in general form by assembly of virus components at the plasma membrane followed by budding and release of virus particles into the extracellular matrix to spread infection. The process of forming new virus particles requires complex interactions between viral and cellular components …

Delineating ΔNp63α'S Function In Epithelial Cells, Suraj Sakaram

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ΔNp63α, the most physiologically relevant isoform of p63, a p53 family member protein, maintains the proliferative capacity of the basal layer of epithelial tissues and is highly expressed in non-melanoma skin cancer. It has been implicated in a variety of different cancers, including breast, lung, and prostate cancers, however the complex role it plays in disease remains poorly understood. Elucidating ΔNp63α’s regulatory network may provide insight into its role in epithelial tissues and in cancer progression. The objective of this thesis is to investigate ΔNp63α’s upstream regulation in the context of non-melanoma skin cancer as well as expand on ΔNp63α’s …

Species Differences In The Effects Of Tcdd On A Transcriptional Regulatory Region Within The Ig Heavy Chain Gene, Zahra J. Alfaheeda

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The immunoglobulin heavy chain gene (Igh) is regulated by numerous regulatory elements including the 3'Igh regulatory region (3'IghRR). Several transcription factors are involved in modulating the 3'IghRR including NF-B, AP-1, and the aryl hydrocarbon receptor (AhR). The AhR is a ligand-activated transcription factor that mediates the transcription of genes involved in the metabolism of environmental toxicants such as TCDD. TCDD binds AhR and regulates immunoglobulin (Ig) expression in B cells of animal models. This modulation appears to be directly mediated by binding of the AhR to dioxin response elements (DRE) within the 3'IghRR. Structural differences have been found between the …

Comparing Antibody-Coated Immune Beads With Flow Cytometry To Measure Ss-2-Microglubulin+ Murine Spleen Cells, Amal Fahad Alajman

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In diseases like AIDS, Multiple Myeloma, Multiple Sclerosis (MS), Type 1 Diabetes, and Systemic Lupus Erythromatosus the detection of major histocompatibility complex type 1 molecules (MHC 1) can be a helpful component of disease diagnosis and prognosis. Most somatic mammalian cells display to varying degrees major histocompatibility complex (MHC) class 1 antigen on the cell surface. MHC1 molecules consist of a polymorphic alpha a chain and a monomorphic beta chain, beta 2 microglubulin (ß2m). The ß2M composition is fairly constant within a species whereas the alpha chain is not. The polymorphic alpha chain is encoded by an MHC complex gene …

The Impact Of Hsv-1 Infection, Socs1 Peptide, And Socs3 Peptide Mimetic On Cell Viability, Morphology, And Cytoskeleton Proteins Of Unpolarized And Cytokine-Polarized M1 Raw 264.7 Murine Macrophages, Mubarak Almutairi

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Mubarak Huraysan Almutairi. M.S. Department of Microbiology and Immunology, Wright State University, 2016. The Impact of HSV-1 Infection, SOCS1 peptide, and SOCS3 peptide mimetic on Cell Viability, Morphology, and Cytoskeleton Proteins of Unpolarized and Cytokine-Polarized M1 RAW 264.7 Murine Macrophages. The immune response against HSV-1 involves macrophages in both innate and adaptive immunity by limiting HSV-1 replication. In this study, the effects of HSV-1 challenge on cell viability, morphology, and cytoskeletal filament in unpolarized and cytokine-polarized murine RAW 264.7 macrophages at 24 and 48 hours were monitored. Because the distribution of cytoskeleton throughout the cell is critical in cell viability …

Correlating Innate Functional Recovery From Stroke Either With Stem Cell Proliferation And/Or Limb Rehabilitation, Devipriyanka Nagarajan

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In the present study 10-12 month female rats were examined for functional recovery from stroke and this recovery was compared with the stem cell/progenitor cell proliferation in the brain (which was measured by Ki67). The cell proliferation indicated by Ki67 showed a 6 fold increase in control animals compared to the rehabilitation animals. The contralateral functional recovery in control animals were 46.6% and in the rehabilitation animals were 24.5%. The physical rehabilitation was carried out to determine if limb rehabilitation can promote greater functional recovery. The results showed that when the animals were made to over use their impaired limb …

Sequence-Based Methods For Detecting And Evaluating The Human Gut Mycobiome, Mallory J. Suhr, Nabaraj Banjara, Heather E. Hallen-Adams

Department of Food Science and Technology: Faculty Publications

We surveyed the fungal microbiota in 16 fecal samples from healthy humans with a vegetarian diet. Fungi were identified using molecular cloning, 454 pyrosequencing and a Luminex analyte specific reagent (ASR) assay, all targeting the ITS region of the rRNA genes. Fungi were detected in each fecal sample and at least 46 distinct fungal operational taxonomic units (OTUs) were detected, from two phyla — Ascomycota and Basidiomycota. Fusarium was the most abundant genus, followed by Malassezia, Penicillium, Aspergillus, and Candida. Commonly detected fungi such as Aspergillus and Penicillium, as well as known dietary fungi Agaricus bisporus and …

Complete Genome Sequence Of Highly Virulent Porcine Reproductive And Respiratory Syndrome Virus Variants That Recently Emerged In The United States, Aspen M. Workman, Timothy P.L. Smith, Fernando A. Osorio, Hiep L.X. Vu

Nebraska Center for Virology: Faculty Publications

A recent outbreak of particularly virulent disease caused by porcine reproductive and respiratory syndrome virus has occurred in swine herds across the United States. We report here the complete genome sequence of eight viral isolates from four Nebraska herds experiencing an outbreak of severe disease in 2016.

Anaplasma Marginale Actively Modulates Vacuolar Maturation During Intracellular Infection Of Its Tick Vector, Dermacentor Andersoni, Foregivemore Magunda, Chelsea Wright Thompson, David A. Schneider, Susan M. Noh

Biological Sciences Faculty Publications

ABSTRACT Tick-borne transmission of bacterial pathogens in the order Rickettsiales is responsible for diverse infectious diseases, many of them severe, in humans and animals. Transmission dynamics differ among these pathogens and are reflected in the pathogen-vector interaction. Anaplasma marginale has been shown to establish and maintain infectivity within Dermacentor spp. for weeks to months while escaping the complex network of vacuolar peptidases that are responsible for digestion ofthe tick blood meal. How this prolonged maintenance of infectivity in a potentially hostile environment is achieved has been unknown. Using the natural vector Dermacentor andersoni, we demonstrated that A. marginale …

Interactomics-Based Functional Analysis: Using Interaction Conservation To Probe Bacterial Protein Functions, J. Harry Caufield

Theses and Dissertations

The emergence of genomics as a discrete field of biology has changed humanity’s understanding of our relationship with bacteria. Sequencing the genome of each newly-discovered bacterial species can reveal novel gene sequences, though the genome may contain genes coding for hundreds or thousands of proteins of unknown function (PUFs). In some cases, these coding sequences appear to be conserved across nearly all bacteria. Exploring the functional roles of these cases ideally requires an integrative, cross-species approach involving not only gene sequences but knowledge of interactions among their products. Protein interactions, studied at genome scale, extend genomics into the field of …

Investigation Of Anaplasma Phagocytophilum And Anaplasma Marginale Adhesin-Host Cell Interactions, Kathryn S. Hebert

Theses and Dissertations

Anaplasma phagocytophilum and A. marginale are the etiologic agents of bovine anaplasmosis and human granulocytic anaplasmosis, respectively. As obligate intracellular pathogens, binding and entry of host cells is a prerequisite for survival. The molecular events associated with these processes are poorly understood. Identifying the adhesins mediating binding, delineating their key functional domains, and determining the molecular determinants to which they bind not only benefits better understanding of Anaplasma spp. pathobiology, but could also benefit the development of novel approaches for protecting against infection. We previously demonstrated that A. phagocytophilum outer membrane protein A (ApOmpA) is critical for bacterial binding and …

Role Of The Exopolysaccharide Alginate In Adherence To And Inflammation Of Pulmonary Epithelial Cells, Brian E. Crossley

Theses and Dissertations

Pseudomonas aeruginosa (PA) infections in Cystic Fibrosis (CF) patients are not easily cleared due to the conversion from a nonmucoid to a mucoid phenotype. Alginate is an acetylated exopolysaccharide produced by mucoid PA that is responsible for increased resistance to antibiotics, host phagocytic killing, and propagating biofilm formation. Understanding the interaction between PA and host cells is critical to understanding chronic infection and inflammation in CF. In order to investigate this, we used A549 pulmonary epithelial cells and murine alveolar macrophages (MH-S) to examine host response to nonmucoid versus mucoid PA infection. Adhesion assays in A549 pulmonary epithelial cells revealed …

Central, Extraventricular And Atypical Neurocytomas: A Clinicopathologic Study Of 35 Cases From Pakistan Plus A Detailed Review Of The Published Literature, Zubair Ahmad, Nasir Ud Din, Aisha Memon, Mohammad Usman Tariq, Romana Idrees, Sheema Hasan

Department of Pathology and Laboratory Medicine

Background: Central neurocytomas are rare neuronal neoplasms with a favorable prognosis. They are typically located in the lateral ventricles of the brain and mostly histologically correspond to WHO grade II with a Mib 1 labelling index of 2 % and are designated as "atypical neurocytomas.AIM: The aim of our study was to describe the common as well as unusual morphologic features and the role of various immunohistochemical stains in the diagnosis of these rare tumors.MATERIALS AND Methods: We retrieved and reviewed 35 cases diagnosed between 2001 and 2015.Results: Sixty percent of patients were males, and the mean age was 26 …

Errors In Surgical Pathology Reports: A Study From A Major Center In Pakistan, Zubair Ahmad, Romana Idrees, Nasir Uddin, Arsalan Ahmed, Saira Fatima

Department of Pathology and Laboratory Medicine

Background:
Errors in surgical pathology diagnosis can have serious consequences for the patient. Since the final product of a surgical pathology lab is the report, errors can be picked by reviewing reports of cases.AIM: To determine the frequency and types of error in surgical pathology reports of cases signed out in 2014 in a laboratory in Karachi, Pakistan.
Material and Methods:
All surgical pathology reports in which changes were made in the original report after sign out and an amended report was issued were included. Errors included: (1) misinterpretations; (2) missing critical information; (3) erroneous critical information; (4) misidentification; and …

Flaviviruses As A Cause Of Undifferentiated Fever In Sindh Province, Pakistan: A Preliminary Report, Erum Khan, Joveria Q. Farooqi, Kelli L. Barr, Dhani Prakoso, Amna Nasir, Akbar Kanji, Sadia Shakoor, Faisal Riaz Malik, Rumina Hasan, John A. Lednicky

Department of Pathology and Laboratory Medicine

Arboviral diseases are expanding worldwide, yet global surveillance is often limited due to diplomatic and cultural barriers between nations. With human encroachment into new habitats, mosquito-borne viruses are also invading new areas. The actual prevalence of expanding arboviruses is unknown in Pakistan due to inappropriate diagnosis and poor testing for arboviral diseases. The primary objective of this study was to document evidence of flavivirus infections as the cause of undifferentiated fever in Pakistan. Through a cooperative effort between the USA and Pakistan, patient exposure to dengue virus (DENV), West Nile virus (WNV), and Japanese encephalitis virus (JEV) was examined in …

Sporadic Early Onset Colorectal Cancer In Pakistan: A Case- Control Analysis Of Microsatellite Instability, Sabeehuddin Siddique, Kanwal Tariq, Sobia Rafiq, Ahmed Raheem, Rashida Ahmed, Munira Shabbir-Moosajee, Kulsoom Ghias

Department of Pathology and Laboratory Medicine

Background: Early onset sporadic colorectal cancer (CRC) is a biologically and clinically distinct entity hypothesized to exhibit differences in histological features and microsatellite instability (MSI) as compared to typical onset CRC. This study compared the MSI status, mismatch repair enzyme deficiency and clinicopathological features of early onset (aged ≤45 years) with controls (>45 years).
Materials and Methods: A total of 30 cases and 30 controls were analyzed for MSI status using the Bethesda marker panel. Using antibodies against hMLH1, hMSH2 and hMSH6, mismatch repair protein expression was assessed by immunohistochemistry. Molecular characteristics were correlated with clinicopathological features.
Results: …

Toxicity Of Engineered Nanomaterials To Plant Growth Promoting Rhizobacteria, Ricky W. Lewis

Theses and Dissertations--Plant and Soil Sciences

Engineered nanomaterials (ENMs) have become ubiquitous in consumer products and industrial applications, and consequently the environment. Much of the environmentally released ENMs are expected to enter terrestrial ecosystems via land application of nano-enriched biosolids to agricultural fields. Among the organisms most likely to encounter nano-enriched biosolids are the key soil bacteria known as plant growth promoting rhizobacteria (PGPR). I reviewed what is known concerning the toxicological effects of ENMs to PGPR and observed the need for high-throughput methods to evaluate lethal and sublethal toxic responses of aerobic microbes. I addressed this issue by developing high-throughput microplate assays which allowed me …

Tillage And Fertilization Influences On Autotrophic Nitrifiers In Agricultural Soil, Shuang Liu

Theses and Dissertations--Plant and Soil Sciences

Nitrification is a biological oxidation of NH₃ to NO₂^- and then to NO₃^-. Understanding how the nitrifier community responds to agricultural management is essential because the community composition is complex and functional distinction of subgroups occurs. Better managing nitrifiers could benefit the environment by increasing nitrogen (N) fertilizer use efficiency, decreasing NO₃^- leaching, and reducing NO and N₂O emissions. This study examined how long-term N fertilization and tillage influenced nitrifier density, ratios, nitrification rates, and the community structure of ammonia-oxidizing bacteria (AOB), ammonia-oxidizing archaea (AOA), and nitrite-oxidizing bacteria (NOB). The …

Studies On Regulation Of Pqq-Dependent Phosphate Solubilization Among Rhizosphere Dwelling Bacteria, Ran An

Theses and Dissertations--Plant and Soil Sciences

Plant growth promotion can be enhanced by soil- and rhizosphere-dwelling bacteria by a several different methods. One method is by promoting nutrient acquisition from soil. Phosphorus is an essential nutrient that plants obtain from soil, but in many cases it is immobilized in forms that are not available for plant uptake. Bacteria can solubilize insoluble soil phosphates by secreting gluconic acid. This chemical is produced from glucose by the activity of the bacterial enzyme glucose dehydrogenase, which requires a coenzyme called PQQ. In this dissertation, I studied the regulation of the glucose dehydrogenase enzyme and the PQQ coenzyme in the …

Response Of Soil Microbial Communities To Saltwater Intrusion In Tidal Freshwater Wetlands, Chansotheary Dang

Theses and Dissertations

Saltwater intrusion due to global change is expected to have a detrimental effect on the biogeochemistry of tidal freshwater wetlands. Of particular concern is that fact that salinization can alter the role of these ecosystems in the global carbon cycling by causing shifts in microbial metabolism that alter greenhouse gas emissions and increase carbon mineralization rates. However, our understanding of how wetland microbial community dynamics will respond to saltwater intrusion is limited. To address this knowledge gap and increase our understanding of how microbial communities in tidal freshwater wetlands change over time (1, 3, 12, and 49 weeks) under elevated …

Diversity Of The Beneficial Bacterium Oxalobacter Formigenes Isolated From The Human Gut, Karen Shaw

Masters Theses

Oxalobacter formigenes is an anaerobic, oxalate degrading organism that colonizes in the mammalian intestinal tract. Currently, O. formigenes is divided into two groups via PCR of the oxc gene. Pulsed field gel electrophoresis (PFGE) is a technique used to resolve large pieces of DNA. Typically, PFGE is used to identify differences between strains for pathogenic bacteria, but this study sought to use it to further understand the diversity of a probiotic organism. This study aimed to develop a methodology for utilizing PFGE to examine O. formigenes. The methodology involved lysing O. formigenes cells in plugs, first in proteinase K, …

Characterization Of Insertion Sequence Is605 In Halanaerobium Hydrogeniformans, Michael C. Sadler

Masters Theses

"Insertion sequences are the smallest prokaryotic transposable elements. These genes play a significant evolutionary role by promoting genome plasticity. Insertion sequences are highly diverse elements that have largely been uncharacterized. As such, the ability to accurately identify, annotate, and infer genomic impact of insertion sequences is lacking. The study of new insertion sequences contributes knowledge to their annotation and evolution. Halanaerobium hydrogeniformans is a unique organism with an abnormally high number of insertion sequences. A family of insertion sequences, IS200/605, showed several interesting distinctions from other elements in the genome, including severe open reading frame degradation, and was characterized in …

Characterizing The Role Of The Fact Complex In Hiv-1 Infection, Angelica Patricia Lopez

Open Access Theses & Dissertations

The lens epithelium-derived growth factor p75 (LEDGF/p75) is a chromatin-bound protein essential for efficient lentiviral integration. Knockdown of this protein severely affects HIV-1 integration levels and thus itâ??s replication. Genome-wide studies have located LEDGF/p75 inside actively transcribed genes where it mediates lentiviral integration. Although this is clearly established, the molecular mechanisms of LEDGF/p75 in HIV-1 infection remain in part elusive.

Using protein-protein interaction assays, we demonstrated that LEDGF/p75 complexes with a chromatin remodeling complex, Facilitates Chromatin Transcription (FACT), a heterodimer of the structure specific recognition protein 1 (SSRP1) and the human homolog of suppressor of Ty 16 (hSpt16). The FACT …

Defining And Characterizing The Step In The Hiv-1 Viral Life Cycle Affected By Fullerene Derivatives, Zachary Sean Martinez

Open Access Theses & Dissertations

Fullerene derivatives, both C60 and C70, were synthesized and characterized to test their activity against HIV-1 infection. The derivatives tested have previously been reported to inhibit HIV-1 replication and/or in vitro activity of reverse transcriptase. It was found that viruses produced in the presence of low micromolar concentrations of most of these fullerene derivatives exhibit a reduction in viral infectivity greater than 99%. Quantification of virion-associated viral RNA and p24 indicates that RNA packaging and viral production were unremarkable in these viruses. These compounds did not affect infectivity of mature virions, indicating no effect on the early steps of the …