Microbiology Commons^™

Genotyping And Drug Resistance Patterns Of M. Tuberculosis Strains In Pakistan., Mahnaz Tanveer, Zahra Hasan, Amna R Siddiqui, Asho Ali, Akbar Kanji, Solomon Ghebremicheal, Rumina Hasan

Department of Pathology and Laboratory Medicine

Background:The incidence of tuberculosis in Pakistan is 181/100,000 population. However, information about transmission and geographical prevalence of Mycobacterium tuberculosis strains and their evolutionary genetics as well as drug resistance remains limited. Our objective was to determine the clonal composition, evolutionary genetics and drug resistance of M. tuberculosis isolates from different regions of the country.

Methods:

M. tuberculosis strains isolated (2003-2005) from specimens submitted to the laboratory through collection units nationwide were included. Drug susceptibility was performed and strains were spoligotyped.

Results:

Of 926 M. tuberculosis strains studied, 721(78%) were grouped into 59 "shared types", while 205 (22%) were identified …

The Role Of Msa In Staphylococcus Aureus Biofilm Formation, Karthik Sambanthamoorthy, Antony Schwartz, Vijayaraj Nagarajan, Mohamed O. Elasri

Faculty Publications

Background

Staphylococcus aureus is an important pathogen that forms biofilms. The global regulator sarA is essential for biofilm formation. Since the modulator of sarA (msa) is required for full expression of sarA and regulates several virulence factors, we examined the capacity of the msa mutant to form biofilm.

Results

We found that mutation of msa results in reduced expression of sarA in biofilm and that the msa mutant formed a weak and unstable biofilm. The msa mutant is able to adhere to surfaces and begins to form biofilm but fails to mature indicating that the defect of the …

Iron Acquisition In Rhodococcus Erythropolis Strain Igts8: Characterization Of A Mutant Strain That Over Produces Siderophore., Melanie Anne Pratt

Electronic Theses and Dissertations

Iron is an essential nutrient for most bacteria because enzymes like nitrate reductase and cytochromes use it as a cofactor. However, in most aerobic, neutral pH environments, iron is essentially insoluble and not easily available for bacteria to use. Many bacteria respond to this problem by releasing small organic compounds called siderophores that bind and effectively solubilize iron so that it can be transported into the cell for growth. The focus of this study was to learn more about the iron acquisition and especially the transport of iron by the soil bacterium Rhodococcus erythropolis. To fulfill this aim, mutant …

Herpes Simplex Virus Glycoprotein D/Host Cell Surface Interaction Stimulates Chlamydia Trachomatis Persistence Via A Novel Pathway., Jennifer Vanover

Electronic Theses and Dissertations

When presented with certain unfavorable environmental conditions, C. trachomatis reticulate bodies (RBs) enter into a viable, yet noncultivable state called persistence. Two hallmarks of persistent chlamydiae are swollen, aberrantly shaped RBs, as viewed by transmission electron microscopy and a decrease in infectious progeny. Several models of chlamydial persistence have been described, including interferon-γ (IFN-γ), IFN-α, IFN-β, and tumor necrosis factor-α-exposure and nutrient deprivation. Previously, we established an in vitro co-infection model of two of the most common sexually transmitted pathogens in the United States, C. trachomatis and Herpes Simplex Virus-2 (HSV). Data from this tissue culture model indicate that: i) …

A Population Genetic Study Of Middle Eastern Populations Using Dys 458 Microvariants And Cohen Modal Haplotypes, Enass Nabeel Tinah

Theses and Dissertations

A comprehensive population study in the Middle East was conducted using different genetic markers in order to establish a wider genetic profile of the Middle Eastern populations. The main goal of this study was to analyze DNA from samples collected from different locations, and produce genetic motifs and patterns that could be used to identify and distinguish the target populations. This information will allow us to analyze the ancestry of these populations, their interactions through time and space, and the effects these interactions have on the populations' structure. We have collected around 1300 individual samples from different populations in the …

Effects Of Shielding Adenoviral Vectors With Polyethylene Glycol On Vector-Specific And Vaccine-Mediated Immune Responses, Eric A. Weaver, Michael A. Barry

Nebraska Center for Virology: Faculty Publications

Many individuals have been previously exposed to human adenovirus serotype 5 (Ad5). This prior immunity has long been known to hinder its use for gene therapy and as a gene-based vaccine. Given these immunogenicity problems, we have tested whether polyethylene glycol (PEG) can blunt immune effects against Ad5 during systemic and mucosal vaccination. Ad5 vectors were covalently modified with 5-, 20-, and 35-kDa linear PEG polymers and evaluated for their ability to produce immune responses against transgene antigen products and the vector itself. We show that shielding Ad5 with different-sized PEGs generally reduces transduction and primary antibody responses by the …

The Human Immunodeficiency Virus Type 1 Envelope Confers Higher Rates Of Replicative Fitness To Perinatally Transmitted Viruses Than To Nontransmitted Viruses, Xiaohong Kong, John T. West, Hong Zhang, Danielle Shea, Tendai J. M'Soka, Charles Wood

Nebraska Center for Virology: Faculty Publications

Selection of a minor viral genotype during perinatal transmission of human Immunodeficiency virus type 1 (HIV-1) has been observed, but there is a lack of information on the correlation of the restrictive transmission with biological properties of the virus, such as replicative fitness. Recombinant viruses expressing the enhanced green fluorescent protein or the Discosoma sp. red fluorescent (DsRed2) protein carrying the V1 to V5 regions of env from seven mother-infant pairs (MIPs) infected by subtype C HIV-1 were constructed, and competition assays were carried out to compare the fitness between the transmitted and nontransmitted viruses. Flow cytometry was used to …

A Microbiological Survey And Characterization, Andrew Daane

All Theses

In our study, two dairy compost heaps and one vegetable scrap compost heap were constructed on two research farms in Clemson, SC. Samples were taken from each heap from different locations at 8 elected intervals and analyzed for enterococci and VRE counts by spread plating on Bile Esculin agar (BEA) and BEA containing 6 g/ml of vancomycin, respectively. The initial populations of enterococci and VRE in compost were in the range of 6.46-7.43 and 5.36-6.70 log CFU/g, respectively. After 30 days of active composting the average VRE populations declined ca. 4.6, 4.07, 3.97 and 1.6 logs at the top, center, …

Dechlorinating And Iron Reducing Bacteria Distribution In A Trichloroethene Contaminated Aquifer, Carmen Lourdes Yupanqui Zaa

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

The Operable Unit 5 (OU 5) area of Hill Air Force Base currently has two trichloroethene-contaminated groundwater plumes underneath residential areas in Sunset and Clinton, Utah. Bioremediation by biological reductive dechlorination can be an important mechanism for the removal of chlorinated compounds from the plumes. The presence of suitable bacteria to carry on reductive dechlorination is the key in the bioremediation process. The goal of this study was to determine the distribution and population density of the 16S rRNA genes of Bacteria, Dehalococcoides ethenogenes, Desulfuromonas michiganensis, Geobacter spp and Rhodoferax ferrireducens-like bacteria, as well as the functional genes trichloroethene reductive …

A Cell Biology Laboratory Exercise To Study Sub-Cellular Organelles In Drosophila, Meghana Tare, Amit Singh

Biology Faculty Publications

The fast-changing scenario of undergraduate education puts emphasis on introducing students to hands-on techniques as part of their laboratory courses. In order to cater to large numbers of students and the time constraints involved with undergraduate level laboratory courses, there is a need for development of experiments that are cost effective and can be completed in a defined time frame. We have devised a laboratory exercise for teaching cell biology using the Drosophila melanogaster model. Drosophila can be reared in a short period of time in a cost effective manner. We used Drosophila tissue to study the sub-cellular organization of …

Aminoglycosides And Syringomycin E As Fungicides Against Fusarium Graminearum In Head Blight Disease, Yukie Kawasaki

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Fusarium graminearum is one of the most problematic phytopathogens in US agriculture. This fungus causes head blight, foot rot, and damping off on wheat and barley. The infection lowers the grain yield and causes contamination of the grain product with mycotoxins. Effective control measures are lacking, and new fungicides that kill F. graminearum but remain safe and economical to use are needed. Newly synthesized aminoglycosides (JL22, JL38, JL39, JL40, NEOF004, NEOF005), classic aminoglycosides (amikacin, gentamicin, kanamycin A, kanamycin B, neomycin, and ribostamycin), and a lipopeptide, syringomycin E (SRE), were studied to determine their antifungal potential to control F. graminearum. …

Evolution Of Picornaviruses: Impacts Of Recombination And Selection, Nicole Noel Lewis-Rogers

Theses and Dissertations

Picornaviruses are responsible for some of the most common and debilitating diseases affecting humans and animals worldwide. The objectives of this dissertation research were (1) estimate phylogenetic relationships among 11 picornavirus genera and within three species: foot-and-mouth disease virus (FMDV: Aphthovirus) which afflicts cloven-hoofed animals and human rhinovirus A and B (HRV: Enterovirus) which cause the common cold; (2) better understand the impact recombination has on genomic organization and evolution; (3) characterize where positive and purifying selection has occurred in proteins and how selection has influenced phenotype. The dissertation includes four studies. The first chapter provides an overview of the …

Specific Compartmentalization Of Iga Ascs In Mouse Salivary Glands Via Differential Expression Of Chemokines And Chemokine Receptors, Yuet Ching Law

Theses and Dissertations

The mucosal system, which forms a barrier between internal organ systems and the external environment, is frequently exposed to pathogenic microorganisms. Immunoglobulin A (IgA) antibody secreting cells (ASCs) localize in the lamina propria, and produce IgA antibodies which help protect mucosal tissues. The concept of a common mucosal immune system in which IgA ASCs have the ability to populate any mucosal site has been proposed (1, 2). However, recent research has suggested that IgA ASCs primed in different mucosal sites might possess different sets of chemokine receptors, and therefore migrate specifically to particular mucosal locations (3). In this study, the …

Exploring The Mialome Of Ticks: An Annotated Catalogue Of Midgut Transcripts From The Hard Tick, Dermacentor Variabilis (Acari: Ixodidae), Jennifer M. Anderson, Daniel E. Sonenshine, Jesus G. Valenzuela

Biological Sciences Faculty Publications

Background: Ticks are obligate blood feeders. The midgut is the first major region of the body where blood and microbes ingested with the blood meal come in contact with the tick's internal tissues. Little is known about protein expression in the digestive tract of ticks. In this study, for analysis of global gene expression during tick attachment and feeding, we generated and sequenced 1,679 random transcripts (ESTs) from cDNA libraries from the midguts of female ticks at varying stages of feeding.

Results: Sequence analysis of the 1,679 ESTs resulted in the identification of 835 distinct transcripts, from these, a total …

Neutralization-Sensitive R5 Shiv-2873nip Encoding Env From A Infant With Recent Hiv Clade C Infection, Nagadenahalli B. Siddappa, Ruijiang Song, Victor G. Kramer, Agnes-Laurence Chenine, Vijayakumar Velu, Helena Ong, Robert A. Rasmussen, Ricky D. Grisson, Charles Wood, Hong Zhang, Chipepo Kankasa, Rama Rao Amara, James G. Else, Francis J. Novembre, David Montefiori, Ruth M. Ruprecht

Nebraska Center for Virology: Faculty Publications

HIV-C (HIV-C) accounts for >56% of all HIV infections worldwide. To investigate vaccine safety and efficacy in non-human primates, a pathogenic, R5-tropic, neutralization sensitive simian-human immunodeficiency virus (SHIV) encoding HIV-C env would be desirable. We have constructed SHIV-2873Ni, an R5 SHIV encoding a primary pediatric HIV-C env isolated from a 2-month old Zambian infant, who progressed to death within one year of birth. SHIV-2873Ni was constructed using SHIV-1157ipd3N4 (Song et al., J. Viol. 80:8729-38, 2006) as backbone since the latter contains additional NF-κB sites in the long terminal repeats (LTRs) to enhance viral replicative capacity. The parental virus, SHIV-2873Ni, was …

Pseudomonas Aeruginosa-Candida Albicans Interactions: Localization And Fungal Toxicity Of A Phenazine Derivative, Jane Gibson, Arpanah Sood, Deborah A. Hogan

Dartmouth Scholarship

Phenazines are redox-active small molecules that play significant roles in the interactions between pseudomonads and diverse eukaryotes, including fungi. When Pseudomonas aeruginosa and Candida albicans were cocultured on solid medium, a red pigmentation developed that was dependent on P. aeruginosa phenazine biosynthetic genes. Through a genetic screen in combination with biochemical experiments, it was found that a P. aeruginosa-produced precursor to pyocyanin, proposed to be 5-methyl-phenazinium-1-carboxylate (5MPCA), was necessary for the formation of the red pigmentation. The 5MPCA-derived pigment was found to accumulate exclusively within fungal cells, where it retained the ability to be reversibly oxidized and reduced, and its …

Molecular Characterisation Of A Bovine-Like Rotavirus Detected From A Giraffe, Emily Mulherin, Jill Bryan, Marijke Beltman, Luke O'Grady, Eugene Pidgeon, Lucie Garon, Andrew Lloyd, John Bainbridge, Helen O'Shea, Paul Whyte, Séamus Fanning

Department of Biological Sciences Publications

Background

Rotavirus (RV), is a member of the Reoviridae family and an important etiological agent of acute viral gastroenteritis in the young. Rotaviruses have a wide host range infecting a broad range of animal species, however little is known about rotavirus infection in exotic animals. In this paper we report the first characterisation of a RV strain from a giraffe calf.

Results

This report describes the identification and detailed molecular characterisation of a rotavirus strain detected from a 14-day-old Giraffe (Giraffa camelopardalis), presenting with acute diarrhea. The RV strain detected from the giraffe was characterized molecularly as G10P[11]. …

Prevalence And Factors Associated With Intestinal Parasitic Infection Among Children In An Urban Slum Of Karachi., Vikram Mehraj, Juanita Hatcher, Saeed Akhtar, Ghazala Rafique, Mohammad Asim Beg

Department of Pathology and Laboratory Medicine

Background:Intestinal parasitic infections are endemic worldwide and have been described as constituting the greatest single worldwide cause of illness and disease. Poverty, illiteracy, poor hygiene, lack of access to potable water and hot and humid tropical climate are the factors associated with intestinal parasitic infections. The study aimed to estimate prevalence and identify factors associated with intestinal parasitic infections among 1 to 5 years old children residing in an urban slum of Karachi Pakistan. Methods And Principal
Findings:A cross sectional survey was conducted from February to June 2006 in Ghosia Colony Gulshan Town Karachi, Pakistan. A simple random …

Modulation Of The Nf-Κb Pathway By Bordetella Pertussis Filamentous Hemagglutinin, Tzvia Abramson, Hassya Kedem, David A. Relman

Faculty Publications, Biological Sciences

Background
Filamentous hemagglutinin (FHA) is a cell-associated and secreted adhesin produced by Bordetella pertussis with pro-apoptotic and pro-inflammatory activity in host cells. Given the importance of the NF-κB transcription factor family in these host cell responses, we examined the effect of FHA on NF-κB activation in macrophages and bronchial epithelial cells, both of which are relevant cell types during natural infection.

Methodology/Principal Findings
Exposure to FHA of primary human monocytes and transformed U-937 macrophages, but not BEAS-2B epithelial cells, resulted in early activation of the NF-κB pathway, as manifested by the degradation of cytosolic IκBα, by NF-κB DNA binding, and …

Prevalence Of Hcv And Hiv Infections In 2005-Earthquake-Affected Areas Of Pakistan., Saeed Khan, Mohammad A Rai, Adnan Khan, Amber Farooqui, Shahana U Kazmi, Syed H Ali

Department of Biological & Biomedical Sciences

Background: On October 8, 2005, an earthquake of magnitude 7.6 hit the Northern parts of Pakistan. In the post-earthquake scenario, overcrowding, improper sewage disposal, contamination of food and drinking water, hasty surgical procedures, and unscreened blood transfusions to earthquake victims most likely promotes the spread of infections already prevalent in the area. Objective: The objective of the study reported here was to determine the prevalence of Human Immunodeficiency and Hepatitis C viruses (respectively, HIV and HCV) in the earthquake-affected communities of Pakistan. The samples were analyzed 2 months and then again 11 months after the earthquake to estimate the burden …

Viral Vectors In The Research Laboratory: Just How Safe Are They?, Dawn P. Wooley, Kimberly Kay Morris, Robert Mcrae, John C. Trefry

Neuroscience, Cell Biology & Physiology Faculty Publications

No abstract provided.

Identification Of Two Gene Clusters And A Transcriptional Regulator Required For Pseudomonas Aeruginosa Glycine Betaine Catabolism, Matthew J. Wargo, Benjamin S. Szwergold, Deborah A. Hogan

Dartmouth Scholarship

Glycine betaine (GB), which occurs freely in the environment and is an intermediate in the catabolism of choline and carnitine, can serve as a sole source of carbon or nitrogen in Pseudomonas aeruginosa. Twelve mutants defective in growth on GB as the sole carbon source were identified through a genetic screen of a nonredundant PA14 transposon mutant library. Further growth experiments showed that strains with mutations in two genes, gbcA (PA5410) and gbcB (PA5411), were capable of growth on dimethylglycine (DMG), a catabolic product of GB, but not on GB itself. Subsequent nuclear magnetic resonance (NMR) experiments with 1,2-(13)C-labeled choline …

Global Analysis Of Gene Expression Changes During Retinoic Acid-Induced Growth Arrest And Differentiation Of Melanoma: Comparison To Differentially Expressed Genes In Melanocytes Vs Melanoma, Mary H. Estler, Goran Boskovic, James Denvir, Sarah Miles, Donald A. Primerano, Richard M. Niles

Biochemistry and Microbiology

BACKGROUND: The incidence of malignant melanoma has significantly increased over the last decade. Some of these malignancies are susceptible to the growth inhibitory and pro-differentiating effects of all-trans-retinoic acid (RA). The molecular changes responsible for the biological activity of RA in melanoma are not well understood.

RESULTS: In an analysis of sequential global gene expression changes during a 4-48 h RA treatment of B16 mouse melanoma cells, we found that RA increased the expression of 757 genes and decreased the expression of 737 genes. We also compared the gene expression profile (no RA treatment) between non-malignant melan-a mouse melanocytes and …

Entamoeba Histolytica: Lipid Rafts Are Involved In Adhesion Of Trophozoites To Host Extracellular Matrix Components, K. Mittal, B. H. Welter, Lesly A. Temesvari

Publications

Adhesion is an important virulence function for Entamoeba histolytica, the causative agent of amoebic dysentery. Lipid rafts, cholesterol-rich domains, function in compartmentalization of cellular processes. In E. histolytica, rafts participate in parasite–host cell interactions; however, their role in parasite–host extracellular matrix (ECM) interactions has not been explored. Disruption of rafts with a cholesterol extracting agent, methyl-β-cyclodextrin (MβCD), resulted in inhibition of adhesion to collagen, and to a lesser extent, to fibronectin. Replenishment of cholesterol in MβCD-treated cells, using a lipoprotein–cholesterol concentrate, restored adhesion to collagen. Confocal microscopy revealed enrichment of rafts at parasite–ECM interfaces. A raft-resident adhesin, the …

The Cradle Of The Deltaf508 Mutation, Danish Saleheen, Philippe M Frossard

Department of Biological & Biomedical Sciences

Cystic fibrosis (CF) is the most common autosomal recessive disorder caused due to mutation/s in the CFTR gene. The most common mutation in CFTR worldwide is deltaF508 and cystic fibrosis genetic analysis consortium revealed that this mutation is responsible for approximately 66% of all CF chromosomes in the world. Studies looking at the DNA polymorphic haplotypes created by CF linked markers suggest that deltaF508 has a single origin as this mutation has been found associated exclusively with one marker haplotype. Despite a high prevalence of this mutation in CF patients in northern parts of Europe, findings suggest that this mutation …

To Love Your Neighbor: A Christian Perspective On The Study Of Microbiology And Immunology, Joy Doan

Faith Learning Integration Papers

I have been blessed with enough of a sense of adventure to have experienced the awe-inspiring beauty of a rain forest at night, the top of Half Dome at Yosemite National Park, the sheer cliffs and rushing waters of the Narrows at Zion National Park, Plateau Point—which seems suspended in the Grand Canyon, and the top of a 14,000-foot peak in the Rocky Mountains of Colorado. These are the types of places about which one of the characters in Robert Pirsig’s Zen and the Art of Motorcycle Maintenance observes, “This is the hardest stuff in the world to photograph. You …

Metabolic Engineering Of A Thermophilic Bacterium To Produce Ethanol At High Yield, A. Joe Shaw, Kara K. Podkaminer, Sunil G. Desai, John S. Bardsley, Stephen R. Rogers, Philip G. Thorne, David A. Hogsett, Lee R. Lynd

Dartmouth Scholarship

We report engineering Thermoanaerobacterium saccharolyticum, a thermophilic anaerobic bacterium that ferments xylan and biomass-derived sugars, to produce ethanol at high yield. Knockout of genes involved in organic acid formation (acetate kinase, phosphate acetyltransferase, and L-lactate dehydrogenase) resulted in a strain able to produce ethanol as the only detectable organic product and substantial changes in electron flow relative to the wild type. Ethanol formation in the engineered strain (ALK2) utilizes pyruvate:ferredoxin oxidoreductase with electrons transferred from ferredoxin to NAD(P), a pathway different from that in previously described microbes with a homoethanol fermentation. The homoethanologenic phenotype was stable for >150 generations …

Multidimensional Protein Fractionation Using Proteomelab Pf 2d™ For Profiling Amyotrophic Lateral Sclerosis Immunity: A Preliminary Report, Joshua Schlautman, Wojciech Rozek, Robert Stetler, R. Lee Mosley, Howard Gendelman, Pawel Ciborowski

Nebraska Center for Virology: Faculty Publications

Background: The ProteomeLab™ PF 2D platform is a relatively new approach to global protein profiling. Herein, it was used for investigation of plasma proteome changes in amyotrophic lateral sclerosis (ALS) patients before and during immunization with glatiramer acetate (GA) in a clinical trial.
Results: The experimental design included immunoaffinity depletion of 12 most abundant proteins from plasma samples with the ProteomeLab™ IgY-12 LC10 column kit as first dimension separation, also referred to as immuno-partitioning. Second and third dimension separations of the enriched proteome were performed on the PF 2D platform utilizing 2D isoelectric focusing and RP-HPLC with the resulting fractions …

Oriented Cell Growth On Self-Assembled Bacteriophage M13 Thin Films, Jianhua Rong, L. Andrew Lee, Kai Li, Brandon Harp, Charlene M. Mello, Zhongwei Niu, Qian Wang

Faculty Publications

Fibrillar M13 bacteriophages were used as basic building blocks to generate thin films with aligned nanogrooves, which, upon chemical grafting with RGD peptides, guide cell alignment and orient the cell outgrowth along defined directions.

The Bile Response Repressor Brer Regulates Expression Of The Vibrio Cholerae Breab Efflux System Operon, Francisca A. Cerda-Maira, Carol S. Ringelberg, Ronald K. Taylor

Dartmouth Scholarship

Enteric pathogens have developed several resistance mechanisms to survive the antimicrobial action of bile. We investigated the transcriptional profile of Vibrio cholerae O1 El Tor strain C6706 under virulence gene-inducing conditions in the presence and absence of bile. Microarray analysis revealed that the expression of 119 genes was affected by bile. The mRNA levels of genes encoding proteins involved in transport were increased in the presence of bile, whereas the mRNA levels of genes encoding proteins involved in pathogenesis and chemotaxis were decreased. This study identified genes encoding transcriptional regulators from the TetR family (vexR and breR) and …