Microbiology Commons^™

Bioinformatic And Sequence Analysis Of Four Resuscitation Promoting Factor (Rpf) Gene Homologues In Mycobacterium Avium Subsp. Paratuberculosis (Mpt), And Expression Of The Putative Mpt Rpfb In Escherichia Coli., David Dale Hedlund

All Graduate Theses, Dissertations, and Other Capstone Projects

Mycobacterium avium subsp. paratuberculosis (Mpt), the causative agent of Johne's disease (JD), is a global problem in the agricultural industry. It is estimated that 25% of all U.S. dairy herds are JD positive. One obstacle in the management of JD is the lack a sensitive diagnostic test for use during the early stages of infection. Resuscitation promoting factors (Rpf) are proteins that promote the growth of many species of Actinobacteria. If Rpf proteins could enhance the growth of Mpt, the sensitivity of diagnostic fecal culture could be improved, and the impact of JD on the dairy industry would be significantly …

The Immune Response In The Central Nervous System During West Nile Virus Persistence, Barbara Sharon Stewart

Legacy Theses & Dissertations (2009 - 2024)

West Nile virus (WNV) persists in a wide array of hosts ranging from mice to humans. In convalescent humans, WNV RNA persists in urine for up to 6.7 years, and IgM antibody against WNV persists in serum for up to 12 months post-inoculation (p.i.). Previous work using the mouse model demonstrated that WNV persists in central nervous system (CNS) tissues as infectious virus and as RNA for up to 4 months and 6 months p.i., respectively. In this study, we sought to elucidate the mechanism for viral persistence in the CNS using the mouse model. Characterization of the leukocyte infiltrate …

Subtype-Associated Differences In Hiv-1 Reverse Transcription Affect The Viral Replication, Sergey Iordanskiy, Mackenzie Waltke, Yanjun Feng, Charles Wood

Nebraska Center for Virology: Faculty Publications

Background: The impact of the products of the pol gene, specifically, reverse transcriptase (RT) on HIV-1 replication, evolution, and acquisition of drug resistance has been thoroughly characterized for subtype B. For subtype C, which accounts of almost 60% of HIV cases worldwide, much less is known. It has been reported that subtype C HIV-1 isolates have a lower replication capacity than B; however, the basis of these differences remains unclear.

Results: We analyzed the impact of the pol gene products from HIV-1 B and C subtypes on the maturation of HIV virions, accumulation of reverse transcription products, integration …

Enhancement Of Autophagy During Lytic Replication By The Kaposi’S Sarcoma-Associated Herpesvirus Replication And Transcription Activator, Hui-Ju Wen, Zhilong Yang, You Zhou, Charles Wood

Nebraska Center for Virology: Faculty Publications

Autophagy is one of two major degradation systems in eukaryotic cells. The degradation mechanism of autophagy is required to maintain the balance between the biosynthetic and catabolic processes and also contributes to defense against invading pathogens. Recent studies suggest that a number of viruses can evade or subvert the host cell autophagic pathway to enhance their own replication. Here, we investigated the effect of autophagy on the KSHV (Kaposi’s sarcoma-associated herpesvirus) life cycle. We found that the inhibition of autophagy reduces KSHV lytic reactivation from latency, and an enhancement of autophagy can be detected during KSHV lytic replication. In addition, …

Chlorella Viruses Encode Most, If Not All, Of The Machinery To Glycosylate Their Glycoproteins Independent Of The Endoplasmic Reticulum And Golgi, James L. Van Etten, James Gurnon, Giane M. Yanai-Balser, David Dunigan, Michael V. Graves

Nebraska Center for Virology: Faculty Publications

In contrast to all other viruses that use the host machinery located in the endoplasmic reticulum and Golgi to glycosylate their glycoproteins, the large dsDNA-containing chlorella viruses encode most, if not all, of the components to glycosylate their major capsid proteins. Furthermore, all experimental results indicate that glycosylation occurs independent of the endoplasmic reticulum and Golgi. (Review article)

Plasmid Injection And Application Of Electric Pulses Alter Endogenous Mrna And Protein Expression In B16.F10 Mouse Melanomas, L. C. Heller, Y. L. Cruz, B. Ferraro, H. Yang, R. Heller

Bioelectrics Publications

The application of electric pulses to tissues causes cell membrane destabilization, allowing exogenous molecules to enter the cells. This delivery technique can be used for plasmid gene therapy. Reporter gene expression after plasmid delivery with eight representative published protocols was compared in B16.F10 mouse melanoma tumors. This expression varied significantly based on the pulse parameters utilized for delivery. To observe the possible influence of plasmid injection and/or pulse application on endogenous gene expression, levels of stress-related mRNAs 4 and 24 h after delivery were determined by PCR array. Increases in mRNA levels for several inflammatory chemokines and cytokines were observed …

Interactions Of The Cellular Sumoylation System With Influenza A Virus And Its Non-Structural Protein Ns1a (Ns1a), Sangita Pal

Open Access Theses & Dissertations

The most important current anti-influenza weapons, vaccination and antiviral drugs, can be rapidly rendered fully ineffective thanks to the virus's high mutational rate, which produces viruses exhibiting new antigenic properties and structural proteins insensitive to the drug's mechanism of action. One attractive alternative is to develop drugs that modulate the activity of cellular systems either required for viral growth or able to neutralize viral growth. Here we demonstrate that the cellular SUMOylation system, a post-translational modification involving the conjugation of the Small Ubiquitin-like MOdifier (SUMO) to specific protein targets using a Ubiquitin-like enzymatic cascade, interacts closely with influenza virus during …

Phocine Distemper Virus Induced Demyelination In The Spinal Cord Of Infected Harbor Seals (Phoca Vitulina), Thomas Siemens

All Theses And Dissertations

Phocine Distemper Virus (PDV) is a Morbillivirus that has been responsible for the death of approximately 40,000 phocid seals in the last three decades. The most commonly reported pathology is virally induced demyelination of the cerebrum and brainstem which leaves axons bare and limits their conductivity of action potentials. While these conditions have been studied at length, there is little mention of spinal cord involvement. This study aims to determine whether demyelination found in the brainstem continues into the cervical spinal cord. The results indicate that demyelination in the cervical spinal cord does not occur alongside the brainstem. It is …

Isolation And Characterization Of Actinobacteria From Soil Samples Of Signy Island, Antarctica., Pan Shing Yi

Student Works (2010-2019)

In this study, ninety five actinobacterial isolates were cultured from seven soils samples collected on Signy Island, maritime Antarctica. Nine different isolation media were used and the isolates plates were incubated at 15ºC for up to four months. SCN agar serves as the better media for the isolation of actinobacteria. Presence of actinobacteria in the soil samples was also detected using PCR with actinobacteria specific primers. The actinobacterial isolates were characterized based on morphology (Gram-stain, coverslip examination, colour grouping), analysis of DAP isomers, antibacterial activities and by molecular methods (ARDRA, analysis of 16S rRNA sequence and detection of NRPS gene). …

Response Surface Optimization Of Crystal Violet Decolorization By Pellets Of Pyncnoporus Sanguineus., Siti Suhana Sulaiman

Student Works (2010-2019)

Ligninolytic enzymes of white-rot fungi have broad specificities and have been implicated in transformation and mineralization of organo-pollutant with chemical structure similar to lignin e.g. synthetic dyes. This study was carried out to optimize selected parameters for decolorization of Crystal violet by Pycnoporus sanguineus in shake flasks. The parameters studied were initial dye concentration (ppm), agitation speed (rpm) and process time. They were optimized using response surface methodology (RSM). Subsequent to the optimization, the decolorization process was scaled up in stirred tank reactor (STR). Effect of different geometry of impellers on the decolorization process was studied. Curved blade and angled …

Genetic Diversity Among Plasmodium Falciparum Field Isolates In Pakistan Measured With Pcr Genotyping Of The Merozoite Surface Protein 1 And 2, Najia Ghanchi, Andreas Martensson, Johan Ursing, Sana Jafri, Sandor Bereczky, Rabia Hussain, Mohammad Asim Beg

Department of Pathology and Laboratory Medicine

Background:The genetic diversity of Plasmodium falciparum has been extensively studied in various parts of the world. However, limited data are available from Pakistan. This study aimed to establish molecular characterization of P. falciparum field isolates in Pakistan measured with two highly polymorphic genetic markers, i.e. the merozoite surface protein 1 (msp-1) and 2 (msp-2).
Methods:Between October 2005 and October 2007, 244 blood samples from Patients with symptomatic blood-slide confirmed P. falciparum mono-infections attending the Aga Khan University Hospital, Karachi, or its collection units located in Sindh and Baluchistan provinces, Pakistan were collected. The genetic diversity of P. falciparum …

Detection And Molecular Characterisation Of Rotavirus, Adenovirus And Bocavirus Circulating In Southern Ireland, Olivia Cashman

Theses

This study reports the detection and molecular characterisation of group A rotavirus circulating in the south of Ireland in both the bovine and human population. Characterisation of the group A rotavirus was carried out through the extraction of viral nucleic acid in faecal specimens collected from hospitals in the Cork and Waterford area and Cork regional veterinary laboratory from 2006-2009. Other gastroenteritis viruses characterised during this study include adenovirus type 41, and the newly identified gastrointestinal virus, human bocavirus, which was identified for the first time in Ireland during this study. The human bocaviruses identified in children in the south …

Humboldt Journal Of Microbiology: Volume 12, Number 1- May 2010, Press Humboldt

Humboldt Journal of Microbiology

No abstract provided.

Quantitative Pcr Assay For Mycobacterium Pseudoshottsii And Mycobacterium Shottsii And Application To Environmental Samples And Fishes From The Chesapeake Bay, D. T. Gauthier, K. S. Reece, J. Xiao, M. W. Rhodes, H. I. Kator, R. J. Latour, C. F. Bonzek, J. M. Hoenig, W. K. Vogelbein

Biological Sciences Faculty Publications

Striped bass (Morone saxatilis) in the Chesapeake Bay are currently experiencing a very high prevalence of mycobacteriosis associated with newly described Mycobacterium species, Mycobacterium pseudoshottsii and M. shottsii. The ecology of these mycobacteria outside the striped bass host is currently unknown. In this work, we developed quantitative real-time PCR assays for M. pseudoshottsii and M. shottsii and applied these assays to DNA extracts from Chesapeake Bay water and sediment samples, as well as to tissues from two dominant prey of striped bass, Atlantic menhaden (Brevoortia tyrannus) and bay anchovy (Anchoa mitchilli). Mycobacterium …

Increased Perfusion And Angiogenesis In A Hindlimb Ischemia Model With Plasmid Fgf-2 Delivered By Noninvasive Electroporation, B. Ferraro, Y. L. Cruz, M. Baldwin, D. Coppola, R. Heller

Bioelectrics Publications

Gene therapy approaches delivering fibroblast growth factor-2 (FGF-2) have shown promise as a potential treatment for increasing blood flow to ischemic limbs. Currently, effective noninvasive techniques to deliver plasmids encoding genes of therapeutic interest, such as FGF-2, are limited. We sought to determine if intradermal injection of plasmid DNA encoding FGF-2 (pFGF) followed by noninvasive cutaneous electroporation (pFGFE+) could increase blood flow and angiogenesis in a rat model of hindlimb ischemia. pFGFE+ or control treatments were administered on postoperative day 0. Compared to injection of pFGF alone (pFGFE-), delivery of pFGFE+ significantly increased FGF-2 expression for 10 days. Further, the …

Yersinia Pestis Virulence Factor Yopm Undermines The Function Of Distinct Ccr2+Gr1+ Cells In Spleen And Liver During Systemic Plague, Zhan Ye

University of Kentucky Doctoral Dissertations

Yersinia pestis, the infective agent of bubonic and pneumonic plague, is classified as a category A agent of bioterrorism. YopM, a protein toxin of Y. pestis, is necessary for full virulence in a systemic plague mouse model with ambient-temperature grown bacteria. We used this model to identify the cells undermined by YopM. Natural killer (NK) cells were previously reported to be lost in spleen and blood in a YopM-associated way; however, NK cell depletion was found not to occur in liver, and ablation of NK cells had no effect on bacterial growth, indicating that NK cells are redundant …

Phage Lysins - Structure, Genetics And Applications, Jennifer Garry

Theses

Antibiotics are of the utmost importance in the fight against pathogenic bacteria, however, in recent years there has been a continual increase in the development of antibiotic resistance among bacteria. As a result it has become essential to develop new technologies to combat these pathogens. One such technology is the application of phage lysins as antimicrobial agents against pathogenic infections. A promising phage lysin is LysK, a 495 amino-acid protein derived from Staphylococcal Phage K. LysK can lyse a wide range of staphylococcal species including MRSA. It is also capable of killing both live and dead cells increasing its potential …

Biosynthesis Of Exopolysaccharides From Lactic Acid Bacteria And Evaluation Of Their Prebiotic Potential, Colette Ann Forde

Theses

In this study, two strains of lactic acid bacteria (LAB) namely Weissella kimchii F28 and W. confusa MGl were used for the production of exopolysaccharides (EPS) from sucrose. Both strains produced glucan polysaccharides. In the case of W. kimchii F28, the EPS isolation procedure was performed six times, making modifications in order to optimize the yield of EPS. The maximum yield of EPS for this strain was 16 g per litre of culture. In the case of W. confusa MGl, the EPS isolation procedure was followed five times, resulting in a maximum yield of EPS for this strain of 34.4 …

Detection And Molecular Characterization Of Canine Enteric Viruses In Southern Ireland., Susan Mcelligott

Theses

Enteric viruses, such as canine parvovirus (CPV) and canine coronavirus (CCoV), are a major cause of gastroenteritis in dogs. More serious clinical symptoms occur in young pups, or when both of these viruses infect an animal simultaneously. Caliciviruses and rotaviruses can also cause gastroenteritis in dogs, but these viruses, while causing significant problems in other animal species and humans, are considered to be less significant pathogens of canines. However, on occasion, these viruses have been reported to produce severe clinical symptoms such as vomiting, haemorrhagic enteritis and severe dehydration.

Many viral vaccines for canines are formulated using old vaccine viral …

Copper Isotope Fractionation During Surface Adsorption And Intracellular Incorporation By Bacteria, Jesica Urbina Navarrete

Open Access Theses & Dissertations

Copper isotopes may prove to be a useful tool for investigating bacteria-metal interactions recorded in natural waters, soils, and rocks. However, experimental data that constrain Cu isotope fractionation in biologic systems are limited and unclear. In this study we utilized Cu isotopes (65Cu) as a tool to investigate Cu-bacteria interactions, including surface adsorption and intracellular incorporation. Experiments were conducted with individual Gram-positive (Bacillus subtilis) and Gram-negative (Escherichia coli) bacterial species as well as with bacterial consortia from several natural environments. Adsorption experiments were conducted with live or dead cells over the pH range 2.5 to 6. Surface adsorption of Cu …