Microbiology Commons^™

Identifying Essential Viral Genes Through Genomic Engineering, Amber Carroll

Mahurin Honors College Capstone Experience/Thesis Projects

Bacteria developed resistance to penicillin a mere four years after the groundbreaking antibiotic was first mass produced (Casadevall, 2010). Since then, the development of antibiotic-resistant bacteria has steadily risen, causing millions of difficult to treat infections annually. The challenge is to identify ways to combat these menacing microbes. Bacteriophages are viruses that infect bacteria and can potentially be used to eliminate deadly antibiotic-resistant bacteria. The number of sequenced bacteriophage genomes has increased tremendously over the past 10 years, but little is known about the function of most bacteriophage genes. The purpose of this study was to expand our understanding of …

Using Probiotics To Flatten The Curve Of Coronavirus Disease Covid-2019 Pandemic., David Baud, Varvara Dimopoulou Agri, Glenn R Gibson, Gregor Reid, Eric Giannoni

Microbiology & Immunology Publications

No abstract provided.

Evaluation Of Antibody Response Directed Against Porcine Reproductive And Respiratory Syndrome Virus Structural Proteins, Hung Q. Luong, Huong T.L. Lai, Hiep L.X. Vu

Nebraska Center for Virology: Faculty Publications

Luciferase-immunoprecipitation system (LIPS), a liquid phase immunoassay, was used to evaluate antibody responses directed against the structural proteins of PRRSV in pigs that were experimentally infected with virulent PRRSV strains. First, the viral N protein was used as a model antigen to validate the assay. The LIPS results were highly comparable to that of the commercial IDEXX PRRS X3 ELISA. Subsequently, the assay was applied to simultaneously measure antibody reactivity against all eight structural proteins of PRRSV. The highest immunoreactivities were detected against GP3, M, and N proteins while the lowest reactivity was detected against ORF5a protein. Comparative analysis of …

Biotic And Abiotic Factors Influencing Arsenic Biogeochemistry And Toxicity In Fluvial Ecosystems: A Review, Laura Barral-Fraga, María Teresa Barral, Keeley L. Macneill, Diego Martiñá-Prieto, Soizic Morin, María Carolina Rodríguez-Castro, Baigal-Amar Tuulaikhuu, Helena Guasch

School of Natural Resources: Faculty Publications

This review is focused on the biogeochemistry of arsenic in freshwaters and, especially, on the key role that benthic microalgae and prokaryotic communities from biofilms play together in through speciation, distribution, and cycling. These microorganisms incorporate the dominant iAs (inorganic arsenic) form and may transform it to other arsenic forms through metabolic or detoxifying processes. These transformations have a big impact on the environmental behavior of arsenic because different chemical forms exhibit differences in mobility and toxicity. Moreover, exposure to toxicants may alter the physiology and structure of biofilms, leading to changes in ecosystem function and trophic relations. In this …

Microbial Degradation Of Urea Oligomers: Potential Slow Release Nitrogen Fertilizers, Maddy Bygd

Departmental Honors Projects

Urea is the most commonly used nitrogen compound in fertilizers across the world. It provides the most nitrogen at the lowest cost. However, with the size and solubility of this compound, it is easily washed out of the soil. This results in the need to continuously add fertilizer to fields, creating adverse effects on the environment due to nitrogen runoff and leading to increased expenses for farmers. Triuret, a urea oligomer, is a compound with promising potential for use as a nitrogen source, providing a slower release of nitrogen into the soil. It is hypothesized that this compound can be …

Crispr-Cas9 Editing Of Nitrate Transporter Gene, Um03849, In Ustilago Maydis, Luke A. Schroeder, Sunita Khanal, Michael H. Perlin

Undergraduate Arts and Research Showcase

Ustilago maydis, the basidiomycete smut-fungus, can infect and cause tumors in corn plants. For this, mating between compatible haploid cells is important. The mating and subsequent dimorphic transition in U. maydis require starvation for nutrients such as nitrogen, in addition to pheromone-receptor interactions between compatible partners. In this research, the CRISPR-Cas9 gene-editing technique was used to create INDEL mutations (sequence insertion or deletion) in the nitrate transporter gene, um03849, in U. maydis. The gene was edited in mating compatible haploid strains 1/2 and 2/9. The phenotypes were characterized for the um03849 mutants as to growth ability, mating …

The Effects Of Nutrient Availability On Pseudomonas Aeruginosa Mono And Co-Culture Biofilms, Julie T. Nguyen, Deborah R. Yoder-Himes Ph.D., Rhiannon Cecil

Undergraduate Arts and Research Showcase

Cystic Fibrosis (CF) is a genetic disorder characterized by faulty ion channels and result in thick mucus accumulation, especially in lungs. Mucus buildup provides ideal conditions for bacterial infections. Pseudomonas aeruginosa (PA) is the second most prevalent bacterium isolated from people with CF and has a high clinical importance. Most CF pathogens form biofilms which make treatment of infections difficult. Biofilms are clusters of cells attached to a surface enclosed in a structured matrix. These structures are a means to provide shelter for bacteria from the environment, especially antibiotics and the immune system. PA alone can form these biofilms, but …

Biogeography Of American Northwest Hot Spring A/B′-Lineage Synechococcus Populations, Eric D. Becraft, Jason M. Wood, Frederick M. Cohan, David M. Ward

Biology

Previous analyses have shown how diversity among unicellular cyanobacteria inhabiting island-like hot springs is structured relative to physical separation and physiochemical differences among springs, especially at local to regional scales. However, these studies have been limited by the low resolution provided by the molecular markers surveyed. We analyzed large datasets obtained by high-throughput sequencing of a segment of the photosynthesis gene psaA from samples collected in hot springs from geothermal basins in Yellowstone National Park, Montana, and Oregon, all known from previous studies to contain populations of A/B′-lineage Synechococcus. The fraction of identical sequences was greater among springs separated by …

Hiding In Plain Sight: The Globally Distributed Bacterial Candidate Phylum Pauc34f, Michael L. Chen, Eric D. Becraft, Maria Pachladaki, Julia M. Brown, Jessica K. Jarett, Josep M. Gasol, Nikolai V. Ravin, Duane P. Moser, Takuro Nunoura, Gerhard J. Herndl, Tanja Woyke, Ramunas Stepanauskas

Biology

Bacterial candidate phylum PAUC34f was originally discovered in marine sponges and is widely considered to be composed of sponge symbionts. Here, we report 21 single amplified genomes (SAGs) of PAUC34f from a variety of environments, including the dark ocean, lake sediments, and a terrestrial aquifer. The diverse origins of the SAGs and the results of metagenome fragment recruitment suggest that some PAUC34f lineages represent relatively abundant, free-living cells in environments other than sponge microbiomes, including the deep ocean. Both phylogenetic and biogeographic patterns, as well as genome content analyses suggest that PAUC34f associations with hosts evolved independently multiple times, while …

Non-Alcoholic Fatty Liver Disease And The Gut Microbiome: The Effects Of Gut Microbial Metabolites On Nafld Progression In A 2-Organ Human-On-A-Chip Model, Rachel H. Boone

Honors Undergraduate Theses

Using a novel, adipose-liver, two-organ, human-on-a-chip system, the metabolic disease non-alcoholic fatty liver disease was modeled. This model was then used to test the effects of the gut microbiome on NAFLD progression. Two products of the gut microbiome, Trimethylamine-n-oxide and butyrate, were selected as representatives of potentially harmful and potentially beneficial compounds. A dose response, adipocyte and hepatocyte monocultures controls, and HoaC systems were run for 14 days. Through this experimentation, it was found that a dysbiosis of the gut microbiome could be influencing NAFLD progression. Additionally, further development and discovery regarding adipose-liver systems was added to the ongoing conversation …

Characterization Of The Physical And Chemical Effect Of Membrane Disruption And Protein Inhibiting Treatments On E. Coli, Khadijah Wright

Honors Undergraduate Theses

The increase in antibacterial resistance has placed the issue of microbial multi-drug resistance on a global stage (Gurunathan, 2019). This issue poses a threat to human and animal health as well as to the environment (Aslam et al., 2018). It affects not only the efficacy of treatment but also how those treatments are conducted (Friedman, Temkin, & Carmeli, 2016). As a result of this ongoing threat, new treatments that have potent effects on bacteria are necessary. One scientific response to this issue has been the development of multifunctional nanoparticles (NPs)(H. Wang et al., 2018). NPs have the ability to be …

Biofilm Associated Staphylococcus Aureus Viability Is Altered By Burkholderia Cenocepacia, Bridget Wall, Tiffany J. Brandt, Dr. Deborah Yoder-Himes

Undergraduate Arts and Research Showcase

Respiratory failure caused by chronic and recurrent microbial infections is the most common cause of death for people with cystic fibrosis (CF)¹, a disease causing the formation of thick mucus in the lungs². Most bacteria can form biofilms, collections of sessile cells adhered to a surface by a secreted substance. Biofilm-associated cells develop antibiotic resistance at higher rates³. The thicker mucus in CF lungs is extremely difficult to clear via action of the mucociliary escalator and its presence fosters the formation of bacterial biofilms. Staphylococcus aureus and Burkholderia cenocepacia are two pathogens commonly found …

Evaluation Of The Symbiotic Relation Between Endophyte And Poplar Trees Exposed To Landfill Leachate, Catalina Vega Hurtado

Masters Theses

“The contamination of soil and groundwater from leachate leakage from landfill has turned, nowadays, into a global public issue. One of the main concerns about this pollutant is the potential threat to human and ecosystem health. The current research studied endophyte bacteria in association with poplar trees (Populus sp.), as a method of bioremediation of landfill leachates. The objective of the project was to identify treatment strategies that may improve plant performance (survival rates, plant fitness, and degradation efficacy) with the purpose of being implemented in phytoremediation plots, aimed to intercept and treat landfill leachate, before the contamination of watershed. …

Draft Genome Sequence Of Verrucosispora Sp. Strain Cwr15, Isolated From A Gulf Of Mexico Sponge, Sarah J. Kennedy, Eleonora Cella, Mohammad Jubair, Taj Azarian, Bill J. Baker, Lindsey N. Shaw

Molecular Biosciences Faculty Publications

Here, we present the draft genome sequence of Verrucosispora sp. strain CWR15, a bacterial symbiont of a Gulf of Mexico sponge. The genome consists of 35 contigs encoding 5,840 genes. The genome is the basis for future study and presents an underexplored taxonomy and biosynthetic potential.

Mir146a-Mediated Targeting Of Fancm During Inflammation Compromises Genome Integrity, Devakumar Sundaravinayagam, Hye Rim Kim, Tingting Wu, Hyun Hee Kim, Semo Jun, Jeong-Heon Cha, Younghoon Kee, Ho Jin You, Jung-Hee Lee

Molecular Biosciences Faculty Publications

Inflammation is a potent inducer of tumorigenesis. Increased DNA damage or loss of genome integrity is thought to be one of the mechanisms linking inflammation and cancer development. It has been suggested that NF-κB-induced microRNA-146 (miR146a) may be a mediator of the inflammatory response. Based on our initial observation that miR146a overexpression strongly increases DNA damage, we investigated its potential role as a modulator of DNA repair. Here, we demonstrate that FANCM, a component in the Fanconi Anemia pathway, is a novel target of miR146a. miR146a suppressed FANCM expression by directly binding to the 3’ untranslated region of the gene. …

Virus-Induced Changes In Nuclear Proteins And Membranes In Nicotiana Benthamiana Cells, Caleb Mathias

Theses and Dissertations--Plant Pathology

Viruses rely on host proteins to complete their life cycles. This results in alterations to normal cell physiology to processes which benefit viral processes such as replication and movement to other cells. This may involve the relocalization of host proteins away from their original subcellular targets to sites which may benefit the virus, a process that is not as well understood as it relates to the nucleus. To identify nuclear proteins that may be involved in such processes, a library of random Nicotiana benthamiana cDNAs were expressed as GFP fusions in a transgenic marker lines expressing a histone 2B:RFP nuclear …

Investigating The Mechanism Of The Coronavirus Endoribonuclease In Antagonizing Innate Immune Signaling, Matthew Hackbart

Dissertations

Coronaviruses (CoVs) are positive-sense RNa viruses that can emerge from endemic reservoirs and infect zoonotically, causing significant morbidity and mortality. CoVs encode an endoribonuclease (EndoU) that cleaves RNa in biochemical assays, but the target and function of EndoU activity during viral replication was not known. My work focused on characterizing the functions of EndoU during infection. I report that EndoU is an innate immune antagonist. to function as an immune antagonist, EndoU cleaves the 5'-Poly-Uridines from Negative-sense viral RNA, termed PUN RNA, which is the product of polyA-templated RNa synthesis. Using a virus containing an EndoU catalytic-inactive mutation, I detected …

Analysis Of Student Perceptions Of Just-In-Time Teaching Pedagogy In Pharmd Microbiology And Immunology Courses, Charitha Madiraju, Eglis Tellez-Corrales, Henry Hua, Jozef Stec, Andromeda M. Nauli, Deborah M. Brown

Nebraska Center for Virology: Faculty Publications

Just-In-Time Teaching (JiTT) active learning pedagogy is utilized by various disciplines, but its value in a professional pharmacy curriculum has not yet been demonstrated. The purpose of our research study is to implement and evaluate JiTT in a Doctor of Pharmacy (PharmD) program. The impetus in implementing JiTT into a PharmD curriculum was to provide students with an out-of-classroom learning opportunity to enhance knowledge-based skills. The current study summarizes the implementation of JiTT in four distinct instances: two iterations of the required courses “Integrated Microbiology and Virology” (Fall 2016 and Fall 2017) and “Integrated Immunology” (Winter 2016–2017 and Winter 2017–2018). …

Lack Of Cd8+ T-Cell Co-Localization With Kaposi’S Sarcoma- Associated Herpesvirus Infected Cells In Kaposi’S Sarcoma Tumors, Salum J. Lidenge, For Yue Tso, Owen Ngalamika, Jaydeep Kolape, John R. Ngowi, Julius Mwaiselage, Charles Wood, John T. West

Nebraska Center for Virology: Faculty Publications

Despite the close association between Kaposi’s sarcoma (KS) and immune dysfunction, it remains unclear whether tumor infiltrating immune cells (TIIC), by their absence, presence, or dysfunction, are mechanistically correlated with KS pathogenesis. Therefore, their potential capacity to serve as prognostic biomarkers of KS disease progression or control is unclear. Because epidemic-KS (EpKS) occurs with HIV-1 co-infection, it is particularly important to compare TIIC between EpKS and HIV-negative African endemic-KS (EnKS) to dissect the roles of HIV-1 and Kaposi Sarcoma-associated herpesvirus (KSHV) in KS pathogenesis. This cross-sectional study of 13 advanced KS (4 EnKS, 9 EpKS) patients and 3 healthy controls …

Prevalence Of Kaposi's Sarcoma-Associated Herpesvirus And Transfusion-Transmissible Infections In Tanzanian Blood Donors, Salum J. Lidenge, Tara Tran, For Yue Tso, John R. Ngowi, Danielle M. Shea, Julius Mwaiselage, Charles Wood, John T. West

Nebraska Center for Virology: Faculty Publications

Objective: Kaposi's sarcoma-associated herpesvirus (KSHV) is the causative agent for Kaposi's sarcoma (KS), one of the most common cancers in Tanzania. We have investigated KSHV prevalence and factors associated with KSHV infection in Tanzania.

Methods: This is a cross-sectional study of voluntary blood-donors from Dar es Salaam, Tanzania. Plasma was screened for KSHV, HIV-1, HBV, HCV and Treponema pallidum (syphilis). Associations between KSHV sero-status and risk factors were analyzed. Odds ratios (OR) and 95% confidence intervals (CI) are reported to evaluate risk factors of KSHV infection. All tests were 2-tailed, and P-values <0.05 were considered statistically significant.

Results: The overall KSHV seroprevalence was 56.9%. Significantly …