Medicine and Health Sciences Commons^™

Understanding Human Gut Bacterial Interactions And Their Role In Colonization Resistance Against Clostridioides Difficile Using Mini-Bioreactor Array And Caenorhabditis Elegans As Model Systems, Achuthan Ambat

Electronic Theses and Dissertations

Although fecal transplantation has shown to be effective against Clostridioides difficile infection (CDI), key species responsible for colonization resistance still need to be better identified. There needs to be more understanding of the rules governing the development of a healthy microbiome upon transplantation. The bottom-up approach of assembling simple to complex communities in model systems such as bioreactors is one of the ways to approach this problem. Studying synthetic communities with less complexity can yield a system-level understanding of microbial interactions. In the first study, we assembled a synthetic community of 14 bacterial species belonging to the core human microbiota …

Genistein-Enriched Pig Gut Microbiota Library As A Potential Probiotic Consortium, Theresah Amponsah

Electronic Theses and Dissertations

Plant-based diets contain numerous flavonoid compounds that produce antibacterial effects and gut health improvement. Genistein is among the most abundant isoflavones present in a plant-based diet and can be found in high amounts in soy products (up to 1g Kg-1). We describe here a robust method to identify genistein tolerant and metabolizing bacteria in swine gut microbiota and to screen the strains that have antibacterial and immunestimulatory properties. Such strains could be developed as non-antibiotic alternatives to prevent enteric infections in pigs and improve gut immunity. To this end, a mini bioreactorbased system was used to enrich genistein metabolizing bacteria …

Emerging Respiratory Viral Pathogens, April Nelsen

Electronic Theses and Dissertations

Respiratory disease is arguably the most important health concern for the production animal industry [1-3]. Respiratory problems accounted for the highest mortality both in the swine and beef industries [1, 4]. Respiratory pathogens remain the most vital for swine and bovine research and disease monitoring [2, 5-7]. As pathogens, in particular viral pathogens, mutate, novel disease-causing viruses emerge, and there becomes an increasing concern and need for identification with control perimeters. One frequently identified disease syndrome is Porcine Respiratory Disease Complex (PRDC). PRDC is characterized by pneumonia of mixed respiratory infections with contributions from the environment and management practices. The …

Role Of Host Restriction Factors On Porcine Reproductive And Respiratory Syndrome Virus (Prrsv) Replication, Pratik Katwal

Electronic Theses and Dissertations

In this study, the role of IFITM3 on PRRSV replication was studied in vitro by expressing exogenous IFITM3 in MARC-145 cells. An average of 31% reduction in PRRSV N protein expression and an average of 5.4 fold decrease in virus titer in the supernatant were observed in IFITM3 overexpressing cells as compared to vector control cells at 24 hours post infection (hpi). Moreover, there was a positive correlation between interferon- induced IFITM3 up-regulation and reduced PRRSV replication. To determine the role of endogenous IFITM3 in PRRSV replication, siRNA induced knockdown of IFITM3 was employed. RT-PCR validated the successful silencing of …

Production Of Monoclonal And Polyclonal Antibodies Against Prrsv Mlvd23-S Tag Peptide Sequence, Katelyn Bettin

Schultz-Werth Award Papers

Porcine reproductive and respiratory syndrome virus (PRRSV) is an RNA virus which belongs to the family Arteriviridae. Since its emergence in the late 1980’s and early 1990’s, PRRSV has had a major economic effect on the swine industry. The most notable effects of this disease are respiratory distress in piglets and late-term pregnancy failures in sows. To track and control spread of this disease, the industry must be able determine which animals have been naturally infected with PRRSV as this contributes to major disease outbreaks and localized pandemics. While vaccines help control the spread of a disease, they also …

Viral And Immune Studies On Bovine Viral Diarrhea Virus (Bvdv) In Bovine Species, Karim Abdelsalam

Electronic Theses and Dissertations

BVDV is a major pathogen of ruminants that causes immune dysfunction and lead to secondary bacterial infection. The underlying mechanism of this dysfunction is not well understood. Based on previous results, we believe that macrophages play a central role in BVDV-associated immune dysfunction. Previously, we have shown that the infected monocyte-derived macrophage (MDM) supernatant of the high virulent strains of BVDV significantly induced lymphocyte apoptosis. In the current study, we performed experiments to identify the apoptotic factors in the infected MDM supernatants in vitro. Also, we have extended our investigation to include testing the impact of infected macrophages on neutrophil …

Rumen Microbial Culture Library And In Vitro Analysis Of Selected Bacterial Species On Colonization Resistance Against Bovine Enteric Pathogens, Maria Antony

Electronic Theses and Dissertations

The anaerobic habitat of the four chambers in ruminant’s foregut plays a vital role in the fermentation of the food by the active involvement of the residing microbial community. The cultivability and the potential role of rumen bacteria in the field of livestock production, host health and immunity are important to study. The developed anaerobic culturing techniques enable us to culture and screen different rumen bacteria to study the anti-pathogenic effects. Whole genome sequencing and analysis help us to understand the genome level characteristics of the rumen bacterial species individually. Since the majority of the rumen biosphere is still not …

Bovine Cellular And Innate Immune Response To Hemostatic Polysaccharides, Brian Kobylkevich

Electronic Theses and Dissertations

Chronic wounds are longstanding dermal wounds that do not heal in a normal amount of time and can commonly last for over a year. These wounds plague humans and farm animals alike. Wounds are characterized by chronic inflammation and are often painful, difficult to heal, have a high recurrence rate, and are potentially debilitating. Long term wounds that do not fully heal are distressing for both humans and animals, but understandably more resources have been directed towards humans. Some treatment options for humans may be suitable for adaptation into the field of animal care. Chronic lesions such as bovine digital …

Universal Swine Influenza Antigen And Vaccine, Zhao Wang

Electronic Theses and Dissertations

Swine influenza A viruses pose a serious concern for global health and worldwide swine industry considering their evasion from vaccine-induced immune responses. Formulation of effective influenza vaccines can be complicated by both antigenic shift to a different subtype of hemagglutinin (HAs) and antigenic drift within a particular HA subtype. To address these concerns, we employed a systematic approach to determine the antigenic determinants of swine influenza A viruses by focusing on H1 and H3 subtypes. In parallel, we used a cross-disciplinary approach including structural biology and modeling, virology, and immunology to identify swine influenza A virus-derived HA fusion intermediates broadly …

The Effects Of Bovine Viral Diarrhea Virus And Bovine Herpesvirus Type 1 On Monocyte-Derived Dendritic Cells, Jacob Sobraske

Electronic Theses and Dissertations

Bovine viral diarrhea virus (BVDV) and bovine herpesvirus type 1 (BHV-1) are important pathogens that affect cattle. In this in vitro study, the main goal was determining the direct effects of BVDV and BHV-1 infections of monocyte-derived dendritic cells (MDDC) on cytokine mRNA expression. For the BVDV infected MDDC experiments, Holstein Friesian and Brown Swiss calves were included with a breed comparison with cytokine expression. Surface marker expression was also measured in BHV-1-infected MDDCs. For both viruses, a high virulent strain (1373 for BVDV and Cooper for BHV-1) and lower virulent strains (20508 for BVDV and Los Angeles for BHV-1) …

Serological Evidence For The Co-Circulation Of Two Lineages Of Influenza D Virus In Equine Populations Of The Midwest United States, Hunter Theodore Nedland

Electronic Theses and Dissertations

Influenza D virus (IDV) is a newly described lineage of the Orthomyxoviridae virus family that was first isolated from diseased swine in 2011 and has subsequently been detected in cattle around the world in 2014. In addition, serological evidence for IDV infection in humans has been recently established. Despite all the progress, the full range of susceptible hosts for this novel virus has yet to be determined, but includes swine, bovine, small ruminants and human. This study was designed to determine if equine is a possible host to this newly emerging influenza virus. 364 equine serum samples were collected in …

Understanding The Effect Of Bvdv On Innate Immune Response Of Neutrophils, Neelu Singh Thakur

Electronic Theses and Dissertations

Bovine viral diarrhea virus (BVDV) is one of the highly prevalent and economically important diseases of cattle industry worldwide. The two major consequences of this disease are persistent infection and immunosuppression. Several studies have been done to determine the underline mechanisms of BVDV-induced immunosuppression targeting antigen presenting cells, adaptive immune system cells and cytokine gene expression. However, very little research has been done to determine the effect of BVDV on neutrophils. Neutrophils are one of the most abundant while blood cells (WBC) in the peripheral blood, which play a critical role in the innate as well as adaptive immune response. …

Senecavirus A: Epidemiology, Pathogenesis And Infection Dynamics, Lok Raj Joshi

Electronic Theses and Dissertations

Senecavirus A (SVA) is a positive sense single-stranded virus that belongs to the genus Senecavirus within the family Picornaviridae. Senecavirus has been present in US swine population since 1988. However, there have been only a few reports of SVA associated with disease in last two decades. In late 2014 early 2015, a sudden increase in the number of cases of Senecavirus A associated with vesicular disease was observed in Brazil, the US and China. Affected pigs presented vesicles on snout, coronary band, and dewclaw. Additionaly, increased neonatal mortality has been reported in the field. Many aspects of SVA epidemiology, …

Surveillance Of Emerging Livestock Viruses, Faten A. Okda

Electronic Theses and Dissertations

Accurate and rapid diagnostic tests are essential for the management and control of emerging infectious diseases. Therefore, the overall goals of studies presented in this dissertation were to provide improved diagnostic and surveillance capabilities for several emerging viral diseases impacting the livestock industry and to provide insight into appropriate control strategies. Recent, severe outbreaks of porcine epidemic diarrhea virus (PEDV) in Asia and North America highlighted the need for well-validated diagnostic tests for identification of PEDV infected animals and evaluation of their immune status. PEDV was first detected in the U.S. in May 2013 and another swine coronavirus, porcine deltacoronavirus …

Intranasal Immunization Of Pigs With Porcine Reproductive And Respiratory Syndrome Virus Like Particles Plus 2’, 3’ –Cgamp Vaccigrade™ Adjuvant Exacerbates Viremia After Virus Challenge, Alexandria Marie Van Noort

Electronic Theses and Dissertations

Porcine reproductive and respiratory syndrome virus (PRRSV) is an economically devastating virus that causes a persistent infection and spreads quickly. The virus causes abortions in sows and respiratory diseases in young piglets. Current PRRSV vaccines appear to provide limited cross-protection. A virus-like particles (VLPs) vaccine is made up of key PRRSV structural proteins, which could be cross protective among the different strains. We tested PRRSV VLPs as a vaccine candidate with a 2’, 3’-cGAMP VacciGradeTM adjuvant against pigs infected with PRRSV after intranasal immunization to evaluate the immune response and protective efficacy by the VLPs and adjuvant. The VLPs contain …

The Indirect Effect Of Bvdv On Immune Suppression: The Role Of Infected Macrophages In Lymphocyte Apoptosis, Karim Abdelsalam

Electronic Theses and Dissertations

BVDV is an important pathogen of cattle that affects both the dairy and beef industry causing severe economic losses. The main problem of BVDV is the immune suppression that is associated with lymphoid depletion shortly post infection. The mechanism of this immune suppression is not well understood. In this in vitro study, we sought to determine the indirect effect of BVDV infection in macrophages as a key player in the pathogenesis of BVD especially, the apoptotic effect induced on lymphocytes. We infected bovine monocyte-derived macrophages (MDM) with either high or low virulent strains of cytopathogenic or non-cytopathogenic strains of BVDV …

Reagents And Tools To Identify Epizootic Hemorrhagic Disease Virus-2, Meghan Syrstad

Electronic Theses and Dissertations

The purpose of this project was to define monoclonal antibodies against viral proteins from Epizootic Hemorrhagic Disease Virus (EHDV) to be used in quality control of vaccines produced by an industrial partner, and development of tools to identify EHDV. EHDV is an Orbivirus, which also includes Bluetongue Virus and African Horse Sickness, which are all transmitted through arthropod vectors. EHDV causes significant morbidity and mortality in white-tailed deer, but has recently been found to infect cattle. EHDV recently caused significant outbreaks affecting both the farmed and wild cervid industry, however few reagents and tools exist to protect against this disease …

Development And Characterization Of A Recombinant Orf Virus Vector Expressing The Spike Protein Of Porcine Epidemic Diarrhea Virus, Kyle Hain

Electronic Theses and Dissertations

Orf virus (ORFV), the type member of the genus Parapoxvirus of the family Poxviridae, causes orf or cutaneous pustular dermatitis in sheep and goats. ORFV is a ubiquitous virus capable of re-infecting its hosts multiple times over time. ORFV causes a non-systemic, self-limiting disease which is usually restricted to the skin surrounding the virus entry sites. ORFV has evolved several immunomodulatory proteins (IMPs) that evade and/or modulate host immune responses to infection and contribute to virus virulence and disease pathogenesis. Given biological properties and unique immunomodula tor y properties, ORFV has gained significant attention in recent years for its …

Development Of An Indirect Elisa, Blocking Elisa, Fluorescent Microsphere Immunoassay And Fluorescent Focus Neutralization Assay For Serologic Evaluation Of Exposure To North American Strains Of Porcine Epidemic Diarrhea Virus, Faten Okda, Xiaodong Liu, Aaron Singrey, Travis Clement, Julie Nelson, Jane Christopher-Hennings, Eric A. Nelson, Steven Lawson

Veterinary and Biomedical Sciences Faculty Publications

Recent, severe outbreaks of porcine epidemic diarrhea virus (PEDV) in Asia and North America highlight the need for well-validated diagnostic tests for the identification of PEDV infected animals and evaluation of their immune status to this virus. PEDV was first detected in the U.S. in May 2013 and spread rapidly across the country. Some serological assays for PEDV have been previously described, but few were readily available in the U.S. Several U.S. laboratories quickly developed indirect fluorescent antibody (IFA) assays for the detection of antibodies to PEDV in swine serum, indicating prior exposure. However, the IFA has several disadvantages, including …

An Evaluation Of Porcine Epidemic Diarrhea Virus Survival In Individual Feed Ingredients In The Presence Or Absence Of A Liquid Antimicrobial, Scott Dee, Casey Neill, Travis Clement, Aaron Singrey, Jane Christopher-Hennings, Eric A. Nelson

Veterinary and Biomedical Sciences Faculty Publications

Background: Contaminated complete feed and porcine plasma are risk factors for PEDV introduction to farms and a liquid antimicrobial has been proven useful for reducing risk. This study provides information on the survivability of PEDV across common swine feed ingredients in the presence or absence of the liquid antimicrobial.
Results: Eighteen ingredients commonly included in commercial swine diets were selected, including 3 grain sources (corn, soybean meal (SBM), dried distillers grains with solubles (DDGS)), 5 porcine by-products (spray-dried plasma, purified plasma, intestinal mucosa, meat and bone meal and red blood cells (RBCs)), 3 vitamin/trace mineral (VTM) mixes (sow, nursery, finishing), …

Virus-Like Particles Generaged By Expressing Proteins Of Porcine Reproductive And Respiratory Syndrome Virus (Prrsv) Using The Recombinant Baculovirus Expression System, April Anne Nelsen

Electronic Theses and Dissertations

Porcine reproductive and respiratory syndrome virus (PRRSV) is an enveloped, single stranded, positive sense RNA virus and a member of Arteriviridae. Its genome encodes 10 open reading frames for at least 7 structural proteins and 14 non-structural proteins. Membrane (M), Nuclepcapsid (N), and Glycoprotein-5 (GP5) are the major structural proteins of PRRSV, while Envelope (E), Glycoprotein-2 (GP2), Glycoprotein-3 (GP3), and Glycoprotein-4 (GP4) are the minor structural proteins of PRRSV. GP5 induces neutralizing antibodies and forms heterodimers with M, while N is the most immunogenic protein of PRRSV. Previous studies have shown viral structural proteins are able to form virus-like particles …

An Evaluation Of A Liquid Antimicrobial (Sal Curb®) For Reducing The Risk Of Porcine Epidemic Diarrhea Virus Infection Of Naïve Pigs During Consumption Of Contaminated Feed, Scott Dee, Casey Neill, Travis Clement, Jane Christopher-Hennings, Eric A. Nelson

Veterinary and Biomedical Sciences Faculty Publications

Background: Since its initial detection in May 2013, porcine epidemic diarrhea virus (PEDV) has spread rapidly throughout the US swine industry. Recently, contaminated feed was confirmed as a vehicle for PEDV infection of naïve piglets. This research provides in vivo data supporting the ability of a liquid antimicrobial product to reduce this risk.
Results: Sal CURB® (Kemin Industries, Des Moines, IA, USA) is a FDA-approved liquid antimicrobial used to control Salmonella contamination in poultry and swine diets. To test its effect against PEDV, Sal CURB®-treated feed was spiked with a stock isolate of PEDV (Ct = 25.22), which PEDV-naïve piglets …

An Evaluation Of Contaminated Complete Feed As A Vehicle For Porcine Epidemic Diarrhea Virus Infection Of Naïve Pigs Following Consumption Via Natural Feeding Behavior: Proof Of Concept, Scott Dee, Travis Clement, Adam Schelkopf, Joel Nerem, David Knudsen, Jane Christopher-Hennings, Eric A. Nelson

Veterinary and Biomedical Sciences Faculty Publications

Background: Since its initial detection in May 2013, porcine epidemic diarrhea virus (PEDV) has spread rapidly throughout the US swine industry. Initially, contaminated feed was proposed as a risk factor for PEDV; however, data were not available to support this theory. Here we provide proof of concept of this risk by describing a novel means for recovering PEDV-contaminated complete feed material from commercial swine sites and conducting an in vivo experiment to prove its infectivity.

Results: For on-farm detection of PEDV RNA in feed, paint rollers were used to collect material from at-risk feed bins from 3 clinically affected breeding …

Characterization Of A Novel Influenza Virus In Cattle And Swine: Proposal For A New Genus In The Orthomyxoviridae Family, Ben M. Hause, Emily A. Collin, Runxia Liu, Bing Huang, Zizhang Sheng, Wuxun Lu, Dan Wang, Eric A. Nelson, Feng Li

Veterinary and Biomedical Sciences Faculty Publications

We have recently reported the isolation of a novel virus, provisionally designated C/swine/Oklahoma/1334/2011 (C/OK), with 50% overall homology to human influenza C viruses (ICV), from a pig in Oklahoma. Deep RNA sequencing of C/OK virus found a matrix 1 (M1) protein expression strategy that differed from that of ICV. The novelty of C/OK virus prompted us to investigate whether C/OK virus could exist in a nonswine species. Significantly, we found that C/OK virus was widespread in U.S. bovine herds, as demonstrated by reverse transcription (RT)-PCR and serological assays. Genome sequencing of three bovine viruses isolated from two herds in different …

Structural, Antigenic, And Evolutionary Characterizations Of The Envelope Protein Of Newly Emerging Duck Tembusu Virus, Kexoang Yu, Zhi-Zhang Sheng, Bing Huang, Xiuli Ma, Yufeng Li, Xiaoyuam Yuan, Zhuoming Qin, Dan Wang, Suvobrata Chakravarty, Feng Li, Minxun Song, Huaichang Sun

Chemistry and Biochemistry Faculty Publications

Since the first reported cases of ducks infected with a previously unknown flavivirus in eastern China in April 2010, the virus, provisionally designated Duck Tembusu Virus (DTMUV), has spread widely in domestic ducks in China and caused significant economic losses to poultry industry. In this study, we examined in detail structural, antigenic, and evolutionary properties of envelope (E) proteins of six DTMUV isolates spanning 2010–2012, each being isolated from individual farms with different geographical locations where disease outbreaks were documented. Structural analysis showed that E proteins of DTMUV and its closely related flavivirus (Japanese Encephalitis Virus) shared a conserved array …

The Presence Of Alpha Interferon At The Time Of Infection Alters The Innate And Adaptive Immune Responses To Porcine Reproductive And Respiratory Syndrome Virus, Susan L. Brockman, Crystal A. Loving, Eric A. Nelson, Laura C. Miller, Tracy L. Nicholson, Karen B. Register, Marvin J. Grubman, Douglas E. Brough, Marcus E. Kehrli Jr

Veterinary and Biomedical Sciences Faculty Publications

Porcine reproductive and respiratory syndrome (PRRS) is one of the most devastating and costly diseases to the swine industry worldwide. Overall, the adaptive immune response to PRRS virus (PRRSV) is weak, which results in delayed elimination of virus from the host and inferior vaccine protection. PRRSV has been shown to induce a meager alpha interferon (IFN-α) response, and we hypothesized that elevated IFN-α levels early in infection would shorten the induction time and increase elements of the adaptive immune response. To test this, we measured both antibody and cell-mediated immunity in pigs after the administration of a nonreplicating human adenovirus …

Development Of A Fluorescent Microsphere Immunoassay For Detection Of Antibodies Against Porcine Reproductive And Respiratory Syndrome Virus Using Oral Fluid Samples As An Alternative To Serum-Based Assays, Robert J. Langenhorst, Steven Lawson, Apisit Kittawornrat, Jeffery J. Zimmerman, Zhi Sun, Yanhua Li, Jane Christopher-Hennings, Eric A. Nelson, Ying Fang

Veterinary and Biomedical Sciences Faculty Publications

For effective disease surveillance, rapid and sensitive assays are needed to detect antibodies developed in response to porcine reproductive and respiratory syndrome virus (PRRSV) infection. In this study, we developed a multiplexed fluorescent microsphere immunoassay (FMIA) for detection of PRRSV-specific antibodies in oral fluid and serum samples. Recombinant nucleocapsid protein (N) and nonstructural protein 7 (nsp7) from both PRRSV genotypes (type I and type II) were used as antigens and covalently coupled to Luminex fluorescent microspheres. Based on an evaluation of 488 oral fluid samples with known serostatus, the oral fluid-based FMIAs achieved >92% sensitivity and 91% specificity. For serum …

The Lack Of An Inherent Membrane Targeting Signal Is Responsible For The Failure Of The Matrix (M1) Protein Of Influenza A Virus To Bud Into Virus-Like Particles, Dan Wang, Aaron Harmon, Jing Jin, David H. Francis, Jane Christopher-Hennings, Eric A. Nelson, Ronald C. Montelaro, Feng Li

Veterinary and Biomedical Sciences Faculty Publications

The matrix protein (M1) of influenza A virus is generally viewed as a key orchestrator in the release of influenza virions from the plasma membrane during infection. In contrast to this model, recent studies have indicated that influenza virus requires expression of the envelope proteins for budding of intracellular M1 into virus particles. Here we explored the mechanisms that control M1 budding. Similarly to previous studies, we found that M1 by itself fails to form virus-like-particles (VLPs). We further demonstrated that M1, in the absence of other viral proteins, was preferentially targeted to the nucleus/perinuclear region rather than to the …

Antibiotic-Mediated Inhibition Of Porcine Reproductive And Respiratory Syndrome Virus (Prrsv) Infection: A Novel Quinolone Function Which Potentiates The Antiviral Cytokine Response In Marc-145 Cells And Pig Macrophages, William A. Cafruny, Richard G. Duman, Raymond R. Rowland, Eric A. Nelson, Grace H. Wong

Veterinary and Biomedical Sciences Faculty Publications

Porcine reproductive and respiratory syndrome virus (PRRSV) is an economically significant agent for which there currently are no effective treatments. Development of antiviral agents for PRRSV as well as many other viruses has been limited by toxicity of known antiviral compounds. In contrast, antibiotics for non-virus microbial infections have been widely useful, in part because of their acceptable toxicity in animals. We report here the discovery that the quinolone containing compound Plasmocin™, as well as the quinolones nalidixic acid and ciprofl oxacin, have potent anti-PRRSV activity in vitro. PRRSV replication was inhibited by these antibiotics in both cultured MARC-145 cells …