Veterinary Microbiology and Immunobiology Commons^™

Rapid Typing Of Mannheimia Haemolytica Major Genotypes 1 And 2 Using Maldi-Tof Mass Spectrometry, John Dustin Loy, Michael L. Clawson

School of Veterinary and Biomedical Sciences: Faculty Publications

Genotype 2 M. haemolytica predominantly associate over genotype 1 with the lungs of cattle with respiratory disease and ICEs containing antimicrobial resistance genes. Distinct protein masses were detected by MALDI-TOF MS between genotype 1 and 2 strains. MALDI-TOF MS could rapidly differentiate genotype 2 strains in veterinary diagnostic laboratories.

A Parapoxviral Virion Protein Targets The Retinoblastoma Protein To Inhibit Nf-Κb Signaling, Ponnuraj Nagendraprabhu, Sushil Khatiwada, Sabal Chaulagain, Gustavo A. Delhon, Daniel L. Rock

School of Veterinary and Biomedical Sciences: Faculty Publications

Poxviruses have evolved multiple strategies to subvert signaling by Nuclear Factor κB (NF-κB), a crucial regulator of host innate immune responses. Here, we describe an orf virus (ORFV) virion-associated protein, ORFV119, which inhibits NF-κB signaling very early in infection (≤30 min post infection). ORFV119 NF-κB inhibitory activity was found unimpaired upon translation inhibition, suggesting that virion ORFV119 alone is responsible for early interference in signaling. A C-terminal LxCxE motif in ORFV119 enabled the protein to interact with the retinoblastoma protein (pRb) a multifunctional protein best known for its tumor suppressor activity. Notably, experiments using a recombinant virus containing an ORFV119 …

Efficacy Of Urtoxazumab (Tma-15 Humanized Monoclonal Antibody Specific For Shiga Toxin 2) Against Post-Diarrheal Neurological Sequelae Caused By Escherichia Coli O157:H7 Infection In The Neonatal Gnotobiotic Piglet Model, Rodney A. Moxley, David H. Francis, Mizuho Tamura, David B. Marx, Kristina Santiago-Mateo, Mojun Zhao

School of Veterinary and Biomedical Sciences: Faculty Publications

Enterohemorrhagic Escherichia coli (EHEC) is the most common cause of hemorrhagic colitis and hemolytic uremic syndrome in human patients, with brain damage and dysfunction the main cause of acute death. We evaluated the efficacy of urtoxazumab (TMA-15, Teijin Pharma Limited), a humanized monoclonal antibody against Shiga toxin (Stx) 2 for the prevention of brain damage, dysfunction, and death in a piglet EHEC infection model. Forty-five neonatal gnotobiotic piglets were inoculated orally with 3 x 10⁹ colony-forming units of EHEC O157:H7 strain EDL933 (Stx1⁺, Stx2⁺) when 22–24 h old. At 24 h post-inoculation, piglets were intraperitoneally …

Cytolytic Toxin Production By Staphylococcus Aureus Is Dependent Upon The Activity Of The Protoheme Ix Farnesyltransferase, Emily Stevens, Maisem Laabei, Stewart Gardner, Greg A. Somerville, Ruth C. Massey

School of Veterinary and Biomedical Sciences: Faculty Publications

Staphylococcus aureus is a medically important pathogen with an abundance of virulence factors that are necessary for survival within a host, including the production of cytolytic toxins. The regulation of toxin production is mediated by the Agr quorum sensing system, and a poorly defined post-exponential growth phase signal independent of Agr. As part of a recent genome wide association study (GWAS) to identify novel loci that alter the expression of cytolytic toxins, a polymorphism in the cyoE gene, which encodes a protoheme IX farnesyltransferase, was identified. This enzyme is essential for processing heme into the electron transport chain for use …

Branched Chain Α-Ketoacid Dehydrogenase Kinase 111–130, A T Cell Epitope That Induces Both Autoimmune Myocarditis And Hepatitis In A/J Mice, Bharathi Krishnan, Chandirasegara Massilamany, Rakesh H. Basavalingappa, Arunakumar Gangaplara, Guobin Kang, Qingsheng Li, Francisco A. Uzal, Jennifer L. Strande, Gustavo A. Delhon, Jean-Jack Riethoven, David J. Steffen, Jay Reddy

School of Veterinary and Biomedical Sciences: Faculty Publications

Introduction: Organ-specific autoimmune diseases are believed to result from immune responses generated against self-antigens specific to each organ. However, when such responses target antigens expressed promiscuously in multiple tissues, then the immune-mediated damage may be wide spread.

Methods: In this report, we describe a mitochondrial protein, branched chain α-ketoacid dehydrogenase kinase (BCKD_k) that can act as a target autoantigen in the development of autoimmune inflammatory reactions in both heart and liver.

Results: We demonstrate that BCKD_k protein contains at least nine immunodominant epitopes, three of which, BCKD_k 71–90, BCKD_k 111–130 and BCKD_k 141–160, …

Β1-Adrenergic Receptor Contains Multiple IaK And IeK Binding Epitopes That Induce T Cell Responses With Varying Degrees Of Autoimmune Myocarditis In A/J Mice, Rakesh H. Basavalingappa, Chandirasegaran Massilamany, Bharathi Krishnan, Arunakumar Gangaplara, Rajkumar A. Rajasekaran, Muhammad Z. Afzal, Jean-Jack Riethoven, Jennifer L. Strande, David J. Steffen, Jay Reddy

School of Veterinary and Biomedical Sciences: Faculty Publications

Myocarditis/dilated cardiomyopathy (DCM) patients can develop autoantibodies to various cardiac antigens and one major antigen is β1-adrenergic receptor (β₁AR). Previous reports indicate that animals immunized with a β₁AR fragment encompassing, 197–222 amino acids for a prolonged period can develop DCM by producing autoantibodies, but existence of T cell epitopes, if any, were unknown. Using A/J mice that are highly susceptible to lymphocytic myocarditis, we have identified β₁AR 171–190, β₁AR 181–200, and β₁AR 211–230 as the major T cell epitopes that bind major histocompatibility complex class II/IA^k or IE^{k …}

The Effect Of Feed Form On Diet Digestibility And Cecal Parameters In Rabbits, Isabella Corsato Alvarenga, Charles Gregory Aldrich, Micah Kohles

School of Veterinary and Biomedical Sciences: Faculty Publications

Fifteen New Zealand rabbits were randomly assigned to one of 3 dietary treatment groups of 5 animals each and fed pelleted, extruded, or muesli diets in a completely randomized design experiment. Rabbits were placed in individual cages with ad libitum access to water and food for 45 days acclimation followed by 30 days experimental period. Feed intake of rabbits fed pelleted and extruded diets was greater (p < 0.05) than rabbits fed the muesli diet (125.6 and 130.4 vs. 91.9 g/d), but weight change and feed efficiency were not affected by treatment. Diet digestibility among the treatments was inconsistent when comparing results obtained from total fecal collection and AIA (please define) as an internal marker. Rabbits fed extruded and pelleted diets had lower (p < 0.05) cecal pH (6.42 and 6.38 vs. 7.02, respectively), and higher (p < 0.05) production of SCFA (18.5 and 19.0 vs. 11.7 mM, respectively) than those fed muesli. The fermentation products from rabbits fed pelleted and extruded diets had a greater proportion of butyrate and less propionate than rabbits fed muesli. The results of this study indicate that the basal dietary composition had a greater impact on diet utilization and cecal fermentation than food form.

Mechanistic Understanding Of N-Glycosylation In Ebola Virus Glycoprotein Maturation And Function, Bin Wang, Yujie Wang, Dylan A. Frabutt, Xihe Zhang, Xiaoyu Yao, Dan Hu, Zhuo Zhang, Chaonan Liu, Shimin Zheng, Shi-Hua Xiang, Yong-Hui Zheng

School of Veterinary and Biomedical Sciences: Faculty Publications

The Ebola virus (EBOV) trimeric envelope glycoprotein (GP) precursors are cleaved into the receptor-binding GP₁ and the fusion-mediating GP₂ subunits and incorporated into virions to initiate infection. GP₁ and GP₂ form heterodimers that have 15 or two N-glycosylation sites (NGSs), respectively. Here we investigated the mechanism of how N-glycosylation contributes to GP expression, maturation, and function. As reported before, we found that, although GP₁ NGSs are not critical, the two GP₂ NGSs, Asn⁵⁶³ and Asn⁶¹⁸, are essential for GP function. Further analysis uncovered that Asn⁵⁶³ and Asn^{618 …}

Pathogenesis, Molecular Genetics, And Genomics Of Mycobacterium Avium Subsp. Paratuberculosis, The Etiologic Agent Of Johne’S Disease, Govardhan Rathnaiah, Denise K. Zinniel, John P. Bannantine, Judith R. Stabel, Yrjo T. Grohn, Michael T. Collins, Raul G. Barletta

School of Veterinary and Biomedical Sciences: Faculty Publications

Mycobacterium avium subsp. paratuberculosis (MAP) is the etiologic agent of Johne’s disease in ruminants causing chronic diarrhea, malnutrition, and muscular wasting. Neonates and young animals are infected primarily by the fecal–oral route. MAP attaches to, translocates via the intestinal mucosa, and is phagocytosed by macrophages. The ensuing host cellular immune response leads to granulomatous enteritis characterized by a thick and corrugated intestinal wall. We review various tissue culture systems, ileal loops, and mice, goats, and cattle used to study MAP pathogenesis. MAP can be detected in clinical samples by microscopy, culturing, PCR, and an enzyme-linked immunosorbent assay. There are commercial …

Experimental Infection Of Conventional Nursing Pigs And Their Dams With Porcine Deltacoronavirus, Sarah Vitosh-Sillman, John Dustin Loy, Bruce W. Brodersen, Clayton L. Kelling, Alan R. Doster, Christina L. Topliff, Eric A. Nelson, Jianfa Bai, Erin Schirtzinger, Elizabeth Poulsen, Barbara Meadors, Joseph Anderson, Benjamin Hause, Gary Anderson, Richard Hesse

School of Veterinary and Biomedical Sciences: Faculty Publications

Porcine deltacoronavirus (PDCoV) is a newly identified virus that has been detected in swine herds of North America associated with enteric disease. The aim of this study was to demonstrate the pathogenicity, course of infection, virus kinetics, and aerosol transmission of PDCoV using 87 conventional piglets and their 9 dams, including aerosol and contact controls to emulate field conditions. Piglets 2–4 days of age and their dams were administered an oronasal PDCoV inoculum with a quantitative real-time reverse transcription (qRT)-PCR quantification cycle (Cq) value of 22 that was generated from a field sample having 100% nucleotide identity to USA/Illinois121/2014 determined …

Membrane Insertion For The Detection Of Lipopolysaccharides: Exploring The Dynamics Of Amphiphile-In-Lipid Assays, Loreen R. Stromberg, Nicolas W. Hengartner, Kirstie L. Swingle, Rodney A. Moxley, Steven W. Graves, Gabriel A. Montaño, Harshini Mukundan

School of Veterinary and Biomedical Sciences: Faculty Publications

Shiga toxin-producing Escherichia coli is an important cause of foodborne illness, with cases attributable to beef, fresh produce and other sources. Many serotypes of the pathogen cause disease, and differentiating one serotype from another requires specific identification of the O antigen located on the lipopolysaccharide (LPS) molecule. The amphiphilic structure of LPS poses a challenge when using classical detection methods, which do not take into account its lipoglycan biochemistry. Typically, detection of LPS requires heat or chemical treatment of samples and relies on bioactivity assays for the conserved lipid A portion of the molecule. Our goal was to develop assays …

Development Of A Novel Self-Propagating Prssv-Vsv G Hybrid Replicon As A Vector For Inducing Broad Prrsv Protection, Asit K. Pattnaik

School of Veterinary and Biomedical Sciences: Faculty Publications

Throughout successive cycles of Pork Check-off funding, our laboratories (Pattnaik’s and Osorio’s labs) have consistently produced new and fundamental information on: (i) the immunologic mechanisms that are important for protection against PRRSV infection, (ii) the structural basis for induction of PRRSV-neutralizing antibodies which are significant for conferring protective immunity, (iii) the possibility of producing a rationally-designed new generation DIVA vaccine that would offer more efficacious protection, and (iv) the presence of conserved B- and T-cell epitopes in the structural and nonstructural proteins (NSPs) of PRRSV.

The Pseudorabies Virus Protein, Pul56, Enhances Virus Dissemination And Virulence But Is Dispensable For Axonal Transport, Gina R. Daniel, Patricia J. Sollars, Gary E. Pickard, Gregory A. Smith

School of Veterinary and Biomedical Sciences: Faculty Publications

Neurotropic herpesviruses exit the peripheral nervous system and return to exposed body surfaces following reactivation from latency. The pUS9 protein is a critical viral effector of the anterograde axonal transport that underlies this process. We recently reported that while pUS9 increases the frequency of sorting of newly assembled pseudorabies virus particles to axons from the neural soma during egress, subsequent axonal transport of individual virus particles occurs with wild-type kinetics in the absence of the protein. Here, we examine the role of a related pseudorabies virus protein, pUL56, during neuronal infection. The findings indicate that pUL56 is a virulence factor …

Large Genomic Differences Between Moraxella Bovoculi Isolates Acquired From The Eyes Of Cattle With Infectious Bovine Keratoconjunctivitis Versus The Deep Nasopharynx Of Asymptomatic Cattle, Aaron M. Dickey, John D. Loy, James L. Bono, Timonthy P. L. Smith, Mike D. Apley, Brian V. Lubbers, Keith D. Dedonder, Sarah F. Capik, Robert L. Larson, Brad J. White, Jochen Blom, Carol G. Chitko-Mckown, Michael L. Clawson

School of Veterinary and Biomedical Sciences: Faculty Publications

Moraxella bovoculi is a recently described bacterium that is associated with infectious bovine keratoconjunctivitis (IBK) or “pinkeye” in cattle. In this study, closed circularized genomes were generated for seven M. bovoculi isolates: three that originated from the eyes of clinical IBK bovine cases and four from the deep nasopharynx of asymptomatic cattle. Isolates that originated from the eyes of IBK cases profoundly differed from those that originated from the nasopharynx of asymptomatic cattle in genome structure, gene content and polymorphism diversity and consequently placed into two distinct phylogenetic groups. These results suggest that there are genetically distinct strains of M. …

Autoimmunity In Picornavirus Infections, Chandirasegara Massilamany, Andreas Koenig, Jay Reddy, Sally Huber, Iwona Buskiewicz

School of Veterinary and Biomedical Sciences: Faculty Publications

Enteroviruses are small, non-enveloped, positive-sense singlestrand RNA viruses, and are ubiquitously found throughout the world. These viruses usually cause asymptomatic or mild febrile illnesses, but have a propensity to induce severe diseases including type 1 diabetes and pancreatitis, paralysis and neuroinflammatory disease, myocarditis, or hepatitis. This pathogenicity may result from induction of autoimmunity to organ-specific antigens. While enterovirus-triggered autoimmunity can arise from multiple mechanisms including antigenic mimicry and release of sequestered antigens, the recent demonstration of T cells expressing dual T cell receptors arising as a natural consequence of Theiler’s virus infection is the first demonstration of this autoimmune mechanism.

Genome-Wide Search For Host Association Factors During Ovine Progressive Pneumonia Virus Infection, Jesse Thompson, Fangrui Ma, Meghan Quinn, Shi-Hua Xiang

School of Veterinary and Biomedical Sciences: Faculty Publications

Ovine progressive pneumonia virus (OPPV) is an important virus that causes serious diseases in sheep and goats with a prevalence of 36% in the USA. Although OPPV was discovered more than half of a century ago, little is known about the infection and pathogenesis of this virus. In this report, we used RNA-seq technology to conduct a genome-wide probe for cellular factors that are associated with OPPV infection. A total of approximately 22,000 goat host genes were detected of which 657 were found to have been significantly up-regulated and 889 down-regulated at 12 hours post-infection. In addition to previously known …

From Crystal Structure Of Α-Conotoxin Gic In Complex With Ac-Achbp To Molecular Determinants Of Its High Selectivity For Α3Β2 Nachr, Bo Lin, Manyu Xu, Xiaopeng Zhu, Yong Wu, Xi Liu, Dongting Zhangsun, Yuanyan Hu, Shi-Hua Xiang, Igor E. Kasheverov, Victor I. Tsetlin, Xinquan Wang, Sulan Luo

School of Veterinary and Biomedical Sciences: Faculty Publications

Acetylcholine binding proteins (AChBPs) are unique spatial homologs of the ligand-binding domains of nicotinic acetylcholine receptors (nAChRs), and they reproduce some pharmacological properties of nAChRs. X-ray crystal structures of AСhBP in complex with α-conotoxins provide important insights into the interactions of α-conotoxins with distinct nAChR subtypes. Although considerable efforts have been made to understand why α-conotoxin GIC is strongly selective for α3β2 nAChR, this question has not yet been solved. Here we present the structure of α-conotoxin GIC in complex with Aplysia californica AChBP (Ac-AChBP) at a resolution of 2.1 Å. Based on this co-crystal structure complemented with molecular docking …

Presence Of Pathogenic Escherichia Coli Is Correlated With Bacterial Community Diversity And Composition On Pre-Harvest Cattle Hides, Jessica Chopyk, Ryan M. Moore, Zachary Dispirito, Zachary R. Stromberg, Gentry L. Lewis, David G. Renter, Natalia Cernicchiaro, Rodney A. Moxley, K. Eric Wommack

School of Veterinary and Biomedical Sciences: Faculty Publications

Background: Since 1982, specific serotypes of Shiga toxin-producing Escherichia coli (STEC) have been recognized as significant foodborne pathogens acquired from contaminated beef and, more recently, other food products. Cattle are the major reservoir hosts of these organisms, and while there have been advancements in food safety practices and industry standards, STEC still remains prevalent within beef cattle operations with cattle hides implicated as major sources of carcass contamination. To investigate whether the composition of hide-specific microbial communities are associated with STEC prevalence, 16S ribosomal RNA (rRNA) bacterial community profiles were obtained from hide and fecal samples collected from a large …

On And Off Retinal Ganglion Cells Differentially Regulate Serotonergic And Gabaergic Activity In The Dorsal Raphe Nucleus, Ting Zhang, Lu Huang, Li Zhang, Minjie Tan, Mingliang Pu, Gary E. Pickard, Kwok -Fai So, Chaoran Ren

School of Veterinary and Biomedical Sciences: Faculty Publications

The dorsal raphe nucleus (DRN), the major source of serotonergic input to the forebrain, receives excitatory input from the retina that can modulate serotonin levels and depressive-like behavior. In the Mongolian gerbil, retinal ganglion cells (RGCs) with alpha-like morphological and Y-like physiological properties innervate the DRN with ON DRN-projecting RGCs out numbering OFF DRN-projecting RGCs. The DRN neurons targeted by ON and OFF RGCs are unknown. To explore retino-raphe anatomical organization, retinal afferents labeled with Cholera toxin B were examined for association with the postsynaptic protein PSD-95. Synaptic associations between retinal afferents and DRN serotonergic and GABAergic neurons were observed. …

Whole Genomic Sequence Analysis Of Bacillus Infantis: Defining The Genetic Blueprint Of Strain Nrrl B-14911, An Emerging Cardiopathogenic Microbe, Chandirasegara Massilamany, Akram Mohammed, John Dustin Loy, Tanya Purvis, Bharathi Krishnan, Rakesh H. Basavalingappa, Christy M. Kelley, Chittibabu Guda, Raúl G. Barletta, Etsuko N. Moriyama, Timothy P.L. Smith, Jay Reddy

School of Veterinary and Biomedical Sciences: Faculty Publications

Background: We recently reported the identification of Bacillus sp. NRRL B-14911 that induces heart autoimmunity by generating cardiac-reactive T cells through molecular mimicry. This marine bacterium was originally isolated from the Gulf of Mexico, but no associations with human diseases were reported. Therefore, to characterize its biological and medical significance, we sought to determine and analyze the complete genome sequence of Bacillus sp. NRRL B-14911.

Results: Based on the phylogenetic analysis of 16S ribosomal RNA (rRNA) genes, sequence analysis of the 16S-23S rDNA intergenic transcribed spacers, phenotypic microarray, and matrix-assisted laser desorption ionization time-offlight mass spectrometry, we propose that this …

Erratum To: Association Of Autophagy In The Cell Death Mediated By Dihydrotestosterone In Autoreactive T Cells Independent Of Antigenic Stimulation, Ting Jia, Anandhan Annadurai, Chandirasegara Massilamany, Rajkumar A. Rajasekaran, Rodrigo Franco, Jay Reddy

School of Veterinary and Biomedical Sciences: Faculty Publications

No abstract provided.

Localization Of Cd8 T Cell Epitope Within Cardiac Myosin Heavy Chain-Α334-352 That Induces Autoimmune Myocarditis In A/J Mice, Chandirasegara Massilamany, Arunakumar Gangaplara, Rakesh H. Basavalingappa, Rajkumar A. Rajasekaran, Vahid Khalilzad-Sharghi, Zhongji Han, Shadi F. Othman, David Steffen, Jay Reddy

School of Veterinary and Biomedical Sciences: Faculty Publications

Background—Cardiac myosin heavy chain-α (Myhc), an intracellular protein expressed in the cardiomyocytes, has been identified as a major autoantigen in cardiac autoimmunity. In our studies with Myhc_334-352-induced experimental autoimmune myocarditis in A/J mice (H-2a), we discovered that Myhc_334-352, supposedly a CD4 T cell epitope, also induced antigen-specific CD8 T cells that transfer disease to naïve animals.

Methods and Results—In our efforts to identify the CD8 T cell determinants, we localized Myhc_338-348 within the full length-Myhc_334-352, leading to four key findings. (1) By acting as a dual epitope, Myhc_338-348 induces both …

Prevalence And Level Of Enterohemorrhagic Escherichia Coli In Culled Dairy Cows At Harvest, Zachary R. Stromberg, Gentry L. Lewis, Sharif S. Aly, Terry W. Lehenbauer, Joseph M. Bosilevac, Natalia Cernicchiaro, Rodney A. Moxley

School of Veterinary and Biomedical Sciences: Faculty Publications

The primary objective of this study was to determine the prevalence and level of enterohemorrhagic Escherichia coli (EHEC) O26, O45, O103, O111, O121, and O145 (collectively EHEC-6) plus EHEC O157 in fecal, hide, and preintervention carcass surface samples from culled dairy cows. Matched samples (n=300) were collected from 100 cows at harvest and tested by a culture-based method and two molecular methods: NeoSEEK STEC (NS) and Atlas STEC EG2 Combo. Both the culture and NS methods can be used to discriminate among the seven EHEC types (EHEC-7), from which the cumulative prevalence was inferred, whereas the Atlas method can discriminate …

Visualizing Herpesvirus Procapsids In Living Cells, Oana Maier, Patricia J. Sollars, Gary E. Pickard, Gregory A. Smith

School of Veterinary and Biomedical Sciences: Faculty Publications

A complete understanding of herpesvirus morphogenesis requires studies of capsid assembly dynamics in living cells. Although fluorescent tags fused to the VP26 and pUL25 capsid proteins are available, neither of these components is present on the initial capsid assembly, the procapsid. To make procapsids accessible to live-cell imaging, we made a series of recombinant pseudorabies viruses that encoded green fluorescent protein (GFP) fused in frame to the internal capsid scaffold and maturation protease. One recombinant, a GFP-VP24 fusion, maintained wild-type propagation kinetics in vitro and approximated wild-type virulence in vivo. The fusion also proved to be well tolerated in …

Cross-Sectional Analysis Of Salivary Cortisol And Cyathostome Infestation In Horses, Danielle L. Keating

Undergraduate Honors Thesis Projects

With an increase in anthelmintic resistance and decreased efficacy of many commercial dewormers, understanding factors that contribute to parasite infestations in horses is integral to their management. The goal of this study was to look at the potential relationship between parasites and stress response by evaluating salivary cortisol levels and cyathostome egg shedding levels. Using a sample size of n = 200 horses from the state of Ohio, fecal and saliva samples were collected from each horse. Fecal egg counts were performed for each horse with a modified Stoll method, and saliva samples were tested with an enzyme-linked immunosorbent assay …

A Synthetic Porcine Reproductive And Respiratory Syndrome Unprecedented Levels Of Heterologous Protection, Hiep Vu, Fangrui Ma, William W. Laegreid, Asit K. Pattnaik, David Steffen, Alan R. Doster, Fernando A. Osorio

School of Veterinary and Biomedical Sciences: Faculty Publications

Current vaccines do not provide sufficient levels of protection against divergent porcine reproductive and respiratory syndrome virus (PRRSV) strains circulating in the field, mainly due to the substantial variation of the viral genome. We describe here a novel approach to generate a PRRSV vaccine candidate that could confer unprecedented levels of heterologous protection against divergent PRRSV isolates. Using a set of 59 non-redundant, full genome sequences of type-2 PRRSV, a consensus genome (designated as PRRSV-CON) was these 59 PRRSV full genome sequences, followed by selecting the most common nucleotide found at each position of the alignment. Next, the synthetic PRRSV-CON …

Overview Of Rhabdo- And Filoviruses, Asit K. Pattnaik, Michael A. Whitt

School of Veterinary and Biomedical Sciences: Faculty Publications

Enveloped viruses with a negative-sense, single-stranded monopartite RNA genome have been classified into the order Mononegavirales. Five families of viruses that constitute the order are: Rhabdoviridae, Filoviridae, Paramyxoviridae, Bornaviridae and Nyamiviridae. Members of these families possess a helical nucleocapsid core containing the viral genome and a host-derived lipid envelope containing viral proteins. This introductory chapter provides a brief overview of the Rhabdoviridae and the Filoviridae, the two families of viruses that are the subject of this book. Many members of these two families are highly significant human and animal pathogens. The rationale and goal of the book is to …

Host Cell Functions In Vesicular Stomatitis Virus Replication, Phat X. Dinh, Anshuman Das, Asit K. Pattnaik

School of Veterinary and Biomedical Sciences: Faculty Publications

Vesicular stomatitis virus (VSV), the prototypic rhabdovirus, has been used as an excellent paradigm for understanding the mechanisms of virus replication, pathogenesis, host response to virus infection and also for myriads of studies on cellular and molecular biology. Biochemical studies as well as high-throughput genomics, proteomics, and chemical approaches have revealed a plethora of cellular factors and pathways that regulate replication of VSV. These factors include those that support virus replication and also those that restrict its replication. This chapter discusses the role(s) of many of these host cell factors and pathways involved in VSV replication. Although mechanistic understanding of …

New Tools To Convert Bacterial Artificial Chromosomes To A Self-Excising Design And Their Application To A Herpes Simplex Virus Type 1 Infectious Clone, Alexsia L. Richards, Patricia J. Sollars, Gregory A. Smith

School of Veterinary and Biomedical Sciences: Faculty Publications

Background: Infectious clones are fundamental tools for the study of many viruses, allowing for efficient mutagenesis and reproducible production of genetically-defined strains. For the large dsDNA genomes of the herpesviridae, bacterial artificial chromosomes have become the cloning vector of choice due to their capacity to house full-length herpesvirus genomes as single contiguous inserts. Furthermore, while maintained as plasmids in Escherichia coli, the clones can be mutated using robust prokaryotic recombination systems. An important consideration in the design of these clones is the means by which the vector backbone is removed from the virus genome upon delivery into mammalian cells. …

Resolving Bovine Viral Diarrhea Virus Subtypes From Persistently Infected U.S. Beef Calves With Complete Genome Sequence, Aspen M. Workman, Michael P. Heaton, Gregory P. Harhay, Timothy P.L. Smith, Dale M. Grotelueschen, David Sjeklocha, Bruce Brodersen, Jessica L. Petersen, Carol G. Chitko-Mckown

School of Veterinary and Biomedical Sciences: Faculty Publications

Bovine viral diarrhea virus (BVDV) is classified into 2 genotypes, BVDV-1 and BVDV-2, each of which contains distinct subtypes with genetic and antigenic variation. To effectively control BVDV by vaccination, it is important to know which subtypes of the virus are circulating and how their prevalence is changing over time. Accordingly, the purpose of our study was to estimate the current prevalence and diversity of BVDV subtypes from persistently infected (PI) beef calves in the central United States. Phylogenetic analysis of the 5′-UTR (5′ untranslated region) for 119 virus strains revealed that a majority (82%) belonged to genotype 1b, and …