Performing Department
Microbiology & Immunology
Non Technical Summary
Infectious disease causes considerable loss for livestock producers by reducing production of animal units and by reduced sales because of food safety concerns. The Microbiology and Immunology (MBI) Department is the only research unit in Montana focused on animal health, particularly on the study of infectious diseases of cattle, bison and sheep. New faculty members joining MBI are required to initiate new research projects. In addition, other faculty not on Montana Agricultural Experiment Station (MAES) funding may be hired to develop new short­ term projects. These projects are in support of the respective missions of MAES and MBI. Funding from this departmental project is to be used by these scientists to generate data to initiate new MAES projects and/or enhance existing MAES project(s) take advantage of unique opportunities and increase competitiveness for grant funding from federal agencies, such as the USDA. Support is also provided to maintain and operate departmental research facilities.Examples, of newly awarded MAES and/or initiated projects that may receive funds from this project include:New MAES projects1. Studies of alpha-herpes viruses on animal health (Taylor). Several members of viral family alpha-herpesvirinae are pathogens that have a wide and pervasive impact on animal health, agricultural practices and economic productivity. Viruses in this group include Equine Herpesvirus-1 (EHV-1), Bovine Herpesvirus-1 (BHV-1) and Suid Herpesvirus-1, also known as Aujesky's disease or Pseudorabies virus (PRV). All three viruses share the hallmarks of neurological infections, long periods of latency that complicate vaccination and control efforts, and the capacity to elicit severe disease. Despite years of research, we understand surprisingly little about the pathways that regulate neuronal transmission of viral infection. We have previously found that neuronal spread of infection involves limited numbers of infectious particles transmitted between cells. These limitations have uncharacterized effects on how these viruses interact with host immune responses and adapt to evolutionary pressures imposed by vaccination. For the state of Montana, the impact of BHV-1 and EHV-1 on cattle and horse production and transport is emergent. Infections with BHV-1 are widespread, primarily associated with infectious bovine rhinotracheitis and a strong risk factor in shipping fever disease and bovine respiratory disease complex. Severe economic losses related to BHV-1 disease are due to reduced productivity and calf abortion or developmental deformity. In unvaccinated herds, seroprevalence can reach as high as 75-80%. Vaccination efforts can reduce overt disease but it cannot fully prevent infection or eliminate spontaneous transmission. Similarly, EHV-1 can be controlled through effective vaccine and quarantine efforts, but recent cases of EHV-1 in Montana, occurring in 2011 and 2013, can be traced to transport of horses between states for competitive events. Together, BHV-1 and EHV-1 remain viruses of concern among agricultural producers both for the state of Montana as well as nationally.Our objective is to identify the molecular factors that regulate PR V infection and spread within an infected host. We aim to answer the critical questions of how exclusion affects virions and what genes are responsible. To this end, we propose experiments that will quantify PRV virion entry, assess the impact of IFN activation and signaling on SIE and replication and characterize the transcriptional responses that correlate with SIE and IFN induction. Preliminary data presented in this proposal suggests that SIE blocks viral entry. Additionally, our preliminary work points to differential effect of antiviral transcription factors needed for IFN signaling. Finally, we present preliminary evidence that differential transcriptional responses correlate with SIE induction. From our preliminary findings, we hypothesize that early transcriptional changes to viral and cellular genes, initiated by the virus, alter the cell's susceptibility to superinfection.Studies of infectious disease Spillover (P/owright). The objectives in this new project are to develop models that integrate the many processes that determine whether spillover transmission will occur, spanning spatial scales from host cells to landscapes, and to parameterize these models with existing data and data collected in the field. Our models aim to predict how the series of barriers to spillover interact to determine the risk of spillover and onward transmission. In particular, we are interested in the emergent properties of spillover. That is, we wish to investigate the patterns that arise when many non-linearly linked, and dynamic processes are combined. Our novel conceptual and quantitative framework should also reveal how past increases in spillover rates have arisen from environmental and social changes that altered the width and alignment of gaps in barriers. Our framewor k will identify the barriers most amenable to intervention and will provide the operationalized models needed to guide prediction and prevention measures against emergence of known pathogens. These models can also be used to rapidly assess the risks to human and livestock health associated with newly discovered pathogens. Moreover, these models should provide an organ izing principle for a new, integrative body of theory and transdiscipl inary spillover investigation.
Animal Health Component
100%
Research Effort Categories
Basic
75%
Applied
25%
Developmental
(N/A)
Goals / Objectives
Infectious disease causes considerable loss for livestock producers by reducing production of animal units and by reduced sales because of food safety concerns. The Microbiology and Immunology (MBI) Department is the only research unit in Montana focused on animal health, particularly on the study of infectious diseases of cattle, bison and sheep. New faculty members joining MBI are required to initiate new research projects. In addition, other faculty not on Montana Agricultural Experiment Station (MAES) funding may be hired to develop new short­ term projects. These projects are in support of the respective missions of MAES and MBI. Funding from this departmental project is to be used by these scientists to generate data to initiate new MAES projects and/or enhance existing MAES project(s) take advantage of unique opportunities and increase competitiveness for grant funding from federal agencies, such as the USDA. Support is also provided to maintain and operate departmental research facilities.
Project Methods
MBI scientists utilize state-of-the-art molecular approaches to address basic and applied problems in infectious diseaseresearch. These research programs require laboratories, large and small animal facilities, clinics, and modern researchequipment, such as flow cytometers, DNA sequencers, and genomics analysis facilities. The study of agents, such as B. abortusrequire BSL-3 facilities approved for Select Agent work. Specific methods and procedures utilized are dependent upon programtype and necessary protocols. New or existing faculty members must develop an understanding of Montana and regional issuesand a more in-depth understanding of existing research programs, prior to developing their own MAES project proposals. Allresearch will have appropriate IBC Protocol Numbers and IACUC Protocol approvals.1. Molecular pathogenesis of West Nile Virus. West Nile virus (WNV) is an emergent neuroinvasive pathogen that causessevere illness in a wide range of vertebrates, including horses. In recently published work, we reported a restriction that limitsthe number of alphaherpes virions infecting neurons (Taylor et al., 2012). A series of experiments that aim to identify ifneuroinvasive spread of WNV is restricted in the number of virions transmitted between cells and characterize the cellularantiviral signaling activated neuronal infection of WNV will be done. By characterizing neuroinvasive spread of WNV, we willbegin to understand how neuroinvasive viruses are controlled during zoonotic infections and understand the impacts on viralpopulation biology and pathogenesis. IBC Protocol Numbers: 032-2014 (WNV), 08-2015 (recombinant Baculovirus) and IACUCProtocol: 2013-282. Genome editing through use of CRISPR technology. Cas9 is a new RNA-guide technology that permits rapid and precisemanipulation of plant and animal genomes. This new gene knockout system is revolutionizing molecular biology and the value ofthis technology is currently estimated in the billions. However, this technique has been primarily restricted to generating geneknockouts (i.e. search and delete functions). Knockouts are critical for determining the biological function of a specific gene, butthe genetic basis of many plant and animal diseases are already known and the next major advance will be to design methodsthat facilitate precise repair of defective genes (i.e. search and replace functions). Here we propose lentiviral delivery of Cas9 togenerate a genome-wide knockout library in human cells that will identify novel pathways that enhance Homology DirectedRepair (HDR) of double-stranded DNA breaks. We anticipate that many of the human genes involved in HDR will haveconserved functions in other animals and plants. Results from this screen will provide fundamental new insight into DNA repairmechanisms, and provide a foundation for designing the next generation of tools for "correcting" genetic disorders andengineering Ag related products for resistance to environmental stress, and improving food security for the rapidly expandingpoppopulation. IBC Protocol 024-2014.Mucosal immunology in pigs (NIH pilot grant peer-review). This study seeks to obtain pilot data to evaluate the currentzoonotic threat and the mechanisms of gastric pathogenesis for human Helicobacter suis (H. suis) infection. H. suis is astomach-dwelling bacterium naturally found in pigs that has been associated with gastric lesions including gastric cancer inhumans. The overall hypothesis is that H. suis is a relevant zoonotic pathogen that disrupts human gastric epithelial cellfunction by triggering inflammatory signaling. In summary, our study will elucidate inflammatory pathways induced by H. suisand will determine whether there is a need for further research into zoonotic H. suis infection. IBC protocol 042-2014.4. New vaccine development for Coxiella and Brucella (new project). Using novel virus-like particles, unique vaccine constructsare being developed for effective T cell immune responses against Coxiella burnetii and Brucella sps. Vaccine constructs willbe used to immunize animals, prior to challenge with with C. burnetii or Brucella sps. Proof of principle studies will be done inmice under BSL-3 containment. Immune reponses against each construct will also be measured in cattle as our main livestockmodel system. Here animals will be immunized with different doses of vaccine and then antibody titers and antigen-specific Tcell responses measured at different times, post vaccination.5. Funding will also be provided for new faculty who develop animal health related projects. Possible projects include diseaseecology of pneumonia in bighorn sheep and domestic sheep, and scouring diseases in cattle.