1a.Objectives (from AD-416)
The objective of this cooperative research project is to determine the local expression of various bovine cytokines in virus infected skin cells by usint in-situ hybridization to monitor expression of viral genes.

1b.Approach (from AD-416)
The cooperator, Dr. Brown, has developed a series of methodologies to detect virus infection and cytokine responses in livestock using in-situ hybridization and immunohistochemistry. We will take advantage of samples obtained at PIADC from experimentally infected cattle and analyze their cytokine response at Dr. Brown's laboratory. A graduate student will carry out the work both at PIADC and at Dr. Brown's laboratory.

3.Progress Report
This project documents research conducted under a specific cooperative agreement between ARS and the University of Georgia. Additional details of this research can be found in the report for the in-house associated project, 1940-32000-047-00D, "Pathogenesis and Genomics of Foot-and-Mouth Disease and Vesicular Stomatitis Viruses", NP 103, Animal Health. This agreement is related to objective 3, "Assess the Role of Insect Vector Transmission on the Pathogenesis of VSV." The project aims to investigate the pathogenesis of vesicular stomatitis in cattle, specifically how the virus may be invading different cells, moving through the body, and causing clinical disease. We are also investigating possible methods of transmission, specifically, the role of flies in the establishment of the virus at the bite site. The planned experiments involve the inoculation of healthy adult cattle with the virus via different methods and collection of tissue, with application of advanced pathologic techniques on the collected tissue, to elucidate mechanisms of pathogenesis. In FY 2007, a pilot study was conducted utilizing scarification or fly bite to infect cattle. Preliminary results include the presence of abundant replication virus at the site of scarification and fly bite and evidence of virus replication within macrophage-type cells in the draining lymph nodes. Major accomplishments over the life of this project cannot be reported as of yet, the results of bench work is still being processed. In addition to infection of additional animals and collection of tissues to expand the amount of data we are accumulating, we will also explore Laser Capture Microdissection as a technique for use in delineating functions of specific cells in the upcoming year. We have contacted investigators at the NIH who have graciously agreed to host Dr. Janildo Reis for training in the fall of 2007. At NIH, he will receive training on the same type of equipment that is present at the Plum Island Animal Disease Center. This collaborative research project is monitored through site visits of UGA collaborators to PIADC, conference calls and email.