Location: Arthropod-borne Animal Diseases Research2011 Annual Report
1a. Objectives (from AD-416)
Maintain and further develop the USDA-Agricultural Research Service (ARS), Arthropod-Borne Animal Disease Research Unit (ABADRU) exotic bluetongue virus (BTV), Rift Valley fever virus (RVFV), and vesicular stomatitis virus (VSV) strain collections that can be used to support genetic and molecular epidemiologic investigations and diagnostic test development, and 2) Characterize the susceptibility, pathogenesis, and virus-vector-host interactions of BTV, RVFV, and VSV in mammalian and invertebrate cell cultures and hosts.
1b. Approach (from AD-416)
BTV, RVFV, and VSV are pathogens affecting livestock that require an integrated investigative approach involving serology, epidemiology, molecular biology, entomology and veterinary science. ARS-ABADRU and Kansas State University scientists will collaboratively investigate epidemiological aspects of the pathogens in cell culture and conduct molecular genetic investigations. Approaches include developing diagnostic tests of exotic and endemic strains of both viruses in high containment facilities. Staff will maintain collections, generate virus preparations by in vitro growth in cell culture, and purify RNA from the virus preparations. Animal infection studies will be conducted with BTV and VSV. The role of Culicoides sonorensis midge saliva and its affect on establishment of infection, replication and persistence will be evaluated. Bite rates, viral transmission, infestation rates and pathogenesis will be measured by methods including clinical disease observations, virus isolation/plaque assay, polymerase chain reaction, ELISA, necropsy, histopathology and immunohistochemistry.
3. Progress Report
Current ABADRU research staff members (6) were provided training by Biosecurity Research Institute (BRI) research support staff (Biosafety, Security, Information Technology, Facility Operations) in biosafety and operational policies and procedures for research programs to be conducted in the BRI. ABADRU research staff members were provided training by Kansas State University in Institutional Animal Care and Use, Institutional Biosafety Committee, and Export Control policies and procedures. All modules were completed. BRI Biosafety staff and Laboratory Coordinators worked with ABADRU research staff members to develop biosafety manuals and standard operating procedures specific to the current research activities. Regulatory compliance reviews of proposed ABADRU research projects by the K-State Institutional Biosafety Committee and Institutional Animal Care and Use Committee have been conducted. Approvals have been completed prior to initiation of all research activities. Permits and Select Agent Approval Status: BRI Biosafety staff worked with ABADRU research staff to submit applications to the Centers for Disease Control and USDA, APHIS for storage and use of select agent pathogens in research. The application process included current specific project, facility, and safety and security program descriptions, as well as personnel security risk assessments. All 6 ABADRU staff members have satisfied training and registration requirements for individual Select Agent Security Risk Assessment (SRA) and have received SRA approval. BRI staff facilitated all required associated site inspections and follow up (CDC March 2010, CDC and USDA, APHIS Oct 2010, APHIS May 2010). • Received Select Agent approval (August, 2010) and USDA, APHIS permit (October, 2010) for transport of Exotic Bluetongue Virus from Colorado State University and storage at the BRI. The virus collection is now securely stored in the BRI (November, 2010). • Received Select Agent approval (May, 2011) and USDA APHIS permit (June, 2011) for laboratory (in vitro) work with Exotic Bluetongue Virus. Laboratory set-up in designated BSL-3 space has begun and is supported with BRI use fees. • Received USDA, APHIS permit for MP12 vaccine strain of Rift Valley fever virus.