2012 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.
Current ABADRU research staff members (10) 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. These training modules include: cyber security, physical security, biosafety, biocontainment, select agent rules, respiratory protection, chemical hygiene/lab safety, hazardous waste awareness, and fire safety that are directly associated with protocols, procedures, practices and policies of the BRI. All modules were completed. All ABADRU staff are entered into training records management.
ABADRU research staff members were provided new or refresher training as needed by Kansas State University in Institutional Animal Care and Use, Institutional Biosafety Committee, and Export Control policies and procedures. All modules were completed.
Additional BRI activities and accomplishments included physical walk-thru of facilities and review of procedures associated with multiple research program planning meetings for projected laboratory and animal space use and ongoing research project support. This included Rift Valley fever virus (RVFV)-MP12 research with lambs, calves, and mice as well as exotic bluetongue virus (BTV) lab work.
BRI Biosafety staff and Laboratory Coordinators worked with ABADRU research staff members to develop biosafety manuals and standard operating procedures specific to the current and future research activities.
The arthropod containment space for exotic BTV transmission studies was made ready in November of 2011 and inspected for approvals by Select Agent regulatory officials (see below) to enable ABADRU to continue Culicoides vector transmission work in the BRI.
Two ABADRU scientists were given Adjunct Faculty status in the Department of Diagnostic Medicine and Pathobiology (Ruder) or the Department of Entomology (Nayduch) at Kansas State University.
ABADRU representation to K-State Institutional Biosafety Committee and Institutional Animal Care and Use Committees continues. Several IBC and IACUC approvals have been processed and approved through the full committee, including: MP12 mouse antibody production, wild type Rift valley fever lab work, and exotic BTV lab work.
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.
USDA, ABADRU research project occupy large animal holding spaces, small animal vivarium spaces and biocontainment laboratories of the BRI.
KSU and ABADRU jointly wrote, acted and produced an animal handling training video entitled “Working with Livestock in Biocontainment” which has been posted by ABSA and can be found at http://www.absa.org/resanimal.html.
Permits, Select Agent approval, and research 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 10 ABADRU staff members have satisfied training and registration requirements for individual Select Agent Security Risk Assessment (SRA) and 9 have received SRA approval (1 newly hired researcher is pending SRA approval). BRI staff facilitated all required associated site inspections and follow up (CDC, USDA and DHS January 2012).
Laboratory (in vitro) work with Exotic Bluetongue Virus in designated BSL-3 space is now ongoing and is supported with BRI use fees.
Rift Valley Fever Virus lab and animal research (lambs, calves, and mice) is ongoing and is supported by BRI lab use fees animal and laboratory work.
Received USDA, APHIS permit for Schmallenberg virus May 2012. Work will be conducted in the BRI with the support of BRI lab use fees.
Following submission of an application for Rift Valley Fever Virus (animal and laboratory work) in FY11, a Select Agent inspection was conducted in January 2012 and approvals are pending.
Following submission of an application for Exotic Bluetongue Virus insect transmission studies in FY11, a Select Agent inspection was conducted in January 2012 and approvals are pending.
ADODR is directly involved in performance of the research and also monitors activities to evaluate research progress through site visits, meeting at conferences, email and phone calls.