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Avian - 2001
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| Project Contact: | Vivek Kapur | Funding: | $87,000 |
| Shirin Munir |
District:
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| • The Problem • Mid-Year Progress Report • |
Respiratory diseases of turkeys are responsible for more than $35 million in losses to Minnesota’s turkey producers each year. This is a small but significant slice of a Minnesota poultry industry that employs more than 25,000 people and annually contributes more than $1 billion to Minnesota’s economy.
A respiratory disease caused by an avian pneumovirus (APV) has been spreading across the turkey production areas of Minnesota. It has caused losses of more than $20 million to Minnesota turkey growers since first diagnosed in 1996. This disease has not been reported in any other major turkey producing state. Thus, one of the greatest potential risks of this disease is the increased condemnation of birds and a ban of birds from Minnesota for foreign markets.
This year’s Rapid Response funding continues to build on emergency funding appropriated by the State Legislature in FY98 to:
Using this knowledge, a plan is being developed to constrain the disease from spreading to uninfected flocks.
January 2001
Background: The overall goal of this project is to develop recombinant vaccines for the control and prevention of infections in turkeys by two of the most important respiratory pathogens, avian pneumovirus (APV) and Pasteurella multocida. Remarkable and progress has been made towards the achievement of this goal and the project is on budget and thought to be ahead of schedule.
Sequencing of APV genome and molecular epidemiologic analysis of APV recovered from Minnesota turkey flocks: We have determined the complete nucleotide sequence of five of the eight genes of APV (N, P, M, F, and M2), including the viral leader sequence. Results indicate also, that vaccine against a single isolate may provide cross-protection against all subgroups of APV.
Strong evidence for the importance of APV fusion protein in protection against APV infection: Preliminary results suggest that a vaccine derived from the F protein has great potential to be highly effective in blocking virus infection.
The APV/US N gene has been cloned and expressed in E. coli: We have successfully expressed the APV/US N gene in E. coli. Studies for evaluating the immunogenic potential of N protein for protection against APV are underway. A highly sensitive N protein based ELISA was also developed to detect APV antibodies in serum samples of infected turkeys.
Recombinant FPV carrying the APV/US F and N gene sequences have been constructed: We have generated recombinant clones carrying the F and N protein genes. Purification of these recombinants is in progress. Investigations involving genetic and phenotypic characterization of recombinants and evaluation of protective efficacy will follow.
Identification of differentially expressed genes induced by APV infection: DNA sequencing was used to facilitate identification of major viral and host genes involved in APV infections. We believe that this insight will eventually be highly useful in designing vaccines and immunological targets to enhance vaccine induced immunity.
The P. multocida genome has been completely sequenced and annotated: We have completely characterized the 2,257,487bp circular genome of an avian isolate of Pasteurella multocida, identifying 2,104 open reading frames in the genome. Many of the putative genes identified represent important virulence factor orthologs (genes with a common ancestry but but which have evolved to become different). Studies are underway to characterize these factors to aid in vaccine development.
Methods for analyzing bacterial gene expression using microarrays have been standardized: DNA microarray technology has been standardized and P. multocida differential gene expression is being studied to facilitate better understanding of the molecular events involved in pathogenesis and virulence that are essential for the development of improved diagnostic tools and vaccine antigens. Several studies are in progress to help identify targets for novel vaccine or antimicrobial agent development.