2007 Annual Report
1a.Objectives (from AD-416)
The Johne's Disease Integrated Program (JDIP) is a multi-institutional USDA program that has the major objective of combating Johne's Disease. This will be done through focused research on several key areas in Johne's Disease. The program is designed such that there are four main research projects and four scientific cores. The principle investigators receiving subcontract funds (John Bannantine and Judy Stabel) are directing two of the scientific cores entitled, Genomics, Antibodies & Proteomics (GAP) and Animal Models & Facilities (AMF). The primary goal of the GAP core is to develop and distribute tools, reagents, and protocols that leverage recent technological advances in genomics and proteomics for members of the JDIP community and stakeholders. The primary goal of the AMF core is to develop and validate animal models and to provide uniform care and maintenance of animals utilized as experimental models for investigations conducted within JDIP.
1b.Approach (from AD-416)
The GAP core will develop an oligonucleotide array representing all genes in M. paratuberculosis (Map). This core will also produce a clone set of the coding sequences in Map. It will also make available transposon mutants for JD research. The AMF core will perform two specific tasks. (1) Assemble and maintain a current listing of available BL-2 certified animal facilities at various JDIP sponsored institutions and (2) establish calf-challenge models for JD research including study of the immune response to virulent Map and candidate vaccines.
Biosafety certification pending.
This report serves to document research conducted under a reimbursable agreement between ARS and the University of Minnesota. Additional details of research can be found in the report for the parent project 3625-32000-093 “Genomic and Immunologic Characteristics of Johne’s Disease”. This project (funded by USDA-CSREES) is being done in collaboration with the University of Minnesota, and is part of a larger consortium of researchers from around the country working on nearly all aspects of Johne’s Disease. The tasks covered by the subcontract awarded to NADC include the development of a clone library of M. avium subspecies paratuberculosis (MAP) genes and purified proteins, development of a MAP oligonucleotide microarray, and development of an improved animal model of Johne’s Disease.
Over 300 purified MAP proteins have been produced and made available to researchers. These proteins have been utilized both in-house as well as by other investigators in 14 different institutions. The proteins are being used for a variety of applications, including diagnostics and immunological assays.
Over 400 microarrays have been produced and used either in-house or by researchers at 7 different institutions. The microarrays have been used for comparing the genomes of MAP isolates from different host animals as well as examining MAP gene expression in response to host cells and other environmental conditions. Additionally, NADC has provided training on the use of the microarrays to researchers from several institutions. The availability of an oligonucleotide microarray has enabled researchers to pursue new lines of research that were previously inaccessible.
A cannulated calf model of MAP infection is currently being evaluated in animals housed at NADC in collaboration with researchers at Washington State University. Samples obtained from these animals will be evaluated in order to determine if this model will be useful for future Johne’s Disease research. Potential applications of an experimental model of Johne’s Disease include vaccine trials and studies into host response to infection.
Activities related to this agreement are monitored by several mechanisms. A monthly teleconference between project and core directors is held in addition to a weekly teleconference amongst members of the executive committee. An annual meeting is sponsored and attended by JDIP participants. Annual progress reports are also submitted by the project and core leaders.