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Genome Sequencing Of Ovine Isolates Of Mycobacterium Avium Subspecies Paratuberculosis Offers Insights Into Host Association, John Bannantine, Chia-Wei Wu, Chungyi Hsu, Shiguo Zhou, David C. Schwartz, Darrell O. Bayles, Michael L. Paustian, David P. Alt, Srinand Sreevatsan, Vivek Kapur, Adel M. Talaat

United States Department of Agriculture-Agricultural Research Service / University of Nebraska-Lincoln: Faculty Publications

Background: The genome of Mycobacterium avium subspecies paratuberculosis (MAP) is remarkably homogeneous among the genomes of bovine, human and wildlife isolates. However, previous work in our laboratories with the bovine K-10 strain has revealed substantial differences compared to sheep isolates. To systematically characterize all genomic differences that may be associated with the specific hosts, we sequenced the genomes of three U.S. sheep isolates and also obtained an optical map.

Results: Our analysis of one of the isolates, MAP S397, revealed a genome 4.8 Mb in size with 4,700 open reading frames (ORFs). Comparative analysis of the MAP S397 isolate showed …

Identification And Characterization Of A Spore-Like Morphotype In Chronically Starved Mycobacterium Avium Subsp. Paratuberculosis Cultures, Elise A. Lamont, J. P. Bannantine, Anibal Armien, Don Sanjiv Ariyakumar, Suelee Robbe-Austerman

United States Department of Agriculture-Agricultural Research Service / University of Nebraska-Lincoln: Faculty Publications

Mycobacteria are able to enter into a state of non-replication or dormancy, which may result in their chronic persistence in soil, aquatic environments, and permissive hosts. Stresses such as nutrient deprivation and hypoxia provide environmental cues to enter a persistent state; however, a clear definition of the mechanism that mycobacteria employ to achieve this remains elusive. While the concept of sporulation in mycobacteria is not novel, it continues to spark controversy and challenges our perceptions of a non-replication. We investigated the potential role of sporulation in one-year old broth cultures of Mycobacterium subsp. paratuberculosis (MAP). We show that dormant cultures …

Immunogenicity And Protective Efficacy Of The Mycobacterium Avium Subsp. Paratuberculosis Attenuated Mutants Against Challenge In A Mouse Model, Jenn-Wei Chen, Syed M. Faisal, Subhash Chandra, Sean P. Mcdonough, Maria A.S. Moreira, Joy Scaria, Chao-Fu Chang, J. P. Bannantine, Bruce Akey, Yung-Fu Chang

United States Department of Agriculture-Agricultural Research Service / University of Nebraska-Lincoln: Faculty Publications

Johne’s disease (JD), caused by Mycobacterium avium subsp. paratuberculosis (MAP), results in serious economic losses worldwide especially in cattle, sheep and goats. To control the impact of JD on the animal industry, an effective vaccine with minimal adverse effects is urgently required. In order to develop an effective vaccine, we used allelic exchange to construct three mutant MAP strains, leuD, mpt64 and secA2. The mutants were attenuated in a murine model and induced cytokine responses in J774A.1 cell. The leuD mutant was the most obviously attenuated of the three constructed mutant strains. Our preliminary vaccine trial in mice demonstrated different …

Evaluation Of Protection In A Mouse Model After Vaccination With Mycobacterium Avium Subsp. Paratuberculois Protein Cocktails, J.R. Stabel, A. Barnhill, J. P. Bannantine, Y.F. Chang, M.A. Osman

United States Department of Agriculture-Agricultural Research Service / University of Nebraska-Lincoln: Faculty Publications

Whole-cell vaccines successfully reduce signs of clinical disease and fecal shedding of Mycobacterium avium subsp. paratuberculosis (MAP), however, these vaccines have some limitations. The present study was conducted to identify MAP proteins that might be candidates for the development of an improved vaccine. MAP proteins were screened for immunogenicity in naturally infected cattle and selected based upon reactivity in the interferon-􏰀 (IFN-􏰀) and Western blot assays. Proteins (MAP1087, MAP1204, MAP1272c, and MAP2077c) were arrayed into 4 overlapping cocktails containing 3 proteins each. The efficacy of the proteins within these cocktails as vaccine candidates was evaluated by subcutaneous immunization of mice, …