Progress 09/01/07 to 08/31/11
Outputs
OUTPUTS: In this project, we examined the impact of genomic rearrangements on M. ap antigenicity and virulence using proteomic, lipidomic and whole-genome sequence analyses. Since the start of this project, we identified a target isolates for lipidomic and proteomic analyses. Such analysis identified unique profiles for specific isolates from wildlife and exotic animals as well as from environmental samples collected from dairy herds. Additionally, we sequenced the whole genome of 15 isolates from different animals and from geographical locations. Our analysis allowed the accurate genotyping of M. ap isolates. PARTICIPANTS: Chia-wei Wu was trained on genomics;C. Hsu was trained on microbiology of paratuberculosis TARGET AUDIENCES: Field practitioners, laboratory workers on Johnes disease. Dairymen dairy herd managers. PROJECT MODIFICATIONS: We used whole-genome sequencing instead of DNA microarrays to study genomic diversity. This change was caused by the speed in Next-generation sequencing and their higher accuracy level than DNA-microarrays.
Impacts
The outcomes of our experiments provided us with novel insights into the molecular pathogenesis and evolution of M. ap on a genome-wide scale. More importantly, our study also provide us with a model for disease transmission between animal populations, especially for those animals that live in cohabitates. New leads for developing novel vaccine and diagnostic candidates are currently tested in my research group.
Publications
- -Hsu, C.Y., Wu, C.W, Talaat, A.M.* Genome-wide sequence variations among Mycobacterium avium subspecies paratuberculosis. Front. Microbiol. 2011. 2: Article 236
- -Ghosh P, Hsu C-Y, Alyamani E, Shehata MM, Al-Dubaib MA, Al-Naeem A et al.: Genome-wide Analysis of the Emerging Infection with Mycobacterium avium subspecies paratuberculosis in the Arabian Camels (Camelus dromedarius). PLoS ONE 2012, In Press.
- -Bannantine, J.P., Wu, C.W, Hsu, C.Y., Talaat, A.M.* Genome Sequencing of Ovine Isolates of Mycobacterium avium subspecies paratuberculosis Offers Insights Into Host Association. BMC-Genomics. 2012 (under review).
- -Wu, C.W, Schramm, T.M., Zhou, S., Schwartz, D.C., Talaat, A.M.* Optical mapping of the Mycobacterium avium subspecies paratuberculosis genome. BMC Genomics. 2009. 10: 25.
- -Wynne, J.W., Seemann, T., Bulach, D., Coutts, S.J., Talaat, A.M., Michalski, W.P. Re-sequencing the Mycobacterium avium subsp. paratuberculosis K10 genome: improved annotation and revised genome sequence. J. Bacteriol. 2010. 192: 6319-6320.
- -Bannantine, J.P., Talaat, A.M.* Genomic and Transcriptomic Studies in Mycobacterium avium subspecies paratuberculosis. Vet. Immunol. Immunopath. 2010. 138: 303-311.
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Progress 09/01/09 to 08/31/10
Outputs
OUTPUTS: We communicated our findings through participation in 3 scientific conferences involing researchers and workers in the field of animal research (CRWAD, Dairy and Animal Sciences Joint Annual Meetings, JDIP annual meeting). We also published 3 reports (see below) that describe in details our results. PARTICIPANTS: James Wynne (Collaborator, Australia); John Bananntine (Collaborator, Ames, IA); Torstin Eckstine (Collaborator, Colorado State University). Chia-wei WU (Graduate Student, University of Wisconsin-Madison). Meagan Cooney, (Graduate Student, University of Wisconsin-Madison). TARGET AUDIENCES: Change the understanding of scientists interested in studying how the genome of M. paratuberculosis has evolved to be a pathogenic organism. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
1-Improved our understanding of the genomic organization of M. paratuberculosis, the causative agent of Johne's disease in animals. 2-Improved our understanding of the pathogenesis of Jhone's disease which help up in controlling this important disease. 3-It allowed us to develop better diagnostic PCR against Johne's disease.
Publications
- 2. Bannantine, J.P., Talaat, A.M.* Genomic and Transcriptomic Studies in Mycobacterium avium subspecies paratuberculosis. Vet. Immunol. Immunopath. 2010. 138: 303-311.
- 3. Wynne, J.W., Seemann, T., Bulach, D., Coutts, S.J., Talaat, A.M., Michalski, W.P. Resequencing the Mycobacterium avium subsp. paratuberculosis K10 genome: improved annotation and revised genome sequence. J. Bacteriol. 2010. 192: 6319-6320.
- 1. Wu, C.W, Schramm, T.M., Zhou, S., Schwartz, D.C., Talaat, A.M.* Optical mapping of the Mycobacterium avium subspecies paratuberculosis genome. BMC Genomics. 2009. 10: 25.
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Progress 09/01/08 to 08/31/09
Outputs
OUTPUTS: We presented our findings in 2 different meetings as follow: 1-"Mycobacteriumparatuberculosis K-10 Genome Sequence Revisited," has been accepted for oral presentation at this year's conference in East Lansing, Michigan on April 18th and19th, 2008. 2-Revisiting the Genome Sequence of Mycobacteriumavium subspecies paratuberculosis using Optical Mapping Resolution. Chia-wei Wu,Timothy M. Schramm, David C. Schwartz, and Adel M. Talaat. 2008. USDA-NRI,Investigators meeting on side of the CRWAD meeting, Chicago, Dec. 2008. Also this poster was presented by the Genomic Science Training Program at UW-Madison. PARTICIPANTS: All of our collaborators were from University of Wisconsin-Madison (Dr. David Schwartz's group) and Colorado State University (Dr. Torsten Ecksten). There was an opportunity for a graduate student to be trained in chemical analysis of DNA including optical mapping. Also, the graduate student became very familiar with electron microscopy protocols. TARGET AUDIENCES: Workers in the field of genomics and bacterial pathogenesis. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
We analyzed the lipidome of 11 isolates of M. ap and M. avium using Thin-Layer Chromatography (TLC). Our analysis indicated that strain Env 77 of MAP differs from all other strains of MAP. It does not have Para-LP-01 (system E) or LpX. WC-A-01 and WC-A-02 appear only when grown on plates. In broth it has several highly non-polar lipids in addition, which are not detected in all other strains of MAP. It has Para-LP-02 (maybe as an compensation to the missing Para-LP-01 since they have the same peptide core; however, it is not explainable why LpX is missing [LpX = Para-LP-01 minus methyl ester]). Several strains have Para-LP-02 expressed in detectable amounts: JTC 1285, Env 195, and Env 77 when growing in broth and on plates, while Env 205, Env 411, JTC 1281, Env 210, and K-10 () express it only on plates. Strains Env 164 and Env 474 do not have detectable amounts of Para-LP-02. All strains express PIMs and the PI anchor. Whereas the lower-order PIMs are always strongly present, the higher-order PIMs are in less abundance. Ac2PIM5 is always present whereas Ac1PIM6 is missing from JTC 1281, Env 411, Env 474, and Env 210. Overall, our analysis suggests several lipidomic profiles for the examined isolates. Such differences can be exploited to develop better diagnostic tests for M. ap isolates. Additionally, we will better understand the molecular pathogenesis of Johne's disease.
Publications
- Optical Mapping of the Mycobacterium avium subspecies paratuberculosis Genome.Chia-wei Wu, Timothy M. Schramm, Shiguo Zhou, David C. Schwartz, Adel M. Talaat. 2008. BMC-Genomics (In press).
- A Novel Cell Wall Lipopeptide Is Important for Biofilm Formation and Pathogenicity of Mycobacterium avium subspecies paratuberculosis. Chia-wei Wu, Shelly K. Schmoller, John P. Bannantine, Torsten M. Eckstein, Julie M. Inamine, Michael Livesey, Ralph Albrecht and Adel M. Talaat. 2009. Microbial Pathogenesis (In press)
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Progress 09/01/07 to 08/31/08
Outputs
OUTPUTS: In this project we plan to examine the impact of genomic rearrangements on M. ap antigenicity and virulence using proteomic, lipidomic and immunoblot analyses. Since the start of this project, we identified a target isolates for lipidomic and proteomic analyses. Additionally, we began to take advantage of our library of insertional mutants to examine the role of in vivo-expressed genes in mycobacterial virulence using the murine model of paratuberculosis. Additional genes were identified as virulence factors from such analysis. We are currently working on confirming their role in virulence. PARTICIPANTS: Project Director: Adel M. Talaat, designed the experiments and performed data analysis. Graduate Student: Chia- wei Wu, run experiments and helped with the data analysis. TARGET AUDIENCES: Researchers and Veterinary Practitioners in the field of Johne's Disease. PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
The outcomes of the proposed experiments will provide novel insights into the molecular pathogenesis of Johne's Disease on a genome-wide scale. More importantly, our study will also provide new leads for developing novel vaccine and diagnostic candidates desperately needed to control JD supporting the mission of the animal protection program.
Publications
- Wu, C-W, Schmoller, S.K., Shin, S.J. and Talaat,A.M. Defining the stressome of Mycobacterium avium subspecies paratuberculosis in vitro and in naturally infected cows. J. Bacteriol. 2007. 189: 7877-7886.
- Wu, C-W, Livesey, M., Schmoller, S.K., Manning, E.J.B., Steinberg, H., Davis, W.C., Hamilton, M.J. and Talaat, A. M. Invasion and Persistence of Mycobacterium avium subspecies paratuberculosis During Early Stages of Johnes Disease in Calves. Infect.Immun. 2007. 7: 2110-2119.
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