Source: PENNSYLVANIA STATE UNIVERSITY submitted to
MYCOBACTERIAL DISEASES OF ANIMALS
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
HATCH
Reporting Frequency
Annual
Accession No.
0231815
Grant No.
(N/A)
Cumulative Award Amt.
(N/A)
Proposal No.
(N/A)
Multistate No.
NE-1201
Project Start Date
Oct 1, 2012
Project End Date
Sep 30, 2017
Grant Year
(N/A)
Program Code
[(N/A)]- (N/A)
Project Director
Kapur, VI.
Recipient Organization
PENNSYLVANIA STATE UNIVERSITY
208 MUELLER LABORATORY
UNIVERSITY PARK,PA 16802
Performing Department
Animal Science
Non Technical Summary
JD is a chronic granulomatous inflammatory intestinal disease of ruminants that results from infection with Mycobacterium avium subspecies paratuberculosis (M. paratuberculosis). First identified over a century ago, JD results in more than $200 million in annual losses to the United States (US) dairy industry each year with additional losses incurred by the other species. The growing recognition of M. paratuberculosis infection in wildlife species is also of considerable concern. Similarly, recent evidence of the presence of M. paratuberculosis in retail milk sources is of concern from a milk quality and potential food safety standpoint. JD remains a major concern for producers with very high prevalence rates (68% of all US dairy herds and 95% of those with over 500 cows have at least one JD positive animal. There have been considerable ongoing efforts made to identify knowledge gaps, define research priorities, and develop recommendations for implementing JD control measures in the field. For instance, a report from the National Research Council of the US National Academies of Sciences on JD concluded that JD is a significant animal-health problem whose study and control deserves high priority from the USDA. It was recognized that the problems associated with JD stem from: (i) difficulties in diagnosis because of an unusually long incubation period and a lack of specific and sensitive diagnostic tests for detecting early infections; (ii) a lack of vaccines or other effective measures for infection control; and, (iii) general lack of awareness of the disease and its true economic and animal-health consequences by producers and veterinarians. The goals of this project are to help address some of these unmet needs.
Animal Health Component
25%
Research Effort Categories
Basic
50%
Applied
25%
Developmental
25%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
3114010107010%
3114010108010%
3114010110010%
3114010109010%
3114010106010%
3113310110010%
3113310117010%
3113310109010%
3113410110010%
3113410117010%
Knowledge Area
311 - Animal Diseases;

Subject Of Investigation
3310 - Beef cattle, live animal; 4010 - Bacteria; 3410 - Dairy cattle, live animal;

Field Of Science
1090 - Immunology; 1060 - Biology (whole systems); 1070 - Ecology; 1100 - Bacteriology; 1080 - Genetics; 1170 - Epidemiology;
Goals / Objectives
Epidemiology and Transmission: Objective 1 will focus on understanding the epidemiology and transmission of JD and TB in animals through the application of predictive modeling and assessment of recommended control practices. Diagnostics: Objective 2 will seek to develop and implement new generations of diagnostic tests for JD and TB. Biology and Pathogenesis: Objective 3 will focus on improving our understanding of biology and pathogenesis of Mycobacterial diseases, as well as the host response to infection, and, Vaccine: Objective 4 will focus on development of programs to create and evaluate and develop new generations of vaccines for JD and TB. As described in the sections below, under the auspices of this multi-state initiative, we propose specific research projects to help achieve each of the 4 objectives and include a strong education and extension plan. We envision many of the projects to be crosscutting in nature (i.e. cut across objectives and/or address both diseases) that will together help address the major animal, human, and societal issues surrounding detection and control of mycobacterial diseases in animals. It is important to note that our research objectives are closely linked and coordinated with our education, extension and outreach plan.
Project Methods
Working in collaboration with other investigators who are part of this multi-state initiative, we anticipate participating in investigations that include the development of improved methods for the rapid, specific, sensitive, and cost-efficient diagnosis of JD infected animals. In addition, we envision studies of the basic biology of the causative organisms of JD and TB and their interaction with the host. Specifically, we anticipate studies that will employ state-of-the art microbiological, molecular biology, genomic, proteomic, metabolomic, immunology, and or bioinformatic approaches. Finally, it is well recognized that defining the host genetic, cellular and molecular events associated with susceptibility to JD is essential for the development of candidate vaccines and host genetic selection for resistance. Specifically, our group will participate in the programs that compare the efficacy of candidate vaccines in animal models of infection. We hypothesize that live attenuated vaccines are likely to elicit a protective response superior to the response elicited by currently available killed vaccines. Pennsylvania Agricultural Experiment Station investigators will take a leadership role in helping coordinate some of these studies, but may not directly participate in all of the bove Taken together, we anticipate that these investigations will reveal important insights into the immune response of animals to mycobacterial infections, as well as lead to the identification and evaluation of candidate vaccines.

Progress 10/01/12 to 09/30/17

Outputs
Target Audience: Scientists, veterinarians, producers, regulatory agencies, educators, policy makers, and the general public. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?This project encompassed approximately 15 faculty members from a variety of institutions across the globe. We estimate two graduate students and/or postdoctoral fellows were trained and received professional development for each faculty member, for a total of 30 students. How have the results been disseminated to communities of interest?The results have been demonstrated via the publications listed in the "Products" section of this progress report, and through an MDA conference that was funded through USDA project #PEN04538, Conference on Mycobacterial Diseases in Animals. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Professor Kapur's laboratory at the PAES has contributed to studies under Objectives 2 (Diagnostics) and 4 (Vaccines) where considerable progress has been made. For Objective 2, studies for antigen discovery in early infection of Johne's disease in cattle, three studies were carried out: Study 1. MTB protein microarray identified candidate sero-reactive antigens for early detection of MAP infection. Since current ELISA methods in diagnosis of Johne's disease have low sensitivity and early infection cannot be detected by using commercially available ELISA methods. In order to identify antigens for early detection of MAP infection, we conducted MTB protein microarray studies. The pilot study was initially conducted with MTB and MAP protein microarrays to screen a small number of serum samples from cows with clinical, sub-clinical MAP infection and negative control. The result showed good correlation of seroreactivities between MTB and MAP protein arrays and confirmed that the MTB array had utility as a screening tool for identifying candidate antigens for Johne's disease diagnostics [1]. Following the pilot study, we selected 180 well characterized serum samples for screening of MTB protein microarrays that contain more than 4,000 expressed MTB proteins covering more than 95% of MTB proteome [2]. The 180 serum samples were stratified into 4 groups: samples were from cows with tests negative from uninfected herds (NL, n=30); with tests negative from infected herds (NH, n=30); with fecal tests positive, serological tests negative (F+E-, n=60); and with both fecal test and ELISA positive (F+E+, n=60) based on previous ELISA and fecal tests. The screening of MTB protein microarray recognized 740 reactive proteins and a total 47 candidate proteins were identified from these reactive proteins with analyses of group comparison and ordinal logistic regression models. The candidate proteins that were significantly reactive include 12 in the NH group and 23 in the F+E- groups, suggesting potential utility for the early detection of MAP infected animals. Of these 47 antigens, only 6 of them have been studied in previous studies. Next, the diagnostic utility of four MAP orthologs (MAP1569, MAP2942c, MAP2609, and MAP1272c) was assessed and reveal moderate to high diagnostic sensitivities (range 48.3% to 76.7%) and specificity (range 96.7% to 100%), with a combined 88.3% sensitivity and 96.7% specificity. Taken together, the results of our analyses have identified several candidate MAP proteins of potential utility for the early detection of MAP infection, as well individual t of MAP proteins that may serve as the foundation for the next generation of well-defined serological diagnosis of JD in cattle. Results from this work has been published [2]. Study 2. Application of MAP protein arrays to confirm and expand the candidate antigen pool. We developed and utilized a MAP protein microarray with more than 800 expressed recombinant MAP proteins on the same set of serum samples [3]. Based on group comparison, ordinal logistic regression models, and ROC curve analysis, additional 50 candidate antigens were identified. Majority of candidate antigens identified in MAP arrays have not been recognized as sero-reactive in previous studies. Based on sub-cellular localization analysis, more than half of these are predicted to represent membrane or extracellular proteins that are likely to play a role in antibody responses to pathogens. Also of note is that more than a quarter of these proteins are classified as "hypothetical protein" that include some proteins unique to MAP. Taken together, the MAP protein microarray study greatly expands the number of potential candidates to evaluate for the early serological detection of MAP infection in animals that are known to be exposed and even infected, but are diagnosed as "negative" based on current commercially available ELISA assays. Manuscript is in the preparation process [3]. Study 3. Validation of selected candidate antigens with multiplex-bead based immunoassays. In order to validate the antigens identified in array studies for their use in diagnostics of MAP infection, we developed a color-coded fluorescent beads based multiplex (Luminex®) assays using both serum and milk samples from infected and control cows [4]. MAP recombinant proteins were coupled with beads and 180 serum and 90 milk samples from NL, F+E- and F+E+ groups were screened using beads coupled antigens. Among 4 antigens, except MAP1272c, MAP1569, MAP2942c, and MAP2609 were recognized by antibodies from serum samples in the F+E- group (p < 0.05), while antibody reactivities to each of 4 antigens were highest in the F+E+ group (p < 0.01). Moreover, milk antibodies recognized each of 4 antigens in both F+E- and F+E+ groups (p < 0.01) and correlated well with serum response (p < 0.01). The antibody reactivity to MAP recombinant proteins has not been evaluated before this study. This study demonstrated that, not only recombinant proteins recognized with significant antibody reactivity in milk during early MAP infection, the milk samples showed even higher sensitivity and specificity as compared with corresponding serum samples. Considering that obtaining of milk from an animal by a non-invasive manner is easier, low cost and more animal-owner friendly, it is important to develop and validate a milk based assays for rapid detection of MAP infection in dairy animals. It is important to note that the candidate proteins evaluated in this study represented only a small subset of candidate antigens identified from our previous MTB and MAP protein array studies and the screening of additional recombinant MAP proteins will be done in future studies. A manuscript describing our results has been submitted [4].

Publications

  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Li L, Bannantine JP, Campo JJ, Randall A, Grohn YT, Katani R, Schilling M, Radzio-Basu J, Kapur V. Identification of sero-reactive antigens for the early diagnosis of Johne's disease in cattle. PLoS One. 2017 Sep 1; 12(9): e0184373. doi: 10.1371/journal.pone.0184373. eCollection 2017. PubMed PMID: 28863177; PubMed Central PMCID: PMC5581170.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Bannantine JP, Campo JJ, Li L, Randall A, Pablo J, Praul CA, Raygoza Garay JA, Stabel JR, Kapur V. Identification of Novel Seroreactive Antigens in Johne's Disease Cattle by Using the Mycobacterium tuberculosis Protein Array. Clin Vaccine Immunol. 2017 Jul 5;24(7). pii: e00081-17. doi: 10.1128/CVI.00081-17. Print 2017 Jul. PubMed PMID: 28515134; PubMed Central PMCID: PMC5498720.
  • Type: Journal Articles Status: Submitted Year Published: 2017 Citation: Li L, Bannantine JP, Campo JJ, Randall A, Grohn YT, Schilling M, et al. Identification of sero-diagnostic antigens for the early diagnosis of Johnes using MAP protein microarray
  • Type: Journal Articles Status: Submitted Year Published: 2017 Citation: Li L, Wagner B, Freer H, Schilling M, Bannantine JP, Campo JJ, et al. Early Detection of Mycobacterium avium subsp. paratuberculosis infection in cattle with multiplex-bead based immuonassays


Progress 10/01/15 to 09/30/16

Outputs
Target Audience:Scientists, veterinarians, producers, regulatory agencies, educators, policy makers, and the general public. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?This project encompassed approximately 15 faculty members from a variety of institutions across the globe. We estimate two graduate students and/or postdoctoral fellows were trained and received professional development for each faculty member, for a total of 30 students. How have the results been disseminated to communities of interest?The results have been demonstrated via the publications listed in the "Products" section of this progress report, and through an MDA conference that was funded through USDA project #PEN04538, Conference on Mycobacterial Diseases in Animals. What do you plan to do during the next reporting period to accomplish the goals?The mission of Mycobacterial Diseases of Animals (MDA) is to facilitate the development of shared research and to address the health impacts and the economic losses from bTB and JD. We will: focus on continued development of mathematical models of JD and tuberculosis (TB) transmission dynamics; use sequence repeats and/or single nucleotide-based typing approaches to differentiate between MAP and M. bovis; develop methods for the early detection of MAP- and M. bovis­ infected animals, including newer generations of molecular, serological and microbiological assays with greater sensitivity and specificity; employ state-of-the art microbiological, molecular biology, genomic, proteomic, metabolomic, immunology, and or bioinformatic approaches to carryout studies investigating mechanisms of pathogen invasion of host cells and tissue, using state-of the art methods in mycobacteriology, cell biology, and genomics; develop candidate vaccines, identify genes and markers associated with susceptibility of animals to mycobacterial infection, and define the cellular and molecular events associated with development of immune responses to MAP and M. bovis in cattle. All the objectives of MDA are closely related to the area of extension and education, and we anticipate regular communications with stakeholders to share information as well as ensure that we are able to meet producers' needs.

Impacts
What was accomplished under these goals? Obj.1 Dr. Yrjo Grohn conducts experiments on the integrated approach to understand the pathways through which pathogen transmission occurs at all levels in an ecosystem.The study focuses on identifying evolutionary factors affecting the ecology and transmission of important slow transmission mycobacteria.The project takes advantage of data, which are already available as well as collecting new data in a longitudinal manner in the US and the UK. The goal of this project is to develop a quantitative methodology for incorporating WGS data into bacterial transmission models for infectious diseases incorporating ecology, economics, molecular biology, and epidemiology. Once the economics components have been implemented and tested, the model will be used to evaluate various MAP control mechanisms, including various test and cull strategies, maintaining a closed herd, and vaccination.The information on agent-to-agent transmission will aid in improving the assessment accuracy and allow a more complete understanding of halting the transmission of infection between animals under various controls. Obj. 2Vivek Kapur, John Bannantine, and Yrjo Grohn are investigating identifying biomarkers for early MAP infection. The team has developed a protein microarray with individual recombinant MAP proteins as a tool for sero-diagnostic antigen discovery by studying the dynamics of the humoral immune response in a baby goat model of infection from JDIP/MDA vaccine project. A protein microarray consisting of recombinant MAP proteins was constructed and used to identify reactive antigens in serum samples from experimentally MAP-infected and neg. control goats. Our preliminary analysis has resulted in the identification of ~50 MAP recombinant proteins that are sero-reactive in infected animals. The preliminary analyses suggest that individual MAP recombinant proteins were reactive even in MAP-infected goats that remained serologically neg. on commercially available MAP ELISA testing, suggesting that the protein microarray approach may enable the identification of antigens that are sero-reactive even during the early stages of infection. The results of these investigations were presented at the 2016 ICP in France. Obj. 3 Dr. Bannantine's lab is investigating the unknown proteins encoded by the genome of MAP, and is taking a step back to describe some fundamental aspects of two proteins that they have been studying for years.These proteins, which contain NlpC/P60 domains, are being studied primarily because they are strong antigens and thus could be used to develop a diagnostic test for JD.The focus is on the function of these proteins.Through crystal structure determination and modification of a single amino acid in the proteins, they are able to determine that both proteins have a similar structure, but one of the proteins lack enzyme activity. They developed a peptidoglycan-binding assay to demonstrate this lack of activity. This discovery may suggest a broader functional repertoire for NlpC/P60 domain-containing proteins than simply peptidoglycan hydrolases, and was presented at the ICP meeting in France and published in the journal Protein Science 2016 vol. 25:840-851. Dr. Paul Coussens' research group analyzed Immune phenotyping data for all cows collected through 2015. Sample subjects were 256 JD-negative cows and 91 positive cows collected from 8 Michigan commercial operations. The analysis included the mean relative percent of CD4+, CD8+ or γδ T cells expressing CD25. Based on the responder statuses, each cow was grouped into one of four total response phenotypes. Response phenotypes were as follows: non-response, with no T cell subtype exhibiting a ≥ 5% change in CD25 expression; both T cell response, with both classical CD4+ or CD8+ T cell subtypes and γδ T cells exhibiting a ≥ 5% change in CD25 expression; classical T cell response, with only classical CD4+ or CD8+ T cells exhibiting a ≥ 5% change in CD25 expression; γδ T cell response, with only γδ T cells exhibiting a ≥ 5% change in CD25 expression. Samples were collected from 36 previously collected, to repeat immune phenotyping and confirm stability of the previously identified response phenotypes.The data were presented at the 13th ICP in France. Sample collections for animals enrolled in the "Calf Study" continued.The purpose of this objective is to follow animals over time on farms with established MAP contamination and to correlate JD's infection with genotype and immune response phenotype. Blood was first sampled from these animals at 1-3 weeks of age for collection of plasma, buffy coats for DNA extraction, and peripheral blood mononuclear cells for immune phenotyping.The sample collections were repeated and consisted of: 37 8-12 month old heifers and 87 2-year-old first calf-cows. Samples were collected from 53 mature new cows identified as JD's milk ELISA+ and age- and group-matched neg. controls from two farms. Samples were collected for immune phenotyping and DNA extraction, and plasma was collected for diagnostics and future assays. Diagnostic data collected are currently being summarized and prepared to send to all participating producers. Obj. 4 Dr. Luiz Bermudez's research group has been working on the oral immunization as a strategy for affecting the course of JD's disease.MAP, the cause of JD's disease, infects the host through the gastrointestinal tract.The infectious phenotype of MAP originates either in the milk or in the intestinal tract of diseased ruminants.The bacterial surface proteomic was obtained after exposure to milk, and the differently expressed surface proteins were analyzed by bioinformatics. Proteins considered important for binding the intestinal mucosa were cloned and purified and anti-proteins antibodies produced. Mice were then infected with MAP orally with concomitant oral administration of the antibodies.Vaccinated animals showed significant protection against challenge. The strategy is currently being investigated in cows. Extension and education,Dr. Ken Olson's accomplishments: World Dairy Expo - Met with editors of dairy trade publications, including one from Mexico, to share information about the MDA MI activities including preliminary information on the development of strategies to control bTB in the developing world, information on our educational module JD-RAP http://jdrap.info/. The WMMB, the US DFR, and the NCDPA also provided space so we could have information available for the ~75,000 attendees. Joint Annual Meeting - An interest session on JD's and bTB was hosted for attendees at the Joint Annual Meeting of the ADSA and the ASAS to introduce animal and dairy science researchers, extension and industry professionals to the MDA. USAHA- An MDA display made information about MDA, the upcoming annual meeting. Presentations were made at the JD Committee and meetings of state, extension and federal veterinarians. An overview of the current US JD Program Efforts was presented by at the 2016 ICP in France. Workshop on accelerating bovine Tuberculosis Control in Developing Countries With funding from Bill & Melinda Gates Foundation, UGA, Cornell, and PSU, a Workshop on accelerating bTB Control in Developing Countries was conducted on December 8-10, 2015 in Rabat, Morocco. The workshop was co-chaired by Vivek Kapur, Martin Vodermeier, Yrjo Grohn, and Fred Quinn. The workshop brought together a diverse group of leading bTB investigators, which worked with policy makers and funding agency representatives to develop a shared vision and strategic framework for the implementation of bTB control programs in developing countries in which the disease is endemic in livestock, humans, and wildlife.

Publications

  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Al-Mamun, M. A., Smith, R. L., Schukken, Y. H. and Grohn, Y. T. (2016) Modeling of Mycobacterium avium subsp. paratuberculosis dynamics in a dairy herd: An individual based approach. J Theor Biol 408:105-117.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Bannantine, J. P., Lingle, C. K., Adam, P. R., Ramyar, K. X., McWhorter, W. J., Stabel, J. R., Picking, W. D. and Geisbrecht, B. V. (2016) NlpC/P60 domain-containing proteins of Mycobacterium avium subspecies paratuberculosis that differentially bind and hydrolyze peptidoglycan. Protein Sci 25:840-851.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Beaver, A., Cazer, C. L., Ruegg, P. L., Grohn, Y. T. and Schukken, Y. H. (2016) Implications of PCR and ELISA results on the routes of bulk-tank contamination with Mycobacterium avium ssp. paratuberculosis. J Dairy Sci 99: 1391-1405.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Beaver, A., Ruegg, P. L., Grohn, Y. T. and Schukken, Y. H. (2016) Comparative risk assessment for new cow-level Mycobacterium avium ssp. paratuberculosis infections between 3 dairy production types: Organic, conventional, and conventional-grazing systems. J Dairy Sci.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Capsel, R. T., Thoen, C. O., Reinhardt, T. A., Lippolis, J. D., Olsen, R., Stabel, J. R. and Bannantine, J. P. (2016) Composition and Potency Characterization of Mycobacterium avium subsp. paratuberculosis Purified Protein Derivatives. PLoS One 11:e0154685
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Espejo, L. A., Zagmutt, F. J., Groenendaal, H., Munoz-Zanzi, C. and Wells, S. J. (2015) Evaluation of performance of bacterial culture of feces and serum ELISA across stages of Johne's disease in cattle using a Bayesian latent class model. J Dairy Sci 98:8227-8239.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Franklin, R. K., Marcus, S. A., Talaat, A. M., KuKanich, B. K., Sullivan, R., Krugner-Higby, L. A. and Heath, T. D. (2015) Correction to: ''A Novel Loading Method for Doxycycline Liposomes for Intracellular Drug Delivery: Characterization of In Vitro and In Vivo Release Kinetics and Efficacy in a J774A.1 Cell Line Model of Mycobacterium smegmatis Infection". Drug Metab Dispos 43: 1805.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Ghosh, P., Shippy, D. C. and Talaat, A. M. (2015) Superior protection elicited by live-attenuated vaccines in the murine model of paratuberculosis. Vaccine 33: 7262-7270.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Hempel, R. J., Bannantine, J. P. and Stabel, J. R. (2016) Transcriptional Profiling of Ileocecal Valve of Holstein Dairy Cows Infected with Mycobacterium avium subsp. Paratuberculosis. PLoS One 11:e0153932.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Krueger, L. A., Beitz, D. C., Humphrey, S. B. and Stabel, J. R. (2016) Gamma delta T cells are early responders to Mycobacterium avium ssp. paratuberculosis in colostrum-replete Holstein calves. J Dairy Sci 99: 9040-9050.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Kugadas, A., Lamont, E. A., Bannantine, J. P., Shoyama, F. M., Brenner, E., Janagama, H. K. and Sreevatsan, S. (2016) A Mycobacterium avium subsp. paratuberculosis Predicted Serine Protease Is Associated with Acid Stress and Intraphagosomal Survival. Front Cell Infect Microbiol 6:85.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Li, L., Katani, R., Schilling, M. and Kapur, V. (2016) Molecular Epidemiology of Mycobacterium avium subsp. paratuberculosis on Dairy Farms. Annu Rev Anim Biosci 4:155-176.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Magombedze, G., Eda, S. and Koets, A. (2016) Can Immune Response Mechanisms Explain the Fecal Shedding Patterns of Cattle Infected with Mycobacterium avium Subspecies paratuberculosis? PLoS One 11: e0146844.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Magombedze, G., Eda, S. and Stabel, J. (2015) Predicting the Role of IL-10 in the Regulation of the Adaptive Immune Responses in Mycobacterium avium Subsp. paratuberculosis Infections Using Mathematical Models. PLoS One 10:e0141539.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Marcus, S. A., Sidiropoulos, S. W., Steinberg, H. and Talaat, A. M. (2016) CsoR Is Essential for Maintaining Copper Homeostasis in Mycobacterium tuberculosis. PLoS One 11:e0151816.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Marcus, S. A., Steinberg, H. and Talaat, A. M. (2015) Protection by novel vaccine candidates, Mycobacterium tuberculosis DeltamosR and DeltaechA7, against challenge with a Mycobacterium tuberculosis Beijing strain. Vaccine 33:5633-5639.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: McDaniel, M. M., Krishna, N., Handagama, W. G., Eda, S. and Ganusov, V. V. (2016) Quantifying Limits on Replication, Death, and Quiescence of Mycobacterium tuberculosis in Mice. Front Microbiol 7:862.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Krueger, L. A., Reinhardt, T. A., Beitz, D. C., Stuart, R. L. and Stabel, J. R. (2016) Effects of fractionated colostrum replacer and vitamins A, D, and E on haptoglobin and clinical health in neonatal Holstein calves challenged with Mycobacterium avium ssp. paratuberculosis. J Dairy Sci 99: 2884-2895.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: McDonald, J. L., Bailey, T., Delahay, R. J., McDonald, R. A., Smith, G. C. and Hodgson, D. J. (2016) Demographic buffering and compensatory recruitment promotes the persistence of disease in a wildlife population. Ecol Lett 19:443-449.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Mitachi, K., Sharma Gautam, L. N., Rice, J. H., Eda, K., Wadhwa, A., Momotani, E., Hlopak, J. P., Eda, S. and Kurosu, M. (2016) Structure determination of lipopeptides from Mycobacterium avium subspecies paratuberculosis and identification of antigenic lipopeptide probes. Anal Biochem 505:29-35.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Otsubo, S., Cossu, D., Eda, S., Otsubo, Y., Sechi, L. A., Suzuki, T., Iwao, Y., Yamamoto, S., Kuribayashi, T. and Momotani, E. (2015) Seroprevalence of IgG1 and IgG4 class antibodies against Mycobacterium avium subsp. paratuberculosis in Japanese population. Foodborne Pathog Dis 12:851-856.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Peterz, M., Butot, S., Jagadeesan, B., Bakker, D. and Donaghy, J. (2016) Thermal Inactivation of Mycobacterium avium subsp. paratuberculosis in Artificially Contaminated Milk by Direct Steam Injection. Appl Environ Microbiol 82:2800-2808.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Pomeroy B, Sipka A, Klaessig S, Schukken Y (2016) Longitudinal characterization of bovine monocyte-derived dendritic cells from mid-gestation into subsequent lactation reveals nadir in phenotypic maturation and macrophage-like cytokine profile in late gestation. J Reprod Immunol 118:1-8.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Rathnaiah, G., Bannantine, J. P., Bayles, D. O., Zinniel, D. K., Stabel, J. R., Grohn, Y. T. and Barletta, R. G. (2016) Analysis of Mycobacterium avium subsp. paratuberculosis Mutant Libraries Reveals Loci-dependent Transposition Biases and Strategies to Novel Mutant Discovery. Microbiology.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Ribeiro-Lima, J., Carstensen, M., Cornicelli, L., Forester, J. D. and Wells, S. J. (2016) Patterns of Cattle Farm Visitation by White-Tailed Deer in Relation to Risk of Disease Transmission in a Previously Infected Area with Bovine Tuberculosis in Minnesota, USA. Transbound Emerg Dis.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Roussey, J. A., Oliveira, L. J., Langohr, I. M., Sledge, D. G. and Coussens, P. M. (2016) Regulatory T cells and immune profiling in johne's disease lesions. Vet Immunol Immunopathol.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Shaughnessy, R. G., Farrell, D., Riepema, K., Bakker, D. and Gordon, S. V. (2015) Analysis of Biobanked Serum from a Mycobacterium avium subsp paratuberculosis Bovine Infection Model Confirms the Remarkable Stability of Circulating miRNA Profiles and Defines a Bovine Serum miRNA Repertoire. PLoS One 10:e0145089.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Slater, N., Mitchell, R. M., Whitlock, R. H., Fyock, T., Pradhan, A. K., Knupfer, E., Schukken, Y. H. and Louzoun, Y. (2016) Impact of the shedding level on transmission of persistent infections in Mycobacterium avium subspecies paratuberculosis (MAP). Vet Res 47:38.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Smith, R. L., Grohn, Y. T., Pradhan, A. K., Whitlock, R. H., Van Kessel, J. S., Smith, J. M., Wolfgang, D. R. and Schukken, Y. H. (2016) The effects of progressing and nonprogressing Mycobacterium avium ssp. paratuberculosis infection on milk production in dairy cows. J Dairy Sci 99:1383-1390.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Smith, R. L., Schukken, Y. H. and Grohn, Y. T. (2015) A new compartmental model of Mycobacterium avium subsp. paratuberculosis infection dynamics in cattle. Prev Vet Med 122: 298-305.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Ribeiro-Lima, J., Schwabenlander, S., Oakes, M., Thompson, B. and Wells, S. J. (2016) Risk profiling of cattle farms as a potential tool in risk-based surveillance for Mycobacterium bovis infection among cattle in tuberculosis-free areas. J Am Vet Med Assoc 248:1404-1413.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Wanzala, S. I., Nakavuma, J., Travis, D. A., Kia, P., Ogwang, S. and Sreevatsan, S. (2015) Draft Genome Sequences of Mycobacterium bovis BZ 31150 and Mycobacterium bovis B2 7505, Pathogenic Bacteria Isolated from Archived Captive Animal Bronchial Washes and Human Sputum Samples in Uganda. Genome Announc 3.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Winthrop, K., Rivera, A., Engelmann, F., Rose, S., Lewis, A., Ku, J., Bermudez, L. and Messaoudi, I. (2016) A Rhesus Macaque Model of Pulmonary Nontuberculous Mycobacterial Disease. Am J Respir Cell Mol Biol 54:170-176.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Wolf, T. M., Sreevatsan, S., Singer, R. S., Lipende, I., Collins, A., Gillespie, T. R., Lonsdorf, E. V. and Travis, D. A. (2016) Noninvasive Tuberculosis Screening in Free-Living Primate Populations in Gombe National Park, Tanzania. Ecohealth 13:139-144.


Progress 10/01/14 to 09/30/15

Outputs
Target Audience:Scientists, veterinarians, producers, regulatory agencies, educators, policy makers, and the general public. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?This project encompassed approximately 15 faculty members from a variety of institutions across the globe. We estimate two graduate students and/or postdoctoral fellows were trained and received professional development for each faculty member, for a total of 30 students. How have the results been disseminated to communities of interest?The results have been demonstrated via the publications listed in the "Products" section of this progress report, and through an MDA conference that was funded through USDA project #PEN04538, Conference on Mycobacterial Diseases in Animals. What do you plan to do during the next reporting period to accomplish the goals?The mission of Mycobacterial Diseases of Animals (MDA) is to facilitate the development of shared research and to address the health impacts and the economic losses from bTB and JD. We will: focus on continued development of mathematical models of JD and tuberculosis (TB) transmission dynamics; use sequence repeats and/or single nucleotide-based typing approaches to differentiate between MAP and M. bovis; develop methods for the early detection of MAP- and M. bovis-infected animals, including newer generations of molecular, serological and microbiological assays with greater sensitivity and specificity; employ state-of-the art microbiological, molecular biology, genomic, proteomic, metabolomic, immunology, and or bioinformatic approaches to carryout studies investigating mechanisms of pathogen invasion of host cells and tissue, using state-of the art methods in mycobacteriology, cell biology, and genomics; develop candidate vaccines, identify genes and markers associated with susceptibility of animals to mycobacterial infection, and define the cellular and molecular events associated with development of immune responses to MAP and M. bovis in cattle. All the objectives of MDA are closely related to the area of extension and education, and we anticipate regular communications with stakeholders to share information as well as ensure that we are able to meet producers' needs.

Impacts
What was accomplished under these goals? In the extension and education objective, Dr. Ken Olson (outreach coordinator for Johne's Disease Integrated Program JDIP) has attended several DC meetings with USDA leadership, partner organizations, and congressional staff, as well as taking part in priority setting at USDA stakeholder meetings. He has presented educational information at producer meetings, including: World Dairy Expo, NIAG, USDA, USAHA/AAVLD (providence, RI), JAM (Italy), and DMI events. In the biology and pathogenesis objective, Dr. Bermudez (Oregon state) has worked on the development of in vitro model systems to address important pathogenic mechanisms of Mycobacterium avium subsp. paratuberculosis (MAP) associated with the survival in ruminants. Dr. Bermudez's lab has developed a couple models that have provided knowledge of benefit to the study of the disease. In one of the models, they have established a sequential infection system from intestinal epithelial cells (first step in the infection) through macrophages and back to epithelial cells (last step on the disease). In the second model system, they used Acanthamoeba castellanii as a surrogate of bovine macrophages and Alamar Blue as a measure of metabolic activity to screen a transposon library of MAP for attenuation. Dr. Adel Talaat (University of Wisconsin-Madison) has worked on the ecology of bovine tuberculosis (bTB) in Egypt, with extensive of field and laboratory testing of bTB. Dr. John Bannantine has investigated the expression of two crucial molecules in T cell function, ZAP-70 (zeta-chain-associated protein of 70kDa) and CTLA-4 (cytotoxic T-lymphocyte antigen-4), in cows naturally infected with MAP. The study indicates changes in crucial signaling molecules in animals infected with MAP. Another study shows MAP proteins activate the Mitogen-activated protein kinase (MAPK) p38 pathway, and play a role in survival of this organism within bovine macrophages. In the host genetics objective, Dr. Luiz Bermudez, Dr. Paul Cousens, and Dr. John Bannantine have investigated the pathogenic mechanisms of MAP and the host responses to Johne's disease (JD). The results from evaluation of the ileal tissue from cows diagnosed with JD indicated that MAP alters its lipid composition during intracellular infection and acquires a pro-inflammatory phenotype, which likely is associated with the inflammatory phase of JD. In the vaccine objective, Dr. Luiz Bermudez (University of Nebraska Lincoln, NE) has been working on construction and screening of a comprehensive bank of 13,536 MAP K-10 Tn5367 mutants for phenotypes related to virulence determinants that could be applied to vaccine and diagnostic assay development. The constructed library is especially useful for identification of genes involved in MAP pathogenesis. Dr. Adel Talaat (University of Wisconsin-Madison, Madison) also has evaluated two new, live attenuated vaccine candidates (ΔmosR and ΔechA7), which show promise and have elicited an immune response similar to Bacille de Calmette et Guerin (BCG) vaccine. In the epidemiology and transmission of mycobacterial diseases objective, Dr. Yrjo Grohn (Cornell) and Dr. Ynte schukken (Cornell) conducted 10-years of longitudinal infection data on MAP in the Northeastern United States, and collected fecal and serum samples in order to understand the pathobiology and epidemiology of MAP infections in dairy herds. The evaluation was conducted on the prevalence of MAP infection and infection patterns. Dr. Yrjo Grohn also has investigated the dynamics of host-pathogen interactions in bovine paratuberculosis using several within- and between-host mathematical modeling of MAP infections.

Publications

  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Amin AS, Hsu CY, Darwish SF, Ghosh P, AbdEl-Fatah EM, Behour TS, et al. Ecology and genomic features of infection with Mycobacterium avium subspecies paratuberculosis in Egypt. Microbiology. 2015;161(Pt 4):807-18. doi: 10.1099/mic.0.000051. PubMed PMID: 25667007.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Babrak L, Danelishvili L, Rose SJ, Bermudez LE. Microaggregate-associated protein involved in invasion of epithelial cells by Mycobacterium avium subsp. hominissuis. Virulence. 2015:0. Epub 2015/08/08. doi: 10.1080/21505594.2015.1072676. PubMed PMID: 26252358.
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Babrak L, Danelishvili L, Rose SJ, Kornberg T, Bermudez LE. The environment of "Mycobacterium avium subsp. hominissuis" microaggregates induces synthesis of small proteins associated with efficient infection of respiratory epithelial cells. Infect Immun. 2014;83(2):625-36. Epub 2014/11/26. doi: IAI.02699-14 [pii]. 10.1128/IAI.02699-14. PubMed PMID: 25422262; PubMed Central PMCID: PMC4294266.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Bannantine JP, Stabel JR, Laws E, MC DC, Souza CD. Mycobacterium avium Subspecies paratuberculosis Recombinant Proteins Modulate Antimycobacterial Functions of Bovine Macrophages. PLoS One. 2015;10(6):e0128966. Epub 2015/06/16. doi: 10.1371/journal.pone.0128966. PONE-D-15-13970 [pii]. PubMed PMID: 26076028; PubMed Central PMCID: PMC4468122.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Bannantine JP, Talaat AM. Controlling Johne's disease: vaccination is the way forward. Front Cell Infect Microbiol. 2015;5:2. doi: 10.3389/fcimb.2015.00002. PubMed PMID: 25653925; PubMed Central PMCID: PMCPMC4301021.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Bermudez LE, Danelishvili L, Babrack L, Pham T. Evidence for genes associated with the ability of Mycobacterium avium subsp. hominissuis to escape apoptotic macrophages. Front Cell Infect Microbiol. 2015;5:63. Epub 2015/09/18. doi: 10.3389/fcimb.2015.00063. PubMed PMID: 26380226; PubMed Central PMCID: PMC4548235.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Bermudez LE, Meek L. Mefloquine and Its Enantiomers Are Active against Mycobacterium tuberculosis In Vitro and in Macrophages. Tuberc Res Treat. 2015;2014:530815. Epub 2015/01/13. doi: 10.1155/2014/530815. PubMed PMID: 25580293; PubMed Central PMCID: PMC4279124.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Danelishvili L, Bermudez LE. Mycobacterium avium MAV_2941 mimics phosphoinositol-3-kinase to interfere with macrophage phagosome maturation. Microbes Infect. 2015;17(9):628-37. Epub 2015/06/06. doi: S1286-4579(15)00105-7 [pii]. 10.1016/j.micinf.2015.05.005. PubMed PMID: 26043821; PubMed Central PMCID: PMC4554883.
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Danelishvili L, Stang B, Bermudez LE. Identification of Mycobacterium avium genes expressed during in vivo infection and the role of the oligopeptide transporter OppA in virulence. Microb Pathog. 2014;76:67-76. Epub 2014/09/24. doi: S0882-4010(14)00142-9 [pii]. 10.1016/j.micpath.2014.09.010. PubMed PMID: 25245008; PubMed Central PMCID: PMC4250378.
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: de Silva K, K MP, D JB, A CP, R JW. CD4(+) T-cells, gammadelta T-cells and B-cells are associated with lack of vaccine protection in Mycobacterium avium subspecies paratuberculosis infection. Vaccine. 2014;33(1):149-55. Epub 2014/12/03. doi: S0264-410X(14)01501-1 [pii]. 10.1016/j.vaccine.2014.10.082. PubMed PMID: 25444806.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Everman JL, Bermudez LE. Antibodies against invasive phenotype-specific antigens increase Mycobacterium avium subspecies paratuberculosis translocation across a polarized epithelial cell model and enhance killing by bovine macrophages. Front Cell Infect Microbiol. 2015;5:58. Epub 2015/08/25. doi: 10.3389/fcimb.2015.00058. PubMed PMID: 26301206; PubMed Central PMCID: PMC4528203.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Everman JL, Eckstein TM, Roussey J, Coussens P, Bannantine JP, Bermudez LE. Characterization of the inflammatory phenotype of Mycobacterium avium subspecies paratuberculosis using a novel cell culture passage model. Microbiology. 2015;161(7):1420-34. Epub 2015/05/10. doi: 10.1099/mic.0.000106. mic.0.000106 [pii]. PubMed PMID: 25957310.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Everman JL, Ziaie NR, Bechler J, Bermudez LE. Establishing Caenorhabditis elegans as a model for Mycobacterium avium subspecies hominissuis infection and intestinal colonization. Biol Open. 2015;4(10):1330-5. Epub 2015/09/26. doi: bio.012260 [pii]. 10.1242/bio.012260. PubMed PMID: 26405050; PubMed Central PMCID: PMC4610217.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Franklin RK, Marcus SA, Talaat AM, KuKanich BK, Sullivan R, Krugner-Higby LA, et al. A Novel Loading Method for Doxycycline Liposomes for Intracellular Drug Delivery: Characterization of In Vitro and In Vivo Release Kinetics and Efficacy in a J774A.1 Cell Line Model of Mycobacterium smegmatis Infection. Drug Metab Dispos. 2015;43(8):1236-45. doi: 10.1124/dmd.115.063602. PubMed PMID: 26033620; PubMed Central PMCID: PMCPMC4518064.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Ganusov VV, Klinkenberg D, Bakker D, Koets AP. Evaluating contribution of the cellular and humoral immune responses to the control of shedding of Mycobacterium avium spp. paratuberculosis in cattle. Vet Res. 2015;46:62. Epub 2015/06/21. doi: 10.1186/s13567-015-0204-1. 10.1186/s13567-015-0204-1 [pii]. PubMed PMID: 26092254; PubMed Central PMCID: PMC4474352.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Godden SM, Wells S, Donahue M, Stabel J, Oakes JM, Sreevatsan S, et al. Effect of feeding heat-treated colostrum on risk for infection with Mycobacterium avium ssp. paratuberculosis, milk production, and longevity in Holstein dairy cows. J Dairy Sci. 2015;98(8):5630-41. Epub 2015/06/16. doi: S0022-0302(15)00412-9 [pii]. 10.3168/jds.2015-9443. PubMed PMID: 26074228.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Koets AP, Grohn YT. Within- and between-host mathematical modeling of Mycobacterium avium subspecies paratuberculosis (MAP) infections as a tool to study the dynamics of host-pathogen interactions in bovine paratuberculosis. Vet Res. 2015;46:60. doi: 10.1186/s13567-015-0205-0. PubMed PMID: 26092284; PubMed Central PMCID: PMCPMC4474448.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Groenendaal H, Zagmutt FJ, Patton EA, Wells SJ. Cost-benefit analysis of vaccination against Mycobacterium avium ssp. paratuberculosis in dairy cattle, given its cross-reactivity with tuberculosis tests. J Dairy Sci. 2015;98(9):6070-84. Epub 2015/06/29. doi: S0022-0302(15)00429-4 [pii]. 10.3168/jds.2014-8914. PubMed PMID: 26117348.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Kincaid VA, London N, Wangkanont K, Wesener DA, Marcus SA, Heroux A, et al. Virtual Screening for UDP-Galactopyranose Mutase Ligands Identifies a New Class of Antimycobacterial Agents. ACS Chem Biol. 2015;10(10):2209-18. doi: 10.1021/acschembio.5b00370. PubMed PMID: 26214585
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Koets AP, Eda S, Sreevatsan S. The within host dynamics of Mycobacterium avium ssp. paratuberculosis infection in cattle: where time and place matter. Vet Res. 2015;46:61. Epub 2015/06/21. doi: 10.1186/s13567-015-0185-0. 10.1186/s13567-015-0185-0 [pii]. PubMed PMID: 26092382; PubMed Central PMCID: PMC4473847.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Leite FL, Eslabao LB, Pesch B, Bannantine JP, Reinhardt TA, Stabel JR. ZAP-70, CTLA-4 and proximal T cell receptor signaling in cows infected with Mycobacterium avium subsp. paratuberculosis. Vet Immunol Immunopathol. 2015;167(1-2):15-21. Epub 2015/07/15. doi: S0165-2427(15)00143-9 [pii]. 10.1016/j.vetimm.2015.06.017. PubMed PMID: 26163934.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Leite FL, Reinhardt TA, Bannantine JP, Stabel JR. Envelope protein complexes of Mycobacterium avium subsp. paratuberculosis and their antigenicity. Vet Microbiol. 2015;175(2-4):275-85. Epub 2014/12/17. doi: S0378-1135(14)00521-5 [pii] . 10.1016/j.vetmic.2014.11.009. PubMed PMID: 25500374.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Louzoun Y, Mitchell R, Behar H, Schukken Y. Two state model for a constant disease hazard in paratuberculosis (and other bovine diseases). Vet Res. 2015;46:67. Epub 2015/06/21. doi: 10.1186/s13567-015-0189-9. 10.1186/s13567-015-0189-9 [pii]. PubMed PMID: 26092587; PubMed Central PMCID: PMC4474326.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Marcus SA, Steinberg H, Talaat AM. Protection by novel vaccine candidates, Mycobacterium tuberculosis DeltamosR and DeltaechA7, against challenge with a Mycobacterium tuberculosis Beijing strain. Vaccine. 2015;33(42):5633-9. doi: 10.1016/j.vaccine.2015.08.084. PubMed PMID: 26363381; PubMed Central PMCID: PMCPMC4600678.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Mitchell RM, Schukken Y, Koets A, Weber M, Bakker D, Stabel J, et al. Differences in intermittent and continuous fecal shedding patterns between natural and experimental Mycobacterium avium subspecies paratuberculosis infections in cattle. Vet Res. 2015;46:66. Epub 2015/06/21. doi: 10.1186/s13567-015-0188-x. 10.1186/s13567-015-0188-x [pii]. PubMed PMID: 26092571; PubMed Central PMCID: PMC4474556.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Montezano D, Meek L, Gupta R, Bermudez LE, Bermudez JC. Flux Balance Analysis with Objective Function Defined by Proteomics Data-Metabolism of Mycobacterium tuberculosis Exposed to Mefloquine. PLoS One. 2015;10(7):e0134014. Epub 2015/07/29. doi: 10.1371/journal.pone.0134014. PONE-D-15-02647 [pii]. PubMed PMID: 26218987; PubMed Central PMCID: PMC4517854.
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Rathnaiah G, Lamont EA, Harris NB, Fenton RJ, Zinniel DK, Liu X, et al. Generation and screening of a comprehensive Mycobacterium avium subsp. paratuberculosis transposon mutant bank. Front Cell Infect Microbiol. 2014;4:144. doi: 10.3389/fcimb.2014.00144. PubMed PMID: 25360421; PubMed Central PMCID: PMCPMC4197770.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Rose SJ, Babrak LM, Bermudez LE. Mycobacterium avium Possesses Extracellular DNA that Contributes to Biofilm Formation, Structural Integrity, and Tolerance to Antibiotics. PLoS One. 2015;10(5):e0128772. Epub 2015/05/27. doi: 10.1371/journal.pone.0128772. PONE-D-15-08025 [pii]. PubMed PMID: 26010725; PubMed Central PMCID: PMC4444313.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Schukken YH, Whitlock RH, Wolfgang D, Grohn Y, Beaver A, VanKessel J, et al. Longitudinal data collection of Mycobacterium avium subspecies Paratuberculosis infections in dairy herds: the value of precise field data. Vet Res. 2015;46:65. Epub 2015/06/21. doi: 10.1186/s13567-015-0187-y. 10.1186/s13567-015-0187-y [pii]. PubMed PMID: 26092492; PubMed Central PMCID: PMC4474331.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Wolf TM, Mugisha L, Shoyama FM, O'Malley MJ, Flynn JL, Asiimwe B, et al. Noninvasive test for tuberculosis detection among primates. Emerg Infect Dis. 2015;21(3):468-70. Epub 2015/02/20. doi: 10.3201/eid2103.140052. PubMed PMID: 25695329; PubMed Central PMCID: PMC4344255.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Wolf TM, Sreevatsan S, Singer RS, Lipende I, Collins A, Gillespie TR, et al. Noninvasive Tuberculosis Screening in Free-Living Primate Populations in Gombe National Park, Tanzania. Ecohealth. 2015. Epub 2015/10/01. doi: 10.1007/s10393-015-1063y. 10.1007/s10393-015-1063-y [pii]. PubMed PMID: 26419483.


Progress 10/01/13 to 09/30/14

Outputs
Target Audience: Scientists, veterinarians, producers, regulatory agencies, educators, policy makers, and the general public. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? This project encompassed approximately 15 faculty from a variety of institutions across the globe. We estimate two graduate students and/or postdoctoral fellows were trained and received professional development for each faculty member, for a total of 30 students. How have the results been disseminated to communities of interest? The results have been demonstrated via the publications listed in this progress report and through an MDA conference, which was funded through another USDA project #4538, Conference on Mycobacterial Diseases in Animals. What do you plan to do during the next reporting period to accomplish the goals? Our main mission is to facilitate the development of shared research to address the health impacts and the economic losses from bTB and JD. We will focus on our main objectives of understanding the epidemiology and transmission of JD and TB in animals, and perform research on the new generations of diagnostic tests for both diseases by understanding the biology and pathogenesis of the diseases. Further emphasis will be made in the area of extension and education, and extended communication with stakeholders to address the producers' needs. We also aim to develop shared core resources to support innovative research and include more training activities for students.

Impacts
What was accomplished under these goals? In extension and education objective, Dr. Julie Smith (University of Vermont) conducted a survey among producers asking for feedback regarding knowledge and information on Johne's Disease (JD) and bovine tuberculosis (bTB). Among other results, she found out that not many producers have knowledge of ways to motivate action to control both diseases. Also, Dr. Jeannette McDonald (University of Wisconsin-Madison) has established a goal-oriented program to create a convenient risk assessment tool, provide education and as-needed, just-in-time assistance; and simplify the management and testing plan; make it easy to collect data, and establish accountability. A website was developed (http://iramp.izsler.it/), and the data was linked to a national database along with automatic calculations and analysis, and printable summaries for producers. In the biology and pathogenesis objective, Dr. Bermudez (Oregon state) has worked on the development of in vitro model systems to address important pathogenic mechanisms of Mycobacterium avium ss. paratuberculosis (MAP) associated with the survival in ruminants. Dr. Bermudez‘s lab has developed a couple models that have provided knowledge of benefit to the study of the disease. In one of the models, they have established a sequential infection system from intestinal epithelial cells (first step in the infection) through macrophages and back to epithelial cells (last step on the disease). In the second model system, they used Acanthamoeba castellanii as a surrogate of bovine macrophages and Alamar Blue as a measure of metabolic activity, to screen a transposon library of MAP for attenuation. Dr. Adel Talaat (University of Wisconsin-Madison) has worked on the ecology of bTB in Egypt with extensive of field and laboratory testing of bTB. In the host genetics objective, Dr. Holly Neibergs (Washington State University) and her research group have performed a genome-wide association study to identify possible causal variants for MAP tissue infection in cattle. The result led to the identification of a 70 kb region on Bos taurus (BTA) 3 as being associated with MAP tissue infection, indicating that at least one functional variant located within a 70 kb region of BTA3 may be a causal variant that results in susceptibility to MAP tissue infection, leading to Johne’s disease. In the vaccine objective, Dr. Fred Quinn (Univ. of Georgia) has been working on programs to develop and evaluate new generations of vaccines for bTB, and the next generation of vaccine candidates, including mucosal vaccines, that could be examined more accurately for disease prevention. In the epidemiology and transmission of mycobacterial diseases objective, Dr. Yrjo Grohn (Cornell) and Dr. Scott Wells (University of Minnesota) have done extensive work on the mathematical model of transmission using Baysian Evolutionary Analysis model.

Publications

  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Bradner, L., et al., Chemical decontamination with N-acetyl-L-cysteine-sodium hydroxide improves recovery of viable Mycobacterium avium subsp. paratuberculosis organisms from cultured milk. J Clin Microbiol, 2013. 51(7):2139-46.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Bradner, L., et al., Optimization of hexadecylpyridinium chloride decontamination for culture of Mycobacterium avium subsp. paratuberculosis from milk. J Clin Microbiol, 2013. 51(5):1575-7
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Cazer, C.L., et al., Associations between Mycobacterium avium subsp. paratuberculosis antibodies in bulk tank milk, season of sampling and protocols for managing infected cows. BMC Vet Res, 2013. 9:p. 234.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Cho, S., et al., Herd-level risk factors associated with fecal shedding of Shiga toxin-encoding bacteria on dairy farms in Minnesota, USA. Can Vet J. Jul 2013; 54(7):693697.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: de Silva, K., et al., Can early host responses to mycobacterial infection predict eventual disease outcomes? Prev Vet Med, 2013. 112(3-4):203-12.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: de Silva, K., et al., Apoptosis of lymph node and peripheral blood cells in ovine Johne's disease. Vet Immunol and Immunopathol, 2013. 156(1-2):82-90.
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Ahlstrom, C., H.W. Barkema, and J. De Buck, Improved short-sequence-repeat genotyping of Mycobacterium avium subsp. paratuberculosis by using matrix-assisted laser desorption ionization-time of flight mass spectrometry. Appl Environ Microbiol, 2014. 80(2):534-9.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Bannantine, J.P., et al., How does a Mycobacterium change its spots? Applying molecular tools to track diverse strains of Mycobacterium avium subspecies paratuberculosis. Let Appl Microbiol, 2013. 57(3):165-73.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Bannantine, J.P., et al., High-impact animal health research conducted at the USDA's National Animal Disease Center. Vet Microbiol, 2013. 165(3-4):224-33.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Faisal, S.M., et al., Evaluation of a Mycobacterium avium subsp. paratuberculosis leuD mutant as a vaccine candidate against challenge in a caprine model. CVI, 2013. 20(4):572-81.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Faisal, S.M., et al., Evaluation of a Salmonella vectored vaccine expressing Mycobacterium avium subsp. paratuberculosis antigens against challenge in a goat model. PLoS ONE, 2013. 8(8):e70171.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Fecteau, M.E., et al., Prevalence of Mycobacterium avium subsp. paratuberculosis fecal shedding in alpacas presented to veterinary hospitals in the United States. J Vet Intern Med/American College of Veterinary Internal Medicine, 2013. 27(5):1228-33.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Espejo, L.A., et al., Effect of delayed exposure of cattle to Mycobacterium avium subsp paratuberculosis on the development of subclinical and clinical Johne's disease. Am J Vet Res, 2013. 74(10):1304-10.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Fecteau, M.E., et al., Persistence of Mycobacterium avium subsp. paratuberculosis in soil, crops, and ensiled feed following manure spreading on infected dairy farms. Can Vet J, 2013. 54(11):1083-5.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Gurung, R.B., et al., Antigenicity of recombinant maltose binding protein-Mycobacterium avium subsp. paratuberculosis fusion proteins with and without factor Xa cleaving. CVI, 2013. 20(12):1817-26.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Hughes, V., et al., Interferon gamma responses to proteome-determined specific recombinant proteins: potential as diagnostic markers for ovine Johne's disease. Vet Immunol Immunopathol, 2013. 155(3):197-204.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Khalifeh, M.S. and J.R. Stabel, Clinical disease upregulates expression of CD40 and CD40 ligand on peripheral blood mononuclear cells from cattle naturally infected with Mycobacterium avium subsp. paratuberculosis. CVI, 2013. 20(8):1274-82.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Knust, B., et al., Evaluation of the effects of a killed whole-cell vaccine against Mycobacterium avium subsp paratuberculosis in three herds of dairy cattle with natural exposure to the organism. J Am Vet Med Assoc, 2013. 242(5):663-9.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Koop, G., et al., Risk factors for subclinical intramammary infection in dairy goats in two longitudinal field studies evaluated by Bayesian logistic regression. Prev Vet Med, 2013. 108(4):304-12.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Kupper, J.D., H.R. Brandt, and G. Erhardt, Genetic association between NOD2 polymorphism and infection status by Mycobacterium avium ssp. paratuberculosis in German Holstein cattle. Animal genetics, 2013. 45(1):114-116.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Lamont, E.A., W.W. Xu, and S. Sreevatsan, Host-Mycobacterium avium subsp. paratuberculosis interactome reveals a novel iron assimilation mechanism linked to nitric oxide stress during early infection. BMC Genomics, 2013. 14:694.
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Lee, S.J., et al., The Mycobacterium avium subsp. Paratuberculosis protein MAP1305 modulates dendritic cell-mediated T cell proliferation through Toll-like receptor-4. BMB reports, 2014. Feb;47(2):115-20.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Leite, F.L., et al., Comparison of fecal DNA extraction kits for the detection of Mycobacterium avium subsp. paratuberculosis by polymerase chain reaction. J Vet Diagn Invest: Official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc, 2013. 25(1):27-34.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Lu, Z., et al., Invasion and transmission of Salmonella Kentucky in an adult dairy herd using approximate Bayesian computation. BMC Vet Res, 2013. 9(1):245.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Lu, Z., et al., Using vaccination to prevent the invasion of Mycobacterium avium subsp. paratuberculosis in dairy herds: a stochastic simulation study. Prev Vet Med, 2013. 110(3-4):335-45.
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Magombedze, G., S. Eda, and V.V. Ganusov, Competition for antigen between Th1 and Th2 responses determines the timing of the immune response switch during Mycobaterium avium subspecies paratuberulosis infection in ruminants. PLoS Computational Biology, 2014. 10(1):e1003414.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Rosa, F.B., et al., Pathology in practice. Johne's disease. J Am Vet Med Assoc, 2013. 242(12):1655-7.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Malley, T.J., et al., Implementation of statistical tools to support identification and management of persistent Listeria monocytogenes contamination in smoked fish processing plants. J Food Prot, 2013. 76(5):796-811.
  • Type: Websites Status: Published Year Published: 2013 Citation: Malone, A.N., et al., Early Weight Development of Goats Experimentally Infected with Mycobacterium avium subsp. paratuberculosis. PLoS ONE, 2013. 8(12):p.e84049.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: McNamara, M., et al., Surface-exposed proteins of pathogenic mycobacteria and the role of cu-zn superoxide dismutase in macrophages and neutrophil survival. Proteome Science, 2013. 11(1):45.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Mortier, R.A., et al., Evaluation of age-dependent susceptibility in calves infected with two doses of Mycobacterium avium subspecies paratuberculosis using pathology and tissue culture. Vet Res, 2013. 44:94.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Pithua, P., et al., Is an individual calving pen better than a group calving pen for preventing transmission of Mycobacterium avium subsp paratuberculosis in calves? Results from a field trial. Res Vet Sci, 2013. 95(2):398-404.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Plain, K.M., et al., High-Throughput Direct Faecal PCR Assay to detect Mycobacterium avium subspecies paratuberculosis in sheep and cattle. J Clin Microbiol, 2013. Mar;52(3):745-57.
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Rose, S.J. and L.E. Bermudez, Mycobacterium avium biofilm attenuates mononuclear phagocyte function by triggering hyperstimulation and apoptosis during early infection. Infect Immun, 2014. 82(1):405-12.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Salgado, M., et al., Evaluation of PMS-PCR technology for detection of Mycobacterium avium subsp. paratuberculosis directly from bovine fecal specimens. Vet Microbiol, 2013. 167(3-4):725-8.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Schwabenlander, M.D., et al., A case of chronic wasting disease in a captive red deer (Cervus elaphus). J Vet Diagn Invest, 2013. 25(5):573-6.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Silva, C.A., et al., Interaction of Mycobacterium leprae with human airway epithelial cells: adherence, entry, survival, and identification of potential adhesins by surface proteome analysis. Infect Immun, 2013. 81(7):2645-59.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Sorge, U.S., S. Kurnick, and S. Sreevatsan, Detection of Mycobacterium avium subspecies paratuberculosis in the saliva of dairy cows: a pilot study. Vet Microbiol, 2013. 164(3-4):383-6.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Smith, R.L., et al., Development of a model to simulate infection dynamics of Mycobacterium bovis in cattle herds in the United States. J Am Vet Med Assoc, 2013. 243(3):411-23.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Smith, R.L., et al., Minimization of bovine tuberculosis control costs in US dairy herds. Prev Vet Med, 2013. 112(3-4):266-75.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Souza, C., et al., Mannosylated lipoarabinomannans from Mycobacterium avium subsp. paratuberculosis alters the inflammatory response by bovine macrophages and suppresses killing of Mycobacterium avium subsp. avium organisms. PLoS ONE, 2013. 8(9):e75924.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Stabel, J.R., et al., Disparate host immunity to Mycobacterium avium subsp. paratuberculosis antigens in calves inoculated with M. avium subsp. paratuberculosis, M. avium subsp. avium, M. kansasii, and M. bovis. CVI, 2013. 20(6):848-57.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Strawn, L.K., et al., Landscape and meteorological factors affecting prevalence of three food-borne pathogens in fruit and vegetable farms. Appl Environ Microbiol, 2013. 79(2):588-600.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Strawn, L.K., et al., Risk factors associated with Salmonella and Listeria monocytogenes contamination of produce fields. Appl Environ Microbiol, 2013. 79(24):7618-27.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Stringer, L.A., et al., Bayesian estimation of the sensitivity and specificity of individual fecal culture and Paralisa to detect Mycobacterium avium subspecies paratuberculosis infection in young farmed deer. J Vet Diagn Invest, 2013. 25(6):759-64.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Thirunavukkarasu, S., et al., In vivo and in vitro expression pattern of Toll-like receptors in Mycobacterium avium subspecies paratuberculosis infection. Vet Immunol Immunopathol, 2013. 156(1-2):20-31.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Volkova, V.V., et al., Modelling dynamics of plasmid-gene mediated antimicrobial resistance in enteric bacteria using stochastic differential equations. Sci Rep, 2013. 3:2463.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Wadhwa, A., et al., Use of ethanol extract of Mycobacterium bovis for detection of specific antibodies in sera of farmed red deer (Cervus elaphus) with bovine tuberculosis. BMC Vet Res, 2013. 9(1):256.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Wang, J., et al., Proteomic analysis and immune properties of exosomes released by macrophages infected with Mycobacterium avium. Microbe Infect, 2014 Apr;16(4):283-91.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Whittington, R.J., et al., Development and validation of a liquid medium (M7H9C) for routine culture of Mycobacterium avium subsp. paratuberculosis to replace modified Bactec 12B medium. J Clin Microbiol, 2013. 51(12):3993-4000.


Progress 10/01/12 to 09/30/13

Outputs
Target Audience: Scientists, veterinarians, producers, regulatory agencies, educators, policy makers, and the general public. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest? Through peer-reviewed publications, the annual meeting and conference, and the jdip.org website. What do you plan to do during the next reporting period to accomplish the goals? We plan to continue to make progress in the areas of Map diagnostics, genomics, and vaccine development, as well as lead the coordination of the MDA consortium, including hosting the annual conference.

Impacts
What was accomplished under these goals? The Mycobacterial Diseases of Animals (MDA) project continues to grow and advance towards achieving its overall objectives by bringing together flexible, functional and highly productive integrated research teams and helping establish core facilities that are interdisciplinary and multi-institutional. MDA continues to actively facilitate the community development of an integrated and coordinated program with research efforts concentrated on strategies to manage, prevent and control the disease; a translational pipeline of new diagnostic tests; vaccine candidates; and the formulation of an outstanding education and training program. Additionally, MDA provides the community with strong communication outlets and extension plans through our regular conference calls and the annual conference of researchers held in association with the Conference for Research Workers in Animal Diseases in Chicago, IL. In addition, our studies have helped elucidated genomic mechanisms for strain diversification and the identification of new targets for the development of novel immune-diagnostics. The Pennsylvania Ag Experiment Station (AES) continues to play a coordinating role to help execute and accomplish multiple services, programs and activities to reach the community as part of the MDA project. Some examples includes the maintenance of a website (www.jdip.org), an annual conference and extension meeting, weekly teleconferences with Executive Committee (EC), and monthly teleconferences with SAB members.

Publications

  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Bannantine, J. P., Li, L. L., Sreevatsan, S., and Kapur, V. (2013). How does a Mycobacterium change its spots? Applying molecular tools to track diverse strains of Mycobacterium avium subspecies paratuberculosis. Lett Appl Microbiol. 57:165-173.
  • Type: Journal Articles Status: Published Year Published: 2012 Citation: Bannantine, J. P., Wu, C. W., Hsu, C., Zhou, S., Schwartz, D. C., Bayles, D. O., Paustian, M. L., Alt, D. P., Sreevatsan, S., Kapur, V., and Talaat, A. M. (2012). Genome sequencing of ovine isolates of Mycobacterium avium subspecies paratuberculosis offers insights into host association. BMC genomics 13, 89.
  • Type: Journal Articles Status: Published Year Published: 2012 Citation: Bannantine, J. P., Lingle, C. K., Stabel, J. R., Ramyar, K. X., Garcia, B. L., Raeber, A. J., Schacher, P., Kapur, V., and Geisbrecht, B. V. (2012). MAP1272c encodes an NlpC/P60 protein, an antigen detected in cattle with Johnes disease. Clinical and vaccine immunology: 19, 1083-1092.