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1

Cocito, C., P. Gilot, M. Coene, M. de Kesel, P. Poupart, and P. Vannuffel. "Paratuberculosis." Clinical Microbiology Reviews 7, no. 3 (July 1994): 328–45. http://dx.doi.org/10.1128/cmr.7.3.328.

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Paratuberculosis (Johne's disease) is a chronic, wasting, widespread mycobacteriosis of ruminants. It involves extensive mycobacterial shedding, which accounts for the high contagiousness, and ends with a fatal enteritis. Decreases in weight, milk production, and fertility produce severe economic loss. The DNA of the etiological agent (Mycobacterium paratuberculosis) has a base composition (66 to 67% G+C) within the range of that of mycobacteria (62 to 70% G+C), a size (4.4 x 10(6) to 4.7 x 10(6) bp) larger than that of most pathogenic mycobacteria (2.0 x 10(6) to 4.2 x 10(6) bp), and a high relatedness (> 90%) to Mycobacterium avium DNA. However, the DNAs of the two organisms can be distinguished by restriction fragment length polymorphism analysis. M. paratuberculosis genes coding for a transposase, a cell wall-associated protein (P34), and two heat shock proteins have been cloned and sequenced. Nucleic acid probes (two of which are species specific) are used, after PCR amplification, for M. paratuberculosis identification in stools and milk. As in leprosy, with disease progression, cellular immune reactions decrease and humoral immune reactions increase. Cutaneous testing with sensitins, lymphocyte proliferation assays, and cytokine tests are used to monitor cellular immune reactions in paratuberculosis, but these tests lack specificity. Complement fixation, immunodiffusion, and enzymometric tests based on antibodies to M. paratuberculosis extracts, to mycobacterial antigen complex A36, to glycolipids, and to proteins help identify affected cattle but are not species specific. The carboxyl-terminal portion of the 34-kDa cell wall-associated A36 protein (P34) carries species-specific B-cell epitopes and is the basis for an enzyme-linked immunosorbent assay. Diagnostic tests for paratuberculosis are also used in Crohn's disease, a chronic human ileitis mimicking Johne's disease, in which isolates identified as M. paratuberculosis have been found.
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2

Waters, W. R., B. J. Nonnecke, M. V. Palmer, S. Robbe-Austermann, J. P. Bannantine, J. R. Stabel, D. L. Whipple, et al. "Use of Recombinant ESAT-6:CFP-10 Fusion Protein for Differentiation of Infections of Cattle by Mycobacterium bovis and by M. avium subsp. avium and M. avium subsp. paratuberculosis." Clinical Diagnostic Laboratory Immunology 11, no. 4 (July 2004): 729–35. http://dx.doi.org/10.1128/cdli.11.4.729-735.2004.

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ABSTRACT Immunological diagnosis of Mycobacterium bovis infection of cattle is often confounded by cross-reactive responses resulting from exposure to other mycobacterial species, especially Mycobacterium avium. Early secretory antigenic target 6 (ESAT-6) and culture filtrate protein 10 (CFP-10) are dominant gamma interferon (IFN-γ)-inducing antigens of tuberculous mycobacteria, and they are absent from many environmental nontuberculous mycobacteria. Because M. avium exposure is the primary confounding factor in the diagnosis of M. bovis-infected animals, in vitro responses to a recombinant ESAT-6:CFP-10 (rESAT-6:CFP-10) fusion protein by blood leukocytes from cattle naturally exposed to M. avium or experimentally challenged with Mycobacterium avium subsp. avium or Mycobacterium avium subsp. paratuberculosis were compared to responses by M. bovis-infected cattle. Responses to heterogeneous mycobacterial antigens (i.e., purified protein derivatives [PPDs] and whole-cell sonicates [WCSs]) were also evaluated. Tumor necrosis factor alpha (TNF-α), IFN-γ, and nitric oxide responses by M. bovis-infected cattle to rESAT-6:CFP-10 exceeded (P < 0.05) the corresponding responses by cattle naturally sensitized to M. avium. Experimental infection with M. bovis, M. avium, or M. avium subsp. paratuberculosis induced significant (P < 0.05) IFN-γ and nitric oxide production to WCS and PPD antigens, regardless of the mycobacterial species used for the preparation of the antigen. Responses to homologous crude antigens generally exceeded responses to heterologous antigens. Nitric oxide and IFN-γ responses to rESAT-6:CFP-10 by blood leukocytes from M. bovis-infected calves exceeded (P < 0.05) the corresponding responses of noninfected, M. avium-infected, and M. avium subsp. paratuberculosis-infected calves. Despite the reported potential for secretion of immunogenic ESAT-6 and CFP-10 proteins by M. avium and M. avium subsp. paratuberculosis, it appears that use of the rESAT-6:CFP-10 fusion protein will be useful for the detection of tuberculous cattle in herds with pre-existing sensitization to M. avium and/or M. avium subsp. paratuberculosis.
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3

Laranja-da-Fonseca, Luis Femando, Alexandre Azevedo Oliva, Christian Campos Pereira, and Marcos Veiga dos Santos. "Doença de Johne: uma doença emergente em rebanhos leiteiros brasileiros." Revista de Educação Continuada em Medicina Veterinária e Zootecnia do CRMV-SP 3, no. 2 (July 1, 2000): 30–39. http://dx.doi.org/10.36440/recmvz.v3i2.3336.

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A Paratuberculose ou Doença de Johne é uma doença infecto-contagiosa, causada pelo Mycobacterium paratuberculosis, que se caracteriza por um processo inflamatório granulomatoso no intestino dos ruminantes domésticos e selvagens, determinando redução na digestibilidade dos alimentos, com consequente queda na produção de leite. Os animais infectados geralmente apresentam diarreia progressiva e perda de peso, Ainda que a literatura nacional apresente descrições da ocorrência de paratuberculose como casos isolados, não existem dados sobre a ocorrência da paratuberculose em rebanhos leiteiros no Brasil. Em estudo recente que avaliou a presença de anticorpos anti-M. paratuberculosis em rebanhos leiteiros do Estado de São Paulo, LARANJA-DA-FONSECA et al. (1999) identificaram que dos 403 animais amostrados, 153 (37,9%) apresentaram anticorpos anti-Mycobacterium paratuberculosis e, das 20 fazendas amostradas, 19 (95%) tiveram pelo menos um animal positivo, existindo assim a necessidade de levantamentos epidemiológicos da doença, afim de que se possa avaliar o real impacto da paratuberculose nos rebanhos brasileiros.
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4

Ionina, S. V. "Cultivation of mycobacterium paratuberculosis." Siberian Herald of Agricultural Science 49, no. 2 (May 22, 2019): 64–69. http://dx.doi.org/10.26898/0370-8799-2019-2-8.

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The paper presents a new solid growth medium for the cultivation of Mycobacterium paratuberculosis consisting of organic and inorganic ingredients. The study of diagnostic informative value and effectiveness of solid growth media used for cultivation of Mycobacterium Paratuberculosis was carried out in the laboratory conditions. Extract from birch-wood ash of 3% concentration and a growth stimulant of biological origin, peat oxide, were introduced as a mineral salt bases into the developed medium. When constructing the test medium, Lоwenstein–Jensen egg growth medium with the addition of mycobactin, which is an extract from Mycobacterium. phlei and contains substances necessary for the nutrition and reproduction of Mycobacterium paratuberculosis on artificial nutrient media, was used as an analogue. A test on compatibility and solubility of the components was done in distilled water in accordance with the generally accepted guidelines. The duration of observation ranged from 60 to 90 days. A comparison was made between the time of appearance of the primary and intensive growth of mycobacteria of paratuberculosis on the experimental medium and the Lоwenstein–Jensen control medium with mycobactin. Colonies of primary and intensive growth of standardized M. paratuberculosis strain and M. paratuberculosis isolate obtained from the cattle biomaterial on experimental egg growth medium appeared 3-7 days faster than on Lоwenstein–Jensen control medium with mycobactin. When inoculating biomaterial from cattle (lymph nodes and intestine), the primary growth of M. paratuberculosis on the experimental medium was noted 7 days earlier than on the control one, and the intensive growth was 3 days earlier. The experimental growth medium is cheaper and simpler to prepare than Lоwenstein–Jensen control medium with mycobactin, whose preparation is a rather laborious technological process.
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5

Paustian, Michael L., Vivek Kapur, and John P. Bannantine. "Comparative Genomic Hybridizations Reveal Genetic Regions within the Mycobacterium avium Complex That Are Divergent from Mycobacterium avium subsp. paratuberculosis Isolates." Journal of Bacteriology 187, no. 7 (April 1, 2005): 2406–15. http://dx.doi.org/10.1128/jb.187.7.2406-2415.2005.

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ABSTRACT Mycobacterium avium subsp. paratuberculosis is genetically similar to other members of the Mycobacterium avium complex (MAC), some of which are nonpathogenic and widespread in the environment. We have utilized an M. avium subsp. paratuberculosis whole-genome microarray representing over 95% of the predicted coding sequences to examine the genetic conservation among 10 M. avium subsp. paratuberculosis isolates, two isolates each of Mycobacterium avium subsp. silvaticum and Mycobacterium avium subsp. avium, and a single isolate each of both Mycobacterium intracellulare and Mycobacterium smegmatis. Genomic DNA from each isolate was competitively hybridized with DNA from M. avium subsp. paratuberculosis K10, and open reading frames (ORFs) were classified as present, divergent, or intermediate. None of the M. avium subsp. paratuberculosis isolates had ORFs classified as divergent. The two M. avium subsp. avium isolates had 210 and 135 divergent ORFs, while the two M. avium subsp. silvaticum isolates examined had 77 and 103 divergent ORFs. Similarly, 130 divergent ORFs were identified in M. intracellulare. A set of 97 ORFs were classified as divergent or intermediate in all of the nonparatuberculosis MAC isolates tested. Many of these ORFs are clustered together on the genome in regions with relatively low average GC content compared with the entire genome and contain mobile genetic elements. One of these regions of sequence divergence contained genes homologous to a mammalian cell entry (mce) operon. Our results indicate that closely related MAC mycobacteria can be distinguished from M. avium subsp. paratuberculosis by multiple clusters of divergent ORFs.
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6

Rowan, Neil J., Scott J. MacGregor, John G. Anderson, Douglas Cameron, and Owen Farish. "Inactivation of Mycobacterium paratuberculosis by Pulsed Electric Fields." Applied and Environmental Microbiology 67, no. 6 (June 1, 2001): 2833–36. http://dx.doi.org/10.1128/aem.67.6.2833-2836.2001.

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ABSTRACT The influence of treatment temperature and pulsed electric fields (PEF) on the viability of Mycobacterium paratuberculosiscells suspended in 0.1% (wt/vol) peptone water and in sterilized cow's milk was assessed by direct viable counts and by transmission electron microscopy (TEM). PEF treatment at 50°C (2,500 pulses at 30 kV/cm) reduced the level of viable M. paratuberculosis cells by approximately 5.3 and 5.9 log10 CFU/ml in 0.1% peptone water and in cow's milk, respectively, while PEF treatment of M. paratuberculosisat lower temperatures resulted in less lethality. Heating alone at 50°C for 25 min or at 72°C for 25 s (extended high-temperature, short-time pasteurization) resulted in reductions ofM. paratuberculosis of approximately 0.01 and 2.4 log10 CFU/ml, respectively. TEM studies revealed that exposure to PEF treatment resulted in substantial damage at the cellular level to M. paratuberculosis.
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7

Coussens, Paul M. "Mycobacterium paratuberculosisand the bovine immune system." Animal Health Research Reviews 2, no. 2 (December 2001): 141–62. http://dx.doi.org/10.1079/ahrr200134.

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AbstractMycobacterium aviumsubspeciesparatuberculosis(M. paratuberculosis) is the causative agent of Johne’s disease, a deadly intestinal ailment of ruminants. Johne’s disease is of tremendous economic importance to the worldwide dairy industry, causing major losses due to reduced production and early culling of animals. A highly controversial but developing link between exposure toM. paratuberculosisand human Crohn’s disease in some individuals has led to the suggestion thatM. paratuberculosisis also a potential food safety concern. As with many other mycobacteria,M. paratuberculosisis exquisitely adapted to survival in the host, despite aggressive immune reactions to these organisms. One hallmark of mycobacteria, includingM. paratuberculosis, is their propensity to infect macrophages. Inside the macrophage,M. paratuberculosisinterferes with the maturation of the phagosome by an unknown mechanism, thereby evading the host’s normal first line of defense against bacterial pathogens. The host immune system begins a series of attacks againstM. paratuberculosis-infected macrophages, including the rapid deployment of activated γδ T cells, CD4+T cells and cytolytic CD8+T cells. These cells interact with the persistently infected macrophage and with each other through a complex network of cytokines and receptors. Despite these aggressive efforts to clear the infection,M. paratuberculosispersists and the constant struggle of the immune system leads to pronounced damage to the intestinal epithelial cells. Enhancing our ability to control this important and tenacious pathogen will require a deeper understanding of howM. paratuberculosisinterferes with macrophage action, the cell types involved in the immune response, the cytokines these cells use to communicate, and the host genetic factors that control the response to infection.
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8

Hostetter, J., R. Kagan, and E. Steadham. "Opsonization Effects on Mycobacterium avium subsp. paratuberculosis-Macrophage Interactions." Clinical Diagnostic Laboratory Immunology 12, no. 6 (June 2005): 793–96. http://dx.doi.org/10.1128/cdli.12.6.793-796.2005.

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ABSTRACT High antibody titers in ruminants infected with Mycobacterium avium subsp. paratuberculosis correlates with disease progression. Effects of humoral responses during mycobacterial infection are not completely understood. This study suggests that activation status may be an important factor in determining macrophage ability to limit proliferation of opsonized M. avium subsp. paratuberculosis.
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9

Barukcic, Ilija. "Mycobacterium Avium Subspecies Paratuberculosis." Modern Health Science 1, no. 1 (June 12, 2018): p19. http://dx.doi.org/10.30560/mhs.v1n1p19.

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Objective: This systematic review assesses the causal relationship between Mycobacterium avium subspecies paratuberculosis (MAP) and Crohn’s disease (CD). Methods: A systematic review and meat-analysis of some impressive PCR based studies is provided aimed to answer among other questions the following question. Is there a cause effect relationship between Mycobacterium avium subspecies paratuberculosis and Crohn’s disease? The method of the conditio per quam relationship was used to proof the hypothesis whether the presence of Mycobacterium avium subspecies paratuberculosis guarantees the presence of Crohn’s disease. In other words, if Crohn’s disease is present, then Mycobacterium avium subspecies paratuberculosis is present too. The mathematical formula of the causal relationship k was used to proof the hypothesis, whether there is a cause effect relationship between Mycobacterium avium subspecies paratuberculosis and Crohn’s disease. Significance was indicated by a p-value of less than 0.05. Result: The studies analyzed (number of cases and controls N=1076) were able to provide evidence that Mycobacterium avium subspecies paratuberculosis is a necessary condition (a conditio sine qua non) and sufficicent conditions of Crohn’s disease. Furthermore, the studies analyzed provide impressive evidence of a cause-effect relationship between Mycobacterium avium subspecies paratuberculosis and Crohn’s disease. Conclusion: Mycobacterium avium subspecies paratuberculosis is the cause of Crohn’s disease (k=+0,377468824, p value < 0.0001).
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10

Dupont, Chris, Keith Thompson, Cord Heuer, Brigitte Gicquel, and Alan Murray. "Identification and characterization of an immunogenic 22 kDa exported protein of Mycobacterium avium subspecies paratuberculosis." Journal of Medical Microbiology 54, no. 11 (November 1, 2005): 1083–92. http://dx.doi.org/10.1099/jmm.0.46163-0.

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An exported 22 kDa putative lipoprotein was identified in an alkaline phosphatase gene fusion library of Mycobacterium avium subsp. paratuberculosis and expressed in Mycobacterium smegmatis. The full nucleic acid sequence of the gene encoding P22 was determined and the ORF was cloned into a mycobacterial expression vector, enabling full-length P22 to be produced as a C-terminal polyhistidine-tagged protein in M. smegmatis. N-terminal sequencing of the recombinant protein confirmed cleavage of a signal sequence. Native P22 was detected in culture supernatants and cell sonicates of M. avium subsp. paratuberculosis strain 316F using rabbit antibody raised to recombinant P22. Investigation of the presence of similar genes in other mycobacterial species revealed that the gene was present in Mycobacterium avium subsp. avium and similar genes existed in Mycobacterium intracellulare and Mycobacterium scrofulaceum. Database searches showed that P22 belonged to the LppX/LprAFG family of mycobacterial lipoproteins also found in Mycobacterium leprae and in members of the Mycobacterium tuberculosis complex. P22 shared less than 75 % identity to these proteins. Recombinant P22 was able to elicit interferon-gamma secretion in blood from eight of a group of nine sheep vaccinated with a live attenuated strain of M. avium subsp. paratuberculosis (strain 316F) compared to none from a group of five unvaccinated sheep. Antibody to P22 was detected by Western blot analysis in 10 out of 11 vaccinated sheep, in two out of two clinically affected cows and in 11 out of 13 subclinically infected cows.
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11

Corneli, Sara, Laura Corte, Luca Roscini, Antonella Di Paolo, Claudia Colabella, Linda Petrucci, Giulio Severi, Monica Cagiola, and Piera Mazzone. "Spectroscopic Characterization of Bovine, Avian and Johnin Purified Protein Derivative (PPD) with High-Throughput Fourier Transform InfraRed-Based Method." Pathogens 8, no. 3 (August 29, 2019): 136. http://dx.doi.org/10.3390/pathogens8030136.

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Tuberculins purified protein derivatives (PPDs) are obtained by precipitation from heat treated mycobacteria. PPDs are used in diagnosis of mycobacterial infections in humans and animals. Bovine PPD (PPDB) is obtained from Mycobacterium bovis (Mycobacterium tuberculosis complex), while Avian PPD (PPDA) and Johnin PPD (PPDJ) are extracted, respectively, from Mycobacterium avium and M. avium subsp. paratuberculosis (M. avium complex). PPDB and PPDA are used for bovine tuberculosis diagnosis, while PPDJ is experimentally used in the immunodiagnosis of paratuberculosis. Although PPDs date back to the 19th Century, limited knowledge about their composition is currently available. The goal of our study was to evaluate Fourier Transform InfraRed (FTIR) spectroscopy as a tool to differentiate PPDB, PPDA, and three PPDJs. The results highlighted that the three PPDs have specific profiles, correlated with phylogenetic characteristics of mycobacteria used for their production. This analysis is eligible as a specific tool for different PPDs batches characterization and for the assessment of their composition. The entire PPD production may be efficiently controlled, since the N content of each preparation is related to IR spectra, with a reference spectrum for each PPD and a standardized analysis protocol.
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12

Stanley, Emma C., Richard J. Mole, Rebecca J. Smith, Sarah M. Glenn, Michael R. Barer, Michael McGowan, and Catherine E. D. Rees. "Development of a New, Combined Rapid Method Using Phage and PCR for Detection and Identification of Viable Mycobacterium paratuberculosis Bacteria within 48 Hours." Applied and Environmental Microbiology 73, no. 6 (January 26, 2007): 1851–57. http://dx.doi.org/10.1128/aem.01722-06.

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ABSTRACT The FASTPlaqueTB assay is an established diagnostic aid for the rapid detection of Mycobacterium tuberculosis from human sputum samples. Using the FASTPlaqueTB assay reagents, viable Mycobacterium avium subsp. paratuberculosis cells were detected as phage plaques in just 24 h. The bacteriophage used does not infect M. avium subsp. paratuberculosis alone, so to add specificity to this assay, a PCR-based identification method was introduced to amplify M. avium subsp. paratuberculosis-specific sequences from the DNA of the mycobacterial cell detected by the phage. To give further diagnostic information, a multiplex PCR method was developed to allow simultaneous amplification of either M. avium subsp. paratuberculosis or M. tuberculosis complex-specific sequences from plaque samples. Combining the plaque PCR technique with the phage-based detection assay allowed the rapid and specific detection of viable M. avium subsp. paratuberculosis in milk samples in just 48 h.
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13

Shin, Sung Jae, and Michael T. Collins. "Thiopurine Drugs Azathioprine and 6-Mercaptopurine Inhibit Mycobacterium paratuberculosis Growth In Vitro." Antimicrobial Agents and Chemotherapy 52, no. 2 (December 10, 2007): 418–26. http://dx.doi.org/10.1128/aac.00678-07.

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ABSTRACT The in vitro susceptibility of human- and bovine-origin Mycobacterium paratuberculosis to the thioupurine drugs 6-mercaptopurine (6-MP) and azathioprine (AZA) was established using conventional plate counting methods and the MGIT 960 ParaTB culture system. Both 6-MP and AZA had antibacterial activity against M. paratuberculosis; isolates from Crohn's disease patients tended to be more susceptible than were bovine-origin isolates. Isolates of Mycobacterium avium, used as controls, were generally resistant to both AZA and 6-MP, even at high concentrations (≥64.0 μg/ml). Among rapidly growing mycobacteria, Mycobacterium phlei was susceptible to 6-MP and AZA whereas Mycobacterium smegmatis strains were not. AZA and 6-MP limited the growth of, but did not kill, M. paratuberculosis in a dose-dependent manner. Anti-inflammatory drugs in the sulfonamide family (sulfapyridine, sulfasalazine, and 5-aminosalycilic acid [mesalamine]) had little or no antibacterial activity against M. paratuberculosis. The conventional antibiotics azithromycin and ciprofloxacin, used as control drugs, were bactericidal for M. paratuberculosis, exerting their killing effects on the organism relatively quickly. Simultaneous exposure of M. paratuberculosis to 6-MP and ciprofloxacin resulted in significantly higher CFU than use of ciprofloxacin alone. These data may partially explain the paradoxical response of Crohn's disease patients infected with M. paratuberculosis to treatment with immunosuppressive thiopurine drugs, i.e., they do not worsen with anti-inflammatory treatment as would be expected with a microbiological etiologic pathogen. These findings also should influence the design of therapeutic trials to evaluate antibiotic treatments of Crohn's disease: AZA drugs may confound interpretation of data on therapeutic responses for both antibiotic-treated and control groups.
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14

Rosseels, Valérie, Sylvie Marché, Virginie Roupie, Marc Govaerts, Jacques Godfroid, Karl Walravens, and Kris Huygen. "Members of the 30- to 32-Kilodalton Mycolyl Transferase Family (Ag85) from Culture Filtrate of Mycobacterium avium subsp. paratuberculosis Are Immunodominant Th1-Type Antigens Recognized Early upon Infection in Mice and Cattle." Infection and Immunity 74, no. 1 (January 2006): 202–12. http://dx.doi.org/10.1128/iai.74.1.202-212.2006.

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ABSTRACT The characterization of protective antigens is essential for the development of an effective, subunit-based vaccine against paratuberculosis. Surface-exposed and secreted antigens, present abundantly in mycobacterial culture filtrate (CF), are among the well-known protective antigens of Mycobacterium tuberculosis and Mycobacterium bovis. Culture filtrate, prepared from Mycobacterium avium subsp. paratuberculosis ATCC 19698 grown as a surface pellicle on synthetic Sauton medium, was strongly and early recognized in experimentally infected B6 bg/bg beige mice and cattle, as indicated by elevated spleen cell gamma interferon (IFN-γ) secretion and lymphoproliferative responses of peripheral blood mononuclear cells, respectively. Strong proliferative and ex vivo IFN-γ responses against antigen 85 (Ag85) complex (a major protein component from M. bovis BCG culture filtrate) could be detected in cattle as early as 10 weeks after oral M. avium subsp. paratuberculosis infection. Synthetic peptides from the Ag85A and Ag85B components of this complex were strongly recognized, whereas T-cell responses were weaker against peptides from the Ag85C protein. A promiscuous T-cell epitope spanning amino acids 145 to 162 of Ag85B (identical sequence in M. bovis and M. avium subsp. paratuberculosis) was identified in experimentally infected cattle. Finally, young calves, born from cows with confirmed paratuberculosis, demonstrated proliferative responses to purified, recombinant Ag85A and Ag85B from M. avium subsp. paratuberculosis. These results indicate that the M. avium subsp. paratuberculosis Ag85 homologues are immunodominant T-cell antigens that are recognized early in experimental and natural infection of cattle.
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Wu, Chia-wei, Shelly K. Schmoller, Sung Jae Shin, and Adel M. Talaat. "Defining the Stressome of Mycobacterium avium subsp. paratuberculosis In Vitro and in Naturally Infected Cows." Journal of Bacteriology 189, no. 21 (August 10, 2007): 7877–86. http://dx.doi.org/10.1128/jb.00780-07.

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ABSTRACT Mycobacterium avium subsp. paratuberculosis causes an enteric infection in cattle, with a great impact on the dairy industry in the United States and worldwide. Characterizing the gene expression profile of M. avium subsp. paratuberculosis exposed to different stress conditions, or shed in cow feces, could improve our understanding of the pathogenesis of M. avium subsp. paratuberculosis. In this report, the stress response of M. avium subsp. paratuberculosis on a genome-wide level (stressome) was defined for the first time using DNA microarrays. Expression data analysis revealed unique gene groups of M. avium subsp. paratuberculosis that were regulated under in vitro stressors while additional groups were regulated in the cow samples. Interestingly, acidic pH induced the regulation of a large number of genes (n = 597), suggesting the high sensitivity of M. avium subsp. paratuberculosis to acidic environments. Generally, responses to heat shock, acidity, and oxidative stress were similar in M. avium subsp. paratuberculosis and Mycobacterium tuberculosis, suggesting common pathways for mycobacterial defense against stressors. Several sigma factors (e.g., sigH and sigE) were differentially coregulated with a large number of genes depending on the type of each stressor. Subsequently, we analyzed the virulence of six M. avium subsp. paratuberculosis mutants with inactivation of differentially regulated genes using a murine model of paratuberculosis. Both bacterial and histopathological examinations indicated the attenuation of all gene mutants, especially those selected based on their expression in the cow samples (e.g., lipN). Overall, the employed approach profiled mycobacterial genetic networks triggered by variable stressors and identified a novel set of putative virulence genes. A similar approach could be applied to analyze other intracellular pathogens.
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16

Speer, C. A., M. Cathy Scott, John P. Bannantine, W. Ray Waters, Yasuyuki Mori, Robert H. Whitlock, and Shigetoshi Eda. "A Novel Enzyme-Linked Immunosorbent Assay for Diagnosis of Mycobacterium avium subsp. paratuberculosis Infections (Johne's Disease) in Cattle." Clinical and Vaccine Immunology 13, no. 5 (May 2006): 535–40. http://dx.doi.org/10.1128/cvi.13.5.535-540.2006.

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ABSTRACT Enzyme-linked immunosorbent assays (ELISAs) for the diagnosis of Johne's disease (JD), caused by Mycobacterium avium subsp. paratuberculosis, were developed using whole bacilli treated with formaldehyde (called WELISA) or surface antigens obtained by treatment of M. avium subsp. paratuberculosis bacilli with formaldehyde and then brief sonication (called SELISA). ELISA plates were coated with either whole bacilli or sonicated antigens and tested for reactivity against serum obtained from JD-positive and JD-negative cattle or from calves experimentally inoculated with M. avium subsp. paratuberculosis, Mycobacterium avium subsp. avium, or Mycobacterium bovis. Because the initial results obtained from the WELISA and SELISA were similar, most of the subsequent experiments reported herein were performed using the SELISA method. To optimize the SELISA test, various concentrations (3.7 to 37%) of formaldehyde and intervals of sonication (2 to 300 s) were tested. With an increase in formaldehyde concentration and a decreased interval of sonication, there was a concomitant decrease in nonspecific binding by the SELISA. SELISAs prepared by treating M. avium subsp. paratuberculosis with 37% formaldehyde and then a 2-s burst of sonication produced the greatest difference (7×) between M. avium subsp. paratuberculosis-negative and M. avium subsp. paratuberculosis-positive serum samples. The diagnostic sensitivity and specificity for JD by the SELISA were greater than 95%. The SELISA showed subspecies-specific detection of M. avium subsp. paratuberculosis infections in calves experimentally inoculated with M. avium subsp. paratuberculosis or other mycobacteria. Based on diagnostic sensitivity and specificity, the SELISA appears superior to the commercial ELISAs routinely used for the diagnosis of JD.
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17

Rosseels, Valérie, Virginie Roupie, Denise Zinniel, Raúl G. Barletta, and Kris Huygen. "Development of Luminescent Mycobacterium avium subsp. paratuberculosis for Rapid Screening of Vaccine Candidates in Mice." Infection and Immunity 74, no. 6 (June 2006): 3684–86. http://dx.doi.org/10.1128/iai.01521-05.

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ABSTRACT Mycobacterium avium subsp. paratuberculosis is a slowly growing mycobacterial species, requiring 6 to 8 weeks of culture before colonies can be counted visually. Here, we describe the development of luminescent M. avium subsp. paratuberculosis expressing luxAB genes of Vibrio harveyi and its use for vaccine testing in an experimental mouse model, replacing fastidious CFU counting by rapid luminometry.
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18

KLANICOVA, B., I. SLANA, H. VONDRUSKOVA, M. KAEVSKA, and I. PAVLIK. "Real-Time Quantitative PCR Detection of Mycobacterium avium Subspecies in Meat Products." Journal of Food Protection 74, no. 4 (April 1, 2011): 636–40. http://dx.doi.org/10.4315/0362-028x.jfp-10-332.

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The aim of this work was to examine various purchased meat products and to find out if any traces of Mycobacterium avium subsp. avium, M. avium subsp. hominissuis, and M. avium subsp. paratuberculosis could be detected in these samples. Analysis of the meat products (raw, cooked, and fermented) was performed using a real-time quantitative PCR (qPCR) method for the detection of specific insertion sequences: duplex qPCR for the detection of IS900 specific for M. avium subsp. paratuberculosis, and triplex qPCR for the detection of IS901 specific for Mycobacterium avium subsp. avium and IS1245 specific for M. avium subsp. hominissuis. Of the 77 analyzed meat samples, 17 (22%) were found to contain M. avium subsp. paratuberculosis DNA, 4 (5%) samples contained Mycobacterium avium subsp. avium DNA, and in 12 (16%) samples M. avium subsp. hominissuis DNA was detected. The concentration of M. avium subsp. paratuberculosis and M. avium subsp. hominissuis DNA in some meat products exceeded 104 genomes per g. Culture examination of these mycobacterial subspecies was negative. By analyzing a range of meat products, we have provided evidence to support the hypothesis that M. avium is present in everyday commodities sold to the general public
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19

Bannantine, John P., Thomas J. Radosevich, Judith R. Stabel, Srinand Sreevatsan, Vivek Kapur, and Michael L. Paustian. "Development and Characterization of Monoclonal Antibodies and Aptamers against Major Antigens of Mycobacterium avium subsp. paratuberculosis." Clinical and Vaccine Immunology 14, no. 5 (March 7, 2007): 518–26. http://dx.doi.org/10.1128/cvi.00022-07.

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ABSTRACT Specific antibodies, available in unlimited quantities, have not been produced against Mycobacterium avium subsp. paratuberculosis, the bacterium that causes Johne's disease (JD). To fill this gap in JD research, monoclonal antibodies (MAbs) against M. avium subsp. paratuberculosis were produced from BALB/c mice immunized with a whole-cell extract of M. avium subsp. paratuberculosis. A total of 10 hybridomas producing MAbs to proteins ranging from 25 to 85 kDa were obtained. All MAbs showed some degree of cross-reactivity when they were analyzed against a panel of whole-cell protein lysates comprising seven different mycobacterial species. The MAbs were characterized by several methods, which included isotype analysis, specificity analysis, epitope analysis, reactivity in immunoblot assays, and electron microscopy. The identities of the antigens that bound to two selected MAbs were determined by screening an M. avium subsp. paratuberculosis lambda phage expression library. This approach revealed that MAb 9G10 detects MAP1643 (isocitrate lyase) and that MAb 11G4 detects MAP3840 (a 70-kDa heat shock protein), two proteins present in high relative abundance in M. avium subsp. paratuberculosis. The epitopes for MAb 11G4 were mapped to the N-terminal half of MAP3840, whereas MAb 9G10 bound to the C-terminal half of MAP1643. Aptamers, nucleic acids that bind to specific protein sequences, against the hypothetical protein encoded by MAP0105c were also generated and tested for their binding to M. avium subsp. paratuberculosis as well as other mycobacteria. These detection reagents may be beneficial in many JD research applications.
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20

Sung, Nackmoon, and Michael T. Collins. "Thermal Tolerance of Mycobacterium paratuberculosis." Applied and Environmental Microbiology 64, no. 3 (March 1, 1998): 999–1005. http://dx.doi.org/10.1128/aem.64.3.999-1005.1998.

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ABSTRACT D values (decimal reduction time; the time required to kill 1 log concentration of bacteria) were determined for both human and bovine strains (Dominic, Ben, BO45, and ATCC 19698) ofMycobacterium paratuberculosis in 50 mM lactate solution (pH 6.8) and in milk at four temperatures (62, 65, 68, and 71°C). Viable M. paratuberculosis organisms were quantified by a radiometric culture method (BACTEC). Thermal death curves for theM. paratuberculosis strains tested were generally linear, with R 2 of ≥0.90, but a few curves (R 2, 0.80 to 0.90) were better described by a quadratic equation. The human strains (Dominic and Ben) had similarD values in milk and in lactate solution. However,D values for the bovine strains (BO45 and ATCC 19698) were significantly different depending on the menstruum. Dvalues for low-passage clinical strains (Dominic, Ben, and BO45) were lower than those of the high-passage laboratory strain (ATCC 19698). The D value based on pooled data for clinical strains ofM. paratuberculosis in milk at 71°C (D 71°C) was 11.67 s. PooledD 62°C, D 65°C, andD 68°C of clinical M. paratuberculosis strains in milk were 228.8, 47.8, and 21.8 s, respectively. The Z value (the temperature required for the decimal reduction time to traverse 1 log cycle) of clinical strains in milk was 7.11°C. The D values of clumped and singleM. paratuberculosis cells were not significantly different. The D values of all M. paratuberculosis strains tested were considerably higher than those published forListeria, Salmonella, and Coxiellaspp. and estimated for Mycobacterium bovis, indicating thatM. paratuberculosis is more thermally tolerant. This study supports the premise that M. paratuberculosis may survive high-temperature, short-time pasteurization when the initial organism concentration is greater than 101 cells/ml.
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21

Konté, M. "La paratuberculose. Diagnostic d'un premier cas chez un bovin d'importation au Sénégal." Revue d’élevage et de médecine vétérinaire des pays tropicaux 41, no. 2 (February 1, 1988): 147–48. http://dx.doi.org/10.19182/remvt.8714.

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Le diagnostic de paratuberculose est porté pour la première fois au Sénégal sur un bovin importé de France. Les circonstances d’isolement de Mycobacterium paratuberculosis sont rapportées. Les particularités culturales du germe sont évoquées et discutées. L’auteur conclut en invoquant la nécessité d’appliquer les mesures de police sanitaire arrêtées en la matière.
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22

LORENCOVA, ALENA, PETRA VASICKOVA, JITKA MAKOVCOVA, and IVA SLANA. "Presence of Mycobacterium avium Subspecies and Hepatitis E Virus in Raw Meat Products." Journal of Food Protection 77, no. 2 (February 1, 2014): 335–38. http://dx.doi.org/10.4315/0362-028x.jfp-13-252.

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Meat and meat products may be the source of various pathogenic and potentially pathogenic agents for humans. We ascertained the occurrence of Mycobacterium avium subsp. paratuberculosis, subsp. avium, and subsp. hominissuis, and hepatitis E virus in retail raw meat products. The DNA of at least one of the target M. avium subspecies was detected in 26 (29.2%) of 89 analyzed samples of meat products. Fourteen (15.7%), 1 (1.1%), and 17 (19.1%) samples contained the DNA of Mycobacterium avium subsp. paratuberculosis, subsp. avium, and subsp. hominissuis, respectively. The number of mycobacterial cells per gram of meat products determined by real-time quantitative PCR ranged from 1.15 × 102 to 6.97 × 103. Mycobacterium chitae and Mycobacterium nonchromogenicum were isolated from three (3.4%) samples. Culture examination was not positive for any M. avium subspecies. Hepatitis E virus RNA was not detected in any of the samples.
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23

Weiss, Douglas J., Oral A. Evanson, Andreas Moritz, Ming Qi Deng, and Mitchell S. Abrahamsen. "Differential Responses of Bovine Macrophages to Mycobacterium avium subsp. paratuberculosis and Mycobacterium avium subsp. avium." Infection and Immunity 70, no. 10 (October 2002): 5556–61. http://dx.doi.org/10.1128/iai.70.10.5556-5561.2002.

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ABSTRACT Mycobacterium avium subsp. paratuberculosis and Mycobacterium avium subsp. avium are antigenically and genetically similar organisms; however, they differ in their virulence for cattle. M. avium subsp. paratuberculosis causes a chronic intestinal infection leading to a chronic wasting disease termed paratuberculosis or Johne's disease, whereas M. avium subsp. avium causes only a transient infection. We compared the response of bovine monocyte-derived macrophages to ingestion of M. avium subsp. paratuberculosis and M. avium subsp. avium organisms by determining organism survival, superoxide and nitric oxide production, and expression of the cytokines tumor necrosis factor alpha (TNF-α), gamma interferon (IFN-γ), interleukin-8 (IL-8), IL-10, IL-12, and granulocyte-monocyte colony-stimulating factor (GM-CSF). Unlike M. avium subsp. paratuberculosis, macrophages were able to kill approximately half of the M. avium subsp. avium organisms after 96 h of incubation. This difference in killing efficiency was not related to differences in nitric oxide or superoxide production. Compared to macrophages activated with IFN-γ and lipopolysaccharide, macrophages incubated with M. avium subsp. paratuberculosis showed greater expression of IL-10 and GM-CSF (all time points) and IL-8 (72 h) and less expression of IL-12 (72 h), IFN-γ (6 h), and TNF-α (6 h). When cytokine expression by macrophages incubated with M. avium subsp. paratuberculosis was compared to those of macrophages incubated with M. avium subsp. avium, M. avium subsp. paratuberculosis-infected cells showed greater expression of IL-10 (6 and 24 h) and less expression of TNF-α (6 h). Therefore, the combination of inherent resistance to intracellular degradation and suppression of macrophage activation through oversecretion of IL-10 may contribute to the virulence of M. avium subsp. paratuberculosis in cattle.
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24

D'Haese, Eva, Iris Dumon, Hadewig Werbrouck, Valerie De Jonghe, and Lieve Herman. "Improved detection of Mycobacterium paratuberculosis in milk." Journal of Dairy Research 72, S1 (July 14, 2005): 125–28. http://dx.doi.org/10.1017/s0022029905001226.

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At present there is no rapid microbiological method for the detection of viable Mycobacterium paratuberculosis in milk. By combining an extensive milk sample pretreatment with solid phase cytometry as the detection technique we were able to demonstrate viable mycobacterial cells in 50 ml of artificially contaminated pasteurized milk in less than one working day.
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25

Homuth, Matthias, Peter Valentin-Weigand, M. Rohde, and Gerald-F. Gerlach. "Identification and Characterization of a Novel Extracellular Ferric Reductase from Mycobacterium paratuberculosis." Infection and Immunity 66, no. 2 (February 1, 1998): 710–16. http://dx.doi.org/10.1128/iai.66.2.710-716.1998.

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ABSTRACT A novel extracellular mycobacterial enzyme was identified in the ruminant pathogen Mycobacterium paratuberculosis. The enzyme was capable of mobilizing iron from different sources such as ferric ammonium citrate, ferritin, and transferrin by reduction of the metal. The purified reductase had a calculatedM r of 17,000, was sensitive to proteinase K treatment, and had an isoelectric point of pH 9. Analysis of the amino acid composition revealed glycine, serine, asparagine (or aspartic acid), and glutamine (or glutamic acid) as the most frequently occurring residues. Enzymatic activity was highest at 37°C and between pH 5 and 10. The calculated Km andV max for ferric ammonium citrate were 0.213 mM and 0.345 mM min−1 mg−1, respectively. Using a specific antireductase antibody in immunoelectron microscopy, we were able to detect the enzyme associated with intracellular mycobacteria in naturally M. paratuberculosis-infected bovine tissue. We propose that the reductase of M. paratuberculosisrepresents an alternative strategy of mycobacteria to mobilize ferric iron and discuss its potential role in bacterial evasion of intracellular defense mechanisms.
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26

Cerf, O., and M. W. Griffiths. "Mycobacterium paratuberculosis heat resistance." Letters in Applied Microbiology 30, no. 4 (April 2000): 341–42. http://dx.doi.org/10.1046/j.1472-765x.2000.00718.x.

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27

Eastwood, M. A., C. P. Choudari, M. Macintyre, A. J. Richardson, and C. S. Stewart. "Antibodies to Mycobacterium paratuberculosis." Gut 34, no. 9 (September 1, 1993): 1291. http://dx.doi.org/10.1136/gut.34.9.1291.

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28

Valentin-Weigand, P., and K. M. Moriarty. "Mycobacterium paratuberculosis binds fibronectin." Research in Microbiology 143, no. 1 (January 1992): 75–79. http://dx.doi.org/10.1016/0923-2508(92)90036-n.

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29

Volpe, Rosario, Thomas Fett, Dominique Cassart, Jacques Godfroid, and Annick Linden. "Mixed Mycobacterium avium subspecies avium and M avium subspecies paratuberculosis infection in a wild red deer (Cervus elaphus) in Belgium." Veterinary Record Case Reports 8, no. 2 (June 2020): e001130. http://dx.doi.org/10.1136/vetreccr-2020-001130.

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This report describes a mixed Mycobacterium avium subspecies avium and M avium subspecies paratuberculosis infection in a free-ranging red deer (Cervus elaphus). The gross presentation was consistent with clinical paratuberculosis as previously reported in red deer and for other ruminants, with poor body condition, diarrhoea and mesenteric lymphadenitis. However, this animal presented unusual lung lesions, with necrosis and calcification similar to those reported for Mycobacterium bovis infection in wild and domestic ruminants. A mixed M avium subspecies avium and M avium subspecies paratuberculosis was shown by quantitative PCR (qPCR) performed on digestive tract samples. In addition, M avium subspecies avium was also detected in the respiratory tract (lungs and bronchial lymph nodes) by qPCR. These results were confirmed by classical bacteriology. Lesions induced by those mycobacteria cannot be differentiated from M bovis lesions. This point is particularly important with regards to increasing interactions between livestock and wild animals.
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30

Castellanos, Elena, Alicia Aranaz, Katherine A. Gould, Richard Linedale, Karen Stevenson, Julio Alvarez, Lucas Dominguez, Lucia de Juan, Jason Hinds, and Tim J. Bull. "Discovery of Stable and Variable Differences in the Mycobacterium avium subsp. paratuberculosis Type I, II, and III Genomes by Pan-Genome Microarray Analysis." Applied and Environmental Microbiology 75, no. 3 (December 1, 2008): 676–86. http://dx.doi.org/10.1128/aem.01683-08.

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ABSTRACT Mycobacterium avium subsp. paratuberculosis is an important animal pathogen widely disseminated in the environment that has also been associated with Crohn's disease in humans. Three M. avium subsp. paratuberculosis genomotypes are recognized, but genomic differences have not been fully described. To further investigate these potential differences, a 60-mer oligonucleotide microarray (designated the MAPAC array), based on the combined genomes of M. avium subsp. paratuberculosis (strain K-10) and Mycobacterium avium subsp. hominissuis (strain 104), was designed and validated. By use of a test panel of defined M. avium subsp. paratuberculosis strains, the MAPAC array was able to identify a set of large sequence polymorphisms (LSPs) diagnostic for each of the three major M. avium subsp. paratuberculosis types. M. avium subsp. paratuberculosis type II strains contained a smaller genomic complement than M. avium subsp. paratuberculosis type I and M. avium subsp. paratuberculosis type III genomotypes, which included a set of genomic regions also found in M. avium subsp. hominissuis 104. Specific PCRs for genes within LSPs that differentiated M. avium subsp. paratuberculosis types were devised and shown to accurately screen a panel (n = 78) of M. avium subsp. paratuberculosis strains. Analysis of insertion/deletion region INDEL12 showed deletion events causing a reduction in the complement of mycobacterial cell entry genes in M. avium subsp. paratuberculosis type II strains and significantly altering the coding of a major immunologic protein (MPT64) associated with persistence and granuloma formation. Analysis of MAPAC data also identified signal variations in several genomic regions, termed variable genomic islands (vGIs), suggestive of transient duplication/deletion events. vGIs contained significantly low GC% and were immediately flanked by insertion sequences, integrases, or short inverted repeat sequences. Quantitative PCR demonstrated that variation in vGI signals could be associated with colony growth rate and morphology.
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31

Ssekitoleko, Judah, Lonzy Ojok, Ahmed Abd El Wahed, Joseph Erume, Ahmad Amanzada, ElSagad Eltayeb, Kamal H. Eltom, and Julius Boniface Okuni. "Mycobacterium avium subsp. paratuberculosis Virulence: A Review." Microorganisms 9, no. 12 (December 19, 2021): 2623. http://dx.doi.org/10.3390/microorganisms9122623.

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To propose a solution for control of Mycobacterium avium subsp. paratuberculosis (MAP) infections in animals as well as in humans, and develop effective prevention, diagnostic and treatment strategies, it is essential to understand the molecular mechanisms of MAP pathogenesis. In the present review, we discuss the mechanisms utilised by MAP to overcome the host defense system to achieve the virulence status. Putative MAP virulence genes are mentioned and their probable roles in view of other mycobacteria are discussed. This review provides information on MAP strain diversity, putative MAP virulence factors and highlights the knowledge gaps regarding MAP virulence mechanisms that may be important in control and prevention of paratuberculosis.
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32

Svastova, P., I. Pavlik, and M. Bartos. "Rapid differentiation of Mycobacterium avium subsp. avium and Mycobacterium avium subsp. paratuberculosis by amplification of insertion element IS901." Veterinární Medicína 47, No. 5 (March 30, 2012): 117–21. http://dx.doi.org/10.17221/5814-vetmed.

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The aim of this study was to examine the specificity of primers designed to detect the insertion element IS901 commonly used in differentiation of Mycobacterium avium complex strains. This study shows that one of these primers non-specifically anneals to a sequence inside insertion element IS900, specific IS of M. avium subsp. paratuberculosis and to another sequence flanking this element. The resulting non-specific amplicon can be a product of amplification from some M. avium subsp. paratuberculosis strains and can simulate the presence of insertion element IS901 in these strains. However size difference between specific and non-specific amplicons allows such false-positive results to be distinguished. In addition the single PCR allows a rapid and simple differentiation between IS901+ M. avium subsp. avium and M. avium subsp. paratuberculosis strains.
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33

Lázaro Sales, Mariana, Érica Bravo Sales, Andréa Alencar Padilha, Omara Tereza Vianello Pereira, and Antônio Augusto Fonseca Junior. "Desenvolvimento de uma PCR em tempo real para diagnóstico de Mycobacterium avium subespécie paratuberculosis." Revista Acadêmica Ciência Animal 11, no. 1 (January 15, 2013): 97. http://dx.doi.org/10.7213/academica.7760.

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A paratuberculose, ou Doença de Johne, é uma doença crônica degenerativa que incide nos ruminantes domésticos. O agente etiológico Mycobacterium avium subsp. paratuberculosis (MAP) é um bacilo de crescimento lento pertencente ao complexo Mycobacterium avium (MAI). Nos EUA, a enfermidade acarreta grandes prejuízos econômicos. No Brasil, são poucos os relatos de casos da doença e não se conhece a real situação epidemiológica da paratuberculose bovina. O objetivo desse trabalho foi padronizar um teste de Reação em Cadeia da Polimerase (PCR) em tempo real para auxiliar na confirmação de casos da enfermidade.A técnica foi testada com DNA de diversas espécies do gênero Mycobacterium e avaliada quanto à sensibilidade e eficiência. Uma vez atestada sua confiabilidade, a metodologia foi aplicada para confirmar um caso suspeito de paratuberculose bovina. Foi enviada para análise no Laboratório Nacional Agropecuário/MG (Lanagro/MG) amostra de linfonodo mesentérico e alça intestinal de um bovino da raça holandesa com suspeita de paratuberculose. A amostra foi processada e inoculada para crescimento em meio Herrold’s com micobactina. A combinação entre a técnica do isolamento bacteriano em meio apropriado juntamente com a PCR em tempo real foi capaz de diagnosticar a presença da micobactéria no animal. Esse foi o primeiro caso de paratuberculose confirmado em um laboratório oficial do Ministério da Agricultura.
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34

Olsen, Ingrid, Liv J. Reitan, Gudmund Holstad, and Harald G. Wiker. "Alkyl Hydroperoxide Reductases C and D Are Major Antigens Constitutively Expressed by Mycobacterium aviumsubsp. paratuberculosis." Infection and Immunity 68, no. 2 (February 1, 2000): 801–8. http://dx.doi.org/10.1128/iai.68.2.801-808.2000.

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ABSTRACT Antigens characteristic for Mycobacterium aviumsubspecies paratuberculosis were identified by crossed immunoelectrophoresis (CIE) and by absorbing out cross-reactive antigens by using a polyclonal and polyvalent Mycobacterium avium subspecies avium antiserum. Two antigens were present in M. avium subsp. paratuberculosis and not detected in Mycobacterium avium subsp.avium. They were identified as antigens 17 and 20 in a CIE reference system for M. avium subsp.paratuberculosis antigens. Purified antigen 20 was identified as alkyl hydroperoxide reductase C (AhpC) while the N-terminal part of purified antigen 17 showed 80% homology with alkyl hydroperoxide reductase D (AhpD) of Mycobacterium tuberculosis. AhpC had a nonreduced mobility in sodium dodecyl sulfate-polyacrylamide gel electrophoresis corresponding to a molecular mass of 45 kDa and is probably a homodimer linked with disulfide bridges in its native form. AhpD had a mobility corresponding to 19 kDa. Monospecific rabbit antiserum against AhpC and AhpD reacted with 9 strains of M. avium subsp. paratuberculosis but not with 20 other mycobacterial strains except for aMycobacterium gordonae strain, against which a weak cross-reactive band was produced. Goats experimentally infected withM. avium subsp. paratuberculosis had strong gamma interferon (IFN-γ) responses toward both AhpC and AhpD, and they also had antibodies against AhpC. The ability of AhpC and AhpD to induce IFN-γ production shows that these proteins potentially could be used in future vaccines or in diagnostic assays. These results further show that AhpC and AhpD are immunologically important proteins which are constitutively and highly expressed in M. aviumsubsp. paratuberculosis without the bacteria being submitted to oxidative stress and that the specificities of antigens can be a matter of different levels of protein expression in various species as well as distinct structural differences.
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35

HRUSKA, K., M. BARTOS, P. KRALIK, and I. PAVLIK. "Mycobacterium avium subsp. paratuberculosisin powdered infant milk: paratuberculosis in cattle – the public health problem to be solved." Veterinární Medicína 50, No. 8 (March 28, 2012): 327–35. http://dx.doi.org/10.17221/5631-vetmed.

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Fifty one products of dried milk baby food purchased from 10 producers from seven countries available on the Czech market have been tested. IS900, the specific fragments for Mycobacterium avium subsp. paratuberculosis (MAP) have been detected using PCR in 25 samples (49.0 %) and fragment f57 by real time PCR in 18 samples (35.3%). These results correspond to the epidemiological situation in Europe and are not unexpected. Paratuberculosis in cattle was almost unknown in the Czech Republic until 1990. An increase in the number of cows with paratuberculosis found in slaughterhouses and the incidence of Crohn&rsquo;s disease in the last decade is evident. The possible risk of MAP dead cells or bacterial structures in food is discussed in respect to autoimmune Crohn&rsquo;s disease. The national programmes of paratuberculosis control and certification of paratuberculosis-free herds should be strongly supported to decrease the risk for children and other people under higher risk. Producers should use MAP free milk for baby food production on a voluntary basis.
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36

Wu, Chia-wei, Michael Livesey, Shelly K. Schmoller, Elizabeth J. B. Manning, Howard Steinberg, William C. Davis, Mary Jo Hamilton, and Adel M. Talaat. "Invasion and Persistence of Mycobacterium avium subsp. paratuberculosis during Early Stages of Johne's Disease in Calves." Infection and Immunity 75, no. 5 (February 12, 2007): 2110–19. http://dx.doi.org/10.1128/iai.01739-06.

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ABSTRACT Infection with Mycobacterium avium subsp. paratuberculosis causes Johne's disease in cattle and is a serious problem for the dairy industry worldwide. Development of models to mimic aspects of Johne's disease remains an elusive goal because of the chronic nature of the disease. In this report, we describe a surgical approach employed to characterize the very early stages of infection of calves with M. avium subsp. paratuberculosis. To our surprise, strains of M. avium subsp. paratuberculosis were able to traverse the intestinal tissues within 1 h of infection in order to colonize distant organs, such as the liver and lymph nodes. Both the ileum and the mesenteric lymph nodes were persistently infected for months following intestinal deposition of M. avium subsp. paratuberculosis despite a lack of fecal shedding of mycobacteria. During the first 9 months of infection, humoral immune responses were not detected. Nonetheless, using flow cytometric analysis, we detected a significant change in the cells participating in the inflammatory responses of infected calves compared to cells in a control animal. Additionally, the levels of cytokines detected in both the ileum and the lymph nodes indicated that there were TH1-type-associated cellular responses but not TH2-type-associated humoral responses. Finally, surgical inoculation of a wild-type strain and a mutant M. avium subsp. paratuberculosis strain (with an inactivated gcpE gene) demonstrated the ability of the model which we developed to differentiate between the wild-type strain and a mutant strain of M. avium subsp. paratuberculosis deficient in tissue colonization and invasion. Overall, novel insights into the early stages of Johne's disease were obtained, and a practical model of mycobacterial invasiveness was developed. A similar approach can be used for other enteric bacteria.
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37

Grant, Irene R., Hywel J. Ball, and Michael T. Rowe. "Isolation of Mycobacterium paratuberculosis from Milk by Immunomagnetic Separation." Applied and Environmental Microbiology 64, no. 9 (September 1, 1998): 3153–58. http://dx.doi.org/10.1128/aem.64.9.3153-3158.1998.

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ABSTRACT An immunomagnetic separation (IMS) technique was developed to facilitate selective isolation of Mycobacterium paratuberculosis cells from milk. Rabbit polyclonal antibodies against radiation-killed intact M. paratuberculosiscells were produced and used to coat sheep anti-rabbit immunoglobulin G (IgG) type M-280 Dynabeads. The rabbit anti-M. paratuberculosis IgG-coated beads (IMB) reacted strongly with laboratory strains of M. paratuberculosis as determined by slide agglutination, and microscopic examination confirmed that M. paratuberculosis cells attached to the IMB. The IMB were found to have a maximum binding capacity of 104 to 105 CFU of M. paratuberculosis. Studies showed that IMS selectively recoveredM. paratuberculosis from inoculated milk containing as few as 10 CFU of M. paratuberculosis per ml when 10 μl of IMB (ca. 106 beads) was added to 1 ml of milk and the preparation was incubated for 30 min at room temperature with gentle agitation. Larger volumes of milk (10 and 50 ml) were centrifuged and resuspended in 1 ml of phosphate-buffered saline–0.05% Tween 20 prior to IMS in order to increase the sensitivity of the method. Currently, primary isolation ofM. paratuberculosis from a milk sample relies on chemical decontamination, followed by culturing on Herrold’s egg yolk medium, which must be incubated at 37°C for up to 18 weeks. The potential value of our IMS method is as an aid for rapid detection ofM. paratuberculosis in milk when it is used in conjunction with end point detection methods, such as IS900PCR or an enzyme-linked immunosorbent assay.
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38

Berger, Sven, Dominik Hinz, John P. Bannantine, and J. Frank T. Griffin. "Isolation of High-Affinity Single-Chain Antibodies against Mycobacterium avium subsp. paratuberculosis Surface Proteins from Sheep with Johne's Disease." Clinical and Vaccine Immunology 13, no. 9 (September 2006): 1022–29. http://dx.doi.org/10.1128/cvi.00163-06.

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ABSTRACT Johne's disease, caused by infection with Mycobacterium avium subsp. paratuberculosis, causes significant economic losses to the livestock farming industry. Improved investigative and diagnostic tools—necessary to understand disease processes and to identify subclinical infection—are much sought after. Here, we describe the production of single-chain antibodies with defined specificity for M. avium subsp. paratuberculosis surface proteins. Single-chain antibodies (scFv) were generated from sheep with Johne's disease by cloning heavy-chain and lambda light-chain variable regions and expressing these in fusion with gene III of filamentous phages. Two scFv clones (designated SurfS1.2 and SurfS2.2) were shown to be immunoreactive against M. avium subsp. paratuberculosis surface targets by flow cytometry, and immunoblotting identified specificity for a 34-kDa proteinase-susceptible determinant. Both antibodies were cross-reactive against Mycobacterium avium subsp. avium but nonreactive against Mycobacterium bovis or Mycobacterium phlei cells and were shown to be capable of enriching M. avium subsp. paratuberculosis cells by a factor of approximately 106-fold when employed in magnetic bead separation of mixed Mycobacterium sp. cultures. Further, magnetic bead separation using SurfS1.2 and SurfS2.2 was capable of isolating as few as 103 M. avium subsp. paratuberculosis cells from ovine fecal samples, indicating the diagnostic potential of these reagents. Finally, inclusion of SurfS1.2 or SurfS2.2 in in vitro broth culture with M. avium subsp. paratuberculosis indicated that surface binding activity did not impede bacterial growth, although colony clumping was prevented. These results are discussed in terms of the potential use of single-chain phage display monoclonal antibodies as novel diagnostic reagents.
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39

Marri, Pradeep Reddy, John P. Bannantine, Michael L. Paustian, and G. Brian Golding. "Lateral gene transfer in Mycobacterium avium subspecies paratuberculosis." Canadian Journal of Microbiology 52, no. 6 (June 1, 2006): 560–69. http://dx.doi.org/10.1139/w06-001.

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Lateral gene transfer is an integral part of genome evolution in most bacteria. Bacteria can readily change the contents of their genomes to increase adaptability to ever-changing surroundings and to generate evolutionary novelty. Here, we report instances of lateral gene transfer in Mycobacterium avium subsp. paratuberculosis, a pathogenic bacteria that causes Johne's disease in cattle. A set of 275 genes are identified that are likely to have been recently acquired by lateral gene transfer. The analysis indicated that 53 of the 275 genes were acquired after the divergence of M. avium subsp. paratuberculosis from M. avium subsp. avium, whereas the remaining 222 genes were possibly acquired by a common ancestor of M. avium subsp. paratuberculosis and M. avium subsp. avium after its divergence from the ancestor of M. tuberculosis complex. Many of the acquired genes were from proteobacteria or soil dwelling actinobacteria. Prominent among the predicted laterally transferred genes is the gene rsbR, a possible regulator of sigma factor, and the genes designated MAP3614 and MAP3757, which are similar to genes in eukaryotes. The results of this study suggest that like most other bacteria, lateral gene transfers seem to be a common feature in M. avium subsp. paratuberculosis and that the proteobacteria contribute most of these genetic exchanges.Key words: mycobacteria, M. avium subsp. paratuberculosis, lateral gene transfer, unique genes, phylogeny.
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40

Suharti, Ika, Ni Luh Putu Ika Mayasari, and Fachriyan Hasmi Pasaribu. "Dekontaminasi Mycobacterium avium subspecies paratuberculosis pada feses menggunakan beberapa jenis desinfektan." Jurnal Sain Veteriner 36, no. 1 (October 15, 2018): 46. http://dx.doi.org/10.22146/jsv.26849.

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Paratuberculosis or Johne’s Disease is a granulomatous enteritis chronic disease of domestic and wild ruminants caused by infection of Mycobacterium avium subspecies paratuberculosis. The disease commonly infects dairy cattle with clinical signs of chronic diarrhea, decreasing body weight, low milk production, oedema, anemia and occasionally infertility. The basic procedure in order to control Paratuberculosis in farms is to do a good and proper handling of animal faecal. Disinfection of animal environments such as pens, faecal, sewerage and sewage are important in prevention of transmission of this disease. The purpose of this research is to determine specific disinfectan and dosage for Mycobacterium avium subspecies paratuberculosis decontamination in cattle feces so it can be applied as disease control measures. Cow's feces were contaminated with MAP 105CFU/ml and treated with ammonium quartener, phenolic and formaldehyde disinfectant doses 10%, 15% and 20%. The effectiveness of the disinfectant was tested based on MAP identification using Löwenstein-Jensen culture medium and nested Polymere Chain Reaction(PCR). The results showed 15% and 20% doses of formaldehyde disinfectants efective to decontaminate Mycobacterium avium subspecies paratuberculosis in catle feces.
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41

Davis, William C., Gaber S. Abdellrazeq, Asmaa H. Mahmoud, Kun-Taek Park, Mahmoud M. Elnaggar, Gaetano Donofrio, Victoria Hulubei, and Lindsay M. Fry. "Advances in Understanding of the Immune Response to Mycobacterial Pathogens and Vaccines through Use of Cattle and Mycobacterium avium subsp. paratuberculosis as a Prototypic Mycobacterial Pathogen." Vaccines 9, no. 10 (September 26, 2021): 1085. http://dx.doi.org/10.3390/vaccines9101085.

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Lack of understanding of the immune response to mycobacterial pathogens has impeded progress in development of vaccines. Infection leads to development of an immune response that controls infection but is unable to eliminate the pathogen, resulting in a persistent infection. Although this puzzle remains to be solved, progress has been made using cattle as a model species to study the immune response to a prototypic mycobacterium, Mycobacterium a. paratuberculosis (Map). As chronicled in the review, incremental advances in characterizing the immune response to mycobacteria during the last 30 years with increases in information on the evolution of mycobacteria and relA, a gene regulating the stringent response, have brought us closer to an answer. We provide a brief overview of how mycobacterial pathogens were introduced into cattle during the transition of humankind to nomadic pastoralists who domesticated animals for food and farming. We summarize what is known about speciation of mycobacteria since the discovery of Mybacterium tuberculsis Mtb, M. bovis Mbv, and Map as zoonotic pathogens and discuss the challenges inherent in the development of vaccines to mycobacteria. We then describe how cattle were used to characterize the immune response to a prototypic mycobacterial pathogen and development of novel candidate vaccines.
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42

Chiodini, R. J. "Abolish Mycobacterium paratuberculosis strain 18." Journal of Clinical Microbiology 31, no. 7 (1993): 1956–58. http://dx.doi.org/10.1128/jcm.31.7.1956-1958.1993.

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43

Erasmus, D. L., T. C. Victor, P. J. van Eeden, V. Falck, and P. van Helden. "Mycobacterium paratuberculosis and Crohn's disease." Gut 36, no. 6 (June 1, 1995): 942. http://dx.doi.org/10.1136/gut.36.6.942-b.

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44

Lund, B. M., C. W. Donnelly, and A. Rampling. "HEAT RESISTANCE OF MYCOBACTERIUM PARATUBERCULOSIS." Letters in Applied Microbiology 31, no. 2 (August 2000): 184–85. http://dx.doi.org/10.1046/j.1365-2672.2000.00784.x.

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45

Tanaka, K., M. Wilks, P. J. Coates, M. J. Farthing, J. A. Walker-Smith, and S. Tabaqchali. "Mycobacterium paratuberculosis and Crohn's disease." Gut 32, no. 1 (January 1, 1991): 43–45. http://dx.doi.org/10.1136/gut.32.1.43.

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46

Woo, Seng-Ryong, Raúl G. Barletta, and Charles J. Czuprynski. "Extracellular ATP Is Cytotoxic to Mononuclear Phagocytes but Does Not Induce Killing of Intracellular Mycobacterium avium subsp. paratuberculosis." Clinical and Vaccine Immunology 14, no. 9 (July 18, 2007): 1078–83. http://dx.doi.org/10.1128/cvi.00166-07.

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ABSTRACT Mycobacterium avium subsp. paratuberculosis is the etiologic agent of Johne's disease, a chronic granulomatous enteritis in ruminants. ATP has been reported to induce cell death of macrophages and killing of Mycobacterium species in human and murine macrophages. In this study we investigated the short-term effect of ATP on the viability of M. avium subsp. paratuberculosis-infected bovine mononuclear phagocytes and the bacilli within them. Addition of 5 mM ATP to M. avium subsp. paratuberculosis-infected bovine monocytes resulted in 50% cytotoxicity of bovine monocytes at 24 h. Addition of 2′(3′)-O-(4-benzoylbenzoyl) ATP triethylammonium salt (Bz-ATP), which is a longer-lived ATP homologue and purinergic receptor agonist, significantly increased the uptake of YO-PRO, which is a marker for membrane pore activation by P2X receptors. Addition of Bz-ATP also stimulated lactate dehydrogenase release and caspase-3 activity in infected bovine monocytes. Neither ATP nor Bz-ATP reduced the survival of M. avium subsp. paratuberculosis in bovine mononuclear phagocytes. Likewise, addition of ATP or Bz-ATP was cytotoxic to murine macrophage cell lines (RAW 264.7 and J774A.1 cells) but did not affect the intracellular survival of M. avium subsp. paratuberculosis, nor were the numbers of viable Mycobacterium avium subsp. avium or Mycobacterium bovis BCG cells altered in bovine mononuclear phagocytes or J774A.1 cells following ATP or Bz-ATP treatment. These data suggest that extracellular ATP does not induce the killing of intracellular M. avium subsp. paratuberculosis in bovine mononuclear phagocytes.
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47

Ayele, W. Y., M. Macháčková, and I. Pavlík. "The transmission and impact of paratuberculosis infection in domestic and wild ruminants." Veterinární Medicína 46, No. 7–8 (January 1, 2001): 205–24. http://dx.doi.org/10.17221/7878-vetmed.

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Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) infects domestic cattle, sheep, goats, deer, camelids and wild ruminants leading to chronic enteritis known as paratuberculosis (Johne&rsquo;s disease). The infection is chronic, progressive and unresponsive to treatment. Most infected animals do not develop clinical disease but may excrete the bacteria. Clinically sick animals suffer emaciation and in some species diarrhoea, followed by eventual death. During the course of the disease, excretion of M. paratuberculosis in faeces and milk occurs, and the organism spreads through the blood and lymph vessels of infected animals to multiple internal organs. The infection disseminates to both the female and male reproductive organs. Though M. paratuberculosis is not classified as a human pathogen, current opinions on the possible role of this mycobacteria in public health is discussed. This article attempts to review the ways and circumstances by which M. paratuberculosis is transmitted within an animal population and the importance of the disease on animal production. Published reports concerning the transmission and epidemiology of the disease are reviewed herein, and preventive and control measures are summarised.
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48

DIMARELI-MALLI (Ζ. ΔΗΜΑΡΕΛΛΗ-ΜΑΛΛΗ), Z., and C. SARRIS (Κ. ΣΑΡΡΗΣ). "Possible association between Mycobacterium paratuberculosis infection and Crohn's disease in human." Journal of the Hellenic Veterinary Medical Society 48, no. 2 (January 31, 2018): 57. http://dx.doi.org/10.12681/jhvms.15794.

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Crohn's disease is a granulomatous ileocolitis of humans, of unknown aetiology, which generally manifests itself during the prime of life. The chronic, progressive clinical course and histological findings are consistent wiht a mycobacterial aetiology. Evidence supporting a pathogenic role for a mycobacterium has become available only in the last decade with the isolation of this microorganism from Crohn's disease tissue. M. paratuberculosis, which is the causative agent of Johne's disease in animals, has been identified in patients with Crohn's disease by PCR and DNA hybridisation techniques. It has been shown that isolates of M. paratuberculosis from Crohn's disease are indentical with pathogenic strains in ruminants.
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49

Dhand, Navneet K., Jenny-Ann L. M. L. Toribio, and Richard J. Whittington. "Adsorption of Mycobacterium avium subsp. paratuberculosis to Soil Particles." Applied and Environmental Microbiology 75, no. 17 (June 26, 2009): 5581–85. http://dx.doi.org/10.1128/aem.00557-09.

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ABSTRACT Attachment of Mycobacterium avium subsp. paratuberculosis to soil particles could increase their availability to farm animals, as well as influence the transportation of M. avium subsp. paratuberculosis to water sources. To investigate the possibility of such attachment, we passed a known quantity of M. avium subsp. paratuberculosis through chromatography columns packed with clay soil, sandy soil, pure silica, clay-silica mixture, or clay-silica complexes and measured the organisms recovered in the eluent using culture or quantitative PCR. Experiments were repeated using buffer at a range of pH levels with pure silica to investigate the effect of pH on M. avium subsp. paratuberculosis attachment. Linear mixed-model analyses were conducted to compare the proportional recovery of M. avium subsp. paratuberculosis in the eluent between different substrates and pH levels. Of the organisms added to the columns, 83 to 100% were estimated to be retained in the columns after adjustment for those retained in empty control columns. The proportions recovered were significantly different across different substrates, with the retention being significantly greater (P < 0.05) in pure substrates (silica and clay-silica complexes) than in soil substrates (clay soil and sandy soil). However, there were no significant differences in the retention of M. avium subsp. paratuberculosis between silica and clay-silica complexes or between clay soil and sandy soil. The proportion retained decreased with increasing pH in one of the experiments, indicating greater adsorption of M. avium subsp. paratuberculosis to soil particles at an acidic pH (P < 0.05). The results suggest that under experimental conditions M. avium subsp. paratuberculosis adsorbs to a range of soil particles, and this attachment is influenced by soil pH.
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50

Foddai, Antonio, Christopher T. Elliott, and Irene R. Grant. "Maximizing Capture Efficiency and Specificity of Magnetic Separation for Mycobacterium avium subsp. paratuberculosis Cells." Applied and Environmental Microbiology 76, no. 22 (September 17, 2010): 7550–58. http://dx.doi.org/10.1128/aem.01432-10.

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ABSTRACT In order to introduce specificity for Mycobacterium avium subsp. paratuberculosis prior to a phage amplification assay, various magnetic-separation approaches, involving either antibodies or peptides, were evaluated in terms of the efficiency of capture (expressed as a percentage) of M. avium subsp. paratuberculosis cells and the percentage of nonspecific binding by other Mycobacterium spp. A 50:50 mixture of MyOne Tosylactivated Dynabeads coated with the chemically synthesized M. avium subsp. paratuberculosis-specific peptides biotinylated aMp3 and biotinylated aMptD (i.e., peptide-mediated magnetic separation [PMS]) proved to be the best magnetic-separation approach for achieving 85 to 100% capture of M. avium subsp. paratuberculosis and minimal (<1%) nonspecific recovery of other Mycobacterium spp. (particularly if beads were blocked with 1% skim milk before use) from broth samples containing 103 to 104 CFU/ml. When PMS was coupled with a recently optimized phage amplification assay and used to detect M. avium subsp. paratuberculosis in 50-ml volumes of spiked milk, the mean 50% limit of detection (LOD50) was 14.4 PFU/50 ml of milk (equivalent to 0.3 PFU/ml). This PMS-phage assay represents a novel, rapid method for the detection and enumeration of viable M. avium subsp. paratuberculosis organisms in milk, and potentially other sample matrices, with results available within 48 h.
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