Journal articles on the topic 'Paratuberculosis'
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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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Tamboli, Cyrus P. "A Hypothesis for Explaining the Geographical Distribution of Crohn’s Disease." Canadian Journal of Gastroenterology 10, no. 3 (1996): 173–77. http://dx.doi.org/10.1155/1996/319469.
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The etiology of Crohn’s disease (CD) remains unknown, although there is epidemiological evidence supporting an environmental influence. Recent molecular techniques, including polymerase chain reaction, have renewed interest in a possible etiological role ofMycobacterium paratuberculosis, which has been isolated from a number of CD patients. The organism causes a chronic enteritis in animals called paratuberculosis, a condition with many clinical and pathological similarities to CD. This review compares the epidemiology of paratuberculosis in animals with the epidemiology of CD in humans. There is considerable overlap of regions with high prevalences of paratuberculosis and CD. This finding adds support to the implication ofM paratuberculosisin the etiology of CD.
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Zavgorodniy, A. I., S. A. Pozmogova, N. V. Goncharova, M. V. Kalashnyk, and V. V. Bilushko. "The study of epizootic sera obtained from ruminant animals in complement fixation test (CFT) with the use of Paratuberculous antigen." Veterinary Medicine: inter-departmental subject scientific collection, no. 105 (August 7, 2019): 41–45. http://dx.doi.org/10.36016/vm-2019-105-8.
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The paper presents results of the study of epizootic blood sera in the complement fixation test (CFT) with paratuberculous antigen. Blood sera were sampled from the cattle and goats. The antigen was produced from the culture filtrate of Mycobacterium avium subspecies paratuberculosis (MAP) in the laboratory for tuberculosis study. The aim of the present study was to clarify the epizootic situation concerning Johne’s disease among the dairy cattle in different regions of Ukraine. To achieve this aim the blood sera from cattle and goats were collected from farms in different regions of Ukraine. Those sera samples were studied in the complement fixation test with the use of paratuberculous antigen that was produced from the culture filtrate of MAP. The above mentioned blood sera were collected from the cattle that had positive allergic reactions on the use of tuberculin (PPD) for mammals. Those animals belonged to the free from tuberculosis and paratuberulosis milk farms. The study of obtained samples of blood sera was conducted in the accordance with the methodological guidelines “Laboratory diagnostics of paratuberculosis” (shutter. NMR FEFU pr. No. 1, dated December 19, 2014). There were studied 1098 blood sera samples from cattle. In addition to this, investigation was conducted on 24 samples of blood sera from goats. As the result of conducted study it was found that 17 samples of blood sera contained specific antibodies against MAP (serum solution 1:10). These blood sera collected from the cattle belonging to 4 farms in Poltava, Donetsk and Khmelnitsky regions. Along with this it was obtained 9 uncertain results in compliment fixation test that was conducted between paratuberculous antigen (ACF) and blood sera from those 4 farms. The results of monitoring studies indicate that M. avium subsp. paratuberculosis pathogen circulates in studied farms. This can lead to the complication of the epizootic situation regarding paratuberculosis and contribute to the spreading of this pathogen to other free from MAP infection farms. There are no anti-paratuberculosis antibodies in blood serum from goats. It is necessary to conduct annual monitoring serological studies of productive dairy cattle and imported animals in order to clarify and control epizootic situation concerning paratuberculosis on the territory of Ukraine
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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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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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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’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’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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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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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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Cocito, C., P. Gilot, M. Coene, M. de Kesel, P. Poupart, and P. Vannuffel. "Paratuberculosis." Clinical Microbiology Reviews 7, no. 3 (1994): 328–45. http://dx.doi.org/10.1128/cmr.7.3.328-345.1994.
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Mota, Rinaldo A., Paulo V. Peixoto, Elise M. Yamasaki, Elizabeth S. de Medeiros, Mateus M. da Costa, Rodolfo M. Peixoto, and Marilene F. Brito. "Ocorrência de paratuberculose em búfalos (Bubalus bubalis) em Pernambuco." Pesquisa Veterinária Brasileira 30, no. 3 (March 2010): 237–42. http://dx.doi.org/10.1590/s0100-736x2010000300008.
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A paratuberculose (doença de Johne) é uma das doenças de maior importância econômica para ruminantes em vários países e pode representar uma ameaça ao desenvolvimento da pecuária brasileira. É uma doença infecto-contagiosa que provoca enterocolite granulomatosa crônica, incurável e de difícil controle, cujo agente é o Mycobacterium avium subsp. paratuberculosis (MAP). Descreve-se a ocorrência de paratuberculose em um rebanho de búfalos no Estado de Pernambuco, Brasil. Não foi encontrado registro, na literatura, da ocorrência de paratuberculose em búfalos no país. De 100 búfalos, cinco mostravam sinais clínicos característicos da doença. À necropsia de dois animais as lesões estavam restritas ao intestino delgado com evidente espessamento da mucosa, aumento de linfonodos mesentéricos e vasos linfáticos proeminentes e dilatados. À microscopia, observaram-se na mucosa do intestino, infiltrado inflamatório granulomatoso com numerosos macrófagos epitelióides e células gigantes de Langhans, além de bacilos álcool-ácido resistentes (BAAR) visualizados através da coloração de Ziehl-Neelsen (ZN). Nos linfonodos mesentéricos, havia espessamento da cápsula e marcada inflamação granulomatosa. O exame direto pela técnica de ZN para pesquisa do bacilo em esfregaços de fezes, raspado de mucosa intestinal e imprint de linfonodos mesentéricos resultou positivo. A PCR IS900 específico de linfonodo mesentérico e mucosa intestinal revelou amplificação de um fragmento de aproximadamente 110pb, confirmada pela comparação com outras sequências de M. avium subsp. paratuberculosis disponíveis no GenBank.
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Patel, Dilip, Lia Danelishvili, Yoshitaka Yamazaki, Marta Alonso, Michael L. Paustian, John P. Bannantine, Lisbeth Meunier-Goddik, and Luiz E. Bermudez. "The Ability of Mycobacterium avium subsp. paratuberculosis To Enter Bovine Epithelial Cells Is Influenced by Preexposure to a Hyperosmolar Environment and Intracellular Passage in Bovine Mammary Epithelial Cells." Infection and Immunity 74, no. 5 (May 2006): 2849–55. http://dx.doi.org/10.1128/iai.74.5.2849-2855.2006.
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ABSTRACT Mycobacterium avium subsp. paratuberculosis is the cause of Johne's disease in cattle and other ruminants. M. avium subsp. paratuberculosis infection of the bovine host is not well understood; however, it is assumed that crossing the bovine intestinal mucosa is important in order for M. avium subsp. paratuberculosis to establish infection. To examine the ability of M. avium subsp. paratuberculosis to infect bovine epithelial cells in vitro, Madin-Darby bovine kidney (MDBK) epithelial cells were exposed to M. avium subsp. paratuberculosis. It was observed that bacteria can establish infection and replicate within MDBK cells. M. avium subsp. paratuberculosis also has been reported to infect mammary tissue and milk, and we showed that M. avium subsp. paratuberculosis infects bovine mammary epithelial cells (MAC-T cell line). Using polarized MAC-T cell monolayers, it was also determined that M. avium subsp. paratuberculosis crosses apical and basolateral surfaces with approximately the same degree of efficiency. Because M. avium subsp. paratuberculosis can be delivered to the naïve host by milk, it was investigated whether incubation of M. avium subsp. paratuberculosis with milk has an effect on invasion of MDBK cells. M. avium subsp. paratuberculosis exposed to milk entered epithelial cells with greater efficiency than M. avium subsp. paratuberculosis exposed to broth medium or water (P < 0.01). Growth of M. avium subsp. paratuberculosis within MAC-T cells also resulted in augmented ability to subsequently infect bovine MDBK cells (P < 0.001). Microarray analysis of intracellular M. avium subsp. paratuberculosis RNA indicates the increased transcription of genes which might be associated with an invasive phenotype.
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Secott, T. E., T. L. Lin, and C. C. Wu. "Mycobacterium avium subsp. paratuberculosis Fibronectin Attachment Protein Facilitates M-Cell Targeting and Invasion through a Fibronectin Bridge with Host Integrins." Infection and Immunity 72, no. 7 (July 2004): 3724–32. http://dx.doi.org/10.1128/iai.72.7.3724-3732.2004.
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ABSTRACT Efficient attachment and ingestion of Mycobacterium avium subsp. paratuberculosis by cultured epithelial cells requires the expression of a fibronectin (FN) attachment protein homologue (FAP-P) which mediates FN binding by M. avium subsp. paratuberculosis. Invasion of Peyer's patches by M. avium subsp. paratuberculosis occurs through M cells, which, unlike other intestinal epithelial cells, express integrins on their luminal faces. We sought to determine if the interaction between FAP-P of M. avium subsp. paratuberculosis and soluble FN enabled targeting and invasion of M cells by M. avium subsp. paratuberculosis in vivo via these surface integrins. Wild-type and antisense FAP-P mutant M. avium subsp. paratuberculosis strains were injected alone or coinjected with blocking peptides or antibodies into murine gut loops, and immunofluorescence microscopy was performed to assess targeting and invasion of M cells by M. avium subsp. paratuberculosis. Nonopsonized M. avium subsp. paratuberculosis preferentially invaded M cells in murine gut loops. M-cell invasion was enhanced 2.6-fold when M. avium subsp. paratuberculosis was pretreated with FN. Invasion of M cells by the antisense FAP-P mutant of M. avium subsp. paratuberculosis was reduced by 77 to 90% relative to that observed for the control strains. Peptides corresponding to the RGD and synergy site integrin recognition regions of FN blocked M. avium subsp. paratuberculosis invasion of M cells by 75 and 45%, respectively, whereas the connecting segment 1 peptide was noninhibitory. Antibodies against the α5, αV, β1, and β3 integrin subunits inhibited M-cell invasion by 52 to 73%. The results indicate that targeting and invasion of M cells by M. avium subsp. paratuberculosis in vivo is mediated primarily by the formation of an FN bridge formed between FAP-P of M. avium subsp. paratuberculosis and integrins on M cells.
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Tasara, T., and R. Stephan. "Development of an F57 Sequence-Based Real-Time PCR Assay for Detection of Mycobacterium avium subsp. paratuberculosis in Milk." Applied and Environmental Microbiology 71, no. 10 (October 2005): 5957–68. http://dx.doi.org/10.1128/aem.71.10.5957-5968.2005.
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ABSTRACT A light cycler-based real-time PCR (LC-PCR) assay that amplifies the F57 sequence of Mycobacterium avium subsp. paratuberculosis was developed. This assay also includes an internal amplification control template to monitor the amplification conditions in each reaction. The targeted F57 sequence element is unique for M.avium subsp. paratuberculosis and is not known to exist in any other bacterial species. The assay specificity was demonstrated by evaluation of 10 known M. avium subsp. paratuberculosis isolates and 33 other bacterial strains. The LC-PCR assay has a broad linear range (2 × 101 to 2 ×106 copies) for quantitative estimation of the number of M. avium subsp. paratuberculosis F57 target copies in positive samples. To maximize the assay's detection sensitivity, an efficient strategy for isolation of M. avium subsp. paratuberculosis DNA from spiked milk samples was also developed. The integrated procedure combining optimal M. avium subsp. paratuberculosis DNA isolation and real-time PCR detection had a reproducible detection limit of about 10 M. avium subsp. paratuberculosis cells per ml when a starting sample volume of 10 ml of M. avium subsp. paratuberculosis-spiked milk was analyzed. The entire process can be completed within a single working day and is suitable for routine monitoring of milk samples for M. avium subsp. paratuberculosis contamination. The applicability of this protocol for naturally contaminated milk was also demonstrated using milk samples from symptomatic M. avium subsp. paratuberculosis-infected cows, as well as pooled samples from a dairy herd with a confirmed history of paratuberculosis.
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Patton, Elisabeth A. "Paratuberculosis Vaccination." Veterinary Clinics of North America: Food Animal Practice 27, no. 3 (November 2011): 573–80. http://dx.doi.org/10.1016/j.cvfa.2011.07.004.
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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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Bermudez, Luiz E., Mary Petrofsky, Sandra Sommer, and Raúl G. Barletta. "Peyer's Patch-Deficient Mice Demonstrate That Mycobacterium avium subsp. paratuberculosis Translocates across the Mucosal Barrier via both M Cells and Enterocytes but Has Inefficient Dissemination." Infection and Immunity 78, no. 8 (May 24, 2010): 3570–77. http://dx.doi.org/10.1128/iai.01411-09.
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ABSTRACT Mycobacterium avium subsp. paratuberculosis, the agent of Johne's disease, infects ruminant hosts by translocation through the intestinal mucosa. A number of studies have suggested that M. avium subsp. paratuberculosis interacts with M cells in the Peyer's patches of the small intestine. The invasion of the intestinal mucosa by M. avium subsp. paratuberculosis and Mycobacterium avium subsp. hominissuis, a pathogen known to interact with intestinal cells, was compared. M. avium subsp. paratuberculosis was capable of invading the mucosa, but it was significantly less efficient at dissemination than M. avium subsp. hominissuis. B-cell knockout (KO) mice, which lack Peyer's patches, were used to demonstrate that M. avium subsp. paratuberculosis enters the intestinal mucosa through enterocytes in the absence of M cells. In addition, the results indicated that M. avium subsp. paratuberculosis had equal abilities to cross the mucosa in both Peyer's patch and non-Peyer's patch segments of normal mice. M. avium subsp. paratuberculosis was also shown to interact with epithelial cells by an α5β1 integrin-independent pathway. Upon translocation, dendritic cells ingest M. avium subsp. paratuberculosis, but this process does not lead to efficient dissemination of the infection. In summary, M. avium subsp. paratuberculosis interacts with the intestinal mucosa by crossing both Peyer's patches and non-Peyer's patch areas but does not translocate or disseminate efficiently.
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O'Brien, R., C. G. Mackintosh, D. Bakker, M. Kopecna, I. Pavlik, and J. F. T. Griffin. "Immunological and Molecular Characterization of Susceptibility in Relationship to Bacterial Strain Differences in Mycobacterium avium subsp. paratuberculosis Infection in the Red Deer (Cervus elaphus)." Infection and Immunity 74, no. 6 (June 2006): 3530–37. http://dx.doi.org/10.1128/iai.01688-05.
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ABSTRACT Johne's disease (JD) infection, caused by Mycobacterium avium subsp. paratuberculosis, represents a major disease problem in farmed ruminants. Although JD has been well characterized in cattle and sheep, little is known of the infection dynamics or immunological response in deer. In this study, typing of M. avium subsp. paratuberculosis isolates from intestinal lymphatic tissues from 74 JD-infected animals showed that clinical isolates of M. avium subsp. paratuberculosis from New Zealand farmed red deer were exclusively of the bovine strain genotype. The susceptibility of deer to M. avium subsp. paratuberculosis was further investigated by experimental oral-route infection studies using defined isolates of virulent bovine and ovine M. avium subsp. paratuberculosis strains. Oral inoculation with high (109 CFU/animal) or medium (107 CFU/animal) doses of the bovine strain of M. avium subsp. paratuberculosis established 100% infection rates, compared to 69% infection following inoculation with a medium dose of the ovine strain. The high susceptibility of deer to the bovine strain of M. avium subsp. paratuberculosis was confirmed by a 50% infection rate following experimental inoculation with a low dose of bacteria (103 CFU/animal). This study is the first to report experimental M. avium subsp. paratuberculosis infection in red deer, and it outlines the strong infectivity of bovine-strain M. avium subsp. paratuberculosis isolates for cervines.
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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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Dubash, Kerman, William P. Shulaw, Steen Beth-Nielsen, Harold F. Stills, and Richard D. Slemons. "Evaluation of an Enzyme-Linked Immunosorbent Assay Licensed by the USDA for Use in Cattle for Diagnosis of Ovine Paratuberculosis." Journal of Veterinary Diagnostic Investigation 7, no. 3 (July 1995): 347–51. http://dx.doi.org/10.1177/104063879500700309.
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A commercially available Mycobacterium phlei-absorbed enzyme-linked immunosorbent assay (ELISA) approved to detect antibodies to Mycobacterium paratuberculosis in cattle was evaluated for its applicability in sheep. The potential for interference with ELISA results from cross-reacting antibodies to Corynebacterium pseudotuberculosis was also investigated. Serum samples were randomly selected from a collection of samples obtained in 1986-1991 from 6 infected and 5 noninfected sheep flocks varying in breed, age, and geographic origin. Tests were performed on sera from 27 paratuberculous sheep, confirmed by histopathology, bacteriologic culture, and/or acid-fast staining of ileal mucosal smears, and on sera from 246 noninfected sheep. The optical density of each sample was expressed as a percentage of the optical density of a known positive sheep serum sample tested on the same plate. These values were log-transformed to achieve normality of distribution, and sensitivity and specificity estimates were calculated based on 2 and 3 standard deviations above the mean of the percent positive value (PPV) of the noninfected sheep. A cutoff value of PPV ≥ 55.74 resulted in an estimated sensitivity of 0.48 and a specificity of 0.95. Sera from 10 noninfected sheep with PPV above the cutoff level of 55.74% were absorbed with heat-treated C. pseudotuberculosis organisms in addition to M. phlei antigens. Sera from 14 ELISA-positive paratuberculous sheep and 23 ELISA-negative noninfected sheep were similarly treated, and results were compared. Absorption with C. pseudotuberculosis resulted in a significant decrease in PPV in all 3 groups of sheep sera, but a greater decrease was observed in the noninfected sheep with PPV above the cutoff level when compared with noninfected sheep with PPV below that level. Results of this study suggest that ELISA may be of value in screening sheep flocks for paratuberculosis, but further experimentation is needed to optimize the sensitivity and specificity of the assay. Exposure to C. pseudotuberculosis may confound results obtained by M. phlei-absorbed ELISA for paratuberculosis.
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Hughes, Valerie, John P. Bannantine, Susan Denham, Stuart Smith, Alfredo Garcia-Sanchez, Jill Sales, Michael L. Paustian, Kevin Mclean, and Karen Stevenson. "Immunogenicity of Proteome-Determined Mycobacterium avium subsp. paratuberculosis-Specific Proteins in Sheep with Paratuberculosis." Clinical and Vaccine Immunology 15, no. 12 (October 8, 2008): 1824–33. http://dx.doi.org/10.1128/cvi.00099-08.
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ABSTRACT Mycobacterium avium subsp. paratuberculosis causes paratuberculosis, a chronic granulomatous enteritis. Detecting animals with paratuberculosis infections is difficult because the currently available tools have low sensitivity and lack specificity; these tools are prone to generating spurious positive test results caused by exposure to environmental M. avium complex organisms. To generate candidate antigens for incorporation into a specific test for paratuberculosis, subspecies-specific proteins were determined by proteomic comparison of M. avium subsp. paratuberculosis and M. avium subsp. avium. Analysis was aimed at revealing proteins only expressed (or predominant) in the protein profile of M. avium subspecies paratuberculosis. Two-dimensional gel electrophoresis resolved approximately 1,000 protein spots from each subspecies. Proteome analysis identified protein spots whose expression profile appeared markedly increased in M. avium subsp. paratuberculosis, and 32 were identified by analysis of their tryptic peptide profile by matrix-assisted laser desorption ionization-time of flight analysis. Thirty of these proteins were cloned, and their recombinant proteins were expressed. Ovine paratuberculosis sera were used to assess their immunoreactivity by enzyme-linked immunosorbent assay (ELISA), Western blotting, and dot blot analysis. Seventeen proteins were detected in at least one of the immunoassays, and eleven proteins were detected by ELISA with an optical density in excess of the cutoff of 0.1 in four of six sera tested. The immunoreactivity of these proteins indicates their potential as unique diagnostic antigens for the development of a specific serological detection of paratuberculosis.
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Khalifeh, M. S., and J. R. Stabel. "Effects of Gamma Interferon, Interleukin-10, and Transforming Growth Factor β on the Survival of Mycobacterium avium subsp. paratuberculosis in Monocyte-Derived Macrophages from Naturally Infected Cattle." Infection and Immunity 72, no. 4 (April 2004): 1974–82. http://dx.doi.org/10.1128/iai.72.4.1974-1982.2004.
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ABSTRACT Gamma interferon (IFN-γ) plays a significant role in the control of mycobacterial infections, including Mycobacterium avium subsp. paratuberculosis. However, the contribution of other immunoregulatory cytokines, such as interleukin-10 (IL-10) and transforming growth factor β (TGF-β), in Johne's disease has not been investigated as yet. In this study, we examined the effects of in vivo and in vitro infection with M. avium subsp. paratuberculosis on the production of IFN-γ, IL-10, and TGF-β by peripheral blood mononuclear cells (PBMC). We also examined the effects of exogenous IFN-γ, IL-10, and TGF-β on M. avium subsp. paratuberculosis survival in the cell cultures. PBMC obtained from naturally infected cows, regardless of their disease status, specifically upregulated IL-10 and TGF-β in culture supernatants in response to stimulation with live M. avium subsp. paratuberculosis. Nonstimulated PBMC recovered from subclinically infected animals secreted the lowest levels of TGF-β, but after stimulation with live M. avium subsp. paratuberculosis, TGF-β levels in the culture supernatants increased to levels similar to that produced by PBMC from healthy animals. The numbers of viable M. avium subsp. paratuberculosis recovered from cultures from naturally infected animals were higher than those from healthy cows after in vitro infection with M. avium subsp. paratuberculosis. The addition of exogenous IL-10 and TGF-β to PBMC isolated from healthy cows inhibited the bactericidal activity of these cells as evidenced by the increased number of viable M. avium subsp. paratuberculosis recovered from these cultures compared to cell cultures containing medium alone. These data suggest important immune regulatory roles for IL-10 and TGF-β during infection with M. avium subsp. paratuberculosis that may be directly related to their effects on macrophage activation and killing of M. avium subsp. paratuberculosis.
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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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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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Judge, Johanna, Ilias Kyriazakis, Alastair Greig, David J. Allcroft, and Michael R. Hutchings. "Clustering of Mycobacterium avium subsp. paratuberculosis in Rabbits and the Environment: How Hot Is a Hot Spot?" Applied and Environmental Microbiology 71, no. 10 (October 2005): 6033–38. http://dx.doi.org/10.1128/aem.71.10.6033-6038.2005.
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ABSTRACT Clustering of pathogens in the environment leads to hot spots of diseases at local, regional, national, and international levels. Scotland contains regional hot spots of Johne's disease (caused by Mycobacterium avium subsp. paratuberculosis) in rabbits, and there is increasing evidence of a link between paratuberculosis infections in rabbits and cattle. The spatial and temporal dynamics of paratuberculosis in rabbits within a hot spot region were studied with the overall aim of determining environmental patterns of infection and thus the risk of interspecies transmission to livestock. The specific aims were to determine if prevalence of paratuberculosis in rabbits varies temporally between seasons and whether the heterogeneous spatial environmental distribution of M. avium subsp. paratuberculosis on a large scale (i.e., regional hot spots) is replicated at finer resolutions within a hot spot. The overall prevalence of M. avium subsp. paratuberculosis in rabbits was 39.7%; the temporal distribution of infection in rabbits followed a cyclical pattern, with a peak in spring of 55.4% and a low in summer of 19.4%. Spatially, M. avium subsp. paratuberculosis-infected rabbits and, thus, the risk of interspecies transmission were highly clustered in the environment. However, this is mostly due to the clustered distribution of rabbits. The patterns of M. avium subsp. paratuberculosis infection in rabbits are discussed in relation to the host's socioecology and risk to livestock.
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Secott, T. E., T. L. Lin, and C. C. Wu. "Fibronectin Attachment Protein Is Necessary for Efficient Attachment and Invasion of Epithelial Cells by Mycobacterium avium subsp. paratuberculosis." Infection and Immunity 70, no. 5 (May 2002): 2670–75. http://dx.doi.org/10.1128/iai.70.5.2670-2675.2002.
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ABSTRACT Attachment and ingestion of Mycobacterium avium subsp. paratuberculosis by two epithelial cell lines were enhanced by soluble fibronectin (FN). Peptide blocking of the FN attachment protein (FAP-P) inhibited the internalization of M. avium subsp. paratuberculosis. Disruption of FAP-P expression significantly reduced attachment and ingestion of M. avium subsp. paratuberculosis by T-24 and Caco-2 cells. The results indicate that the interaction between FN and FAP-P facilitates attachment and internalization of M. avium subsp. paratuberculosis by epithelial cells.
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Sharma, M. L., M. Prajapati, and Y. Panth. "Demonstration of Circulating Antibodies of Mycobacterium avium Subspecies paratuberculosis in Cattle of Rupandehi District, Nepal." Nepalese Veterinary Journal 36 (December 1, 2019): 23–29. http://dx.doi.org/10.3126/nvj.v36i0.27749.
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Paratuberculosis, caused by Mycobacterium avium subspp. paratuberculosis, is a chronic intestinal infection of global importance in mainly domestic and wild ruminants. The main objective of the study was to find out the seroprevalence of Paratuberculosis in cattle of Rupandehi district. The research was conducted from October 2016 to December 2016. A total of 184 blood samples were collected from Jugular vein of cattle and tested by Enzyme Linked Immunosorbent Assay (ELISA). The Paratuberculosis Indirect Screening Test Kit was developed by ID. Vet, France. Cattle with history of chronic diarrhoea and emaciation were taken as study population along with other cattle in close association with them. Overall seroprevalence in Rupandehi district was found to be 4.89%. No significant relation of paratuberculosis was found with age, breed, parity, body condition score and location. Higher prevalence was found in cattle of older age and low body condition score. The result of this study reports the presence of bovine paratuberculosis in cattle of Rupandehi district.
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Scandurra, Gabriella M., Geoffrey W. de Lisle, Sonia M. Cavaignac, May Young, R. Pamela Kawakami, and Desmond M. Collins. "Assessment of Live Candidate Vaccines for Paratuberculosis in Animal Models and Macrophages." Infection and Immunity 78, no. 3 (December 28, 2009): 1383–89. http://dx.doi.org/10.1128/iai.01020-09.
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ABSTRACT Mycobacterium avium subsp. paratuberculosis (basonym M. paratuberculosis) is the causative agent of paratuberculosis, a chronic enteritis of ruminants. To control the considerable economic effect that paratuberculosis has on the livestock industry, a vaccine that induces protection with minimal side effects is required. We employed transposon mutagenesis and allelic exchange to develop three potential vaccine candidates, which were then tested for virulence with macrophages, mice, and goats. All three models identified the WAg906 mutant as being the most attenuated, but some differences in the levels of attenuation were evident among the models when testing the other strains. In a preliminary mouse vaccine experiment, limited protection was induced by WAg915, as evidenced by a reduced bacterial load in spleens and livers 12 weeks following intraperitoneal challenge with M. paratuberculosis K10. While we found macrophages and murine models to be rapid and cost-effective alternatives for the initial screening of M. paratuberculosis mutants for attenuation, it appears necessary to do the definitive assessment of attenuation with a ruminant model.
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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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Sung, Nackmoon, and Michael T. Collins. "Variation in Resistance of Mycobacterium paratuberculosis to Acid Environments as a Function of Culture Medium." Applied and Environmental Microbiology 69, no. 11 (November 2003): 6833–40. http://dx.doi.org/10.1128/aem.69.11.6833-6840.2003.
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ABSTRACT Acid resistance of Mycobacterium paratuberculosis was examined as a function of growth conditions (i.e., in vitro growth medium and pH). M. paratuberculosis was cultured in either fatty acid-containing medium (7H9-OADC) or glycerol-containing medium (WR-GD or 7H9-GD) at two culture pHs (pHs 6.0 and 6.8). Organisms produced in these six medium and pH conditions were then tested for resistance to acetate buffer at pHs 3, 4, 5, and 6 at 20°C. A radiometric culture method (BACTEC) was used to quantify viable M. paratuberculosis cell data at various acid exposure times, and D values (decimal reduction times, or the times required to kill a 1-log10 concentration of bacteria) were determined. Soluble proteins of M. paratuberculosis grown under all six conditions were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) to identify proteins that may be associated with acid resistance or susceptibility. The culture medium affected growth rate and morphology: thin floating sheets of cells were observed in 7H9-OADC versus confluent, thick, waxy, and wrinkled pellicles in WR-GD. Culture medium pH affected growth rate (which was highest at pH 6.0), but it had little or no effect on D values for M. paratuberculosis at any test pH. When grown in 7H9-OADC, M. paratuberculosis was more acid resistant at all test pHs (higher D values) than when grown in WR-GD. Glycerol appeared to be the culture medium component most responsible for lower levels of M. paratuberculosis acid resistance. When glycerol was substituted for OADC in the 7H9 medium, D values were significantly lower than those of 7H9-OADC-grown M. paratuberculosis and were approximately the same as those for M. paratuberculosis grown in WR-GD medium. Comparison of the SDS-PAGE protein profiles for M. paratuberculosis cultures grown in 7H9-OADC, WR-GD, or 7H9-GD medium revealed that increased expression of 34.2- and 14.0-kDa proteins was associated with higher levels of acid resistance of M. paratuberculosis grown in 7H9-OADC medium and that 56.6- and 41.3-kDa proteins were associated with lower levels of acid resistance. This is the first report showing that in vitro culture conditions significantly affect growth characteristics, acid resistance, and protein expression of M. paratuberculosis, and the results emphasize the importance of culture conditions for in vitro susceptibility studies.
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Bannantine, John P., Jason F. J. Huntley, Elizabeth Miltner, Judith R. Stabel, and Luiz E. Bermudez. "The Mycobacterium avium subsp. paratuberculosis 35 kDa protein plays a role in invasion of bovine epithelial cells." Microbiology 149, no. 8 (August 1, 2003): 2061–69. http://dx.doi.org/10.1099/mic.0.26323-0.
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Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) enters intestinal epithelial cells of cattle and other ruminants via a mechanism that remains to be fully elucidated. This study showed that a gene encoding the M. paratuberculosis 35 kDa major membrane protein (MMP) is expressed at a higher level in low-oxygen and high-osmolarity conditions that are similar to the environment of the intestine. In addition, cattle with Johne's disease produced antibodies against MMP, suggesting that the protein is present during infection. The gene encoding MMP was cloned and expressed as a fusion protein with the maltose-binding protein (MBP–MMP) in Escherichia coli. Rabbit antisera were raised against a M. paratuberculosis whole-cell sonicate and MMP-specific antibodies were purified from these sera by affinity chromatography. MMP was localized to the surface of M. paratuberculosis by immunoelectron microscopy and by immunoblot analysis of fractionated protein lysates. Both anti-MMP antibodies and MBP–MMP protein inhibited M. paratuberculosis invasion of cultured Madin–Darby bovine kidney cells by 30 %. In similar invasion experiments with M. paratuberculosis incubated in low oxygen tension, these antibodies and protein decreased invasion by 60 %. Collectively, these data show that the 35 kDa MMP is a surface exposed protein that plays a role in invasion of epithelial cells. The authors suggest that the MMP is a virulence factor of M. paratuberculosis that may be important in the initiation of infection in vivo.
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O'Reilly, Ciara E., Lisa O'Connor, Wayne Anderson, Peter Harvey, Irene R. Grant, John Donaghy, Michael Rowe, and Pat O'Mahony. "Surveillance of Bulk Raw and Commercially Pasteurized Cows' Milk from Approved Irish Liquid-Milk Pasteurization Plants To Determine the Incidence of Mycobacterium paratuberculosis." Applied and Environmental Microbiology 70, no. 9 (September 2004): 5138–44. http://dx.doi.org/10.1128/aem.70.9.5138-5144.2004.
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ABSTRACT Over the 13-month period from October 2000 to November 2001 (inclusive), the Food Safety Authority of Ireland (FSAI) carried out surveillance of Irish bulk raw (n = 389) and commercially pasteurized (n = 357) liquid-milk supplies to determine the incidence of Mycobacterium paratuberculosis. The pasteurization time-temperature conditions were recorded for all pasteurized samples. Overall, 56% of whole-milk pasteurized samples had been heat treated at or above a time-temperature combination of 75°C for 25 s. All analyses were undertaken at the Department of Food Science (Food Microbiology) laboratory at Queen's University Belfast. Each milk sample was subjected to two tests for M. paratuberculosis: immunomagnetic separation-PCR (IMS-PCR; to detect the presence of M. paratuberculosis cells, live or dead) and chemical decontamination and culture (to confirm the presence of viable M. paratuberculosis). Overall, M. paratuberculosis DNA was detected by IMS-PCR in 50 (12.9%; 95% confidence interval, 9.9 to 16.5%) raw-milk samples and 35 (9.8%; 95% confidence interval, 7.1 to 13.3%) pasteurized-milk samples. Confirmed M. paratuberculosis was cultured from one raw-milk sample and no pasteurized-milk samples. It is concluded that M. paratuberculosis DNA is occasionally present at low levels in both raw and commercially pasteurized cows' milk. However, since no viable M. paratuberculosis was isolated from commercially pasteurized cows' milk on retail sale in the Republic of Ireland, current pasteurization procedures are considered to be effective.
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Grant, Irene R., Edward I. Hitchings, Alan McCartney, Fiona Ferguson, and Michael T. Rowe. "Effect of Commercial-Scale High-Temperature, Short-Time Pasteurization on the Viability of Mycobacterium paratuberculosis in Naturally Infected Cows' Milk." Applied and Environmental Microbiology 68, no. 2 (February 2002): 602–7. http://dx.doi.org/10.1128/aem.68.2.602-607.2002.
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ABSTRACT Raw cows' milk naturally infected with Mycobacterium paratuberculosis was pasteurized with an APV HXP commercial-scale pasteurizer (capacity 2,000 liters/h) on 12 separate occasions. On each processing occasion, milk was subjected to four different pasteurization treatments, viz., 73�C for 15 s or 25 s with and without prior homogenization (2,500 lb/in2 in two stages), in an APV Manton Gaulin KF6 homogenizer. Raw and pasteurized milk samples were tested for M. paratuberculosis by immunomagnetic separation (IMS)-PCR (to detect the presence of bacteria) and culture after decontamination with 0.75% (wt/vol) cetylpyridinium chloride for 5 h (to confirm bacterial viability). On 10 of the 12 processing occasions, M. paratuberculosis was detectable by IMS-PCR, culture, or both in either raw or pasteurized milk. Overall, viable M. paratuberculosis was cultured from 4 (6.7%) of 60 raw and 10 (6.9%) of 144 pasteurized milk samples. On one processing day, in particular, M. paratuberculosis appeared to have been present in greater abundance in the source raw milk (evidenced by more culture positives and stronger PCR signals), and on this occasion, surviving M. paratuberculosis bacteria were isolated from milk processed by all four heat treatments, i.e., 73�C for 15 and 25 s with and without prior homogenization. On one other occasion, surviving M. paratuberculosis bacteria were isolated from an unhomogenized milk sample that had been heat treated at 73�C for 25 s. Results suggested that homogenization increases the lethality of subsequent heat treatment to some extent with respect to M. paratuberculosis, but the extended 25-s holding time at 73�C was found to be no more effective at killing M. paratuberculosis than the standard 15-s holding time. This study provides clear evidence that M. paratuberculosis bacteria in naturally infected milk are capable of surviving commercial high-temperature, short-time pasteurization if they are present in raw milk in sufficient numbers.
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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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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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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’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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Vidic, B., S. Savic, V. Vidic, M. Jovicin, and N. Prica. "Economic impact of paratuberculosis on milk production." Biotehnologija u stocarstvu 29, no. 2 (2013): 183–91. http://dx.doi.org/10.2298/bah1302183v.
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Paratuberculosis (Johne's disease) is a chronic inflammatory bowel disease, primarily affecting ruminants. The aetiologic agent is Mycobacterium avium subsp. paratuberculosis (MAP). The disease is characterised by persistent diarrhea, weight loss and protein-losing enteropathy. Paratuberculosis can cause significant economic loss in affected herds, as a result of reduced milk yield, increased incidence of mastitis, altered milk constituents, increased somatic cell counts, poor feed conversion, increased susceptibility to disease in general, reduced reproductive efficiency, premature culling and reduced cull cow values. The economic impact of paratuberculosis includes production losses due to sub-clinical and clinical cases, losses due to increased replacement of animals and costs of control measures. Due to the fact that most cases of paratuberculosis are subclinical and precise prevalence data are often lacking, it is difficult to assess the economic consequences of paratuberculosis. For instance, estimates of milk production losses are inconsistent. Some studies found equivalent or even higher milk productions in test-positive animals. Other studies showed losses in test-positive animals of up to 19.5% of the 0 to 305 days-in-milk production, depending on parity.
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Cheng, Zilong, Mengda Liu, Peng Wang, Peng Liu, Meng Chen, Jiandong Zhang, Sidang Liu, and Fangkun Wang. "Characteristics and Epidemiological Investigation of Paratuberculosis in Dairy Cattle in Tai’an, China." BioMed Research International 2020 (March 13, 2020): 1–7. http://dx.doi.org/10.1155/2020/3896754.
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Paratuberculosis, a chronic and sometimes fatal disease of ruminants, is caused by Mycobacterium avium subsp. paratuberculosis (MAP). In this study, we examined paratuberculosis cases among 2–4-year-old dairy cows at farms in Shandong Province, China. Paratuberculosis cases were diagnosed based on clinical symptoms, pathological autopsy, and histopathological inspection. Characteristics of paratuberculosis in the affected dairy cattle included poor body condition, persistent diarrhea, subcutaneous edema, granulomatous ileitis (multibacillary), mesenteric lymphadenitis, and hepatitis. Acid-fast bacilli from fecal specimens and lymphocytes were putatively identified as MAP based on Ziehl-Neelsen staining, then confirmed using polymerase chain reaction-based testing and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analyses. Overall, only one MAP strain was isolated from a herd with symptomatic diarrhea. However, analysis of 586 serum samples from nine herds in Tai’an City revealed that 66.7% of herds and 14.2% of animals were seropositive for MAP. Our findings suggest that paratuberculosis is widely prevalent and therefore a significant threat to the dairy industry in Tai’an City, Shandong Province, China.
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Olsen, Ingrid, Morten Tryland, Harald G. Wiker, and Liv J. Reitan. "AhpC, AhpD, and a Secreted 14-Kilodalton Antigen fromMycobacterium avium subsp.paratuberculosis Distinguish between Paratuberculosis and Bovine Tuberculosis in an Enzyme-Linked Immunosorbent Assay." Clinical Diagnostic Laboratory Immunology 8, no. 4 (July 1, 2001): 797–801. http://dx.doi.org/10.1128/cdli.08.4.797-801.2001.
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ABSTRACT Sera from cattle naturally infected with Mycobacterium avium subsp. paratuberculosis(n = 56) and naturally (n = 4) and experimentally (n = 8) infected withMycobacterium bovis were tested for the presence of antibodies against paratuberculosis antigens. An enzyme-linked immunosorbent assay (ELISA) was established based on absorption ofM. avium subsp. paratuberculosis antigens on a hyperimmune antiserum against M. avium subsp.avium proteins in order to remove cross-reacting antigens. This absorbed-antigen ELISA recognized 66% of animals with paratuberculosis (37 of 56), while none of the animals with naturally occurring bovine tuberculosis (TB) had detectable antibodies. However, the animals with experimental bovine TB also responded in this ELISA. Similar results were found in a commercial ELISA, showing that neither of these tests was able to distinguish between paratuberculosis and bovine TB. The sera were further tested for antibody activities against purified AhpC and AhpD, which are proteins constitutively expressed byM. avium subsp. paratuberculosis, and against a secreted 14-kDa protein present in culture filtrates from theM. avium complex. Elevated antibody levels to AhpC, AhpD, and the 14-kDa antigen were found in 27% (13 of 48), 15% (7 of 48), and 27% (13 of 48), respectively, of the cattle with paratuberculosis. Together these ELISAs were positive with 35% (17 of 48) of the animals. None of the animals with bovine TB had detectable antibodies against any of the purified proteins despite their high levels of cross-reacting antibodies. These results show that purified specific antigens are needed to differentiate between paratuberculosis and bovine TB in ELISA.
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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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DANIELS, M. J., D. HENDERSON, A. GREIG, K. STEVENSON, J. M. SHARP, and M. R. HUTCHINGS. "The potential role of wild rabbits Oryctolagus cuniculus in the epidemiology of paratuberculosis in domestic ruminants." Epidemiology and Infection 130, no. 3 (June 2003): 553–59. http://dx.doi.org/10.1017/s0950268803008471.
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Mycobacterium avium subspecies paratuberculosis, the organism responsible for paratuberculosis in cattle and sheep has been found in wild rabbits (Oryctolagus cuniculus) in the east of Scotland. Few studies have investigated either the level of faecal contamination by rabbits on farms, or the potential infectivity of rabbit excreta. The rate of rabbit faecal contamination deposited and the numbers encountered were estimated for 21 fields on 4 farms with a paratuberculosis problem. 7357±2571 S.E.M. rabbit faecal pellets were deposited per hectare per day and up to 81000 pellets/ha (‘standing crop’) were encountered in October/November 1998. Where access to rabbits was restricted, the standing crop of faeces encountered fell to 22000 pellets/ha.The prevalence of infection with M. a. paratuberculosis was assessed for 83 rabbits from the four farms. M. a. paratuberculosis was isolated from rabbits on all farms with an overall prevalence of 17%. Out of 17 rabbits from which urine was available, M. a. paratuberculosis was isolated from two – the first reported isolation from urine in wild rabbits. The mean number of colony-forming units per gram of infected rabbit faeces was 7·6×105±5·2×105.A relative estimate of the input of M. a. paratuberculosis onto pasture, at the stocking levels found on the four farms, showed that sheep and cattle potentially contributed 4 and 125 times more organisms/ha per day respectively than rabbits. However, rabbits could still contribute millions of M. a. paratuberculosis organisms per ha per day. Existing rabbit control measures on farms may be inadequate in reducing the risk of transmission to livestock.
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Donaghy, J. A., N. L. Totton, and M. T. Rowe. "Persistence of Mycobacterium paratuberculosis during Manufacture and Ripening of Cheddar Cheese." Applied and Environmental Microbiology 70, no. 8 (August 2004): 4899–905. http://dx.doi.org/10.1128/aem.70.8.4899-4905.2004.
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ABSTRACT Model Cheddar cheeses were prepared from pasteurized milk artificially contaminated with high 104 to 105 CFU/ml) and low (101 to 102 CFU/ml) inocula of three different Mycobacterium paratuberculosis strains. A reference strain, NCTC 8578, and two strains (806PSS and 796PSS) previously isolated from pasteurized milk for retail sale were investigated in this study. The manufactured Cheddar cheeses were similar in pH, salt, moisture, and fat composition to commercial Cheddar. The survival of M. paratuberculosis cells was monitored over a 27-week ripening period by plating homogenized cheese samples onto HEYM agar medium supplemented with the antibiotics vancomycin, amphotericin B, and nalidixic acid without a decontamination step. A concentration effect was observed in M. paratuberculosis numbers between the inoculated milk and the 1-day old cheeses for each strain. For all manufactured cheeses, a slow gradual decrease in M. paratuberculosis CFU in cheese was observed over the ripening period. In all cases where high levels (>3.6 log10) of M. paratuberculosis were present in 1-day cheeses, the organism was culturable after the 27-week ripening period. The D values calculated for strains 806PSS, 796PSS, and NCTC 8578 were 107, 96, and 90 days, respectively. At low levels of contamination, M. paratuberculosis was only culturable from 27-week-old cheese spiked with strain 806PSS. M. paratuberculosis was recovered from the whey fraction in 10 of the 12 manufactured cheeses. Up to 4% of the initial M. paratuberculosis load was recovered in the culture-positive whey fractions at either the high or low initial inoculum.
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Spahr, U., and K. Schafroth. "Fate of Mycobacterium avium subsp.paratuberculosis in Swiss Hard and Semihard Cheese Manufactured from Raw Milk." Applied and Environmental Microbiology 67, no. 9 (September 1, 2001): 4199–205. http://dx.doi.org/10.1128/aem.67.9.4199-4205.2001.
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ABSTRACT Raw milk was artificially contaminated with declumped cells ofMycobacterium avium subsp. paratuberculosis at a concentration of 104 to 105 CFU/ml and was used to manufacture model hard (Swiss Emmentaler) and semihard (Swiss Tisliter) cheese. Two different strains of M. avium subsp.paratuberculosis were tested, and for each strain, two model hard and semihard cheeses were produced. The survival of M. avium subsp. paratuberculosis cells was monitored over a ripening period of 120 days by plating out homogenized cheese samples onto 7H10-PANTA agar. In both the hard and the semihard cheeses, counts decreased steadily but slowly during cheese ripening. Nevertheless, viable cells could still be detected in 120-day cheese.D values were calculated at 27.8 days for hard and 45.5 days for semihard cheese. The most important factors responsible for the death of M. avium subsp. paratuberculosisin cheese were the temperatures applied during cheese manufacture and the low pH at the early stages of cheese ripening. Since the ripening period for these raw milk cheeses lasts at least 90 to 120 days, theD values found indicate that 103 to 104 cells of M. avium subsp.paratuberculosis per g will be inactivated.
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SELIM, A., R. HALIM, E. GALILA, and F. HAMOUDA. "Seroprevalence and associated risk factors for bovine paratuberculosis in dairy cattle." Journal of the Hellenic Veterinary Medical Society 72, no. 1 (April 9, 2021): 2647. http://dx.doi.org/10.12681/jhvms.26746.
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Paratuberculosis is an economically important disease in dairy cows and requires continuous surveillance. The study aimed to investigate the seroprevalence of bovine paratuberculosis (Johne’s disease) in one of dairy farm in Egypt. A total of 964 dairy cattle were blood sampled and examined with an ELISA method. One-hundred fifty-five (16.1%) samples reacted positively. The results revealed that age was significantly associated with the prevalence of paratuberculosis in dairy cattle, particularly in animals over 6 years of age. Furthermore, the lactation period, milk yield and pregnancy had non-significant effect on appearance of paratuberculosis in cattle.
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Williams, Stephanie L., N. Beth Harris, and Raúl G. Barletta. "Development of a Firefly Luciferase-Based Assay for Determining Antimicrobial Susceptibility of Mycobacterium aviumsubsp. paratuberculosis." Journal of Clinical Microbiology 37, no. 2 (1999): 304–9. http://dx.doi.org/10.1128/jcm.37.2.304-309.1999.
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Paratuberculosis (Johne’s disease) is a fatal disease of ruminants for which no effective treatment is available. Presently, no drugs against Mycobacterium avium subsp.paratuberculosis (M. paratuberculosis), the causative agent of Johne’s disease, are approved for use in livestock. Additionally, M. paratuberculosis has been linked to a human chronic granulomatous ileitis (Crohn’s disease). To assist in the evaluation of antimicrobial agents with potential activity against M. paratuberculosis, we have developed a firefly luciferase-based assay for the determination of drug susceptibilities. The microorganism used was M. paratuberculosis K-10(pYUB180), a clinical isolate carrying a plasmid with the firefly luciferase gene. The MICs determined by the broth macrodilution method were as follows: amikacin, 2 μg/ml; Bay y 3118, 0.015 μg/ml; clarithromycin, 1.25 μg/ml;d-cycloserine, 25 μg/ml; ethambutol, 20 μg/ml; and rifabutin, 0.5 μg/ml. The strain was resistant to isoniazid and kanamycin. The results obtained by the luciferase assay were identical or fell within 1 doubling dilution. These results suggest that a combination of amikacin, clarithromycin, and rifabutin may be the most efficacious therapy for the treatment of M. paratuberculosis infections and that the use of fluoroquinolone class of antibiotics deserves further consideration. We demonstrate that the luciferase drug susceptibility assay is reliable for M. paratuberculosis and gives results within 7 days, whereas the broth macrodilution method requires 14 days.
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Shin, Sung Jae, Donghee Cho, and Michael T. Collins. "Diagnosis of Bovine Paratuberculosis by a Novel Enzyme-Linked Immunosorbent Assay Based on Early Secreted Antigens of Mycobacterium avium subsp. paratuberculosis." Clinical and Vaccine Immunology 15, no. 8 (June 11, 2008): 1277–81. http://dx.doi.org/10.1128/cvi.00105-08.
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ABSTRACT We previously reported that protein antigens of serodiagnostic potential were more abundant in culture filtrates than cellular extracts from liquid cultures of Mycobacterium avium subsp. paratuberculosis (D. Cho and M. T. Collins, Clin. Vaccine Immunol. 13:1155-1161, 2006). Based on this observation, a novel enzyme-linked immunosorbent assay (ELISA) using antigens secreted by young (early- to mid-log-phase) cultures of M. avium subsp. paratuberculosis JTC303 (a low-passage isolate originating from the ileum of a Holstein bull) in mycobactin-supplemented Watson-Reid medium (pH 6.0) was developed and evaluated using a previously described panel of bovine sera (M. T. Collins et al., Clin. Diagn. Lab. Immunol. 12:685-692, 2005) that included 444 paratuberculosis cases and 412 controls. The new assay, called JTC-ELISA, had a significantly higher diagnostic sensitivity and an equivalent specificity compared to those of five commercial paratuberculosis ELISA kits. By receiver-operating characteristic analysis, the JTC-ELISA had the highest area under the curve of the six assays evaluated. The JTC-ELISA was particularly sensitive at detecting low-level fecal shedders of Mavium subsp. paratuberculosis (40%; the sensitivity of the commercial kits was 20%). The JTC-ELISA works effectively on both serum and milk samples for the detection of cattle with subclinical M. avium subsp. paratuberculosis infections, providing a cost-effective diagnostic tool to support paratuberculosis control programs in cattle herds.
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Harris, N. Beth, Suelee Robbe-Austerman, and Janet B. Payeur. "Effect of Egg Yolk on the Detection of Mycobacterium Avium Subsp. Paratuberculosis Using the ESP II Liquid Culture System." Journal of Veterinary Diagnostic Investigation 17, no. 6 (November 2005): 554–60. http://dx.doi.org/10.1177/104063870501700605.
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Rapid diagnosis of paratuberculosis in infected cattle is important for the successful control of Johne disease within herds. Thus, improving culture methods for Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) will aid in the identification of asymptomatic animals. Egg yolk is a component of the media used for growing M. paratuberculosis, but its requirement as a supplement has not been reported. Using the ESP II liquid culture system, 2 different sources and 5 concentrations (3.3%, 1.6%, 0.8%, 0.4%, and 0%) of egg yolk were analyzed. Egg yolk source did not affect either recovery rate or time to detection, but both parameters were significantly improved when the 3.3% egg yolk concentrations (final volume) were used over media containing no egg yolk. This study also assessed the recovery of M. paratuberculosis from fecal samples that were cultured multiple times using Herrold egg yolk agar (HEY). Specimens containing greater than 70 cfu/g feces could routinely be identified as positive for M. paratuberculosis after only 1 culture attempt, whereas specimens with fewer bacteria were only intermittently positive, even after 5 replicate cultures. Therefore, this study indicates that the sensitivity of the Trek Diagnostic ESP II liquid culture system for M. paratuberculosis is affected by egg yolk concentration and that single culture attempts using HEY solid media may not identify specimens containing low numbers of bacteria.
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Schwarz, D. G. G., I. A. Carvalho, P. A. G. Pietralonga, A. C. S. Faria, and M. A. S. Moreira. "Paratuberculose em pequenos ruminantes domésticos." Arquivos do Instituto Biológico 79, no. 3 (September 2012): 443–52. http://dx.doi.org/10.1590/s1808-16572012000300019.
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Mycobacterium avium subesp. paratuberculosis (MAP) é o agente etiológico da paratuberculose em ruminantes domésticos e silvestres. Em caprinos e ovinos causa uma enterite granulomatosa crônica com emagrecimento progressivo seguido de morte, raramente a diarreia é observada. Nesse aspecto, a doença é considerada uma ameaça mundial aos rebanhos, pois pode permanecer no estádio subclínico por anos, manifestando perdas indiretas na produção animal e na disseminação do agente. Além disso, existem relatos da possível relação de MAP com a doença de Crohn, determinando, assim, a sua relevância na saúde pública. O Brasil não dispõe de dados quantificando as reais perdas produtivas nos rebanhos acometidos pela doença, e poucas informações do comprometimento de caprinos e ovinos no país são relatadas. Assim, este artigo busca revisar a paratuberculose em pequenos ruminantes domésticos.
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de Juan, Luc�a, Julio �lvarez, Beatriz Romero, Javier Bezos, Elena Castellanos, Alicia Aranaz, Ana Mateos, and Lucas Dom�nguez. "Comparison of Four Different Culture Media for Isolation and Growth of Type II and Type I/III Mycobacterium avium subsp. paratuberculosis Strains Isolated from Cattle and Goats." Applied and Environmental Microbiology 72, no. 9 (September 2006): 5927–32. http://dx.doi.org/10.1128/aem.00451-06.
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ABSTRACT Culture is considered the definitive technique for Johne's disease diagnosis, and it is essential for later applications of certain molecular typing techniques. In this study, we have tested four solid media (Herrold's egg yolk medium [HEYM] with sodium pyruvate and mycobactin [HEYMm-SP], HEYM with mycobactin and without sodium pyruvate [HEYMm], Middlebrook 7H11 with mycobactin [Mm], and L�wenstein-Jensen with mycobactin [LJm]) for isolation of Mycobacterium avium subsp. paratuberculosis strains in 319 tissue samples from cattle herds and goat flocks. We have shown that each of the two main groups of M. avium subsp. paratuberculosis (type II and type I/III) has different requirements for growth in the culture media studied. The recommended solid media for isolation of type I/III strains are LJm and Mm, since the combination of both media allowed the recovery of all these strains. The most widespread culture medium, HEYM, is not suitable for the isolation of this group of M. avium subsp. paratuberculosis strains. Regarding the type II strains, HEYMm-SP was the medium where more strains were isolated, but the other three media are also needed in order to recover all type II strains. The incubation period is also related to the strain type. In conclusion, because the type of strain cannot be known in advance of culture, coupled with the fact that cattle and goats can be infected with both groups of strains, we recommend the use of the four solid media and the prolongation of the incubation period to more than 6 months to detect paratuberculous herds/flocks and to determine the true prevalence of the infection.
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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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Whan, Lynne, Hywel J. Ball, Irene R. Grant, and Michael T. Rowe. "Occurrence of Mycobacterium avium subsp. paratuberculosis in Untreated Water in Northern Ireland." Applied and Environmental Microbiology 71, no. 11 (November 2005): 7107–12. http://dx.doi.org/10.1128/aem.71.11.7107-7112.2005.
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ABSTRACT Mycobacterium avium subsp. paratuberculosis is the known cause of Johne's disease of both domestic and wild ruminants and has been implicated as a possible cause of Crohn's disease in humans. The organism is shed in the feces of infected animals and can survive for protracted periods in the environment and hence could be present in catchment areas receiving agricultural runoff. A limited survey was undertaken in Northern Ireland to test for M. avium subsp. paratuberculosis in untreated water entering nine water treatment works (WTWs) over a 1-year period. Three detection methods were employed, viz., immunomagnetic separation-PCR and culture on Herrold's egg yolk medium (HEYM) and BACTEC 12B medium, the latter both supplemented with mycobactins. Of the 192 untreated water samples tested, 15 (8%) tested M. avium subsp. paratuberculosis positive by one or more of the three detection methods. M. avium subsp. paratuberculosis was successfully isolated from eight untreated water samples, three by BACTEC culture and five by culture on HEYM. Although the highest incidence of M. avium subsp. paratuberculosis was found in spring, overall, there was no statistically significant difference between the seasons. No significant correlation was found between numbers of coliforms or fecal coliforms and the presence of M. avium subsp. paratuberculosis. In general, a higher incidence of M. avium subsp. paratuberculosis was found in untreated water entering those WTWs that had a high mean water pH value over the sampling period. This work indicates the need to determine the efficacy of water treatment processes to either kill or remove M. avium subsp. paratuberculosis from untreated water and the possible risks posed by contact with recreational water sources.