scholarly journals Differential proteome analysis of Mycobacterium avium subsp. paratuberculosis grown in vitro and isolated from cases of clinical Johne's disease

Microbiology ◽  
2011 ◽  
Vol 157 (2) ◽  
pp. 557-565 ◽  
Author(s):  
Mathias Weigoldt ◽  
Jochen Meens ◽  
Klaus Doll ◽  
Isabel Fritsch ◽  
Petra Möbius ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Mycobacterium Avium ◽  
Proteome Analysis ◽  
Economic Problem ◽  
Johne's Disease ◽  
Johne’S Disease ◽  
Differentially Expressed ◽  
Associated Proteins ◽  
Mycobacterium Avium Subsp Paratuberculosis

Bovine Johne's disease (paratuberculosis), caused by Mycobacterium avium subspecies paratuberculosis, poses a significant economic problem to the beef and dairy industry worldwide. Despite its relevance, however, pathogenesis of Johne's disease is still only partially resolved. Since mycobacterial membrane proteins expressed during infection are likely to play an important role in pathogenesis, membrane-enriched fractions, namely mucosa-derived membranes (MDM) and culture-derived membranes (CDM), of M. avium subsp. paratuberculosis from three cows with clinical paratuberculosis were investigated. An initial analysis by 2D difference gel electrophoresis (2D DIGE) and MALDI-TOF-MS analysis revealed four differentially expressed proteins with only one predicted membrane protein. Due to this limited outcome, membrane preparations were subjected to a tube–gel trypsin digestion and investigated by using nanoflow-liquid-chromatography-coupled tandem MS. Based on this approach a total of 212 proteins were detected in MDM including 32 proteins of bovine origin; 275 proteins were detected in CDM; 59 % of MDM and CDM proteins were predicted to be membrane-associated. A total of 130 of the proteins were detected in both MDM and CDM and 48 predicted membrane proteins were detected in MDM from at least two cows. Four of these proteins were not detected in CDM, implying differential expression in the host. All membrane-associated proteins, especially the four identified as being differentially expressed, might be relevant targets for further analyses into the pathogenesis of bovine paratuberculosis.

scholarly journals Rapid Expression of Mycobacterium avium subsp. paratuberculosis Recombinant Proteins for Antigen Discovery

2006 ◽  
Vol 14 (1) ◽  
pp. 102-105 ◽  
Author(s):  
Lingling Li ◽  
Shirin Munir ◽  
John P. Bannantine ◽  
Srinand Sreevatsan ◽  
Sagarika Kanjilal ◽  
...  
Keyword(s):  
Recombinant Proteins ◽  
Mycobacterium Avium ◽  
In Vitro Transcription ◽  
Johne's Disease ◽  
Johne’S Disease ◽  
Antigen Discovery ◽  
Diagnostic Assays ◽  
Mycobacterium Avium Subsp Paratuberculosis ◽  
Granulomatous Enteritis

ABSTRACT Mycobacterium avium subsp. paratuberculosis is the causative agent of Johne's disease, a chronic granulomatous enteritis of ruminants and other species. Detection of infection in animals is hampered by the lack of sensitive and specific diagnostic assays. We describe here an approach that utilizes translationally active PCR fragments for the rapid in vitro transcription and translation of recombinant proteins for antigen discovery in M. avium subsp. paratuberculosis. The investigations showed that the MAP1272c protein selectively reacts with sera from Johne's disease-positive cattle and represents an antigen of potential utility in M. avium subsp. paratuberculosis immunodiagnostics.

scholarly journals The collection of lymphatic fluid from the bovine udder and its use for the detection of Mycobacterium avium subsp. paratuberculosis in the cow

2011 ◽  
Vol 24 (1) ◽  
pp. 23-31 ◽  
Author(s):  
Johannes L. Khol ◽  
Pablo J. Pinedo ◽  
Claus D. Buergelt ◽  
Laura M. Neumann ◽  
Walter Baumgartner ◽  
...  
Keyword(s):  
Mycobacterium Avium ◽  
Fluid Collection ◽  
Johne's Disease ◽  
Johne’S Disease ◽  
Mycobacterium Avium Subsp ◽  
Enzyme Linked Immunosorbent Assay ◽  
Lymph Fluid ◽  
Mycobacterium Avium Subsp Paratuberculosis ◽  
Lymphatic Fluid ◽  
Elisa Result

The objective of the current study was to evaluate the feasibility of lymph collection from the bovine udder and to investigate if the lymphatic fluid might be of diagnostic value in cows infected with Mycobacterium avium subsp. paratuberculosis (MAP), the etiologic agent of paratuberculosis. Lymph fluid collection was attempted from 58 cows, and the reactions of the cows as well as the level of difficulty of the procedure were recorded in 56 animals. Lymph samples (51 in total) were tested for the presence of MAP by nested polymerase chain reaction. Collection of the lymphatic fluid caused no or mild signs of discomfort in 94.6% of the cows; in 51.8% of cows, lymphatic fluid was attained on the first attempt, while sample collection was unsuccessful in 12.1%. Mycobacterium avium subsp. paratuberculosis was detected in 43.1% of all lymph samples. The bacterium was present in 66.7% of cows with clinical Johne’s disease, in 42.8% of asymptomatic cows with a positive or suspicious enzyme-linked immunosorbent assay (ELISA) result in blood, and in 38.7% of cows with a negative ELISA result in blood. The present study shows that the procedure was well tolerated by most cows and can easily be performed on farm. The current report of the isolation of MAP from lymph fluid suggests that the present approach could be used for the early detection of Johne’s disease in cattle.

scholarly journals Assessing the Inactivation of Mycobacterium avium subsp.paratuberculosisduring Composting of Livestock Carcasses

2013 ◽  
Vol 79 (10) ◽  
pp. 3215-3224 ◽  
Author(s):  
Victoria L. Tkachuk ◽  
Denis O. Krause ◽  
Tim A. McAllister ◽  
Katherine E. Buckley ◽  
Tim Reuter ◽  
...  
Keyword(s):  
Mycobacterium Avium ◽  
Latency Period ◽  
Johne's Disease ◽  
Johne’S Disease ◽  
Detection Methods ◽  
Cattle Industry ◽  
Content Type ◽  
Plastic Bags ◽  
Long Latency

ABSTRACTMycobacterium aviumsubsp.paratuberculosiscauses Johne's disease (JD) in ruminants, with substantial economic impacts on the cattle industry. Johne's disease is known for its long latency period, and difficulties in diagnosis are due to insensitivities of current detection methods. Eradication is challenging asM. aviumsubsp.paratuberculosiscan survive for extended periods within the environment, resulting in new infections in naïve animals (W. Xu et al., J. Environ. Qual. 38:437-450, 2009). This study explored the use of a biosecure, static composting structure to inactivateM. aviumsubsp.paratuberculosis. Mycobacterium smegmatiswas also assessed as a surrogate forM. aviumsubsp.paratuberculosis. Two structures were constructed to hold three cattle carcasses each. Naturally infected tissues and ground beef inoculated with laboratory-culturedM. aviumsubsp.paratuberculosisandM. smegmatiswere placed in nylon and plastic bags to determine effects of temperature and compost environment on viability over 250 days. After removal, samples were cultured and growth of both organisms was assessed after 12 weeks. After 250 days,M. aviumsubsp.paratuberculosiswas still detectable by PCR, whileM. smegmatiswas not detected after 67 days of composting. Furthermore,M. aviumsubsp.paratuberculosisremained viable in both implanted nylon and plastic bags over the composting period. As the compost never reached a homogenous thermophilic (55 to 65°C) state throughout each structure, anin vitroexperiment was conducted to examine viability ofM. aviumsubsp.paratuberculosisafter exposure to 80°C for 90 days. Naturally infected lymph tissues were mixed with and without compost. After 90 days,M. aviumsubsp.paratuberculosisremained viable despite exposure to temperatures typically higher than that achieved in compost. In conclusion, it is unlikely composting can be used as a means of inactivatingM. aviumsubsp.paratuberculosisassociated with cattle mortalities.

scholarly journals Complete Genome Sequence of the Telford Type S Strain of Mycobacterium avium subsp. paratuberculosis

2019 ◽  
Vol 8 (11) ◽  
Author(s):  
Rudiger Brauning ◽  
Karren Plain ◽  
Milan Gautam ◽  
Tonia Russell ◽  
C. Carolina Correa ◽  
...  
Keyword(s):  
Genome Sequence ◽  
Mycobacterium Avium ◽  
Causative Agent ◽  
Complete Genome Sequence ◽  
Complete Genome ◽  
Johne's Disease ◽  
Johne’S Disease ◽  
Mycobacterium Avium Subsp ◽  
Content Type ◽  
Mycobacterium Avium Subsp Paratuberculosis

Mycobacterium avium subsp. paratuberculosis is the causative agent of Johne’s disease (JD).

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