Proceedings of the 10th International Colloquium on Paratuberculosis

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Bull TJ, McMinn EJ, Sidi-Boumedine K, Skull A, Durkin D, Neild P, Rhodes G, Pickup R, Hermon-Taylor J, 2003. Detection and verification <strong>of</strong> Mycobacterium avium subsp. paratuberculosis in fresh ileocolonic mucosal biopsy specimens from individuals with and without Crohn's disease. J Clin Microbiol, 41, 2915–2923. Chacon O, Bermudez L, Barletta R, 2004. Johne's disease, inflammatory bowel disease, and Mycobacterium paratuberculosis. Ann Rev Microbiol, 58, 329-363. Cheng J, Bull TJ, Dalton P, Cen S, Finlayson C, Hermon-Taylor J, 2005. Mycobacterium avium subspecies paratuberculosis in <strong>the</strong> inflamed gut tissues <strong>of</strong> patients with Crohn’s disease in China and its potential relationship to <strong>the</strong> consumption <strong>of</strong> cow’s milk: a preliminary study. World J Microbiol Biotechnol, 21, 1175-1179. Feller M, Huwiler K, Stephan R, Altpeter E, Shang A, Furrer H, Pfyffer G, Jemmi T, Baumgartner A, Egger M, 2007. Mycobacterium avium subspecies paratuberculosis and Crohn's disease: a systematic review and meta-analysis. Lancet Infect Dis, 7, 607– 613. Hermon-Taylor J, 2001. Protagonist. Mycobacterium avium subspecies paratuberculosis is a cause <strong>of</strong> Crohn’s disease. Gut, 49, 755-756. Quirke P, 2001. Mycobacterium avium subspecies paratuberculosis is a cause <strong>of</strong> Crohn's disease. Gut, 49: 757-760. Romero C, Hamdi A, Valentine JF, Naser SA, 2005. Evaluation <strong>of</strong> surgical tissue from patients with Crohn's disease for <strong>the</strong> presence <strong>of</strong> Mycobacterium avium subspecies paratuberculosis DNA by in situ hybridization and nested polymerase chain reaction, Inflamm Bowel Dis, 11, 116–125. Ryan P, Bennett MW, Aarons S, Lee G, Collins JK, O’Sullivan GC, O’Connell J, Shanahan F, 2002. PCR detection <strong>of</strong> Mycobacterium paratuberculosis in Crohn’s disease granulomas isolated by laser capture microdissection. Gut, 51, 665–670. Scanu AM, Bull TJ, Cannas S, Sanderson JD, Sechi LA, Dettori G, Zanetti S, Hermon-Taylor J, 2007. Mycobacterium avium subspecies paratuberculosis infection in cases <strong>of</strong> irritable bowel syndrome and comparison with Crohn's disease and Johne's disease: common neural and immune pathogenicities. J Clin Microbiol, 45, 3883-3890. Schwartz D, Shafran I, Romero C, Piromalli C, Biggerstaff J, Naser N, Chamberlin W, Naser SA, 2000. Use <strong>of</strong> short-term culture for identification <strong>of</strong> Mycobacterium avium subsp. paratuberculosis in tissue from Crohn's disease patients. Clin Microbiol Infect, 6, 303– 307. Shanahan F, 2002. Crohn’s disease. Lancet, 359, 62-69. 71
#130 Mycobacterium avium subsp. paratuberculosis detection in faeces and tissue samples by quantitative real time PCR and by cultivation Iva Slana, Marija Kaevska, Petr Kralik, Alena Kralova, Ivo Pavlik Veterinary Research Institute, Czech Republic Due to <strong>the</strong> fact that Mycobacterium avium subsp. paratuberculosis (MAP) is primarily an intestinal pathogen, <strong>the</strong> most common matrix that is used for MAP detection by cultivation is faeces (and after slaughter <strong>of</strong> animal is an intestinal mucosa). For culture, it is necessary to perform decontamination to eliminate <strong>the</strong> growth <strong>of</strong> o<strong>the</strong>r bacteria. Although MAP is quite tolerant to sample decontamination procedures, <strong>the</strong> sensitivity <strong>of</strong> culture is reduced. Methods based on PCR are suggested as an alternative tool for <strong>the</strong> diagnosis <strong>of</strong> paratuberculosis. We focused on <strong>the</strong> introduction and optimization <strong>of</strong> an effective and reliable method for <strong>the</strong> DNA isolation <strong>of</strong> MAP from faeces and from tissue samples. The developed techniques <strong>of</strong> <strong>the</strong> DNA isolation are based on a slightly modified commercialy available kit. Detection <strong>of</strong> isolated DNA is made by two independent quantitative real time PCR (qPCR) assays targeting two MAP specific elements: multicopy element IS900 and single copy element F57. Both qPCR assays incorporate an internal amplification control amplified with <strong>the</strong> same primers as <strong>the</strong> targets and <strong>the</strong> identical probe is used in both assays. Faecal and tissue samples <strong>of</strong> different animal species suffering from paratuberculosis were collected over a period <strong>of</strong> last 16 months and tested for <strong>the</strong> presence <strong>of</strong> MAP. More than 1691 faecal samples and 361 tissue samples were examined by qPCR and simultaneously by culture. Based on <strong>the</strong>se results we determine <strong>the</strong> dependence <strong>of</strong> concentration MAP DNA in faecal sample and <strong>the</strong> possibility to detect MAP DNA in tissue sample. 72
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Book of Abstracts
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Proceedings <stron
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Grant Support and Sponsorship <stro
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Scott Wells - Planning Committee Ch
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Monday, August 10, 2009 - All sessi
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Kari Røste Lybeck, Anne Kristine S
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1120-1140 #88: Influence of
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Concurrent Session B - Rm 175 Wille
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Thursday, August 13 Thursday, Augus
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Diagnostics and Genotyping Convenor
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Page 29 and 30: To better determine the</st
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Page 58 and 59: (b) IS900 Nested PCR, using p90-p91
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Page 100 and 101: Table 2. Paratuberculosis PCR resul
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#173 Local and systemic roles for b
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#23 Role of Mycoba
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Host Response and Immunology Poster
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#28 Intestinal MAP Infection via Pe
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#73 Age susceptibility of</
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#95 Role of nitric
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#134 Analysis of <
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#157 Study of <str
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#163 Immunohistochemical expression
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#193 Development of</strong
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#209 Murine macrophages carrying an
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#224 Application of</strong
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Table 1 UK and Cyprus MAP survey Re
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#228 Mycobacterium avium ssp. parat
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#145 MERKAL AWARD LECTURE Multinomi
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#50 Assessment of
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Table 1. Posterior median (CrIs) <s
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#55 Birth clusters of</stro
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from their dam, se
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#88 Influence of b
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#104 Relation between faecal shedde
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Epidemiology Poster Abstracts 109 1
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#22 The Prevalence of</stro
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CONCLUSION IS1311-based nested Ma/M
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#42 Seroprevalence survey o
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#77 Characterisation of</st
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Environmental samples were collecte
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REFERENCES 1) Chiodini RJ, Van Krui
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#113 Effect of soi
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#136 Error-adjusted, variance parti
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Latium Region Viterbo districts RES
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#144 The suitability of</st
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#178 Age structure of</stro
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A statistically significant lower f
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#183 Sero-prevalence of</st
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#218 Johne’s disease in t
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#244 A survey to estimate t
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Control Programs Keynote Lecture Re
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#202 Milk quality assurance for par
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#141 Control of pa
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#198 Use of small
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Control Programs Poster Abstracts 1
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RESULTS Loss of al
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#40 Managing Bovine Johnes Disease
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#74 Economic analysis of</s
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#87 Evaluation of
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separately for each existing herd <
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#98 An inter-laboratory ring-trial
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#120 Paratuberculosis vaccine inter
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The statement provides: • A stand
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COMPLEMENTARY ASPECTS On-Farm Disea
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#168 Paratuberculosis in Iran: past
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#180 Behavioural change in owners <
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the fact that <str
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#182 An integrated risk based appro
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etween 0 to 10, depending on <stron
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#203 Milk quality assurance for par
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#234 Johne’s disease vaccine: a c
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Pathogenomics and MAP Biology Conve
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#45 Identification of</stro
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#117 Adsorption of
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#222 Atypical structural features <
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Pathogenomics and MAP Biology Persp
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#52 Is ‘Indian Bison Type’ Myco
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#69 In vitro susceptibility <strong
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#118 Cloning, expression and purifi
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#176 A reference proteomic pr<stron
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#213 Construction the</stro
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#230 Iron concentration dependent s
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Abstract not available at press tim
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#174 Sharing of
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#177 Associations between CARD15 po
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Perspectives Talk: Public Health My
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A total of 206 qua
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Feller, M., K. Huwiler, R. Stephan,
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In 2008, the Ameri
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#115 Crohn’s disease and ruminant
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International John
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#76 Towards improved reporting stan
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#97 The EU ParaTBTools Project - Th
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#241 US Vaccine Initiative through
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Paratuberculosis ELISA Reliable, si
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PRIONICS AG WAGISTRASSE 27A PHONE +
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