Proceedings of the 10th International Colloquium on Paratuberculosis

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In <strong>the</strong> presence <strong>of</strong> super-shedders in herds, 57.1, 16.7, 20.0, and 70.0% <strong>of</strong> animals o<strong>the</strong>r than identified as super-shedders shed <strong>the</strong> same strain as that <strong>of</strong> contemporary super-shedders on farm A. Although this scenario did not occur on farm B, 100% <strong>of</strong> culture-positive animals on farm C o<strong>the</strong>r than identified as super-shedders shed <strong>the</strong> same strain as that <strong>of</strong> contemporary super-shedders during first, third, and fourth visits to <strong>the</strong> farm. For <strong>the</strong> available tissue data, tissues from about 82% <strong>of</strong> cows o<strong>the</strong>r than super-shedders were culture-positive for MAP, indicating a true infection. For low shedders in three herds, about 65% <strong>of</strong> animals for fecals and 56% for fecal and tissues, shared only <strong>the</strong> same strain as super-shedders. On farm C, animals 491, 493, and 495 were purchased from o<strong>the</strong>r sources; animal 491 was truly infected, which may be a case <strong>of</strong> adult infection. In few occasions we found multiple strains in different samples during <strong>the</strong>ir life time in some animals. It was indicated that while it is reasonable to speculate that coinfection with multiple genotypes is possible, it was recognized that this may be rare event (Harris et al., 2006). To conclude, taken toge<strong>the</strong>r <strong>the</strong> results <strong>of</strong> MAP strains and observed shedding levels, at least 50% <strong>of</strong> low shedders have <strong>the</strong> same strain as that <strong>of</strong> contemporary super-shedders. The results <strong>of</strong> this study indicate that very few cows had characteristics <strong>of</strong> a possible pass-through animal; many more cows were truly infected. Sharing <strong>of</strong> <strong>the</strong> same strain <strong>of</strong> low shedders with <strong>the</strong> contemporary super-shedders suggests that low shedders may be infected as adults by <strong>the</strong> super-shedders. The use <strong>of</strong> SSR strain-typing combined with observed shedding levels provided a unique opportunity to get a better insight into herd infection dynamics <strong>of</strong> MAP. ACKNOWLEDGEMENTS This project was supported in part by <strong>the</strong> USDA-Agricultural Research Service (Agreements. 58-1265-3-155, 58-1265-3-156, 58-1265-3-158, 58-1265-4-020, and 58-1265-8-064) for <strong>the</strong> Regional Dairy Quality Management Alliance (RDQMA) and <strong>the</strong> Johne’s Disease Integrated Program (JDIP, USDA contract 45105). We acknowledge JDIP for providing scholarship award (USDA-CSREES-NRA Award No 2008-55620-18710) to attend and present this study at 10 th <strong>International</strong> <strong>Colloquium</strong> on Paratuberculosis (ICP) meeting at Minneapolis, MN, USA. REFERENCES Amonsin A, Li LL, Zhang Q, Bannantine JP, Motiwala AS, Sreevatsan S, Kapur V, 2004. Multilocus short sequence repeat sequencing approach for differentiating among Mycobacterium avium subsp. paratuberculosis strains. J. Clin. Microbiol, 42, 1694–1702. Harris NB, Payeur JB, Kapur V, Sreevatsan S, 2006. Short-sequence-repeat analysis <strong>of</strong> Mycobacterium avium subsp. paratuberculosis and Mycobacterium avium subsp. avium isolates collected from animals throughout <strong>the</strong> United States reveals both stability <strong>of</strong> loci and extensive diversity. J. Clin. Microbiol, 42, 2970–2973. Möbius P, Luyven G, Hotzel H, Köhler H, 2008. High genetic diversity among Mycobacterium avium subsp. paratuberculosis strains from German cattle herds shown by combination <strong>of</strong> IS900 restriction fragment length polymorphism analysis and mycobacterial interspersed repetitive unit-variable-number tandem-repeat typing. J. Clin. Microbiol, 46, 972-981. Pradhan AK, Van Kessel JS, Karns JS, Wolfgang DR, Hovingh E, Nelen KA, Smith JM, Whitlock RH, Fyock T, Ladely S, Fedorka-Cray PJ, Schukken YH, 2009. Dynamics <strong>of</strong> endemic infectious diseases <strong>of</strong> animal and human importance on three dairy herds in <strong>the</strong> nor<strong>the</strong>astern United States. J. Dairy Sci, 92, 1811–1825. Whitlock RH, Sweeney RW, Fyock TL, Smith J, 2005. MAP super-shedders: ano<strong>the</strong>r factor in <strong>the</strong> control <strong>of</strong> Johne’s disease. <strong>Proceedings</strong> <strong>of</strong> <strong>the</strong> 8 th <strong>International</strong> <strong>Colloquium</strong> <strong>of</strong> Paratuberculosis, Abstr, p164. 33
#105 MIRU-VNTR genotyping <strong>of</strong> Mycobacterium avium subspecies paratuberculosis revealed Typespecific differences at two component regulatory systems Elena Castellanos, Beatriz Romero, Sabrina Rodriguez, Javier Bezos, Lucia de Juan, Ana Mateos, Lucas Dominguez, Alicia Aranaz Animal Health Surveillance Center (VISAVET). Complutense University <strong>of</strong> Madrid (UCM). Animal Health Department. Veterinary School. UCM, Spain In this report, we genotyped a collection <strong>of</strong> Spanish Mycobacterium avium subspecies paratuberculosis isolates with a combination <strong>of</strong> six mycobacterial interspersed repetitive units (MIRU) and variable number tandem repeats (VNTR) loci; MIRU-2, MIRU-3 (Bull et al., 2003), VNTR-25, VNTR-32, VNTR-292 (Thibault et al., 2007) and VNTR-259 (this study). This research forms part <strong>of</strong> <strong>the</strong> EU project ParaTBtools. For this purpose we screened 70 M. a. paratuberculosis Type II and III isolates, obtained from domestic livestock (goats and cattle) and two isolates recovered from wild animal species (fallow deer- Dama dama- and mouflon- Ovis orientalis musimon) throughout 22 different Spanish localities in a nine-year period. The selected combination <strong>of</strong> MIRU-VNTR loci differentiated 17 different allelic variants within this population <strong>of</strong> M. a. paratuberculosis isolates, with a resulting discriminatory index (D) <strong>of</strong> 0.82. An interesting feature was <strong>the</strong> M. a. paratuberculosis Type-specific differences observed at <strong>the</strong> number <strong>of</strong> repeats at MIRU-3, located between genes SenX3 and RegX3, components <strong>of</strong> a regulatory system with a role in <strong>the</strong> virulence <strong>of</strong> Mycobacterium tuberculosis (Magdalena et al., 1998). In addition, M. a. paratuberculosis Type-specific number <strong>of</strong> tandem repeats was also obtained for <strong>the</strong> novel locus VNTR-259, which as in <strong>the</strong> case <strong>of</strong> MIRU-3, was located between a two component regulatory system, AfsQ1/AfsQ2, which has a role in <strong>the</strong> regulation <strong>of</strong> <strong>the</strong> antibiotic production strategies in Streptomyces (Shu et al., 2009). References: Bull TJ, Sidi-Boumedine K, McMinn EJ, Stevenson K, Pickup R, Hermon-Taylor J, 2003. Mycobacterial interspersed repetitive units (MIRU) differentiate Mycobacterium avium subspecies paratuberculosis from o<strong>the</strong>r species <strong>of</strong> <strong>the</strong> Mycobacterium avium complex. Mol. Cell Probes 17, 157-164. Magdalena J, Supply P, Locht C, 1998. Specific differentiation between Mycobacterium bovis BCG and virulent strains <strong>of</strong> <strong>the</strong> Mycobacterium tuberculosis complex. J. Clin. Microbiol. 36, 2471-2476. Shu D, Chen L, Wang W, Yu Z, Ren C, Zhang W, Yang S, Lu Y, Jiang W, 2009. AfsQ1-Q2-sigQ is a pleiotropic but conditionally required signal transduction system for both secondary metabolism and morphological development in Streptomyces coelicolor. Appl. Microbiol. Biotechnol. 81, 1149-1160. Thibault VC, Grayon M, Boschiroli ML, Hubbans C, Overduin P, Stevenson K, Gutierrez MC, Supply P, Biet F, 2007. New variable-number tandem-repeat markers for typing Mycobacterium avium subsp. paratuberculosis and M. avium strains: comparison with IS900 and IS1245 restriction fragment length polymorphism typing. J. Clin. Microbiol. 45, 2404-2410. 34
Page 1 and 2: Book of Abstracts
Page 4 and 5: Proceedings <stron
Page 6 and 7: Grant Support and Sponsorship <stro
Page 8 and 9: Scott Wells - Planning Committee Ch
Page 10 and 11: Monday, August 10, 2009 - All sessi
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Page 18 and 19: Thursday, August 13 Thursday, Augus
Page 20: Diagnostics and Genotyping Convenor
Page 23 and 24: #229 Detecting Johne’s Disease he
Page 25 and 26: Kalis CHJ, Hesselink JW, Barkema HW
Page 27 and 28: MATERIALS AND METHODS Sampling For
Page 29 and 30: To better determine the</st
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Page 33: Fig. 1. Dendogram (showing genetic
Page 37: Diagnostics and Genotyping Poster A
Page 40 and 41: Missouri). Serum from a cow with do
Page 42 and 43: #7 Significance of
Page 44 and 45: RESULTS Of the 327
Page 46 and 47: Direct fecal nested Map PCR testing
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Page 58 and 59: (b) IS900 Nested PCR, using p90-p91
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Page 64 and 65: indexes, compared to each single an
Page 66 and 67: #108 Evaluation of
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Page 76 and 77: could be partly attributed to a sma
Page 78 and 79: #146 Analysis of v
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#191 Potentiating day-old blood sam
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samples from the s
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(MAA) (MAP87 and MAP3776) and 1 fro
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#196 Inter- and intra-subtype genot
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RESULTS In 601 (29%) of</st
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#211 Culture of my
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#242 Application and field validati
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#251 Diagnosis of
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Table 2. Paratuberculosis PCR resul
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#254 Programmes on Paratuberculosis
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Fig. 1. European countries specifyi
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A bulk milk quality assurance progr
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Republic of Lithua
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#276 Elimination of</strong
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other projects and
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Woodbine KA, Schukken YH, Green LE,
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Host Response and Immunology Keynot
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#33 MERKAL AWARD LECTURE No interfe
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#75 Interleukin 17 and interleukin
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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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247
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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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273
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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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