OIE Reference Laboratory Reports s5

Paratuberculosis

OIE Reference Laboratory Reports

Activities in 2011

Name of disease (or topic) for which you are a designated OIE Reference Laboratory: / Paratuberculosis
Address of laboratory / Veterinary Research Institute
Hudcova 70
621 00 Brno
CZECH REPUBLIC
Tel.: / (+420) 5 3333 1601
Fax: / (+420) 5 4121 1229
e-mail address: /
website:
Name (including Title and Position) of Head of Laboratory (Responsible Official): / Professor Ivo Pavlik
Head of the Deparatment
Food and Feed Safety
Name (including Title and Position) of OIE Reference Expert: / Professor Ivo Pavlik
Head of the Deparatment
Food and Feed Safety
Name (including Title and Position) of writer of this report
(if different from above):

Part I: Summary of general activities related to the disease

1. Test(s) in use/or available for the specified disease/topic at your laboratory

Test (Total) / For / Specificity / Total
ELISA () / Antibodies / Paratuberculosis / 1949 blood samples from cattle
Culture HPC-sedimentation () / M. avium subsp. paratuberculosis / Paratuberculosis / 1278 cattle faeces
723 cattle tissues
377 wild ruminant faeces
31 wild ruminant tissues
123 environmental samples
Quantitative Real Time PCR (1996) / M. avium subsp. paratuberculosis / F57 and IS9001 / 226 tissue (8 human, 151 cattle, 65 food, 2 wild ruminants)
1418 faeces (1235 cattle, 131wild ruminants, 1 horse, 1jackal, 1 seal)
26 milk
13 milk filters
37 blood (13 cattle, 24 wild ruminants)
276 environment
PCR Quadruplex (39) / Isolates / Quadruplex system for M.avium species differentiation2 / M. a. paratuberculosis: 5laboratory isolates
16SrDNA PCR (145) / Isolates (84)
Tissue (46)
Environment (8)
Faeces (7) / 16S rRNA PCR system for Mycobacterium sp. and M.avium complex members identification2,3 / 58 M.avium sp.
4 M. intracellulare
60 other atypical mycobacteria4
23 non mycobacteria
IS900 RFLP (0) / Isolates / Standardized method5

2. 1 Specific fragments for M. avium subsp. paratuberculosis; home made technique: Slana, I., Kralik, P., Kralova, A., Pavlik, I.: On-farm spread of Mycobacterium avium subsp. paratuberculosis in raw milk studied by IS900 and F57 competitive real time quantitative PCR and culture examination. International Journal of Food Microbiology, 2008, 128 (2), 250-257.

3. 2 Home made technique for the detection and differentiation of members of the M. avium species: Moravkova, M., Hlozek, P., Beran, V., Pavlik, I., Preziuso, S., Cuteri, V., Bartos, M.: Strategy for the detection and differentiation of Mycobacterium avium species in isolates and heavily infected tissues. Research in Veterinary Science, 2008, 85 (2), 257-264.

4. 3 Wilton, S., Cousins, D.: Detection and identification of multiple mycobacterial pathogens by DNA amplification in a single tube. PCR Methods Applications, 1992, 1, 269-273.

5. 4 Non-identified mycobacterial species other than members of M. avium species, M. avium complex and M.tuberculosis complex.

6. 5 Pavlik, I., Horvathova, A., Dvorska, L., Bartl, J., Svastova, P., Du Maine, R., Rychlik, I.: Standardisation of Restriction fragment length polymorphism for Mycobacterium avium subspecies paratuberculosis. Journal of Microbiological Methods, 1999, 38 (1-2), 155-167.

2. Production and distribution of diagnostic reagents

None

Part II: Activities specifically related to the mandate
of OIE Reference Laboratories

3. International harmonisation and standardisation of methods for diagnostic testing or the production and testing of vaccines

a) Establishment and maintenance of a network with other OIE Reference Laboratories designated for the same pathogen or disease and organisation of regular inter-laboratory proficiency testing to ensure comparability of results

None

b) Organisation of inter-laboratory proficiency testing with laboratories other than OIE Reference Laboratories for the same pathogens and diseases to ensure equivalence of results

None

4. Preparation and supply of international reference standards for diagnostic tests or vaccines

None

5. Research and development of new procedures for diagnosis and control

Kralik, P., Slana, I., Kralova, A., Babak, V., Whitlock, R.H., Pavlik, I.: Development of a predictive model for detection of Mycobacterium avium subsp. paratuberculosis in faeces by quantitative real time PCR. Veterinary Microbiology, 2011, 149, 133-138. doi:10.1016/j.vetmic.2010.10.009

Pribylova, R., Slana, I., Kaevska, M., Lamka, J., Babak, V., Pavlik, I.: The evidence of Mycobacterium avium subsp. paratuberculosis in soil and plant tissues after their exposure to naturally contaminated faeces. Current Microbiology, 2011, 62, 1405-1410.

6. Collection, analysis and dissemination of epizootiological data relevant to international disease control

None

7. Maintenance of a system of quality assurance, biosafety and biosecurity relevant to the pathogen and the disease concerned

Consultations given to the experts in Uruguay, Serbia and Bosnia and Herzegovina.

8. Provision of consultant expertise to OIE or to OIE Member Countries

Under the terms of OIE Reference laboratory two laboratory trainings are provided:

http://www.vri.cz/en/departments/laboratory-oie_reference_laboratory_for_paratuberculosis

The quantification of Mycobacterium avium subsp. paratuberculosis, Mycobacterium avium subsp. avium and Mycobacterium avium subsp. hominissuis in different matrices using real-time qPCR (5 days)

Sequence analysis for identification of mycobacteria (4 days)

9. Provision of scientific and technical training to personnel from other OIE Member Countries

None

10. Provision of diagnostic testing facilities to other OIE Member Countries

None

11. Organisation of international scientific meetings on behalf of OIE or other international bodies

None

12. Participation in international scientific collaborative studies

§ National grants

Animal disease research, prevention and food chain protection, part: Mycobacterial infections of animals. Institutional Research Plan, 1.1.2009-31.12.2013. Ministry of Agriculture, the Czech Republic, No.: MZE 0002716202, Project leader: Prof. MVDr. Jiri Rubes, CSc., VRI Brno, Czech Republic.

Paratuberculosis in the Czech Republic: the study of spread in the ruminants, of the risk of the contamination of the food of animal origin and development of diagnostic methods for rapid eradication and certification program. 1.1.2008-31.12.2012. Ministry of Agriculture, the Czech Republic, No.: MZE QH81065. Project leader: Prof. MVDr. Ivo Pavlik, CSc., VRI Brno, Czech Republic.

Methods of eradication of production diseases of cattle - bovine viral diarrhoea - mucosal disease and paratuberculosis. Research in Agricultural Complex, Ministry of Agriculture, Czech Republic, Acronym: Metlum. Project leader: MVDr. Kamil Kovarcik, PhD., VRI Brno, Czech Republic, 1.1.2010-31.12.2014.

§ European Union Grants:

Center for Advanced Microbiology & Immunology Research in Veterinary Medicine. Acronym: AdmireVet. Registration No. CZ.1.05/2.1.00/01.0006. Regional R&D Centre. Ministry of Education, Youth and sports of the Czech Republic. 1.12.2009-31.12.2013. Co-ordinator: Prof. MVDr. Ivo Pavlík, CSc., VRI Brno, Czech Republic.

13. Publication and dissemination of information relevant to the work of OIE (including list of scientific publications, internet publishing activities, presentations at international conferences)

¡ Presentations at international conferences and meetings

Kaevska, M., Pribylova, R., Slana, I., Kralik, P., Pavlik, I.: Spread of potentially pathogenic mycobacteria in the environment: possible routes of transmission through soil and plants. EPS Hohhot International Natural Sciences Development Forum 2011, August 3-6 Huhhot, China, 64.

Klanicova, B., Seda, J., Slana, I., Pavlik, I.: Quantitative real time PCR and cultivation method for detection of Mycobacterium avium subspecies in sediments of drinking water reservoirs. Annual Congress of the European Society of Mycobacteriology, 26th – 29th June, 2011, Lubeck, Germany, 81.

Kralik, P., Slana, I., Kralova, A., Babak, V., Whitlock, R.H., Pavlik, I.: Cost-efficient detection of Mycobacterium avium subsp. paratuberculosis in faeces by quantitative real time PCR and development of a predictive model for fighting the disease. In: qPCR 2011 Event - 5th international qPCR Symposium & Industrial Exhibition & Application Workshops, 28th March - 1st April 2011, Freising, Germany, 25-26.

Pavlik, I.: Emerging and re-emerging potentially pathogenic mycobacteria – current situation, ecology and impact on animal’s and human’s health. 1st Annual Symposium of Bacteriology and Infection (SBI-2011), July 30th –August 1st, 2011, Peking, China, 143.

Pavlik, I.: Various stages in the life cycle of syrphid flies (Eristalis tenax; Diptera: Syrphidae) and other Diptera as potential mechanical vectors of potentially pathogenic mycobacteria causing infections in pigs and cattle. EPS Hohhot International Natural Sciences Development Forum 2011, August 3-6 Huhhot, China, 58.

¡ Scientific publications in peer-reviewed journals

Hruska, K., Slana, I., Kralik, P., Pavlik, I.: Mycobacterium avium subsp. paratuberculosis in powdered infant milk: F57 competitive real time PCR. Veterinarni Medicina, 2011, 56 (5), 226-230. http://www.vri.cz/docs/vetmed/56-5-226.pdf

Klanicova, B., Vondruskova, H., Slana, I., Kaevska, M., Pavlik, I.: Real-time quantitative PCR detection of Mycobacterium avium subspecies in meat products. Journal of Food Protection, 2011, 74 (4), 636-640.

Pribylova, R., Kralik, P., Donnelly, N., Matiasovic, J., Pavlik, I.: Mycobacterium avium subsp. paratuberculosis and the expression of selected virulence and pathogenesis genes in response to 65 degreee C and pH 2.0. Brazilian Journal of Microbiology, 2011, 42, 807-817.

Pribylova, R., Lamka, J., Kopecna, M., Trcka, I., Moravkova, M., Pavlik, I. Mycobacterial screening of Czech red deer (Cervus Elaphus) populations in overwintering sites, 2004-2006. The Journal of Wildlife Diseases, 2011, 47 (3), 780-783.

Pribylova, R., Slana, I., Kralik, P., Kralova, A., Babak, V., Pavlik, I.: Correlation of Mycobacterium avium subsp. paratuberculosis counts in gastrointestinal tract, muscles of the diaphragm and the masseter of dairy cattle and potential risk for consumers. International Journal of Food Microbiology, 2011, 151, 314-318.

Slana, I., Pribylova, R., Kralova, A., Pavlik, I.: Persistence of Mycobacterium avium subsp. paratuberculosis at a farm-scale biogas plant supplied with manure from paratuberculosis-affected dairy cattle. Applied and Environmetal Microbiology, 2011, 77 (9), 3115-3119.

¡ Other communications

Knowledge dissemination is an important activity of the OIE Reference Laboratories for Paratuberculosis and Avian Tuberculosis, supported by the Veterinary Research Institute, Brno, Czech Republic and Biomedical Technology, Epidemiology and Food Safety Global Network. Databases on paratuberculosis and Crohn’s disease digest were updated in 2011 with a total of 360 new scientific articles (228 and 132, respectively), published in peer reviewed journals and indexed in the Web of Science® (Thomson Reuters) database. Members of the Biomedical Technology, Epidemiology and Food Safety Global Network were informed about new publications by e-mails in weekly intervals. Both databases are available at the web page of the OIE Reference Laboratory for Paratuberculosis, hosted by the Veterinary Research Institute, Brno. Network members from 22 countries (Argentina, Australia, Canada, Czech Republic, Denmark, Egypt, France, Germany, India, Ireland, Italy, Nigeria, Portugal, Saudi Arabia, Slovakia, Slovenia, Spain, Switzerland, The Netherlands, Turkey, UK and USA) requested papers from those on offer. In total, 42 new items of information on mycobacteria were distributed by e-mail to network members as the Centaur Global Network Information and were simultaneously archived at the network web page. This open access system means that all information is available not only for the Centaur Global Network members. Everybody can register as a person, interested in any or all of the 27 fields of interest, one of which is (08) Mycobacterial diseases. In total, 210 e-mail notifications were dispatched in 2011, mostly to distribution lists (27) Scientific information, (23) Education, (05) Zoonoses, (04) Food borne diseases and (08) Mycobacterial diseases. The distance learning course on Good Research Practice was run also in 2011 and the course assigned GRP 2012 has been opened in October 2011 and has to be finished by 31 March 2012. The running course has registered 88articipants from 18 countries (Australia, Bangladesh, Croatia, Czech Republic, Germany, Ethiopia, Finland, India, Japan, Kenya, Nigeria, Republic of Korea, Serbia, South Africa, Spain, Sudan, Sweden and Tunisia). The previous course, GRP 2011, has been running from October 2010 to 31 March 2011. This course had 105 participants from 27 countries (Australia, Bangladesh, Brazil, Cameroon, Czech Republic, Ethiopia, Ghana, India, Iran, Iraq/Kurdistan, Japan, Kenya, Korea, Macedonia, Malaysia, Mongolia, Nepal, Nigeria, Pakistan, Peru, Qatar, Serbia, South Africa, Spain, Sweden, United Kingdom, Zambia).

URL addresses of hyperlinks:
OIE Reference Laboratory for Paratuberculosis,

http://www.vri.cz/en/departments/laboratory-oie_reference_laboratory_for_paratuberculosis

Veterinary Research Institute, Brno

http://www.vri.cz

Biomedical Technology, Epidemiology and Food Safety Global Network (CENTAUR)

http://centaur.vri.cz
registration on line

http://centaur.vri.cz/?page=registration

Database on paratuberculosis

http://www.vri.cz/en/publications/paratuberculosis

Database on Crohn’s disease digest

http://www.vri.cz/en/publications/cd_and_map

OIE Reference Laboratory for Paratuberculosis,

http://www.vri.cz/en/departments/laboratory-oie_reference_laboratory_for_paratuberculosis

distance learning course on Good Research Practice

http://centaur.vri.cz/?page=grp_e-course

GRP 2011 Report

http://centaur.vri.cz/docs/GRP2011/BULLETIN.pdf

Centaur Global Network Information

http://centaur.vri.cz/?page=cgn_information

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Annual reports of OIE Reference Centres, 2011 3