OIE Reference Laboratory Reports
Activities in 2011
Name of disease (or topic) for
which you are a designated OIE
Reference Laboratory:
Address of laboratory
Paratuberculosis
Veterinary Research Institute
Hudcova 70
621 00 Brno
CZECH REPUBLIC
Tel.:
(+420) 5 3333 1601
Fax:
(+420) 5 4121 1229
e-mail address:
pavlik@vri.cz
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):
Annual reports of OIE Reference Centres, 2011
1
Paratuberculosis
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
ELISA ()
Antibodies
Paratuberculosis
1949 blood samples from cattle
Paratuberculosis
1278 cattle faeces
723 cattle tissues
377 wild ruminant faeces
31 wild ruminant tissues
123 environmental samples
M. avium subsp.
paratuberculosis
F57 and IS9001
226 tissue (8 human, 151 cattle,
65 food, 2 wild ruminants)
1418 faeces (1235 cattle,
131 wild ruminants, 1 horse,
1 jackal, 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
16S rDNA PCR (145)
Isolates (84)
Tissue (46)
Environment (8)
Faeces (7)
16S rRNA PCR system
for Mycobacterium sp.
and M. avium complex
members identification2,3
Isolates
Standardized method5
Culture HPC-sedimentation
()
Quantitative Real Time PCR
(1996)
IS900 RFLP (0)
2.
3.
4.
5.
6.
2.
M. avium subsp.
paratuberculosis
Total
M. a. paratuberculosis:
5 laboratory isolates
58 M. avium sp.
4 M. intracellulare
60 other atypical mycobacteria4
23 non mycobacteria
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.
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.
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.
4
Non-identified mycobacterial species other than members of M. avium species, M. avium complex and
M. tuberculosis complex.
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.
Production and distribution of diagnostic reagents
None
2
Annual reports of OIE Reference Centres, 2011
Paratuberculosis
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)
Annual reports of OIE Reference Centres, 2011
3
Paratuberculosis
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.
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Annual reports of OIE Reference Centres, 2011
Paratuberculosis
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.
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., 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.
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.
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 email 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
Annual reports of OIE Reference Centres, 2011
5
Paratuberculosis
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