Bovine tuberculosis
OIE Reference Laboratory Reports
Activities in 2010
Name of disease (or topic) for which you are a designated OIE Reference Laboratory: / Bovine tuberculosisAddress of laboratory: / Agence nationale de sécurité sanitaire de l’alimentation, de l’environnement et du travail (ANSES)/
French Agency for Food, Environmental &
Occupational Health Safety
Laboratoire de santé animale de Maisons-Alfort /
Maisons-Alfort Animal Health Laboratory
Unité Zoonoses Bactériennes /
Bacterial Zoonoses Unit
23, avenue du Général-de-Gaulle
94706 Maisons-Alfort Cedex
France
Tel.: / (+33[0]1) 49.77.13.00
Fax: / (+33[0]1) 49.77.13.44
e-mail address: /
website: / www.anses.fr
Name of Head of Laboratory (Responsible Official): / Dr Pascal Boireau
Name of OIE Reference Expert: / Dr María Laura Boschiroli
Name 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 at your laboratory
Test /M.tuberculosis complex
/ M. avium-intracellulare / Other non-tuberculous mycobacteria / Other samples (negative or contaminated) / Ongoing tests / TotalIsolation and strain identification / M. tuberculosis: 1 / 25 / 6 / 77 / 21 / 130
Strain Identification / M.bovis: 278
M. tuberculosis: 1 / 61 / 142 / 96 / 3 / 581
PCR diagnosis / M.bovis: 15
M.microti: 8
M. tuberculosis: 1
M. tuberculosis complex : 52 / 23 / 53 / 94 / - / 246
Tuberculin control: bovine PPD / 1 / 8
Spoligotyping / 270 / - / - / - / - / 270
MLVA (8 loci) / 500 / - / - / - / - / 500
Agent identification:
Isolation: samples are homogenised and afterwards decontaminated with either 4% NaOH or 4% H2SO4. Egg-based media are used for inoculation (Lowenstein Jensen + Coletsos). These samples were issued from different animal species: farm animals (cattle, goats, pigs, alpaga), pets (cats, dogs, reptiles), wild life (roe deer, red deer, wild boar, badgers, foxes), wild animals in captivity (sea lions, crocodiles, antelopes, fallow deer, kangaroos), ornamental fish, wild, farm and ornamental birds, and environmental samples (earth and water).
Identification: biochemical and cultural characterisation, as well as PCR-based identification are performed. Molecular typing: it is performed by Spoligotyping for all the M tuberculosis complex species and Multiple Locus Variable number tandem repeat Analysis (MLVA) on ETR-A/D, QUB 11a, QUB 11b, QUB 3232, QUB 26) for M. bovis.
2. Production and distribution of diagnostic reagents
Type of reagent / Amount supplied nationally(including for own use) / Amount supplied to other countries
Guinea pig’s Immunisation emulsion (M. bovis AN5) / - / 20 ml
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
Nil
4. Preparation and supply of international reference standards for diagnostic tests or vaccines
Nil
5. Research and development of new procedures for diagnosis and control
Bovine Tuberculosis (BTB) in Wildlife: epidemiological follow up of the first wildlife outbreak in France: BTB infection in wild ungulates (red deer and wild boar) was discovered in 2001 in the Brotonne forest, Normandy. Consequently we have been investigating the infection evolution in free ranging animal populations (red deer, wild boars, roe deer, red foxes and badgers) by studying both bacterial harbouring (bacterial culture on organs presenting or not TB like lesions) but also the disease progress (anatomo-pathological analysis) on these animal populations ever since 2001. The results of the hunting season 2005-2006 demonstrated that control measures originally applied in 2002 (fencing, ban of game artificial feeding, game-carcass removal, etc.) alone were not efficient enough to control dissemination within wildlife and from wild to domestic animals. Due to the severity of the lesions disclosed by deer and their consequent potential role as a reservoir of M.bovis in the forest, it was decided to cull the totality of its population (aprox. 150 animals, hunting season 2006-2007). As for wild boar, eradication of this species is unfeasible due their over-sized population. However, 310 hundred animals and 199 in 2006-2007 and 2007-2008 respectively were analysed (bacterial culture on organs presenting or not TB like lesions, anatomo-pathological analysis) for assessing the infection progression and the possible role of this species as a dead end, a spill over or a real reservoir host not only intra-species but also for other species including cattle. Compared to 2005-2006, when prevalence figures (bacteriology) were of 37% (n= 155), in 2006-2007 we observed that it was lowered to 31% (n = 255), in 2007-2008 to 19 % (n = 199) in 2008-2009 to 11 % (n= 200) and in 2009-2010 only one animal was found infected out of 162. The reduction of the total number of infected animals could be mainly explained as deer, considered the primary source of M.bovis transmission for wild boar, has almost been eradicated from the forest. Furthermore, as the wild boar population has been drastically been reduced due to over-hunting, inter-species transmission might probably have also been reduced.
BTB Epidemiology in France and introduction of new diagnostic tools in the field: the epidemiological situation and the pertinence of regulation texts for controlling bovine TB in France is being evaluated with regards to the entire country context but also in particular regions where BTB rates are still high. The implementation of new tools like PCR or INF-g (BovigamÒ) is now achieved and currently used mainly in the high prevalence region in the contry. This analysis is performed in collaboration with the epidemiology team of the Contagious Infection Unit, Maisons-Alfort Veterinary College, and a working committee composed of members of the Agriculture Ministry, regional veterinary laboratories, veterinary services and local and national farmers´ organisations.
Molecular characterisation of French M. bovis strains by spoligotyping and VNTR for epidemiological follow-up and application in surveillance programmes for bovine tuberculosis: Employed both for cattle and wildlife M.bovis strains giving rise to TB outbreaks, the combination of both molecular typing methods allows us a perfect strain differentiation leading us to a tight epidemiological transmission follow up. We have pursued and enlarged our strain-collection characterisation in order to explain the origin of various outbreaks throughout the country. At present, animal strains are being compared with human strains so as to assess the potential risk of zoonotic transmission of M.bovis in France.
6. Collection, analysis and dissemination of epizootiological data relevant to international disease control
Report to the 2009 OIE questionnaire on wildlife diseases
2009 Annual Report on Zoonoses to the European Commission
7. Provision of consultant expertise to OIE or to OIE Member Countries
Nil
8. Provision of scientific and technical training to personnel from other OIE Member Countries
Boschiroli M L. (2010). Molecular Epidemiology of Mycobacterium bovis: exploiting molecular data. In: Molecular Tools and Epidemiology of Tuberculosis Course (2009-2010), Institut Pasteur, Paris, France, 2010/06/04.
Dr Bernardo Alonso, head of the Bovine tuberculosis and paratuberculosis OIE Reference Laboratory in Buenos Aires, Argentina (june 2010). Molecular diagnosis of bovine tuberculosis and molecular typing by spoligotyping of Mycobacterium bovis trainee. Laboratoire de Santé Animal-Anses, Maisons-Alfort, France, 2010/06/14-18
9. Provision of diagnostic testing facilities to other OIE Member Countries
Nil.
10. Organisation of international scientific meetings on behalf of OIE or other international bodies
Nil.
11. Participation in international scientific collaborative studies
Participation in the collaborative study launched by Noel Smith (VLA, UK) and Robin Skuce (AFBINI, UK) in order to identify particular genetic signatures and characterise M.bovis cattle strains from different regions of the world.
12. 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
1. Hars J, Zanella G, Richomme C, Garin-Bastuji B & Boschiroli M L. Bovine tuberculosis in wildlife in France: an update. 9th EWDA conference, Vlieland, Netherland, 2010/09/13-16.
2. Moyen J L, Faye S, Brugère L, Grewis L, Boschiroli M L & Sellal E. Sensitivity of real time PCR test for Bovine tuberculosis diagnosis. The European Association of Veterinary Laboratory Diagnosticians, First meeting, Lelystad, Netherland, 2010/09/15-17.
- Keck N, Moyen J L, Gueneau E & Boschiroli M L. Ré-emergence de la tuberculose bovine en France: des situations contrastées. Journées Scientifiques de l'Association française des Directeurs et cadres des Laboratoires Vétérinaires Publics d'Analyses, Bordeaux, France, 2010/10/14-15.
- Boschiroli M L, Gueneau E, Keck N & Moyen J L. Nouveaux outils pour le dépistage et le diagnostique de la tuberculose bovine: avantages et limites 25ème Journée Technique du GTV Bourgogne, Autun, France, 2010/10/20.
- Boschiroli M L. Actualité sur les fondamentaux de la tuberculose 25ème Journée Technique du GTV Bourgogne, Autun, France, 2010/10/20.
- Boschiroli M L & Garin-Bastuji B. (2010). Outils disponibles pour le diagnostic de la tuberculose bovine Réunion de travail Tuberculose Bovine. Nouveau plan d'action, DGAl, Pari, 2010/11/03.
¡ Scientific publications in peer-reviewed journals
1. Richomme C, Boschiroli M L, Hars J,Casabianca F & Ducrot C. (2010). Bovine tuberculosis in livestock and wild boar on the Mediterranean island, Corsica. Journal of Wildlife Diseases, 46, 627-31.
2. Keck N, Dutruel H, Smyej F, Nodet M & Boschiroli M L. (2010). Tuberculosis due to Mycobacterium bovis in a Camargue horse. Veterinary Record, 166, 499-500.
3. Panteix G, Gutierrez M C, Boschiroli M L, Rouviere M, Plaidy A, Pressac D, Porcheret H, Chyderiotis G, Ponsada M, Van Oortegem K, Salloum S, Cabuzel S, Banuls A L, Van de Perre P & Godreuil S. (2010). Pulmonary tuberculosis due to Mycobacterium microti: a study of six recent cases in France. J Med Microbiology, 59, 984-9.
4. Godreuil S, Jeziorski E, Banuls A L, Fraisse T, Van de Perre P & Boschiroli M L. (2010). Intrafamilial cluster of pulmonary tuberculosis due to Mycobacterium bovis of the African 1 clonal complex. Journal of Clinical Microbiology, 48, 4680-3.
5. Zhang J, Abadia E, Refregier G, Tafaj S, Boschiroli M L, Guillard B, Andremont A, Ruimy R & Sola C. (2010). Mycobacterium tuberculosis complex CRISPR genotyping: improving efficiency, throughput and discriminative power of 'spoligotyping' with new spacers and a microbead-based hybridization assay. J Med Microbiology, 59, 285-94.
6. Hars J, Richomme C & Boschiroli M L. (2010). La tuberculose bovine dans la faune sauvage en France / Bovine tuberculosis in wildlife in France. Bull. Epidémiol. Santé Anim. Alim, 38 28-31.
7. Hars J, Richomme C & Boschiroli M L. (2010). La tuberculose bovine dans la faune sauvage en France / Bovine tuberculosis in wildlife in France. BEH hors-série.
8. Fediaevsky A, Dufour B, Boschiroli M L & Moutou F. (2010). Bilan de la surveillance de la tuberculose bovine en 2009 :une prévalence globalement faible mais un renforcement de la lutte dans certaines zones. Bull. Epidémiol. Santé Anim. Alim, 40, 3-8.
¡ Other communications
o Consultancy and expertise reports
1. Benet J J,Dufour B,Hars J,Garin-Bastuji B & Boschiroli M L. Rapport d'Expertise Tuberculose Bovine - Côte d'Or (DGAL confidentiel) 2010/10.
2. Benet J J,Dufour B,Hars J,Garin-Bastuji B & Boschiroli M L. Rapport d'Expertise Tuberculose Bovine - Dordogne (DGAL confidentiel) 2010/10.
3. Hars J,Sigaud M,Payne A,Tosi J C,Rolland B,Rambaud T,Game Y,Garin-Bastuji B & Boschiroli M L. Rapport final de l’enquête menée durant la saison de chasse 2009-2010, Programme de Surveillance de la Tuberculose chez les animaux sauvages de la Foret de Brotonne (Seine-Maritime et Eure). Convention MAAP/ONCFS, 30. 2010/12.
o PhD Thesis:
Faye S (2010). Évaluation de nouveaux outils de diagnostic de la tuberculose bovine: conditions d’utilisation d’un test de dosage d’INF-gamma et d’un test PCR IS6110 en temps réel (Soutenance 2010/10/06).
13. Inscription of diagnostic kits on the OIE Register
i) Did you participate in expert panels for the validation of candidate kits for inscription on the OIE Register? If yes, for which kits?
Nil
ii) Did you submit to the OIE candidate kits for inscription on the OIE Register? If yes, for which kits?
Nil
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This laboratory is accredited by the Laboratories Section of the French Committee for Accreditation (COFRAC) in compliance with the ISO/IEC 17025 standard and COFRAC rules of application under the N° 1-1799 for test activities
Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010 5