Bovine tuberculosis
OIE Reference Laboratory Reports
Activities in 2011
Name of disease (or topic) for which you are a designated OIE Reference Laboratory: / Bovine tuberculosisAddress of laboratory: / Animal Health & Veterinary Laboratories Agency Weybridge
Woodham Lane,
New Haw,
Addlestone,
Surrey
KT15 3NB
UNITED KINGDOM
Tel.: / (44.1932) 34 11 11
Fax: / (44.1932) 34 70 46
e-mail address: /
website: / http://www.defra.gov.uk/ahvla/
Name of Head of Laboratory (Responsible Official): / Catherine Brown
Name of OIE Reference Expert: / Prof. Glyn Hewinson
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/topic at your laboratory
Test / For / Specificity / TotalCulture /
M. bovis isolation from cattle
/ 11880Culture / M. bovis isolation from animals other than cattle. / 788
Spoligotyping / M. bovis molecular test from cattle isolates / Specific to direct repeat region of M. bovis genome / 4530
Spoligotyping / M. bovis molecular test for isolates obtained from animals other than cattle / Specific to direct repeat region of M. bovis genome / 175
Spoligotyping / M. bovis molecular test for isolates obtained from isolates submitted from outside the UK / Specific to direct repeat region of M. bovis genome / 255
6-loci VNTR / M. bovis molecular typing test from isolates obtained from cattle and wildlife from Great Britain / Alleles specific for the M.tuberculosis-complex / 4320
IFNg ELISA –
‘Bovigam test’ / M. bovis infection in cattle / Measures comparative gamma interferon responses to bovine and avian tuberculin / 24,827
Antibody ELISA –
‘Brock test’ / M. bovis infection in Eurasian badgers (Meles meles) / M. bovis MPB83 antigen / 28 (test now withdrawn)
Lateral-flow immunoassay (Chembio Brock [TB] Stat-Pak®) / M. bovis infection in Eurasian badgers (Meles meles) / Cocktail of MTB complex antigens, including MPB83 / 956
Lateral-flow immunoassay (Chembio Brock [TB] Stat-Pak®) / M. bovis infection in animals other than badgers (e.g. alpaca, deer, tapir) / Cocktail of MTB complex antigens, including MPB83 / Zoo animals: 12
Alpacas: 415
Lateral Flow immunoassay (Chembio Brock [TB] Stat-pak®) / M. bovis infection in cattle/bovidae / Cocktail of MTB complex antigens, including MPB83 / 0
2. Production and distribution of diagnostic reagents
Type of reagent / Amount supplied nationally(including for own use) / Amount supplied to other countries
ESAT-6, CFP-10, Rv3615c peptides as antigens for Bovigam IGRA and intradermal skin test assays / Spain (200 doses), Mexico (400doses), The Netherlands (400 doses)
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
Liaison between AHVLA, the CRL in Madrid and the NRL in France, has taken place with respect to the standardisation of the culture of TB.
AHVLA took part in an inter laboratory comparison of molecular tests on diagnosis and epidemiology.
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
Discussions only, have taken place between AHVLA and a laboratory at Perigueux, France.
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
Results of a three year field evaluation of an injectable licensed vaccine for TB in badgers were published in 2011. Bacillus Calmette-Guérin (BCG) vaccination of free-living badgers reduced the incidence of positive serological test results (BROCK TB Stat-Pak) by 73.8 per cent.
In the same year we also reported that intramuscular BCG vaccination of captive badgers reduced the progression, severity and excretion of Mycobacterium bovis infection after experimental challenge when given at a dose 10times that prescribed for humans. BCG at the human dose was poorly protective. In common with other species, BCG did not appear to prevent infection of badgers subjected to experimental challenge, but did significantly reduce the overall disease burden.
Two projects are presently on-going on the development and testing of BCG vaccine suitable for oral delivery in bait.
Collaboration with Servicio Regional de Investigación y Desarrollo Agroalimentario, Centro de Biotecnología Animal, Asturias, Spain has been strengthened through visitation of a scientist from this Facility to AHVLA for the purposes of training in badger interferon-gamma testing. This resulted in one joint publication on the state of infection of badgers in Spain with Mycobacterium bovis and Mycobacterium avium complex. The prevalence, distribution and pathology related to infection with Mycobacterium bovis and other mycobacteria were determined in trapped (n=36) and road-killed (n=121) badgers in Spain from 2006 to 2010. The prevalence of M. bovis based on bacteriological culture from road-killed badgers was 8/121 (6.6%) and from trapped badgers was 0/36 (0%). Tuberculosis/M. bovis infection was evident in 15/121 (12.4%) road-killed badgers when bacteriology and histopathology were combined. Mycobacterium avium complex was isolated by culture from the tracheal aspirate of 1/36 (2.8%) trapped badgers and from tissue pools from 8/121 (6.6%) road-killed badgers.
Further work continues at AHVLA through a number of overseas collaborations to evaluate new immunological tests for the diagnosis of TB in badgers. This also includes a project to develop practical methods with which to obtain a diagnostic sample from restrained badgers without the use of general anaesthesia.
Defra-funded projects on cattle vaccines are either ongoing, have been completed or will begin in January 2012. Specific points to highlight in this area:
1) A dossier to support a marketing authorization for BCG in cattle has been submitted to the Veterinary Medicines Directorate, UK.
2) Over the past year AHVLA have been engaged in collecting and analyzing data in support of OIE validation test application of using specific antigens in the Bovigam IGRA assay as DIVA antigen to be used in conjunction with BCG vaccination. The drafting of this application is underway.
3) Mucosal vaccine delivery of virally vectored subunit vaccines was compared in cattle to systemic delivery route. Results demonstrated that equivalent systemic AND local immune responses were induced by both route.
4) Bovine IL-2-specific antibody reagents were developed that can be used for intracellular FACS staining, ELISA and ELISPOT (Whelan et al., PLoS One, 2012). These reagents will enhance our capabilities to study T cell memory and could serve as additional read-out system for diagnostic assays based on cellular immunity.
5) Several new antigens that are recognized in infected animals but not in naive cattle, based on hypoxia-induced (stage-specific) antigens were identified and peptide cocktails formulated (Jones GJ et al., Clin. Vaccine Immunol., 2012).
6) Results of a multinational collaborative study using a serological assay (IDEXX TB assay) were published. The data showed that, as in other serum assays, acceptable sensitivity was dependent on skin test boosting (Waters WR et al, Clin Vaccine Immunol, 2011).
7) Biomarkers correlating with disease that could increase the performance of in vitro assays of cellular immunity have been defined in mice models and validated in cattle (Aranday-Cortes E et al., PLoS One, 2012).
8) A project developing and validating blood-based ante-mortem diagnostic tests (interferon gamma release assay (IGRA), serological assays) to diagnose tuberculosis in camelids has been completed and its results are being analysed at present. This Project was funded by the GB industry.
9) Comparison study of cell-based (interferon-gamma) and serological ante mortem tests for the diagnosis of feline tuberculosis in domestic cats in Great Britain has been published (Rhodes SG et al., Vet Immun Immunopath 2011). Study collaboration between AHVLA and Royal (Dick) School of Veterinary Studies, Edinburgh, UK., National OIE Bovine TB and Paratuberculosis Reference Laboratory, France, Federal Veterinary Office, Switzerland and Chembio Diagnostic Systems Inc., USA.
10) AHVLA has further optimised a tuberculin-type skin test based on defined proteins and peptides, which could lead to a defined ‘tuberculin’, but is also applicable as DIVA skin test.
11) AHVLA has continued its international collaborations through several EU funded grants and also with colleagues in the USA, Mexico, New Zealand and East Africa.
12) AHVLA has initiated a small ring trial of the Bovigam EIA assay in collaboration with the EU Community Reference Laboratory in Madrid, Spain.
13) AHVLA has developed a gamma interferon release assay (IGRA) for tapirs.
6. Collection, analysis and dissemination of epizootiological data relevant to international disease control
Maintenance of www.Mbovis.org for the International standardisation of spoligotype nomenclature.
7. Maintenance of a system of quality assurance, biosafety and biosecurity relevant to the pathogen and the disease concerned
All AHVLA processing laboratories are accredited to ISO:17025. Internal and external auditing occurs regularly to ensure the status is upheld effectively. Each accredited laboratory partakes in routine internal quality control and external quality assurance of the tests.
Internal Safety audits are carried out regularly and Departmental Safety Advisors have been appointed for all TB processing laboratories. Access to the Containment Level 3 (CL3) suites at all sites, is controlled by the use of swipe cards.
8. Provision of consultant expertise to OIE or to OIE Member Countries
AHVLA has maintained collaborative links with ANSES (Agence Nationale de Sécurité Sanitaire, de l’Alimentation, de l’Environnement et du Travail), provided input into the role played by badgers to bovine TB in France, and made recommendations on strategies for controlling TB in wildlife.
9. Provision of scientific and technical training to personnel from other OIE Member Countries
AHVLA hosted a one-week training and discussion visit for representatives of government agencies from South Korea (August 2011)
10. Provision of diagnostic testing facilities to other OIE Member Countries
No activity
11. Organisation of international scientific meetings on behalf of OIE or other international bodies
None organised
12. Participation in international scientific collaborative studies
Towards the end of 2011, a collaborative study was initiated at ANSES to determine the tissue distribution of BCG within badgers given BCG orally. For this study, staff from AHVLA visited and worked at ANSES in order to transfer methods related to oral vaccination, culture, and conduct post mortem examination of vaccinated badgers.
AHVLA continues to contribute to research led by the University College Dublin to investigate the protective response of badgers vaccinated orally with BCG followed by experimental challenge with M. bovis. A new study has been initiated whereby badgers will be fed oral BCG vaccine in bait.
EU project TBSTEP, a consortium aimed at looking at improved control strategies for tuberculosis in animals (including vaccination approaches and improved diagnosis).
EU project NOPERSIST, a consortium of SME and research institutes aimed at improved diagnosis, and treatment of human and animal tuberculosis.
On-going collaborations with researchers in Spain, Mexico and New Zealand on the validation of ante-mortem diagnostic tests for cattle and goats (e.g. defined skin test reagents), as well as on cattle vaccine development (USA, NZ, Ethiopia).
AHVLA is leading an EMIDA consortium (11 partners) on the validation and development of diagnostic tests for mycobacterial diseases. This project has been approved for funding in December 2011.
AHVLA has organised large scale International collaborations for the molecular identification of European 1, European 2, African 1 and African 2 clonal complexes of M. Bovis by spoligotyping and deletion typing.
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
Oral presentations:
“Vaccines for badger TB – how we got to where we are and where we are going”. Gloucestershire Wildlife Trust, Bovine TB, badgers and cattle seminar. Bristol, UK, 2 December 2011.
“Vaccines for badger TB – where are now? (and how we got here)”. Veterinary Research Club 423rd meeting. London, UK, 9 December 2011.
Chambers MA chaired panel discussion and presented his own research work at Goals of vaccination: prevent disease, transmission, or cure? Many Hosts of Mycobacteria: A Comparative Symposium, Florida, USA, 20-23 March 2011.
Vordermeier HM. Plenary oral presentation at Keystone conference in Vancouver (15-20 January 2011): Tuberculosis: Immunology, cell biology and novel vaccination strategies.
Vordermeier HM. Invited plenary oral presentation at Annual Conference of the American Animal Health Association, Buffalo, NY (30.09 – 3.10.2011).
Smith NH. Invited plenary oral presentation. EUMEDNET-TB workshop in Sofia, Bulgaria (Sep. 2011)
Smith NH. Invited presentations at College of Veterinary Medicine, Huazhong Agricultural University (HZAU), Wuhan, China (May, 2011).
¡ Scientific publications in peer-reviewed journals
Balseiro A, Rodríguez O, González-Quirós P, Merediz I, Sevilla IA, Davé D, Dalley DJ, Lesellier S, Chambers MA, Bezos J, Muñoz M, Delahay RJ, Gortázar C, Prieto JM. Infection of Eurasian badgers (Meles meles) with Mycobacterium bovis and Mycobacterium avium complex in Spain. Vet J. 2011;190(2):e21-5.
Lesellier S, Palmer S, Gowtage-Sequiera S, Ashford R, Dalley D, Davé D, Weyer U, Salguero FJ, Nunez A, Crawshaw T, Corner LA, Hewinson RG, Chambers MA. Protection of Eurasian badgers (Meles meles) from tuberculosis after intra-muscular vaccination with different doses of BCG. Vaccine. 2011;29(21):3782-90.
Chambers MA, Rogers F, Delahay RJ, Lesellier S, Ashford R, Dalley D, Gowtage S, Davé D, Palmer S, Brewer J, Crawshaw T, Clifton-Hadley R, Carter S, Cheeseman C, Hanks C, Murray A, Palphramand K, Pietravalle S, Smith GC, Tomlinson A, Walker NJ, Wilson GJ, Corner LA, Rushton SP, Shirley MD, Gettinby G, McDonald RA, Hewinson RG. Bacillus Calmette-Guérin vaccination reduces the severity and progression of tuberculosis in badgers. Proc Biol Sci. 2011278 : 1913-20.
Alvarez J, Bezos J, de Juan L, Vordermeier M, Rodriguez S, Fernandez-de-Mera IG, Mateos A, Dominguez L. Diagnosis of Tuberculosis in Camelids: Old Problems, Current Solutions and Future Challenges. Transbound Emerg Dis. 2012 59:1-10.
Ameni G, Vordermeier M, Firdessa R, Aseffa A, Hewinson G, Gordon SV, Berg S. Mycobacterium tuberculosis infection in grazing cattle in central Ethiopia. Vet J. 2011 188:359-61