1. 13 years in 20 minutes: Badger vaccine development and efficacy Dr Mark A. Chambers TB Science Lead Animal Health and Veterinary Laboratories Agency, U.K.

2. Badger vaccination: ‘another tool in the box’ Badger vaccination could help reduce the prevalence and severity of bovine TB in a badger population and thereby reduce the rate of transmission to cattle From Defra infographic - http://www.flickr.com/photos/defragovuk

3. Why BCG? Safety, cost, efficacy, known effect on preventing disseminated disease Freeze-dried vaccine obtained from SSI, Denmark Intramuscular route (IM) chosen for use  Can be administered in the field without anaesthesia

4. 1.Provide safety data for BCG using captive badgers 2.Determine efficacy of BCG in badgers using experimental challenge 3.Provide safety and preliminary efficacy data in wild badgers 4.Compile registration dossier and deal with questions arising Animal Test Certificate Randomised Badger Culling Trial Development of immunological assays Preparation for field study Safety in captive badgers – GLP Study INJECTABLE BCG Safety/efficacy in wild badgers – GCP field study Efficacy in captive badgers M.A. Prepare licence application dossier

5. 1.Provide safety data for BCG using captive badgers 2.Determine efficacy of BCG in badgers using experimental challenge 3.Provide safety and preliminary efficacy data in wild badgers 4.Compile registration dossier and deal with questions arising Animal Test Certificate Randomised Badger Culling Trial Development of immunological assays Preparation for field study Safety in captive badgers – GLP Study INJECTABLE BCG Safety/efficacy in wild badgers – GCP field study Efficacy in captive badgers M.A. Prepare licence application dossier

6. No fever following vaccination No influence on weight/feeding behaviour Modest swelling at site of vaccination in some badgers Data used to obtain Animal Test Certificate for field evaluation GLP safety study with captive badgers 10x overdose – 2-8x10 8 live bacteria Target dose – 2-8x10 7 live bacteria

7. 1.Provide safety data for BCG using captive badgers 2.Determine efficacy of BCG in badgers using experimental challenge 3.Provide safety and preliminary efficacy data in wild badgers 4.Compile registration dossier and deal with questions arising Animal Test Certificate Randomised Badger Culling Trial Development of immunological assays Preparation for field study Safety in captive badgers – GLP Study INJECTABLE BCG Safety/efficacy in wild badgers – GCP field study Efficacy in captive badgers M.A. Prepare licence application dossier

8. 1.Provide safety data for BCG using captive badgers 2.Determine efficacy of BCG in badgers using experimental challenge 3.Provide safety and preliminary efficacy data in wild badgers 4.Compile registration dossier and deal with questions arising Animal Test Certificate Randomised Badger Culling Trial Development of immunological assays Preparation for field study Safety in captive badgers – GLP Study INJECTABLE BCG Safety/efficacy in wild badgers – GCP field study Efficacy in captive badgers M.A. Prepare licence application dossier

9. Vaccine efficacy studies with captive badgers Two studies completed, in total: –6 unvaccinated –8 vaccinated with 2-8x10 6 live bacteria –9 vaccinated with 2-8x10 7 live bacteria 17 weeks between vaccination and experimental infection All badgers infected with M. bovis by endobronchial installation using a flexible fibrescope Samples taken for immunology & culture at regular intervals throughout Detailed post mortem examination 12 weeks after infection

10. Assessment of vaccine efficacy Size, appearance, and distribution of gross TB- confirmed lesions at PM used to derive score Statistically significant reduction in disease burden when higher dose of vaccine used (also more immunogenic) 10-fold lower dose gave variable levels of protection Even with an effective dose some animals are not protected for reasons not understood Total Visible Lesion Score

11. Proportion of animals excreting M. bovis per treatment group and reduction in % of excreting animals

12. 1.Provide safety data for BCG using captive badgers 2.Determine efficacy of BCG in badgers using experimental challenge 3.Provide safety and preliminary efficacy data in wild badgers 4.Compile registration dossier and deal with questions arising Animal Test Certificate Randomised Badger Culling Trial Development of immunological assays Preparation for field study Safety in captive badgers – GLP Study INJECTABLE BCG Safety/efficacy in wild badgers – GCP field study Efficacy in captive badgers M.A. Prepare licence application dossier

13. 1.Provide safety data for BCG using captive badgers 2.Determine efficacy of BCG in badgers using experimental challenge 3.Provide safety and preliminary efficacy data in wild badgers 4.Compile registration dossier and deal with questions arising Animal Test Certificate Randomised Badger Culling Trial Development of immunological assays Preparation for field study Safety in captive badgers – GLP Study INJECTABLE BCG Safety/efficacy in wild badgers – GCP field study Efficacy in captive badgers M.A. Prepare licence application dossier

14. GCP field study with wild badgers Objective: collect data on safety of IM BCG to wild badgers and maximise the chances of observing protective effect of vaccination Study site: ~55km 2 area of high badger density in England –Approx. 92 social groups defined by bait marking Two trapping sessions per year (2006-2009) –Vaccinated annually but only once per year  1787 trappings, 844 badgers, 519 vaccinates, 325 controls  >80 staff involved in total Badgers trapped and brought to central sampling facility –Micro-chipped and tattooed –Blood and clinical samples taken for immunology and culture –Vaccine injected into ~61% of badgers

15. Safety Excretion of BCG None seen from 3657 clinical samples Temperature monitoring 24 hours after vaccination Temperature fluctuations not attributed to vaccination Swelling at site of vaccination Seen in (22/265) ~8% of badgers but no skin lesions Trial badgers found dead within study area (n = 33) Subjected to detailed post-mortem examination Examination of vaccination site Culture for M. bovis and BCG BCG cultured from LNs of one vaccinated badger only

16. Vaccine effect Badgers were not killed for post-mortem confirmation of TB; relied on three ‘live’ tests 2010: Highly significant reduction (of 74%) in incidence of positive Stat-Pak results at social group level as a result of vaccination –Consistent with protective effect of vaccination, as antibody production positively correlated with extent & severity of TB in humans & badgers

17. Vaccine effect (cont.) 2012: Further analysis of data identifies an indirect protective effect in unvaccinated cubs (‘herd effect’) –Risk of unvaccinated badger cubs testing positive to TB decreased significantly as the proportion of vaccinated individuals in their social group increased (Odds ratio = 0.08) –When more than a third of their social group had been vaccinated, the risk to unvaccinated cubs was reduced by 79% (Odds ratio = 0.21)  Cubs that we have been unable to vaccinate because they are underground might still be protected  Beneficial effect not seen in adults so still need to vaccinate cubs as they emerge and are subject to potentially greater infection challenge Blood tests are not an absolute indicator of protection from disease so the results cannot tell us the degree of vaccine efficacy

18. 1.Provide safety data for BCG using captive badgers 2.Determine efficacy of BCG in badgers using experimental challenge 3.Provide safety and preliminary efficacy data in wild badgers 4.Compile registration dossier and deal with questions arising Animal Test Certificate Randomised Badger Culling Trial Development of immunological assays Preparation for field study Safety in captive badgers – GLP Study INJECTABLE BCG Safety/efficacy in wild badgers – GCP field study Efficacy in captive badgers M.A. Prepare licence application dossier

19. 1.Provide safety data for BCG using captive badgers 2.Determine efficacy of BCG in badgers using experimental challenge 3.Provide safety and preliminary efficacy data in wild badgers 4.Compile registration dossier and deal with questions arising Animal Test Certificate Randomised Badger Culling Trial Development of immunological assays Preparation for field study Safety in captive badgers – GLP Study INJECTABLE BCG Safety/efficacy in wild badgers – GCP field study Efficacy in captive badgers M.A. Prepare licence application dossier

20. Compilation of licensing dossier Bound by European Legislation Takes substantial time and expertise –Government employed an external consultant –4 months to compile and write –82 questions dealt with in total –13 months from start to finish

21. Conclusions Results from wild badgers are encouraging as indirect benefits of vaccination apparent Starting with an existing commercial source of BCG saved time, money, and effort As with BCG in other species, it is not 100% effective in preventing TB in badgers As far as we know BCG does not benefit already infected animals Not all badgers in an area will be trapped and vaccinated – based on experience expect 70% to be trapped Submission of dossier is time-consuming Takes time - 10 years from start to finish But it can be done!

22. An oral badger TB vaccine A vaccine that can be delivered orally is a potentially cheaper and more practical way of vaccinating large numbers of badgers in the wild than an injectable vaccine Steps to licensing an oral vaccine the same as for injectable vaccine But, R&D phase more demanding and no guarantee of success

23. Oral badger vaccine – issues to consider The science –Efficacy Need to produce an oral vaccine formulation that gives consistent protection in badger challenge experiments equivalent to that seen with injectable –Oral vaccines are inherently more variable than injected –Progress Have candidate oral vaccines that can give significant protection to badgers against experimental infection Further work required to show consistency in the levels of protection achieved

24. Oral badger vaccine – issues to consider The science –Deployment Identifying a palatable bait, compatible with live vaccine and the best way and time to deploy it through field studies –Uptake vs cost vs effort –Progress Palatable bait identified Evaluated in the field without vaccine –Aiming for coverage at least as good as with injectable vaccine (est. 70%) –>70% uptake by cubs when deployed down holes but need to improve uptake by adults –Further field work to be undertaken next year

25. Oral badger vaccine – issues to consider Regulatory –Even when we have identified an efficacious licensable vaccine candidate we still have 4-5 years of further work to do to complete regulatory studies, transfer technology to manufacturer and scale up, and get the product licensed Manufacturing process Environmental/operator safety issues –Need labelling, type of packaging Access to non-target species –Safety, implications for cattle exposure Field safety study (GCP) –Expensive, when to initiate? –Progress GLP safety study completed

26. Acknowledgements Funding: –Defra FERA: –Woodchester Park Field Team (now AHVLA) –Stats Team AHVLA –Badger Research Team –TB Diagnostic Laboratory –Pathology Department –Animal Services Unit