Protection and diagnostic interference induced by heat inactivated, phage-inactivated and live vaccine prototypes against animal tuberculosis

Leire Fernández¹Miguel Fuertes¹Maria V. Geijo¹Natalia Elguezabal¹Jose L Serrano-Mestre²Lucía Vázquez-Iniesta²Rafael Prados-Rosales²Lorraine MICHELET³Maria Laura Boschiroli³Bernat Pérez de Val^4,5Gareth Jones⁶Ramon A Juste¹Joseba M Garrido¹Iker A Sevilla^1*

• ¹Departamento de Sanidad Animal, NEIKER-Instituto Vasco de Investigación y Desarrollo Agrario, Basque Research and Technology Alliance (BRTA), Derio, Spain
• ²Department of Preventive Medicine and Public Health and Microbiology, School of Medicine, Universidad Autónoma de Madridsp, Madrid, Spain
• ³Université Paris-Est, Laboratoire de Santé Animale, Unité Zoonoses Bactériennes, Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Maisons-Alfort, France
• ⁴IRTA, Programa de Sanitat Animal, Centre de Recerca en Sanitat Animal (CReSA), Campus de la Universitat Autònoma de Barcelona (UAB), Bellaterra, Spain
• ⁵Unitat mixta d’investigació IRTA-UAB en Sanitat Animal, CReSA, Campus de la UAB, Bellaterra, Spain
• ⁶Department of Bacteriology, Animal and Plant Health Agency (APHA), Surrey, United Kingdom

Vaccination emerges as a promising cost-effective tool to reduce the impact and spread of animal tuberculosis, especially in regions where test-and-slaughter eradication strategy is socioeconomically unfeasible or unfruitful for different reasons, provided it is safe, efficacious and compatible with diagnosis. In this study, we preliminarily evaluated the diagnostic interference (using guinea pigs) and the protective efficacy (using mice) of three heat-inactivated, three phage-inactivated and one live attenuated vaccine prototypes prepared from M. bovis, M. caprae and M. microti. Phage-inactivation killed almost all (96.41-99.92%) bacteria to be included in vaccines and filtering was used to remove the remaining viable cells. All the assayed vaccines induced skin test reactions in response to bovine tuberculin, but they were smaller in the phage-inactivated vaccine groups. All the vaccines were diagnosis-compatible with defined skin test antigens based on ESAT-6, CFP-10 and Rv3615c. In contrast with the rest of prototypes, vaccination with heat- and phage-inactivated M. microti did not prompt the production of detectable anti-MPB70+MPB83 antibodies. Mean bacterial burden was lower in all vaccinated groups in comparison with the control, being significantly reduced in the lungs of the heat-inactivated M. microti and M. caprae and phage-inactivated M. caprae groups. Considering both diagnostic interference and protection collectively, the heat-inactivated M. microti vaccine showed the best performance. Further studies to evaluate these vaccines and to improve phage-driven inactivation are warranted.