To meet the demographic pressure, the livestock industry must intensify production while striving for sustainable management. With the emergence and re-emergence of infectious diseases, including those caused by mycoplasmas, improving animal health and welfare is a key aspect of this challenge. While antibiotic treatments are facing an alarming rate of resistance, vaccines are only available for a very limited number of species and often provide insufficient protection. The RAMbo-V project intends to build on advances in bacterial genome engineering to pave the way for the development of synthetic vaccines against Mycoplasma bovis, a livestock pathogen associated with emerging episodes worldwide. This will be done by adapting and improving genetic tools recently developed in other mycoplasma species and using a high-throughput strategy for large-scale mapping of highly conserved antibody epitopes in the M. bovis pan-proteome. To engineer an avirulent vaccine chassis and secure its genetic stability, virulence factors non-essential for laboratory growth will be deleted from the M. bovis genome together with mobile genetic elements and chromosomal regions contributing to horizontal gene flows. Finally, the surface of M. bovis will be redesigned to allow optimal expression of selected epitopes through the use of a transmembrane carrier and the locking of surface variation mechanisms. By exploring an innovative vaccine strategy, RAMbo-V will have a positive impact on animal production and its dependence on antimicrobials. RAMbo-V will thus contribute to the development of sustainable food systems that manage demographic pressure with environmental, societal, and ethical challenges.< Réduire