Abstract
Histomonas meleagridis, a protozoan parasite responsible for histomonosis (syn. Blackhead disease, histomoniasis), presents an increasing challenge for poultry health, particularly with the ban of licensed prophylactic and treatment options. Recent studies have explored H. meleagridis proteome, exoproteome, and surfaceome, linking molecular data to virulence and in vitro attenuation. Nevertheless, proteins involved in interactions with hosts remain largely unknown. In this study, we conducted immunoproteome analyses to identify key antigens involved in the humoral immune response of the parasite's main hosts, turkeys and chickens. Immunogenic proteins were isolated via immunoprecipitation using sera from chickens and turkeys that were vaccinated with a single attenuated strain and challenged with virulent strains of the protozoan, respectively. Mass spectrometry identified 155 putative H. meleagridis immunogenic proteins, of which 43 were recognized by sera from both hosts. In silico antigenicity screening (VaxElan) identified 33 pan-reactive antigens, with VaxiDL further highlighting 10 as potential vaccine candidates. Comparative analysis revealed host-specific immune responses, with 16 differential immunogenic proteins in chickens (6 specific to virulent and 10 to attenuated preparations) and 19 unique proteins in turkeys, all associated with virulent strains. These results enhance our understanding of H. meleagridis immunogenic protein dynamics and host-pathogen specificities, supporting the development of improved diagnostic tools and potential protective measures against the infection.