The aim of this study was to evaluate the effects of in-ovo vaccination on different incubation days of broiler embryos derived from young and old breeders on incubation indexes, vaccine response, and broiler performance. A number of 20,160 fertile eggs was distributed according to a completely randomized design in a 4 x 2 factorial arrangement (inovo vaccination on 16, 17, 18, or 19 days of incubation, and breeders of 31 or 52 weeks of age), totaling eight treatments with 15 replicates of 168 eggs each. Vaccination procedures and vaccines (strains and doses) were those routinely applied in commercial hatcheries. After hatch, 960 male chicks were housed and distributed according to the same experimental design previously applied in the hatchery. There were hatching losses (p<0.05) when eggs were vaccinated before 18 days of incubation. Greater Marek's disease antibody titers were obtained when the in-ovo vaccination was performed on day 19 of incubation, regardless breeder age. Embryonic age at vaccination did not compromise broiler performance in the field, and the flexibility of embryonic age for in-ovo vaccination can reduces incubation costs.
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