This study evaluates the impact of rabbit hemorrhagic disease virus (RHDV) vaccination on the comfort, exploratory, Ingestive, and physical behaviors of two rabbit breeds Chinchilla and V-line housed individually in cages. Behavioral responses were assessed at baseline (24 hours before vaccination) and at 1, 8, 27 and 72 hours post-vaccination using frequency and duration measures. The results showed that comfort behavior, neither lying nor stretching was observed in any group, while V-line rabbits exhibited a significantly lower frequency of sitting compared to Chinchilla rabbits, although no differences were noted in the duration of sitting. Self-grooming (auto-grooming) behavior was significantly reduced in both frequency and duration at 8 hours post-vaccination across all groups. Exploratory behavior showed increased sniffing duration at 1 hour post-vaccination in V-line rabbits, while the frequency of investigating the enclosure decreased in all groups at 1 and 8 hours compared to control Chinchilla rabbits. Ingestive behavior analysis revealed a significant reduction in feeding duration in vaccinated Chinchilla rabbits at 8 hours post-vaccination and an increase in drinking duration in vaccinated rabbits compared to non-vaccinated counterparts. These changes in ingestive behavior suggest potential physiological responses to vaccination. Physical activity, assessed through hopping, showed no notable differences in jumping, and reduced hopping Overall, the results highlight breed-specific behavioral differences following vaccination, with notable effects on self-grooming, feeding, and hopping activities. These findings suggest that vaccination with RHDV may influence various behavioral patterns, potentially due to immune or stress responses, though breed differences may also play a significant role.