sheep pox (SP) still represents a major threat to animal wealth. Vaccination is the only reliable remedy for such highly infectious and contagious diseases. Chitosan nanoparticles (CNP) could be successfully added to live as well as inactivated vaccines to enhance their immune responses as it exhibits a good delivery system. In this study, CNP was added to maintenance essential media (MEM) with ratios of 3, 15 and 30 μg/ml. Such preparations were overlayed on confluent vero cell sheets for 7 days. CNP augmented MEM was proved to be unharmful for VERO cells. The prementioned concentrations of CNPMEM were mounted the infected sheets of cells that had been inoculated with the Romanian vaccinal strain of sheep pox virus (SPV). Appearance of the cytopathic effect as well as SPV infectivity titers were slightly improved particularly with CNP-MEM that was augmented with 15μg/ml. The former infected culture was used to prepare pilot batch of CNP supplemented sheep pox vaccine. The immune response of the pilot batch was assessed comparing with the conventionally produced SP vaccine by inoculating them into separate groups of susceptible sheep. There was slight increase in the serum neutralizing antibody titers in the group inoculated with the CNP augmented vaccine over the traditional one. It has been deduced that CNP could be used in sheep pox vaccine preparation to enhance the vaccine quality.