This study was applied on 631 sheep with ages ranged from 3month to 2 year, from which 60 animals were suffering from nervous manifestation. A total 45 brain samples were collected from recently dead and emergency slaughtered sheep and send to the lab. For bacteriological and histopathological examination. The bacteriological examination results 22 positive samples for L. monocytogenes(48.8%) with maximum results at 3-6 months of age (57.1%). Moreover, the result showed that the highest percentages for L. monocytogens isolation was at autumn and winter. Histopathological examination revealed that there were micro abscesses in the brain. In the same study, trial for preparation of listeria bacterin from the local strains isolated from the sheep was performed in rabbits as laboratory animal. Blood samples were collected from the rabbits groups for deferential leucocytic count and ELISA technique for detection of antibodies after challenge test. The results showed that there was elevation in antibody titer after vaccination and challenge. In addition, there was elevation in WBCs, Lymphocytes, Monocytes and Granulocytes in challenged group compared with vaccinated group. The results of ELISA were highly statistically significant.