Fowl cholera is an avian bacterial disease caused by Pasteurella multocid which causes economic losses due to mortalities which may reach 100% and loss in body weight. The present work was designed to develop a novel improved local gamma-irradiated inactivated fowl cholera vaccine candi-date, which stimulates vaccinated chickens mucosal immunity with different application routes to determine the most effective one. The antibody titers of vaccinated chickens measured by ELISA was found to be increased 2 weeks post vaccination until the third week, groups G3 and G3B which were vaccinated with the prepared irradiated FC vaccine via the intraocular route and they were the best as they recorded a titer of (399.528). On the fourth week, groups vaccinated by intranasal and oral route revealed positive immune status titers. The non-boosted group G4 was negative. Groups G3, G4B and G3B recorded (501.632, 517.274 and 665.037) respectively. On the fifth week, the antibody titers increased, but declined on the 6th week for groups G1, G2, G2B, G3 and G4 (257.746, 71.1541, 98.118, 395.339 and 230.759) but the titers increased for groups G1B, G3B, and G4B (483.433, 875.387 and 638.955). Complete protection was to the side of (0.3ml/dose) intraocular vaccinated-boosted group. While the commercial formalized vaccine was 90% for boosting group. Groups vaccinated-boosted by the gamma-irradiated intranasal (0.3 ml) vaccine recorded 80%. In conclusion, the study proved that gamma-irradiated I/O administrated vaccine is an effective protective vaccine. Montanide gel ISA 71 is a suitable adjuvant to enhance a protective immune response against FC.

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