Pasteurella multocida is the common cause of Duck septicemia (Pasteurellosis) which affects ducks leading to high economic losses to duck producers. Pasteurella multocida infections are associated with severe, life-threatening systemic disease involving both hemorrhagic pneumonia and septicemia. A cross-sectional study was carried out on 220 samples collected from different breeds of ducks (Mallard, Muscovy and Baladi) from Minufiya governorate, Egypt. Only 16 cases were positive for Pasteurella multocida infection (7.3%). These isolates were confirmed microscopically, biochemically and rapidly by using Vitek2 compact system. The application of PMT-ELISA and mouse lethality test for the 16 isolates of P.multocida serotypes for differentiation between toxigenic and non toxigenic isolates. All tested of 16 isolates showed to be toxigenic by using PMT-ELISA and mouse lethality test. The application of PCR and multiplex PCR for the detection of tox A gene and capsular serotyping of toxigenic isolates of P.multocida respectively. All 16 P.multocida isolates were positive to tox A and capsular type A (100%). This study concluded that P. multocida serve as a major cause of Duck septicemia (Pasteurellosis) which affects ducks leading to high economic losses in poultry industry. Also indicated that the majority of P. multocida serotypes are toxigenic.