Aiming to enhance the immune efficacy of the combined vaccine against newcastle disease and avian influenza, N-[(2-hydroxy-3-trimethylammonium) propyl] chitosan chloride (HTCC) hydrogel microspheres and the blending emulsion of oil with microspheres (MPs) were investigated as vaccine adjuvants. After optimization, the maximized antigen absorption efficiency reached 90% when the antigens were incubated with 2 mg/mL microspheres with the size of 1 ?m for 2 h. In vivo experiments indicated that both the microspheres and blending emulsion significantly enhance the antigen specific HI titers. In detail, the blending emulsion group was superior to the microspheres in the enhancement of HI titers. Most importantly, both the microspheres and blending emulsion could induce robust and long-term cellular immune responses compared to oil adjuvant, and the proliferation levels of peripheral blood mononuclear cells were 1.5 folds to that of the soluble antigen group. These results suggested that the HTCC hydrogel microspheres and blending emulsion would be promising adjuvants for poultry vaccine.