comparative evaluation of montanide adjuvant methods (o/w vs. w/o/w) in experimental neospora caninum vaccine for canine immune response

This study aimed to evaluate the efficacy of a vaccine against neospora caninum using two oil emulsion adjuvant models. neospora caninum, a protozoan parasite, significantly affects canine health, causing neosporosis. native dogs under one year old, susceptible to neosporosis, were selected and tested for seronegativity. study materials included dmem culture medium, vero cell lines, antibiotics, bovine serum, and an attenuated neospora caninum strain. preparations involved phosphate buffer solutions, trypsin solution, tbe buffer, agarose gel, ethidium bromide, and tween phosphate buffer for elisa. pcr detected neospora caninum dna using specific primers. the study involved freezing and passage of vero cell lines, antigen preparation, and vaccine formulation with montanide isa 70 oil adjuvant in o/w and w/o/w formulations. dogs were immunized subcutaneously, and blood samples were collected at 7 and 21 days post-immunization. vero cell line cultures remained clear and contamination-free, with neospora caninum tachyzoites adapting well and destroying vero cells within 7 days. mycoplasma tests confirmed no contamination, and vaccines showed no microbial contamination after 24 hours. blood factor analysis revealed no significant differences pre- and post-immunization, indicating no pathogenicity. liver enzyme levels showed no significant change (p<0.05). immunization with the attenuated strain and montanide oil adjuvant in w/o/w formulation produced a higher antibody titer compared to control and other formulations (p<0.05). different oil adjuvant formulations in the experimental vaccine from attenuated neospora caninum strains in dogs elicit varied immune responses. adjusting the water and oil phase of the vaccine enhances the humoral immune response. immunizing dogs with this strain and adjuvants are a safe and effective method.