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Abstract
This work was designed to study the effect of propolis as immunostimulant on the immune response of fish with or without vaccination using formalized killed vaccine against A.hyDROPhila. A total number of 720 apparently healthy catfish {Clarias grapeinus} were divided into 8 equal groups. Each group (90 fish) Group 1 served as control. Fish of groups 2 - 4 injected with proplis (propolis 10% - propolis 20% - propolis 30%), respectively. Fish of group 5-7 injected with same doses of propolis ( 1.0 ml sterile phosphate buffer) containing the specific concentrations of propolis was given to each fish for a period of 10 days .and vaccinated with formalized- killed vaccine ( propolis 10% + vacc – propolis 20% +vacc – propolis 30% +vacc); respectively. Fish of group 8 vaccinated only with formalized- killed vaccine. The fish were fed minced meat diet through the experiment and kept under observation for two weeks before starting the experiment. The experiment extended for 6 weeks after acclimatization.
Vaccination was done via intra-peritoneal injection using the 0.1 ml of formalin- killed bacterial cells... Blood samples were collected from the caudal vein of five fish weekly from each group into two clean dry tubes to evaluate the plate and passive agglutination tests. Moreover, all groups injected with 0.2 ml of A.hyDROPhila (containing 108 bacteria ml-1). The relative level of protection (RLP) among the challenged fish was determined.
The antibody titer of plate agglutination test at 7th day of experiment was 1 in group 1 and 2 in groups 2,3,4 and 3 in groups 5,8 while the other groups 6,7 was 4.Titer 14th day of experiment was 1 in group 1 and10 in groups,2,3,4,8 and 40 in group 7 while the other groups 5,6 was 20. The titer at 21th day of experiment was 1 in group 1 and 10 in groups2,3,4,8 and 20 in groups 5,6,7,The titer at 28th day of experiment was 1 in group 1 and 10 in groups 2,3,4,8 while the other groups 5,6,7 was 20 . The titer at 35th of experiment 1 in group 1 and 3 in groups 2, 3, 4 while the other groups (5, 6, 7) the titer was 10 in each group..but group 8 was 4. The titer at 42th day of examined fish was 1 in group 1and 3 in groups 2,3,4,8 while the other groups 5, 6, 7 was 4.
The antibody titer of indirect hemagglutination test at 7th day of experiment was 1 in group 1 and 2 in groups 2, 3, 4. While the group 5, 8 was 3 but groups 6, 7 were 4. The titer 14th day of experiment was1in group 1 and 10 in groups 2,3,4,8 and 20 in group 5, 7 and 40 in group 7. The titer at 21th day of experiment was 1 in group 1 and 10 in groups 2, 3, 4, 5, 8 and 20 in group 6 and 40 in group 7. The titer at 28th day of experiment was 1 in group 1 and10 in group 2,3,4,8 and 20 in group 5, 6, 7. The titer at 35th day of experiment was 1 in group 1and 4 in groups 2, 3, 4 while the other groups (5, 6, 7), the titer was 10 but group 8 was 3. The titer at 42th day of the experiment 1 in group 1,groups 2,3,4 was 2 and in groups 5,8 was 3 while the groups 6,7 was 4.
The Comparative between plate agglutination test and indirect Hemagglutination in revealed seams variation in the group 5- 7 at 21th day of experiment but no remarkable changes were observed at 28th day of experiment.
After Bioassay against A.hyDROPhila,The relative level of protection among the challenged fish of all groups (1-8) was 0, 33.34 ,44.45, 55,56, 88.89 ,88.89 ,94.45 and 83.84 for the different groups respectively.The best result was obtained in group seven which injected propolis and vaccine.
Polyacrylamid gel electrophorectic analysis of serum proteins was done on sera of experimental fish it was found that propolis increase serum pre -albumin, albumin and globulins the result was very highly significant increase (P< 0.001) in level of serum of examined fish.