Fig. 2
Determination of serum IgG1, IgG2, IgM, and IgA antibody reactivity against H. somni rHsp60. Box-plot graphs of serum IgG1 (a), IgG2 (b), IgM (c) and IgA (d) antibodies reactivity against H. somni rHsp60 antigen in ELISA. Serum samples were obtained as follows: S1: before the first intranasal immunization (24–48 h of life); S2: two weeks after the first immunization (14–16 days of age); S3: two weeks after the second intranasal immunization (28–30 days of age); S4: two weeks after the third subcutaneous immunization (42–44 days of age); S5: two weeks after S4 (59–61 days of age). Individual study groups were labelled as CpG (vaccine formulation rHsp60 + rOMP40 + CpG), Con (control group), and MPLA (vaccine formulation rHsp60 + rOMP40 + MPLA). Significant differences within individual study groups between consecutive samplings (e.g., CpG S1-S2, S2-S3, S3-S4, S4-S5, etc.) are labelled with the same lowercase letters (a) for p ≤ 0.05. Significant differences within individual study groups between S1 and other samplings (e.g., CpG S1-S2, S1-S3, S1-S4, and S1-S5) are labelled with two asterisks (**) for p ≤ 0.01, or an asterisk (*) for p ≤ 0.05. The serum dilution was 1:50 for IgA, 1:100 for IgG2 and IgM; 1:1000 for IgG1. The median line across the box, lower, and upper boxes indicate the 25th percentile to the 75th percentile, and whisker non-outlier range.
