Role of Pre-Farrow Natural Planned Exposure of Gilts in Shaping the Passive Antibody Response to Rotavirus A in Piglets

Figures

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  Figure 1. NPE administration and sample collection (serum, colostrum, and milk) schedule. NPE administration to gilts, farrowing and weaning are indicated by red, blue and green triangles. Gilts (N = 12 per group) and five piglets per gilt were sampled individually. Control group gilts did not receive NPE.

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  Figure 2. Validation of antigen expression via immunocytometric staining and immunoblotting. (A) HEK293A cells transfected with pcDNA3.1 (+) construct encoding G5 VP7 RVA showing positive anti-His tag staining. (B) HEK293A cells transfected with pcDNA3.1 (+) construct encoding G4 VP7 RVA showing positive anti-His tag staining. (C) Negative control. (D) Purified G5 RVA VP7 (37 kd). (E) Purified G4 RVA VP7 (37 kd). M—protein marker; S—culture supernatant; P—cell pellet; FT—flow through; W—wash fraction; E1, E2, E3 and E4—sequential G4 protein elutes. (F) Purified P[7] and P[23] VP4* (55 kd) protein elutes. (G) Western blot confirmation of the affinity-purified G5, G4, P[7] and P[23] antigens using anti-His monoclonal primary and HRP-tagged anti-mouse IgG secondary antibodies.

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  Figure 3. Kinetics of gilt serum antibody levels against RVA. Progression of serum IgG and IgA levels over time in gilts receiving three (group 1), two (group 2), one (group 3) or no (group 4, control) doses of NPE. Gilt serum IgG levels against G5 (A), G4 (B), P[7] (C) and P[23] (D) RVA antigens. Gilt serum IgA levels against G5 (E), G4 (F), P[7] (G) and P[23] (H) RVA antigens. The horizontal axis represents sample collection time-points (−5 W = 5 weeks pre-farrow; −3 W = 3 weeks pre-farrow; F = at farrowing; +3 W = 3 weeks post-farrow or at weaning). The vertical axis represents geometric mean antibody titers (GMTs). * p < 0.033, ** p < 0.002, *** p < 0.001, mixed effects model.

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  Figure 4. Kinetics of colostrum/milk antibody levels against RVA. Progression of colostrum/milk IgG and IgA levels over time in gilts receiving three (group 1), two (group 2), one (group 3) or no (group 4, control) doses of NPE. Colostrum/milk IgG levels against G5 (A), G4 (B), P[7] (C) and P[23] (D) RVA antigens. Colostrum/milk IgA levels against G5 (E), G4 (F), P[7] (G) and P[23] (H) RVA antigens. The horizontal axis represents sample collection time-points (−5 W = 5 weeks pre-farrow; −3 W = 3 weeks pre-farrow; F = at farrowing; +3 W = 3 weeks post-farrow or at weaning). The vertical axis represents geometric mean antibody titers (GMTs). * p < 0.033, ** p < 0.002, *** p < 0.001, mixed effects model.

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  Figure 5. Kinetics of piglet serum antibody response to RVA. Progression of piglet serum IgG and IgA levels over time in piglets born to gilts receiving three (group 1), two (group 2), one (group 3) or no (group 4, control) doses of NPE. Piglet serum IgG levels against G5 (A), G4 (B), P[7] (C) and P[23] (D) RVA antigens. Piglet serum IgA levels against G5 (E), G4 (F), P[7] (G) and P[23] (H) RVA antigens. The horizontal axis represents sample collection time-points (−5 W = 5 weeks pre-farrow; −3 W = 3 weeks pre-farrow; F = at farrowing; +3 W = 3 weeks post-farrow or at weaning). The vertical axis represents geometric mean antibody titers (GMTs). * p < 0.033, ** p < 0.002, *** p < 0.001, mixed effects model.